Enhancement of n-3 PUFAs utilization for functional meat production in slow-growing Korat chicken: evaluation of characteristics of glucose transporter-targeted lipid nanoparticles

Enhancement of n-3 PUFAs utilization for functional meat production in slow-growing Korat chicken: evaluation of characteristics of glucose transporter-targeted lipid nanoparticles | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Enhancement of n-3 PUFAs utilization for functional meat production in slow-growing Korat chicken: evaluation of characteristics of glucose transporter-targeted lipid nanoparticles Piyaradtana Homyok, Anyanee Kamkaew, Teerapong Yata, Worapapar Treesuppharat, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4761693/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Dec, 2025 Read the published version in Scientific Reports → Version 1 posted 14 You are reading this latest preprint version Abstract The aim of this research was to investigate the synthesis of suitable carrier of nanoparticles for improving the utilization of n-3 polyunsaturated fatty acids (n-3 PUFAs) source in chicken diets. Lipid nanoparticles were successfully prepared with two different n-3 oil sources, tuna and algal oils using hot and high-pressure homogenization method. Four preparations were defined as followed: non-targeting lipid nanoparticles containing tuna oil (TO_NPs), non-targeting lipid nanoparticles containing algal oil (AO_NPs), targeting lipid nanoparticles containing tuna oil (TO_TNPs) and targeting lipid nanoparticles containing algal oil (AO_TNPs). A second study was conducted for the targeting procedure, the treatments as followed: Control, TO_NPs and TO_TNPs. Thirty-three slow-growing chickens were examined during the post-administration kinetic at 2, 4, 8, 12 and 24 h. The physicochemical characteristics of lipid nanoparticles, storage stability and in vivo biodistribution were evaluated. The results showed that the particle diameters of TO_NPs and AO_NPs were 223.7 and 294.4 nm, whereas the particle diameters of TO_TNPs and AO_TNPs were 134.7 and 184.0 nm, respectively. The polydispersity index (PDI) and zeta-potential of nanoparticles showed a good distribution and stability in colloid dispersions, respectively. Moreover, the nanoparticles of the TNPs groups were less susceptible to lipid oxidation than that of the NPs groups during a storage at 4°C. The study of the biodistribution based on the Nile red intensity and the determination of n-3 PUFAs composition in chicken meat confirmed the effectiveness of targeted lipid-based nanoparticles to transport directly fatty acids into the skeletal muscle cells of chicken. Biological sciences/Biochemistry/Lipids/Fats Biological sciences/Biochemistry/Lipids/Fatty acids Biological sciences/Biochemistry/Lipids/Lipid peroxides Biological sciences/Biochemistry/Lipids/Oils Physical sciences/Materials science/Nanoscale materials/Nanoparticles Physical sciences/Nanoscience and technology/Nanobiotechnology/Nanoparticles Biological sciences/Chemical biology/Transporters Physical sciences/Chemistry/Biochemistry/Lipids/Fatty acids Physical sciences/Chemistry/Biochemistry/Lipids/Lipid peroxides Physical sciences/Chemistry/Biochemistry/Lipids/Oils Physical sciences/Nanoscience and technology/Nanomedicine/Imaging techniques and agents n-3 polyunsaturated fatty acids Lipid-based nanoparticles Physiochemical characteristics In vitro storage stability In vivo biodistribution Slow-growing chickens Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Oils containing high amounts of n-3 polyunsaturated fatty acids (n-3 PUFAs) are susceptible to oxidation process [ 1 ]. Many studies have tried to improve the storage stability of these oil sources, and the encapsulation technology was the more efficient [ 2 , 3 ]. Another possibility is to use the nanotechnology to improve the utilization of n-3 PUFAs oil source [ 4 ]. This technology has a high potential, and we investigated its use to enhance the accumulation of n-3 PUFAs in chicken muscles to produce functional meat. Korat chicken, a slow-growing breed that is very popular in Thailand, has been developed as an alternative option catering to the preferences of Thai chicken producers [ 5 ] and could be used to produce functional meat enriched with n-3 PUFAs. Previous studies showed that dietary 4% tuna oil supplementation increased n-3 PUFAs content reaching 19.03% of total fatty acids in Korat chicken meat [ 6 ]. However, n-3 PUFAs sources are susceptible to lipid oxidation that can affect the feed quality [ 7 ]. The protection of these lipids through encapsulation alone is inadequate to enhance the utilization of n-3 PUFAs sources, as a portion of these fatty acids will also be distributed among non-targeted organs such as liver. Different studies showed that lipid nanoparticles could be synthesized by applying the edible oil such as fish oil [ 8 – 10 ]. However, various nanoparticle size, dispersion ability of particles, zeta-potential (the observation of a robust electric potential at the particle surface boundary demonstrates the existence of a potent repulsive force, which contributes significantly to enhancing the overall physical colloidal stability) and high amount of PUFAs including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) might increase the unstable form of particle size during storage [ 9 ] and plain lipid nanoparticles could be degraded by digestive enzymes [ 11 ]. Another important parameter is the ability of nanoparticle surface coating [ 12 ]. Modifying their surface with PEGylating helps to avoid enzyme activity in the digestive tract [ 12 , 13 ]. Moreover, generating a surface with a specific substrate recognized with target transporter/receptor could allow the transport of beneficial substance directly to the target organs [ 14 – 17 ]. Therefore, we investigated the physicochemical characteristics of lipid nanoparticles and their susceptibility to lipid oxidation during storage and their ability to transfer the bioactive compounds (n-3 PUFAs) into the muscles. For feeding animals, the important point to consider is that lipid nanoparticles need to transfer perfectly to the target organ after the passage through the digestive tract and adsorptive cells that depends on the stability of lipid nanoparticles. The aim of our study was to analyze the effects of n-3 PUFAs source, targeting process, and storage temperature and duration on the physicochemical characteristics of lipid nanoparticles, their oxidation susceptibility, and their ability to transfer n-3 PUFAs to chicken meat. Results Physicochemical characteristics of lipid-based nanoparticles containing n-3 PUFAs oils The physical characterization of nanoparticles containing n-3 PUFAs oils (Tuna oil, TO and Algal oil, AO) are presented on Table 1 . The mean particle diameter of the NPs groups was higher than that of the TNPs groups (P ≤ 0.001) whereas tuna oil in both nanoparticle forms had smaller particle diameters than algal oil (P ≤ 0.001). The TO_TNPs group had the highest PDI value (P < 0.001) and the NPs groups had higher zeta-potential values than the TNPs groups (P < 0.001) and the value was highest in AO_NPs group. Table 1 Physical characteristic of n-3 PUFA oils within different type of lipid-nanoparticles 1 . Treatment Average diameter (nm) Polydispersity index Zeta-potential (mV) TO_NPs 223.7 ± 1.01 c 0.315 ± 0.014 b -42.6 ± 0.47 b AO_NPs 294.4 ± 1.83 d 0.261 ± 0.007 a -34.2 ± 0.46 c TO_TNPs 134.7 ± 1.18 a 0.365 ± 0.008 c -48.1 ± 0.68 a AO_TNPs 184.0 ± 1.21 b 0.298 ± 0.010 ab -49.4 ± 0.67 a P-value ≤ 0.001 ≤ 0.001 ≤ 0.001 1 Two different n-3 PUFA oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles. Data are presented as mean ± SE (n = 3). a−c Values in the same column with different superscripts letters are significantly different (P < 0.05). Thermal and chemical properties of lipid-based nanoparticles The melting point of glyceryl monostearate (GMS) a raw material used for nanoparticle synthesis was 76.6 o C, whereas the nanoparticle suspension including TO_NPs, AO_NPs, TO_TNPs and AO_TNPs showed lower endothermic peaks at 46.8, 46.4, 45.3 and 45.0 o C, respectively (Fig. 1 ). Both the n-3 PUFA sources and nanoparticle types changed the melting point in nanoparticle form by lipid nanoparticles composite of algal oil and the targeting groups had the lowest melting points of lipid nanoparticles. The Fourier-transformed infrared spectroscopy (FTIR) analysis of nanomaterials and lipid-based nanoparticles is shown in Fig. 2 . The peak intensity of lipid nanoparticles was more flatten than individual spectra of each material. It can be concluded that tuna crude oil was packed tightly inside the nanoparticles. Due to functional group which is composite in the structure of lipid, triglyceride and fatty acid that could be found in tween 20, alkyl polyglucoside, GMS and tuna oil, the peak intensity decreased after the synthesis into lipid-based nanoparticles. The FTIR spectra show in Fig. 2 , The bands related to hydroxyl group (3499 − 3233 cm − 1 , -OH stretching) [ 18 ]. was found in tween 20, poloxamer, alkyl polyglucoside and GMS. For lipid (2921 − 2850 cm − 1 , asymmetric and symmetric stretching vibration of CH 2 groups in lipid alkyl chain, ν asym CH 2 and ν sym CH 2 ) [ 19 ] and fatty acid (1641 cm − 1 , C = C stretching, 1470–1460 cm − 1 , -CH 3 bending and 1240 − 1207 cm − 1 , -CH 2 bending) [ 20 – 22 ] were found in tween 20, alkyl polyglucoside, and tuna oil. The ester linkages (1741 − 1734 cm − 1 , ester carbonyl group (-C = O)) [ 20 ] was found in GMS, tween 20 and triglyceride in tuna oil but not occurred in alkyl polyglucoside and poloxamer. The cyclic ether (1029 cm − 1 , cyclic ether (-COC) and pyranose ring (1150 cm − 1 , -COC glycosidic bond) [ 23 ] were found in alkyl polyglucoside. The wavelength at 720 cm − 1 indicated that this long chain carbon [ 24 ] was found in GMS, tween 20, alkyl polyglucoside, and tuna oil. In addition, the wavelength ranging at 2921 − 2850 cm − 1 (-CH stretching) and 1500 − 500 cm − 1 (-CH bending and long chain carbon) had a reduced intensity in physical mixture and synthesized of lipid nanoparticles. In vitro storage stability of n-3 PUFA lipid-based nanoparticles Particle size stability The results of the storage stability on particle size of the n-3 PUFA lipid-based nanoparticles are shown on Table 2 . The synthesis of n-3 PUFA source oils in the form of targeted-lipid nanoparticles groups (TO_TNPs and AO_TNPs) can help to maintain the size of the nanoparticles, but the storage conditions were better at low temperature storage (4°C) for 4 weeks, while the particle size of the non-targeted lipid nanoparticles groups changed at a faster rate at all temperature storage (TO_NPs_4°C, TO_NPs_RT, AO_NPs_4°C and AO_NPs_RT). However, during the storage, there were statistically significant changes in nanoparticle size for the AO_NPs_4°C (P < 0.05), AO_NPs_RT (P < 0.01), TO_TNPs_RT (P < 0.05), TO_TNPs_RT (P < 0.001) and AO_TNPs_RT (P < 0.001). During the storage, the change in nanoparticle size was faster at room temperature than at 4°C. In addition, the use of algal oil resulted in changes particle size faster in lipid-based nanoparticles (AO_NPs_4°C, AO_NPs_RT, and AO_TNPs_RT). Table 2 Stability of n-3 PUFA oils within different type of lipid-nanoparticles 1 on particle diameter during storage at room temperature (RT) or 4°C. Treatments Particle diameter (nm) Start Week 1 Week 2 Week 3 Week 4 SEM P-value TO_NPs_4°C 225.23 cB 215.83 cA 233.37 cB 197.03 bA 230.02 cB 4.09 < 0.01 TO_NPs_RT 223.50 cA 233.83 dA 258.83 cB 258.8 cB 268.88 dB 6.02 < 0.05 AO_NPs_4°C 303.53 dAB 328.93 eB 316.87 dA 300.4 dA 320.03 eA 3.62 < 0.05 AO_NPs_RT 299.60 dA 332.17 eB 341.83 dB 358.53 eB 344.78 fB 5.93 < 0.01 TO_TNPs_4°C 136.50 a 137.9 a 139.67 a 142.73 a 139.4 a 1.39 0.853 TO_TNPs_RT 134.00 aA 137.47 aA 131.23 aA 136.5 aA 140.82 aB 1.04 < 0.05 AO_TNPs_4°C 176.7 b 183.7 b 177.57 b 178.8 b 183.51 b 1.21 0.312 AO_TNPs_RT 176.4 bA 187.53 bB 194.00 bB 192.13 bB 195.09 bC 1.92 < 0.001 SEM 12.92 14.89 15.40 15.43 15.23 P-value < 0.001 < 0.001 < 0.001 < 0.001 < 0.001 1 Two different n-3 PUFA oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles. Data are the average ± standard error of the mean (SEM) of three independent replicates (n = 3). For each attribute, different small letters indicate statistically significant (P < 0.05), highly significant differences (P < 0.01) and very highly significant differences (P < 0.001) between different samples at a same time of storage. Different capital letters in a same row indicate very highly significant differences (P < 0.001) over time. Oxidative stability evaluated with peroxidative value The peroxidative values are presented in the Table 3 . The unencapsulated tuna oil (TO_4°C) and tuna oil within lipid nanoparticles were stored at 4°C (TO_NPs_4°C and TO_TNPs_4°C) and had reduced peroxidative values ​​(P < 0.001) compared to the groups kept at room temperature except at the start of the experiment where the values ​​were not different. Moreover, no statistically significant differences in peroxide production in unencapsulated algal oil were found between the two storage temperatures (AO_4°C and AO_RT) from week 1 to week 4. In addition, algal oil within lipid nanoparticles (AO_NPs_4°C, AO_NPs_RT, AO_TNPs_4°C and AO_TNPs_RT at first day) showed a statistically highly significant (P < 0.001) increase in peroxide value after synthesis. The synthesis of oil within nanoparticle form and storage at suitable temperature influenced the peroxidative value reaction. Pre-synthetic and post-synthetic oils in nanoparticle form kept at low temperature (4°C) showed a statistically significant (P < 0.05) slowdown of peroxidative activity by comparison with the storage at room temperature from week 1 to week 3. Oxidative stability evaluated with TBARS value The malondialdehyde (MDA) values are presented on the Table 4 . Synthesizing tuna oil into the targeted-lipid nanoparticle increased the oxidation stability as the amount of MDA was lower in the non-targeted lipid nanoparticles but for algal oil, the reverse was observed. Moreover, storage at low temperature (4°C) slow down the production rate of MDA. The synthesized algal oil within lipid nanoparticles in the experimental groups (AO_NPs_4°C, AO_NPs_RT, AO_TNPs_4°C and AO_TNPs_RT) had a greater increase in MDA than the unsynthesized algal oil (AO_4°C and AO_RT) (P < 0.001). Table 3 Stability of n-3 PUFA oil sources within different type of lipid-nanoparticles 1 on peroxidative value during a storage at room temperature (RT) or 4°C. Treatment Peroxidative value (meq of oxygen/kg fat) Start Week 1 Week 2 Week 3 Week 4 SEM P-value TO_4°C 0.106 dB 0.110 cdB 0.113 cdB 0.079 cA 0.184 cC 0.009 < 0.001 TO_RT 0.113 dA 0.160 fB 0.213 iC 0.153 eB 0.306 dD 0.018 < 0.001 AO_4°C 0.033 aC 0.022 aA 0.018 aA 0.026 aB 0.017 aA 0.002 < 0.001 AO_RT 0.032 aA 0.031 aA 0.032 aA 0.041 abB 0.035 aAB 0.001 < 0.001 TO_NPs_4°C 0.094 cAB 0.108 cBC 0.129 deD 0.119 dCD 0.085 bA 0.005 < 0.001 TO_NPs_RT 0.095 cA 0.130 deB 0.196 hiC 0.211 fC 0.112 bAB 0.013 < 0.001 AO_NPs_4°C 0.113 dA 0.153 fB 0.161 fgB 0.164 eB 0.161 cB 0.006 < 0.001 AO_NPs_RT 0.111 dA 0.210 gCD 0.181 ghBC 0.242 gD 0.159 cB 0.012 < 0.001 TO_TNPs_4°C 0.059 bB 0.064 bB 0.064 cB 0.062 bcB 0.039 aA 0.003 < 0.001 TO_TNPs_RT 0.063 bA 0.116 cdC 0.097 cB 0.107 dBC 0.102 bBC 0.005 < 0.001 AO_TNPs_4°C 0.107 dA 0.099 cA 0.127 dB 0.108 dA 0.103 bA 0.003 < 0.001 AO_TNPs_RT 0.108 dA 0.140 efB 0.149 efBC 0.164 eC 0.114 bA 0.006 < 0.001 SEM 0.005 0.009 0.01 0.011 0.013 P-value < 0.001 < 0.001 < 0.001 < 0.001 < 0.001 1 Two different n-3 oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles. Data are the average ± standard error of the mean (SEM) of three independent replicates (n = 3). For each attribute, different small letters indicate very highly significant over time (P < 0.001) between different samples at a same time of storage. Different capital letters in a same row indicate highly significant differences over time (P < 0.001). Table 4 Stability of n-3 PUFA oil sources within different type of lipid-nanoparticles 1 on malondialdehyde value during storage at room temperature (RT) or 4°C. Treatment Malondialdehyde value (µmol MDA/kg oil) Start Week 1 Week 2 Week 3 Week 4 SEM P-value TO_4°C 14.01 eC 10.81 dB 7.95 bcA 14.89 dC 13.32 deC 0.69 < 0.001 TO_RT 13.76 eB 14.08 efB 17.97 fC 6.53 cA 17.9 fC 1.13 < 0.001 AO_4°C 1.23 aB 1.53 aC 1.25 aB 0.70 aA 2.15 aD 0.13 < 0.001 AO_RT 1.23 aA 2.36 aC 1.60 aB 1.40 abAB 2.82 aD 0.17 < 0.001 TO_NPs_4°C 10.61 dA 18.88 gC 9.41 cA 14.92 dB 15.65 efB 0.94 < 0.001 TO_NPs_RT 10.61 dA 22.42 iD 12.38 dB 15.72 dC 21.73 gD 1.29 < 0.001 AO_NPs_4°C 9.21 cB 15.38 fC 9.10 cB 6.75 cA 8.51 cB 0.79 < 0.001 AO_NPs_RT 8.88 cA 21.06 hC 21.65 gC 20.75 eC 13.55 deB 1.39 < 0.001 TO_TNPs_4°C 13.70 eC 13.03 eC 6.42 bA 8.58 cB 9.07 cB 0.76 < 0.001 TO_TNPs_RT 13.64 eA 14.58 fA 14.70 eA 21.74 eB 21.68 gB 1.03 < 0.001 AO_TNPs_4°C 4.54 bB 4.31 bB 2.92 aA 3.00 bA 5.64 bC 0.28 < 0.001 AO_TNPs_RT 4.54 bA 7.89 cB 8.15 cB 7.61 cB 11.4c dC 0.61 < 0.001 SEM 0.78 1.14 1.03 1.17 1.09 P-value < 0.001 < 0.001 < 0.001 < 0.001 < 0.001 1 Two different n-3 oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles. Data are the average ± standard error of the mean (SEM) of three independent replicates (n = 3). For each attribute, different small letters indicate very highly significant differences (P < 0.001) between different samples at a same time of storage. Different capital letters in a same row indicate very highly significant differences (P < 0.001) over time. Transfer of lipid nanoparticles to the target organ The Nile red distribution in Pectoralis major sample is shown in Fig. 3 . The Nile red distribution was increased within targeted-lipid nanoparticles (TNPs) at 8 and 12 h after oral gavage by comparison with non-targeted lipid nanoparticles (NPs). Then it decreased to the initial content at 24 h. The results demonstrated that the targeted lipid nanoparticles had the potential to be absorbed into the bloodstream and transferred to the target organ faster than other forms of nanoparticles. Fatty acid composition of breast meat of Korat chicken The fatty acid profile of breast meat of the chickens after oral gavage with the lipid-based nanoparticles is shown on Table 5 . C18:3n3 content in breast meat of TNPs treatment at 4 h was decreased (P 0.05) whereas TNPs treatment at all time point lower than control (P < 0.05). The EPA content in TNPs group at 4 h was higher (P < 0.05) than that in the NPs treatment. The DHA content in TNPs treatment at 4 h was higher (P < 0.05) than that in the NPs. By the way, the highest of DHA content was showed in the TNPs group at 24 h. No significant different in EPA contents at 2, 8, 12 and 24 h, and DHA content at 2, 8 and 12 h compared between NPs and TNPs treatment. C18:2n6 contents no different compared between NPs and TNPs at each time. C20:4n6 contents in TNPs treatment at 24 h higher (P 0.05). C18:1n9 contents in TNPs treatment at 24 h were lowest (P < 0.05) compared to that in NPs and control treatment. Coefficient of efficacy of lipid-nanoparticles The coefficient of efficacy of targeted and non-targeted lipid nanoparticles are showed on Table 6 . TNPs had higher potential to transfer EPA and DHA into the chicken skeletal muscle than the NPs groups. The coefficient of efficacy of TNPs was higher than that of the NPs treatment in each time. At 2 h both NPs and TNPs treatments were able to transfer the n-3 PUFA but after that the TNPs treatment showed higher potential to transfer the n-3 PUFA at 4 and 24 h in TNPs treatment. The n-3 PUFA amount at 8 and 12 h after oral administration was decreased. In contrast, the n-3 PUFA content in NPs treatment was reduced at 4 h then increased slightly until 24 h. Table 5 Fatty acid composition of TO oil within different type of lipid-nanoparticles 1 at 2, 4, 8, 12 and 24 h after oral administration. Treatment Fatty acid composition (% of total fatty acids) C16:0 C18:0 C18:1n9 C18:2n6 C18:3n3 C20:4n6 EPA DHA Control 19.52 9.31 20.33 b 22.94 b 0.35 d 23.21 ab 0.21 ab 1.71 ab NPs at 2h 18.62 9.13 20.23 b 21.01 ab 0.22 abc 26.11 ab 0.20 ab 1.63 ab TNPs at 2h 19.95 9.76 19.52 b 21.69 b 0.24 bc 24.65 ab 0.27 ab 1.85 abc NPs at 4h 20.57 9.84 21.75 b 21.09 ab 0.32 cd 22.47 ab 0.16 a 1.57 a TNPs at 4h 19.76 9.87 18.60 b 20.18 ab 0.20 ab 26.60 ab 0.28 b 2.46 d NPs at 8h 20.76 9.63 20.64 b 22.35 b 0.29 bcd 22.02 ab 0.21 ab 1.82 abc TNPs at 8h 19.91 8.96 20.16 b 20.83 ab 0.22 abc 25.23 ab 0.25 ab 2.40 cd NPs at 12h 20.18 8.55 25.89 bc 22.13 b 0.27 bcd 18.75 a 0.20 ab 1.78 abc TNPs at 12h 19.49 8.38 23.09 bc 21.85 b 0.23 abc 22.38 ab 0.23 ab 2.22 bcd NPs at 24h 18.62 9.24 19.75 b 20.21 ab 0.20 ab 27.77 b 0.23 ab 2.55 d TNPs at 24h 19.72 9.15 14.49 a 17.50 a 0.12 a 35.62 c 0.29 b 3.43 e SEM 0.18 0.12 0.53 0.32 0.01 0.84 0.01 0.10 P-value 0.203 0.058 < 0.001 < 0.01 < 0.001 < 0.001 < 0.05 < 0.001 1 Tuna oil (TO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles. Statistical analysis is based on F-test. Data are the average ± standard error of the mean (SEM) of three independent replicates (n = 3). For each attribute, different small letters indicate statistically significant (P < 0.05), highly significant differences (P < 0.01) and very highly significant differences (P < 0.001). Table 6 Coefficient of transfer efficacy of TO within different type of lipid-nanoparticles 1 at 2, 4, 8, 12 and 24 h after oral administration relative to control. Treatment Coefficient of efficacy (%) EPA DHA Control 0.002 0.013 NPs at 2h 0.62 0.76 TNPs at 2h 2.32 2.27 NPs at 4h -1.41 -1.85 TNPs at 4h 2.55 9.65 NPs at 8h 0.06 0.64 TNPs at 8h 0.80 4.93 NPs at 12h 0.55 2.85 TNPs at 12h 1.65 6.52 NPs at 24h 1.07 9.70 TNPs at 24h 2.38 16.98 1 Tuna oil (TO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles. Discussion By comparison with AO nanoparticles, TO nanoparticles had lower diameter, higher PD index and lower ZP. AO contains a high amount of long-chain unsaturated fatty acids such as DHA. The fatty acid can move to the surface of the nanoparticles, resulting in an enlarged particle size [ 25 ]. It seems to be indicated the n-3 PUFA oil can change the uniformity of the nanoparticle diameter. Moreover, the PD index involved their particle size population. As is obviously in AO nanoparticles lower PD index than TO nanoparticles. A larger PD index value indicated a broad particle size distribution of the formulation whereas a low value of the PD index indicated monodisperse samples [ 26 ]. This implies that AO can maintain the uniformity of particle size better than TO. In reference to the quality of TO contains phospholipids [ 27 ], this compound had potential to lesser particle size [ 28 ] meanwhile can increase zeta-potential value with their negative charged [ 29 ]. TO and AO was applied in this present study is unrefined oil and refined oil, respectively. Unrefined TO contains impurities such as phospholipid, free fatty acid, aldehyde, ketones and pigment [ 30 ] that rapidly activated by pro-oxidant effect more than refined AO result to higher oxidized of TO at the beginning of experiment whereas algal oil could be better potent lipid peroxidation by greater quality, lower oxidative rate and might be prevented lipid oxidation by the carotenoids activity [ 2 , 31 , 32 ]. Thus, the lipid oxidative products were higher for TO than the AO at start of experiment. On the reverse, TO_NPs had lower peroxidative values than AO_NPs and TO_TNPs had lower peroxidative values than AO_TNPs. In contrast, the MDA content of TO_NPs and TO_TNPs was higher than that of AO_NPs and AO_TNPs. The possibility of the increase of peroxide value (PV) due to the sources had high free fatty acid induce lipid peroxidation stage [ 33 ]. AO had higher PV after lipid nanoparticle synthesis indicated that the hydroperoxides was produced continuously with higher rate from these free fatty acids, especially the oxidation of n-3 PUFA [ 34 ] whereas TO had lower oxidized rate which change into the MDA slowly than that AO. By the way, the lower of the MDA content in AO because the changed of the n-3 PUFAs oxidized into the secondary product [ 35 ]. Moreover, the products higher in oxidation may have a low PV when the primary oxidation metabolites are largely metabolized to secondary oxidation metabolites [ 36 ]. Therefore, the MDA for AO in lipid nanoparticles may rapidly shifted to another compound quicker than TO in lipid nanoparticles. By the way, the further study needs to invest the volatile compound of n-3 PUFAs oxidized to confirm the phenomenon. Therefore, tuna oil loaded in lipid nanoparticles was selected to further investigated the potential of the targeting process on the in vivo biodistribution, fatty acid profile, and coefficient of efficacy. The lower melting point of lipid nanoparticles mixtures than that of GMS due to the lipid matrix within the nanoparticles. Oils containing n-3 PUFAs (TO and AO) have liquid lipid properties leading to convert crystalline state to an amorphous state due to the disordered arrangement of molecules [ 8 , 9 ]. More than that, TO in lipid nanoparticles has higher endothermic peak than that algal oil due to undesirable compounds of tuna oil. Unrefined oil or triglyceride are more complex than those of the fatty acids and required more thermal energy [ 37 ]. Moreover, the melting point was affected by the high content in unsaturated fatty acids in the formulation [ 38 ]. The polyunsaturated fatty acids implying a larger space to be accommodated into the polymeric matrix indicate induced a more amorphous structure of the lipid carrier [ 39 ]. Thus, there are reasonable that the endothermic peak in TO treatment was shifted at higher temperature than that AO treatment. The DSC is an effective tool to investigate the melting behavior and crystalline state of nanocarriers and their impact on some properties such as stability of lipid nanoparticles, which is useful for further use in animal feed processing. During the storage at room temperature (RT), the diameter of nanoparticles was higher than that of nanoparticles stored at 4°C particularly after 3 and 4 weeks of storage particularly for NPs treatment whereas there was no effect of temperature storage for TNPs treatment. The diameter of nanoparticles increased with the storage duration. Similarly with literature researches indicated the diameter of lipid nanoparticle was enlarged by storage for long time particularly unmodified-surface nanoparticles [ 28 , 40 , 41 ]. The reason is the aggregation of surfactants between nanoparticle [ 42 ] or the interaction of reactive oxygen species (ROS) with surface of nanoparticle [ 43 ]. In contrast, the modified-surface nanoparticles can resolve the conflict and enhance oxidative stability by preventing pre-oxidation on the surface of the nanoparticle boundary [ 44 , 45 ]. Furthermore, the reduce particle size during storage due to the major compounds such as n-3 PUFAs in the matrix were descended by high temperature [ 46 , 44 ]. The peroxidative value and the MDA level were higher during the storage at RT by comparison with the treatments stored at 4°C. The high temperature can increased the kinetic energy contact of oil with oxygen [ 47 ]. The higher of temperature had increase the rate of hydroperoxide decomposition. The reactivity of redox reactions had greater rates [ 48 ]. The amphiphilic lipid was reacted with the pro-oxidation at the interface and hydroperoxide was produced. The free radicals penetrate through the surface layer and stimulated the oxidation with n-3 PUFAs [ 49 ]. Thus, the study of storage stability with different temperature indicated that the lipid oxidation continuously occurred during of storage particularly under high temperature for unmodified-surface nanoparticles. This should be noted that the storage of n-3 PUFAs in the lipid nanoparticle should be given priority for improving the sustainable quality prolonged enough for further procedures such as the drying process. The smaller nanoparticles formability in TNPs treatments might caused by the alkyl polyglucoside that was not present in the NPs groups. The nanoparticles synthesis with targeting had greater ability to perform lipid nanoparticles [ 50 – 53 ]. The stable of particles size in TNPs groups were indicated that the alkyl polyglucoside which use as the targeting lipid nanoparticles, have protective property to maintain the physical structure during storage. The change of particle size was faster in algal oil group possibly due to high amount of free fatty acids that have a carboxylic group with anionic, and are able to transfer into the surface of nanoparticles [ 54 ]. However, the treatment AO_TNPs_4°C was different because the low temperature condition slowdown the breakdown of triglycerides to free fatty acids, particularly, n-3 PUFAs such as DHA that can affect the particle size [ 25 ]. It is possible that the algal oil group had higher amount of free fatty acids inducing an increase in the nanoparticle size. Although in the previous studies demonstrated that the storage conditions no effect on particle size of nanoparticles [ 8 , 55 – 57 ] contrast with present study shown kept at 4°C and RT are effect diameter of the unmodified surface nanoparticle. This indicated that NPs is unstable form due to may interact with ROS present at the interface or surrounding aqueous phase [ 58 ] such as impurities in crude oil which act as pro-oxidants [ 27 , 59 ]. In addition, the PDI values describe the width or spread of the particles size distribution of the nanoparticles ranged between 0.1 and 0.3 indicating a good stability of the emulsion [ 60 ]. However, the TO_NPs group had the highest PDI value, possibly due to attractive hydrophobic interactions between the particles [ 61 ]. The use of liquid oils such as fish oil can make the nanoparticles more physically stable by reducing the crystalline structure. Therefore, it prevents the incorporation of nanoparticles [ 59 ] and increases PDI value. Moreover, the lower PDI value in AO nanoparticles indicated that is more dispersion in suspension than that TO nanoparticles. The PD Index is based on size of nanoparticles [ 32 ]. The higher of the multiple particle size population show more PDI value while higher uniformity is lower polydisperse result to lower the PD Index [ 58 ]. Moreover, zeta-potential found in all groups were lower than − 30 mV. The NPs groups had lower values than the TNPs groups, (P < 0.001). The synthesis including tween 20, poloxamer and alkyl polyglucoside, resulted in a decrease in the electric potential difference of the nanoparticles [ 62 ]. For the NPs groups, the n-3 PUFA oils source influenced the zeta-potential value. This is due to the movement of carboxylic fatty acids in the region near the outer layer of the nanoparticles. This resulted in more negative charge conversion [ 25 ]. The TNPs groups contained alkyl polyglucoside. When this raw material is used in combination with a poloxamer, it results in a lower voltage difference than that observed for the NPs groups. Moreover, unrefined tuna oil contained phospholipids which are anionic groups [ 63 ] and may have contributed to the high zeta-potential in TO_NPs group. The melting temperature was higher for NPs nanoparticles than for TNPs nanoparticles because of targeting process effect on decreasing particle size. The size of particle became smaller resulting in decreased melting point [ 64 ]. By the way, the surface-modification of lipid nanoparticles could be effect on increase the particle size of nanoparticles [ 14 , 17 ] whereas the particle size was decreased in the present study due to the ratio of targeting surfactant in formulation [ 65 ]. It has been shown that the active substances and raw materials have similar properties, namely the lipid molecules, which affect the form of nanoparticles. The hydroxyl group (3499 − 3233 cm − 1 , -OH stretching) was found in raw materials with properties as an emulsifier, including tween 20 [ 24 ], poloxamer [ 66 ], alkyl polyglucoside [ 67 ] and GMS [ 30 ]. However, after the synthesis, the nanoparticles containing tuna oil had no effect on triglyceride degradation as it did not convert to free fatty acids, which should appear at wavelength of 1711 cm − 1 [ 54 ]. The wavelength at 1095 cm − 1 (-OCC, ether bond) provided by tween 20 and poloxamer implicated in the properties of emulsifier hydrophilic part of the molecule. The parts of raw materials, alkyl polyglucoside occur the wavelength at 1029 cm − 1 indicated the vibration of functional group of cyclic ether (-COC) and Pyranose ring and 1150 cm − 1 (-COC glycosidic bond) [ 23 ]. The peak wavelength was changed towards a lower intensity in the physical mixture and lipid nanoparticles. It indicated that the raw material was integrated into the system before the nanoparticles would be formed. Moreover, the peak that disappeared at wavelength below 1095 cm − 1 in both mixtures was due to the structure of the molecules (including CH-stretching, long chain carbon) that bind together very densely. The nanomaterial changed their position from its own specific properties. The groups that had the hydrophobic properties occurred together inside the particles, while the hydrophilic groups occurred around the periphery of the particles. This phenomenon presented the similar wavelength peak of -OH stretching a functional group found in the surfactant family of tween 20, alkyl polyglucoside, and poloxamer. Moreover, the apparent wavelength in the spectrum at 1741 − 1734 cm − 1 , 1641 cm − 1 and 1150 cm − 1 showed that the nanoparticles contained oil, such as tuna oil, inside the mixtures. The wavelength ranging at 2921 − 2850 cm − 1 (-CH stretching) and 1500 − 500 cm − 1 (-CH bending and long chain carbon) disappeared or had a reduced intensity, indicating that the molecules in the mixture are moving according to the molecular properties mentioned above. These functional groups can be found in fat-containing raw materials, which have nonpolar molecules. This property is more prone to interactions together by noncovalent bonds such as van der Waals binding within particles [ 68 ]. Furthermore, the absence of new peaks in both mixtures showed that during the synthesis, there was no reaction that formed intermolecular covalent bonds [ 56 , 69 ]. The targeted-lipid nanoparticles were able to slow down the peroxidative reaction by comparison with the oil within non-targeted lipid nanoparticles because the surfactants act as protectors avoiding the inside oil to react with outside oxidants. The peroxidative value was significantly different in weekly storage of each treatment. This means that the initial lipid oxidation reaction was dynamic over the time storage and this constantly occurring leads to increase rate of the reaction [ 70 ]. In addition, the amount of peroxide in some week was found to be low. This could mean that the substance has already converted to another molecule, or it may not have happened yet because of the product of initial stage was quickly oxidized into the next stage. However, the industry recommended peroxidative value for fish oil should not exceed 18 meq active O 2 /kg after 1 month storage [ 71 ]. So, the n-3 PUFAs lipid nanoparticles can be kept more than one month because their peroxidative values are lower than the recommended value. By the ways, further study should carry out the long-term storage period to confirm our data on storage stability for commercial products. The MDA content was higher for NPs nanoparticles than for TNPs nanoparticles suggesting a higher susceptibility to oxidation for NPs nanoparticles. May be, the triglycerides were hydrolyzed to free fatty acids with negatively charged carboxylic groups causing deposition in the outer layer of the nanoparticles. The resulting negative ions induced lipid oxidation [ 72 ]. During the synthesis, lipids were heated may be favoring fatty acid oxidation. The measurement of lipid-based nanoparticles by in vivo study indicated that the targeted lipid-based nanoparticles showed the potential to transfer the fatty acids directly into skeletal muscle cells of Korat chickens. Nile red staining showed that FA deposition in muscles was more efficient by using TNPs compared to NPs nanoparticles at 8 h and 12 h after oral gavage with lipid-based nanoparticles. The lipid nanoparticles seem to act as a target carrier specific to the receptors at the microvilli. This phenomenon is confirmed by our previous research [ 73 ]. Specific-transporters such as Sodium-glucose transporter 1 (SGLT1) and glucose transporter (GLUT) 2, 5, 7, etc., serve to accept single molecules such as glucose and galactose into enterocytes [ 74 – 76 ]. For EPA deposition, the amount deposited 4 h after oral gavage was higher with TNPs nanoparticles compared with NPs nanoparticles. There was no difference between treatments for the other time points for EPA deposition. For DHA deposition, the amount deposited between 4 h and 24 h post-gavage was higher with TNPs nanoparticles compared with NPs nanoparticles. TNPs treatment was more efficient than NPs treatment and the efficacy of deposition was higher for DHA than for EPA that could be explained by a higher amount of DHA in nanoparticles compared to the EPA content. The results revealed the potential for both the receptors to function as individual molecules within the cell and to bind to those same molecules when attached to the nanoparticles and introduced into the cell. For example, nanoparticle uptake by microvilli receptor-specific monomolecular such as galactose [ 77 ], L-carnitine [ 51 ] and taurocholic acid [ 52 ] has been studied in digestive tract and muscle cells [ 78 ]. The utilization of receptor-specific molecules enhances the uptake of essential substances into the bloodstream and subsequent delivery to the targeted organs [ 14 , 17 ]. There are indications that intestinal cells possess mechanisms capable of responding to the absorption of active substances in the nanoparticle form, wherein these nanoparticles are transported into the bloodstream through penetration into intestinal cells and subsequently directed towards target cells with specific transporter including GLUT via the endocytosis pathways [ 79 ]. However, intracellular transport is divided into several pathways that affects the delivery of nanoparticles into the bloodstream differently [ 80 ]. Consequently, the comprehension and elucidation of the nanoparticle delivery pathway in functional intestinal cells at the molecular level will enable the identification of the underlying processes involved in nanoparticle transmission to the target organs. This understanding can further facilitate the development of appropriate models utilizing lipid nanoparticles. The coefficient of efficacy in TNPs groups indicated that the targeting process had potential to transfer the n-3 PUFA into target organ. By the way, the slightly decrease of n-3 PUFA amount in this group might due to the entering process of targeting-lipid nanoparticles in the cell via receptor-mediated endocytosis and sorting into lysosome, endocytosis with low pH in endosome make the ligand dissociated from their receptor [ 50 ]. Then, the endosome vesical combined with lysosome for degradation and the cargos were degraded by hydrolases to be single molecules [ 80 ]. Moreover, plain lipid nanoparticles are discriminated and weakened and could be recognized by the reticuloendothelial system [ 81 ]. On the other hand, targeting lipid nanoparticles escape the phagocytosis, resulting in a longer circulation time [ 82 ] and might enhanced the n-3 PUFA accumulation. Further study should be conducted in haematology and serum biochemistry to determine the response of lipid nanoparticles in chicken. Conclusions This study provides conclusive evidence that nanotechnology has the potential to enhance the utilization of n-3 PUFA oil resources through nanoparticle synthesis. However, it is crucial to consider the source of the oil used for synthesis, as it can significantly affects parameters such as nanoparticle size, nanoparticle dispersion, and zeta-potential. Based on the outcomes of thermal and chemical analysis, it is established that n-3 PUFA oils were successfully encapsulated within lipid nanoparticles along with solid lipid, glyceryl monostearate. This was evidenced by the observed reduction in melting points as indicated by DSC analysis. Furthermore, the utilization of n-3 PUFA oil in nanoparticle form exhibited the ability to hinder lipid oxidation, with the rate of oxidation being contingent upon the storage temperature. Finally, the in vivo distribution findings substantiated the efficacy of targeted lipid-based nanoparticles to deposit long-chain n-3 PUFA in the chicken muscles. Material and methods Experimental design for in vitro study For the in vitro study, four preparations were considered, each with three replications. These were compared with free oils (crude tuna oil, TO; feed grade, T. C. Union Agrotech Co. Ltd., Bangkok, Thailand and algal oil, AO (Schizochytrium sp.); Shanghai Wellboost Health Food Co., Ltd, Shanghai, China), and TO and AO loaded non-targeted nanoparticles (NP) and targeted nanoparticles (TNPs), respectively, including TO_NPs, AO_NPs, TO_TNPs and AO_TNPs. The physicochemical characteristics and in vitro storage stability of lipid nanoparticles were investigated. Lipid nanoparticles synthesis The n-3 PUFA loaded lipid-based nanoparticles were prepared by hot and high-pressure homogenization approach (Fig. 4 ) as previously described [ 83 ]. In this method, the lipid phase consisting of GMS (5 g), span 80 (3 g) (Croda, Bangkok, Thailand) and n-3 PUFAs (20 g) (tuna oil or algal oil) was blended and melted above 70°C to form an organic phase. The fatty acid composition of crude tuna oil and algal oil is indicated on Table 7 . Meanwhile, the aqueous phase was prepared by dispersing emulsifying agent, glycerol (2.5 g) Tween 20 (3 g), Poloxamer 188 (2 g) (Croda, Bangkok, Thailand) in distilled water. The aqueous phase was added drop wise to lipid phase at above 70°C with continuous agitation at 300 rpm for 3 min to get a primary emulsion. The pre-emulsion was then homogenized by using high-speed homogenizer (IKA, Altra-Turrac T25, Germany) at 8,000 rpm for 3 min. Then, it was sonicated (Qsonica sonicator, USA) at 30 A pulse on 30 sec and off 10 sec interval for 5 min to obtain n-3 PUFA loaded non-targeted lipid-based nanoparticles (NPs). For targeted lipid-based nanoparticles (TNPs), 2 g of alkyl polyglucoside was blended with organic phase preparation. Subsequently the dispersion was cooled in ice water bath. The prepared dispersion was stored in airtight container at 4°C. Table 7 Fatty acid composition of n-3 PUFA oils (% of total fatty acids) Fatty acid composition TO AO TO loaded in nanoparticles Palmitic Acid (C16:0) 20.28 38.01 29.43 Stearic Acid (C18:0) 5.28 1.47 8.46 Oleic Acid (C18:1n9) 14.16 - 22.64 Linoleic Acid (C18:2n6) 2.84 0.23 7.69 α-Linolenic acid (C18:3n3) 1.68 0.59 2.40 Arachidonic Acid (C20:4n6) 2.52 0.49 4.30 Eicosapentaenoic acid (C20:5n3) 6.98 0.72 3.60 Docosahexaenoic acid (C22:6n3) 20.03 37.63 9.24 SFA 39.15 52.91 43.95 MUFA 24.39 - 28.82 PUFA 36.45 46.96 27.23 n-3 PUFA 30.30 46.01 15.24 n-6 PUFA 6.11 8.02 11.99 n-6/n-3 ratio 0.20 0.17 0.79 SFA Saturated fatty acid, MUFA Monounsaturated fatty acid, PUFA Polyunsaturated fatty acid, n-3 PUFA Omega-3 polyunsaturated fatty acids, n-6 PUFA Omega-6 polyunsaturated fatty acids. 1 Two different n-3 PUFA oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles Physicochemical characteristic analysis A Zetasizer 3000HS (Malvern Instruments, Malvern, UK) was employed to characterize NPs and TNPs for particle diameter, polydispersity index (PDI), and zeta-potential. Thermal and chemical properties of lipid nanoparticles Differential scanning calorimetry (DSC) DSC is a thermo-analytical technique in which the difference in the amount for heat required to increase the temperature of a sample and reference is measured as a function of temperature. Usually, both the sample and reference are maintained at nearly the same temperature throughout the experiment. Generally, the temperature program for a DSC analysis is designed such that the sample holder temperature increases linearly as a function of time. The reference sample should have a well-defined heat capacity over the range of temperatures to be scanned. DSC (Pyris I DSC, Perkin Elmer, USA) was performed on samples of nanoparticles with and without feed or oil ingredient components to determine whether there were any interactions with the core-shell components. To determine the phase transition temperatures of the nanoparticles content n-3 PUFA and GMS, dried samples were placed in open aluminum pans. An empty pan was used as a reference pan compared with sample pan. Baseline correction and data treatment were conducted with OriginPro 9.85 (OriginLab, Northampton, Massachusetts, USA). Fourier-transformed infrared spectroscopy (FTIR) In order to evaluate potential interactions between functionalized ligands and excipients within the lipid nanoparticles, a FTIR spectrum was acquired. The samples, consisting of solid lipid, liquid lipid, glucose-functionalized ligand, surfactant, physical mixture, non-target lipid nanoparticles loaded with tuna oil and algal oil, as well as targeting lipid nanoparticles loaded with tuna oil and algal oil, were meticulously ground with potassium bromide (KBr) to generate an infrared transparent matrix. Subsequently, FTIR scanning was performed using a Shimadzu Corporation FTIR-8400S spectrometer, covering the spectral range of 4,000 to 500 cm − 1 with a resolution of 1.0 cm − 1 . Baseline correction and data treatment were conducted with OriginPro 9.85. In vitro storage stability The assessment of storage stability was conducted following the methodology outlined as previously described [ 83 ] with some modifications. Free oil and oil synthesized within nanoparticles (n = 3) were stored at 4°C in stability cabinets during four weeks. The hydrodynamic diameter (HD) was measured at different time points by dynamic light scattering (DLS) using a Malvern Zetasizer at an angle of 90° at 25°C and the oxidative stability was investigated by measuring PV and TBARS value. In vitro oxidative stability and peroxide value The determination of the primary oxidation products was carried out using the procedure outlined as previously described [ 84 ]. In order to measure the PV, the sample was added to 9.8 mL of a chloroform: methanol mixture (2:1, v/v) and mixed for 5 seconds on a vortex mixer. Subsequently, a solution of ammonium thiocyanate (50 µL) was introduced into the mixture, followed by the addition of iron (II) solution (50 µL). The iron (II) solution was prepared by combining BaCl 2 and FeSO 4 at a final concentration of 0.144 mol L − 1 , resulting in individual concentrations of 0.132, respectively. After 5 min incubation period at room temperature, the absorbance of the samples was measured at 500 nm against a blank (solution without any sample) by a Shimadzu-1800 UV–visible spectrophotometer (Shimadzu, Tokyo, Japan). The quantities of lipid hydroperoxides were determined by employing an external standard curve constructed with cumene hydroperoxide (purity 80%, sigma-Aldrich Co, USA) with concentrations ranging between 0 and 16 µM. In vitro oxidative stability and thiobarbituric acid reactive substances The secondary oxidation products were monitored by quantification of the thiobarbituric acid (TBA) reactive substances (TBARS) as previously described [ 85 ]. In brief, the samples were mixed with 1.8 mL of deionized water and 4.0 mL of TBA solution. TBA solution was prepared by dissolving 15 g of trichloroacetic acid (15% w/v) and 0.375 g of TBA (0.375%) in 100 mL of 0.25 mol L − 1 HCl. In the following, the mixtures were heated in a boiling water bath for 15 min and then cooled to room temperature. Finally, the mixtures were centrifuged (2000 g for 15 min). The intensity of the color created as a result of the reaction between TBA with malondialdehyde (MDA), an important by-product of lipid peroxidation, was measured at 532 nm. The standard curve of 1,1,3,3-tetraethoxypropane was used to determine the MDA concentrations. Experimental design for in vivo study All procedures in the present study were approved by the Ethics Committee on Animal Use of the Suranaree University of technology, Nakhon Ratchasima, Thailand (SUT-IACUC-009/2021). The experiment was conducted at Suranaree University of Technology (SUT) farm. The experimental model employed a completely randomized design, comprising three treatments and three replicates. The treatments consisted of control, tuna oil-loaded non-targeted lipid nanoparticles and tuna oil-loaded targeted lipid nanoparticles. Thirty-three female chickens, aged 56 days with an average weight ranging from 750 to 800 g, were individually housed in cages for a period of 7 days as an adaptation phase prior beginning the experiment. The chickens received commercial diet and water ad libitum for the whole period and withdrawal feeding 12 h before investigation. At 63 days of age, the lipid nanoparticles (2 ml for each chicken) were administered through oral administration at once, utilizing a syringe affixed to a designated feeding tube. After administration, the chickens were euthanized by cervical dislocation at distinct time intervals after gavage, 2, 4, 8, 12, and 24 h (3 chickens per treatment and per time interval). At each time point, all the breast muscle of the chickens was harvested and stored at -25°C for imaging purposes, as well as for the subsequent analysis of fatty acid composition. In vivo imaging of tissue biodistribution The investigation of nanoparticle distribution in muscle samples involved assessing the quantity of nanoparticles deposited in the target organ by utilizing the optical in vivo imaging system (IVIS), which captured the signal emitted by the fluorescent molecules. The fluorescence was observed within the appropriate wavelength range during the absorption (excitation) and emission processes. This methodology bears resemblance to a confocal laser-scanning microscope. However, the IVIS system is specifically designed for detecting signals from larger specimens. In this study, this instrument was employed to validate the presence of lipid nanoparticles within the target organ, the muscle. The In vivo imaging of nanoparticle accumulation in birds was assessed following the methodology as previously described [ 14 ]. The birds were administered NR-NPs or NR-Glu-NPs orally at a dose of 1 mg/kg (equivalent to 3.14 µM, 2 mL) via oral gavage. Samples were eliminate unwanted fluid before quantify the near-infrared fluorescence signal intensities by using an In Vivo Imaging System MS FX PRO (Carestream Health Inc., Rochester, NY, USA) with an excitation band pass filter at 540 nm and an emission at 600 nm. Images were processed using Bruker Molecular Imaging software version 7.1.3.20550 (Bruker, Billerica, MA, USA). Mean intensity was performed to analyze statistics. The Nile red concentration was equivalent in all experiments (n = 3 birds per treatment). Fatty acid composition The lipids were extracted from approximately 5 g of each muscle sample using 90 ml of chloroform: methanol (2:1, v/v) [ 86 ]. After that, the methylation was conducted with around 20 to 25 mg of extracted fat [ 87 ]. The fatty acid methyl esters (FAME) were analyzed using gas chromatography (Hewlett-Packard 7890A; Agilent Technologies, Santa Clara, CA, USA) with a capillary column (SP 2560, Supelco Inc., Bellefonte, PA, USA, 100 m × 0.25 mm i.d., 0.20-µm film thickness) and a flame ionization detector. The carrier gas was helium at a flow rate of 0.95 ml/min. The temperatures of the injector and detector were 260°C. The initial column temperature was 70°C. It raised to 175°C at a rate of 13°C/min, and then to 240°C at a rate of 4°C/min. Compound were identified and quantified by using Masshunter software (v10.0.707.0, Agilent, USA). Coefficient of efficacy (%) was performed to evaluate the potential of lipid nanoparticles using the following formula: Coefficient of efficacy (%) = \(\:\left(\frac{\text{F}\text{A}\text{n}\:\text{i}\text{n}\:\text{s}\text{a}\text{m}\text{p}\text{l}\text{e}\:\text{t}\text{r}\text{e}\text{a}\text{t}\text{e}\text{d}\:-\:\text{F}\text{A}\text{n}\:\text{i}\text{n}\:\text{u}\text{n}\text{t}\text{r}\text{e}\text{a}\text{t}\text{e}\text{d}}{\text{F}\text{A}\text{n}\:\text{a}\text{m}\text{o}\text{u}\text{n}\text{t}\:\text{i}\text{n}\:\text{s}\text{u}\text{s}\text{p}\text{e}\text{n}\text{s}\text{i}\text{o}\text{n}\text{s}\:\text{u}\text{s}\text{e}\text{d}\:}\right)\times\:100\) Statistical Analysis Analysis of variance will be performed by GLM procedure for a completely randomized design using SPSS Version 18.0 (SPSS Inc., Chicago, I11., USA). University Edition with the following statistical model: Yij = µ + τi + εij Where: Yij = the dependent variable, µ = the overall mean, τi = the treatment effect, and εij = the random residual error. Significant differences between treatment means were assessed by Tukey's multiple comparison tests after a significant F-test. The level of statistically significance will establish at P < 0.05. The values for physical characteristic of lipid nanoparticles were expressed as means ± standard error (SE) and the value for storage stability and fatty acid composition were expressed as means ± standard error of mean (SEM), which represents the pooled SEM for the model. Data availability The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. Declarations Ethical statement The study was approved by the Ethics Committee on Animal Use of the Suranaree University of technology, Nakhon Ratchasima, Thailand (SUT-IACUC-009/2021). All methods were performed in accordance with the relevant guidelines and regulations. ARRIVE Guidelines statement This study is reported in accordance with ARRIVE guidelines ( https://arriveguidelines.org ). Competing interests: The authors declare that they have no competing interests. Funding This research project was supported by the National Research Council of Thailand (NRCT) : NRCT5-RGJ63007-093, as well as by Suranaree University of Technology (SUT), Thailand Science Research and Innovation (TSRI), and National Science, Research and Innovation Fund (NSRF) (NRIIS number 160352). Author Contribution W.M. and A.M. conducted conceptualization, funding acquisition, and project administration. P.H., T.Y., A.K., A.M. and W.M. conceived and designed the methodology and investigation. T.Y. performed lipid nanoparticles synthesis. P.H. and W.M. performed and investigated the physicochemical characteristic, in vitro studies and fatty acid composition. P.H., W.T. and A.A. performed and investigated the samples imaging. P.H., T.Y., A.K., E.B., C.B. and W.M. performed data curation and formal analysis. P.H., E.B. and W.M. performed writing – original draft. P.H., E.B., C.B. and W.M. performed writing – review & editing the manuscript. All authors read and approved the final manuscript. Acknowledgments This research project was supported by the National Research Council of Thailand (NRCT) : NRCT5-RGJ63007-093, the Royal Golden Jubilee Ph.D. (RGJ-PHD) Program as well as by Suranaree University of Technology (SUT). The authors also highly appreciate the Center of Excellence on Technology and Innovation for Korat chicken Business Development, SUT and French National Institute for Agriculture, Food, and Environment (INRAE), UMR Biologie des Oiseaux et Aviculture for partly supporting this research. Data Availability The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. References Packer, L., Weber, S. U. & Rimbach, G. Molecular aspects of alpha-tocotrienol antioxidant action and cell signalling. J. Nutr . 131, 369s-373s, https://doi.org/10.1093/jn/131.2.369S (2001). Chang, M. et al. Optimization of cultivation conditions for efficient production of carotenoid-rich DHA oil by Schizochytrium sp. S31. Process Biochemistry. 94, 190–197, https://doi.org/https://doi.org/10.1016/j.procbio.2020.04.007 (2020). Wang, Y., Ghosh, S, & Nickerson M. T. Microencapsulation of flaxseed oil by lentil protein isolate-κ-carrageenan and -ι-carrageenan based wall materials through spray and freeze drying. Molecules. 27, 3195. https://doi.org/10.3390/molecules27103195 (2022). Kharat, M & McClements D. J. Fabrication and characterization of nanostructured lipid carriers (NLC) using a plant-based emulsifier: Quillaja saponin. Food Res. Int. 126, 108601. https://doi.org/10.1016/j.foodres.2019.108601 (2019). Kaewsatuan, P. et al. Comparative proteomics revealed duodenal metabolic function associated with feed efficiency in slow-growing chicken. Poult. Sci. 101, 101824, https://doi.org/10.1016/j.psj.2022.101824 (2022). Hang, T. T. T., Molee, W. & Khempaka, S. Linseed oil or tuna oil supplementation in slow-growing chicken diets: Can their meat reach the threshold of a “high in n-3 polyunsaturated fatty acids” product? J. Appl. Poult. Res. 27, 389–400, https://doi.org/https://doi.org/10.3382/japr/pfy010 (2018). Sanguansri, L. et al. Encapsulation of mixtures of tuna oil, tributyrin and resveratrol in a spray dried powder formulation. Food Funct. 4, 1794–1802, https://doi.org/10.1039/c3fo60197h (2013). Zhu, Q. Y. et al. Preparation of Deep Sea Fish Oil-Based Nanostructured Lipid Carriers with Enhanced Cellular Uptake. J. Nanosci. Nanotechnol. 15, 9539–9547, https://doi.org/10.1166/jnn.2015.10880 (2015). Shahparast, Y., Eskandani, M., Rajaei, A. & Yari Khosroushahi, A. Preparation, Physicochemical Characterization and Oxidative Stability of Omega-3 Fish Oil/alpha-Tocopherol-co-Loaded Nanostructured Lipidic Carriers. Adv. Pharm. Bull. 9, 393–400, https://doi.org/10.15171/apb.2019.046 (2019). Taha, E. et al. Cod liver oil nano-structured lipid carriers (Cod-NLCs) as a promising platform for nose to brain delivery: Preparation, in vitro optimization, ex vivo cytotoxicity & in vivo biodistribution utilizing radioiodinated zopiclone. Int. J. Pharm. X. 5, 100160, https://doi.org/https://doi.org/10.1016/j.ijpx.2023.100160 (2023). Wang, T. & Luo, Y. Biological fate of ingested lipid-based nanoparticles: current understanding and future directions. Nanoscale. 11, 11048–11063, https://doi.org/10.1039/c9nr03025e (2019). Zhang, X., Chen, G., Zhang, T., Ma, Z. & Wu, B. Effects of PEGylated lipid nanoparticles on the oral absorption of one BCS II drug: a mechanistic investigation. Int. J. Nanomedicine. 9, 5503–5514, https://doi.org/10.2147/ijn.S73340 (2014). Li, H., Guissi, N. E. I., Su, Z., Ping, Q. & Sun, M. Effects of surface hydrophilic properties of PEG-based mucus-penetrating nanostructured lipid carriers on oral drug delivery. RSC Advances. 6, 84164–84176, https://doi.org/10.1039/c6ra18724b (2016). Qin, J. J., Wang, W., Sarkar, S. & Zhang, R. Oral delivery of anti-MDM2 inhibitor SP141-loaded FcRn-targeted nanoparticles to treat breast cancer and metastasis. J. Control Release. 237, 101–114, https://doi.org/10.1016/j.jconrel.2016.07.008 (2016). Kou, L. et al. Transporter-Guided Delivery of Nanoparticles to Improve Drug Permeation across Cellular Barriers and Drug Exposure to Selective Cell Types. Front. Pharmacol. 9, 27, https://doi.org/10.3389/fphar.2018.00027 (2018). Krishna, K. V. et al. Design and Biological Evaluation of Lipoprotein-Based Donepezil Nanocarrier for Enhanced Brain Uptake through Oral Delivery. ACS Chem. Neurosci. 10, 4124–4135, https://doi.org/10.1021/acschemneuro.9b00343 (2019). Tunki, L. et al. Modulating the site-specific oral delivery of sorafenib using sugar-grafted nanoparticles for hepatocellular carcinoma treatment. Eur. J. Pharm. Sci. 137, 104978, https://doi.org/10.1016/j.ejps.2019.104978 (2019). Banu, V. & Rajendran, S. Corrosion inhibition of carbon steel by tween 20 self-assembling monolayers. J. Chem. Pharm. Res. 7, 127–132, (2015). Kiefer, J. et al. Vibrational structure of the polyunsaturated fatty acids eicosapentaenoic acid and arachidonic acid studied by infrared spectroscopy. J. Mol. Struct. 965, 121–124, https://doi.org/10.1016/j.molstruc.2009.11.052 (2010). Gholami, H., Yeganeh, H., Burujeny, S. B., Sorayya, M. & Shams, E. Vegetable Oil Based Polyurethane Containing 1,2,3-Triazolium Functional Groups as Antimicrobial Wound Dressing. J. Polym. Environ. 26, 462–473, https://doi.org/10.1007/s10924-017-0964-y (2018). Raeisi, S., Ojagh, S. M., Quek, S. Y., Pourashouri, P. & Salaün, F. Nano-encapsulation of fish oil and garlic essential oil by a novel composition of wall material: Persian gum-chitosan. LWT. 116, 108494, https://doi.org/https://doi.org/10.1016/j.lwt.2019.108494 (2019). Takeungwongtrakul, S., Karnjanapratum, S., Kaewthong, P., Nalinanon, S. Change in fatty acid profile, volatile compounds and FTIR spectra of samrong seed oil during storage. Int. J. Agric. Technol. 16, 475–484, (2020). Md Salim, R., Asik, J. & Sarjadi, M. S. Chemical functional groups of extractives, cellulose and lignin extracted from native Leucaena leucocephala bark. Wood Sci. Technol. 55, 295–313, https://doi.org/10.1007/s00226-020-01258-2 (2021). Ng W, S., Lee, C. S., Cheng, S. F., Chuah, C. H. & Wong, S. F. Biocompatible Polyurethane Scaffolds Prepared from Glycerol Monostearate-Derived Polyester Polyol. J. Polym. Environ. 26, 2881–2900, https://doi.org/10.1007/s10924-017-1175-2 (2018). Caldas, A. R. et al. Omega-3- and Resveratrol-Loaded Lipid Nanosystems for Potential Use as Topical Formulations in Autoimmune, Inflammatory, and Cancerous Skin Diseases. Pharmaceutics. 13, 1202, https://doi.org/10.3390/pharmaceutics13081202 (2021). Mudalige, T. et al. in Nanomaterials for Food Applications (eds Amparo López Rubio, Maria José Fabra Rovira, Marta martínez Sanz, & Laura Gómez Gómez-Mascaraque) 313–353 (Elsevier, 2019). Šimat, V. et al. Production and Refinement of Omega-3 Rich Oils from Processing By-Products of Farmed Fish Species. Foods. 8, 125, https://doi.org/10.3390/foods8040125 (2019). Sreedhar, R., Kumar, V. S., Bhaskaran Pillai, A. K. & Mangalathillam, S. Omega-3 Fatty Acid Based Nanolipid Formulation of Atorvastatin for Treating Hyperlipidemia. Adv. Pharm. Bull. 9, 271–280, https://doi.org/10.15171/apb.2019.031 (2019). Álvarez-Benedicto, E. et al. Optimization of phospholipid chemistry for improved lipid nanoparticle (LNP) delivery of messenger RNA (mRNA). Biomater Sci. 10, 549–559, https://doi.org/10.1039/D1BM01454D (2022). Sathivel, S. Thermal and flow properties of oils from salmon head. J. Am. Oil Chem. Soc. 82, 147–152, https://doi.org/10.1007/s11746-005-1057-6 (2005). Shi, T. Q., Wang, L. R., Zhang, Z. X., Sun, X. M. & Huang, H. Stresses as First-Line Tools for Enhancing Lipid and Carotenoid Production in Microalgae. Front. Bioeng. Biotechnol. 8, 610, https://doi.org/10.3389/fbioe.2020.00610 (2020). Sehl, A. et al. How do algae oils change the omega-3 polyunsaturated fatty acids market?. OCL. 29, 20. https://doi.orFg/10.1051/ocl/2022018 (2022). Tengku Mohamad, T. R. & Birch, E. Physicochemical characterisation and oxidative stability of refined hoki oil, unrefined hoki oil and unrefined tuna oil. Int. J. Food Sci. Technol. 48, 2331–2339, https://doi.org/10.1111/ijfs.12222 (2013). Gaweł, S., Wardas, M., Niedworok, E., Wardas, P., 2004. Dialdehyd malonowy (MDA) jako wskaźnik procesów peroksydacji lipidów w organizmie [Malondialdehyde (MDA) as a lipid peroxidation marker]. Wiad. Lek. 57, 453–455. Nogueira, M., Scolaro, B., Milne, G. & Castro, I. Oxidation products from omega-3 and omega-6 fatty acids during a simulated shelf life of edible oils. LWT. 101, 113–122, https://doi.org/10.1016/j.lwt.2018.11.044 (2018). Ismail, A., Bannenberg, G., Rice, H. B., Schutt, E. & MacKay, D. Oxidation in EPA- and DHA-rich oils: an overview. Lipid Technol. 28, 55–59, https://doi.org/https://doi.org/10.1002/lite.201600013 (2016). Yin H, Purification of fish oils and production of protein powders with EPA and DHA enriched fish oils [Doctoral Dissertations]. [Louisiana, U.S.]. Louisiana State University; 2011. Barros, D., Reed, P., Alves, M., Santos, R. & Oliva, A. Biocompatibility and Antimicrobial Activity of Nanostructured Lipid Carriers for Topical Applications Are Affected by Type of Oils Used in Their Composition. Pharmaceutics . 13, 1950, https://doi.org/10.3390/pharmaceutics13111950 (2021). Lacatusu, I. et al. Ivy leaves extract based – lipid nanocarriers and their bioefficacy on antioxidant and antitumor activities. RSC Advances. 6, 77243–77255, https://doi.org/10.1039/c6ra12016d (2016). Nejadmansouri, M., Hosseini, S. M. H., Niakousari, M. & Yousefi, G. Physicochemical properties and storage stability of ultrasound-mediated WPI-stabilized fish oil nanoemulsions. Food Hydrocoll. 61, 801–811, https://doi.org/10.1016/j.foodhyd.2016.07.011 (2016). Zaky, M. F. et al. Influence of Surface-Modification via PEGylation or Chitosanization of Lipidic Nanocarriers on In Vivo Pharmacokinetic/Pharmacodynamic Profiles of Apixaban. Pharmaceutics. 15, 1668, https://doi.org/10.3390/pharmaceutics15061668 (2023). Khosa, A., Reddi, S. & Saha, R. N. Nanostructured lipid carriers for site-specific drug delivery. Biomed. Pharmacother. 103, 598–613, https://doi.org/10.1016/j.biopha.2018.04.055 (2018). Terada, T., Kulkarni, J. A., Huynh, A., Tam, Y. Y. C. & Cullis, P. Protective Effect of Edaravone against Cationic Lipid-Mediated Oxidative Stress and Apoptosis. Biol. Pharm. Bull. 44, 144–149, https://doi.org/10.1248/bpb.b20-00679 (2021). Meulewaeter, S. et al. Continuous freeze-drying of messenger RNA lipid nanoparticles enables storage at higher temperatures. J. Control. Release. 357, 149–160, https://doi.org/10.1016/j.jconrel.2023.03.039 (2023). Tang, C. H., Chen, H. L. & Dong, J. R. Solid Lipid Nanoparticles (SLNs) and Nanostructured Lipid Carriers (NLCs) as Food-Grade Nanovehicles for Hydrophobic Nutraceuticals or Bioactives. Appl. Sci. 13, 1726, https://doi.org/10.3390/app13031726 (2023). Shah, N. V. et al. Nanostructured lipid carriers for oral bioavailability enhancement of raloxifene: Design and in vivo study. J. Adv. Res. 7, 423–434, https://doi.org/10.1016/j.jare.2016.03.002 (2016). Fadda, A. et al. Innovative and Sustainable Technologies to Enhance the Oxidative Stability of Vegetable Oils. Sustainability. 14, 849, https://doi.org/10.3390/su14020849 (2022). Musakhanian, J., Rodier, J. D. & Dave, M. Oxidative Stability in Lipid Formulations: a Review of the Mechanisms, Drivers, and Inhibitors of Oxidation. AAPS PharmSciTech. 23, 151, https://doi.org/10.1208/s12249-022-02282-0 (2022). Wang, C. et al. Emulsion structure design for improving the oxidative stability of polyunsaturated fatty acids. Compr. Rev. Food Sci. Food Saf. 19, 2955–2971, https://doi.org/10.1111/1541-4337.12621 (2020). Ling, S. S., Yuen, K. H., Magosso, E. & Barker, S. A. Oral bioavailability enhancement of a hydrophilic drug delivered via folic acid-coupled liposomes in rats. J. Pharm. Pharmacol. 61, 445–449, https://doi.org/10.1211/jpp/61.04.0005 (2009). Kou, L. et al. Cotransporting Ion is a Trigger for Cellular Endocytosis of Transporter-Targeting Nanoparticles: A Case Study of High-Efficiency SLC22A5 (OCTN2)-Mediated Carnitine-Conjugated Nanoparticles for Oral Delivery of Therapeutic Drugs. Adv. Healthc. Mater. 6, https://doi.org/10.1002/adhm.201700165 (2017). Tian, C. et al. Improving intestinal absorption and oral bioavailability of curcumin via taurocholic acid-modified nanostructured lipid carriers. Int. J. Nanomedicine. 12, 7897–7911, https://doi.org/10.2147/IJN.S145988 (2017). Tian, C. et al. N-acetyl-L-cysteine functionalized nanostructured lipid carrier for improving oral bioavailability of curcumin: preparation, in vitro and in vivo evaluations. Drug Deliv. 24, 1605–1616, https://doi.org/10.1080/10717544.2017.1391890 (2017). Guillén, M. D. & Cabo, N. Infrared spectroscopy in the study of edible oils and fats. J. Sci. Food Agric . 75, 1–11, https://doi.org/https://doi.org/10.1002 /(SICI)1097-0010(199709)75:13.0.CO;2-R (1997). Averina, E., Müller, R., Popov, D. & Radnaeva, L. Physical and chemical stability of nanostructured lipid drug carriers (NLC) based on natural lipids from Baikal region (Siberia, Russia). Pharmazie . 66, 348–356, https://doi.org/10.1691/ph.2011.0326 (2011). Pornputtapitak, W. et al. Effect of Oil Content on Physiochemical Characteristics of gamma-Oryzanol-Loaded Nanostructured Lipid Carriers. J. Oleo. Sci. 68, 699–707, https://doi.org/10.5650/jos.ess18127 (2019). Saedi, A., Rostamizadeh, K., Parsa, M., Dalali, N. & Ahmadi, N. Preparation and characterization of nanostructured lipid carriers as drug delivery system: Influence of liquid lipid types on loading and cytotoxicity. Chem. Phys. Lipids. 216, 65–72, https://doi.org/10.1016/j.chemphyslip.2018.09.007 (2018). Lúcio, M. et al. Nanostructured Lipid Carriers Enriched Hydrogels for Skin Topical Administration of Quercetin and Omega-3 Fatty Acid. Pharmaceutics . 15, 2078, https://doi.org/10.3390/pharmaceutics15082078 (2023). Cameron-Smith, D., Albert, B. B. & Cutfield, W. S. Fishing for answers: is oxidation of fish oil supplements a problem? J. Nutr. Sci. 4, e36, https://doi.org/10.1017/jns.2015.26 (2015). Awad, T. S., Helgason, T., Weiss, J., Decker, E. A. & McClements, D. J. Effect of Omega-3 Fatty Acids on Crystallization, Polymorphic Transformation and Stability of Tripalmitin Solid Lipid Nanoparticle Suspensions. Cryst. Growth Des. 9, 3405–3411, https://doi.org/10.1021/cg8011684 (2009). Helgason, T., Awad, T. S., Kristbergsson, K., McClements, D. J. & Weiss, J. Influence of Polymorphic Transformations on Gelation of Tripalmitin Solid Lipid Nanoparticle Suspensions. J. Am. Oil Chem. Soc. 85, 501–511, https://doi.org/https://doi.org/10.1007/s11746-008-1219-9 (2008). Surassmo, S., Ruktanonchai, U., Srinuanchai, W. & Yostawonkul, J. Formulation development of plain nanoemulsion based on the influence of surfactant combinations. Chiang Mai J. Sci. 40, 994–999, (2012). Oomah, B. D. Flaxseed as a functional food source. J. Sci. Food Agric. 81, 889–894, https://doi.org/10.1002/jsfa.898 (2001). Akhoond Zardini, A., Mohebbi, M., Farhoosh, R. & Bolurian, S. Production and characterization of nanostructured lipid carriers and solid lipid nanoparticles containing lycopene for food fortification. J. Food Sci. Technol. 55, 287–298, https://doi.org/10.1007/s13197-017-2937-5 (2018). Luo, Q. et al. Ascorbate-conjugated nanoparticles for promoted oral delivery of therapeutic drugs via sodium-dependent vitamin C transporter 1 (SVCT1). Artif. Cells Nanomed. Biotechnol. 46, 198–208, https://doi.org/10.1080/21691401.2017.1417864 (2018). Kannan, K. Compatibility studies of camptothecin with various pharmaceutical excipients used in the development of nanoparticle formulation. Int. J. Pharm. Pharm. Sci. 5, 315–321, (2013). Pajić, N. Z. B. et al. Alkyl polyglucoside vs. ethoxylated surfactant-based microemulsions as vehicles for two poorly water-soluble drugs: physicochemical characterization and in vivo skin performance. Acta Pharm. 67, 415–439, https://doi.org/doi:10.1515/acph-2017-0036 (2017). Aryani, D., Siswodihardjo, S., Soeratri, W., Rahmasari, F. & Sari, D. Experimental development and molecular docking: nanostructured lipid carriers (nlcs) of coenzyme q10 using stearic acid and different liquid lipids as lipid matrix. Int. J. Appl. Pharm. 13, 108–116, https://doi.org/10.22159/ijap.2021v13i1.39890 (2021). Pezeshki, A., Ghanbarzadeh, B., Mohammadi, M., Fathollahi, I. & Hamishehkar, H. Encapsulation of Vitamin A Palmitate in Nanostructured Lipid Carrier (NLC)-Effect of Surfactant Concentration on the Formulation Properties. Adv. Pharm. Bull. 4, 563–568, https://doi.org/10.5681/apb.2014.083 (2014). Palmquist, D. L. & Jenkins, T. C. Challenges with fats and fatty acid methods. J. Anim. Sci. 81, 3250–3254, https://doi.org/10.2527/2003.81123250x (2003). Chang, C. & Nickerson, M. Stability and in vitro release behaviour of encapsulated omega fatty acid-rich oils in lentil protein isolate-based microcapsules. Int. J. Food Sci. Nutr. 69, 1–12, https://doi.org/10.1080/09637486.2017.1336513 (2017). Komaiko, J., Sastrosubroto, A. & McClements, D. Encapsulation of ω-3 fatty acids in nanoemulsion-based delivery systems fabricated from natural emulsifiers: Sunflower phospholipids. Food Chem. 203, 331–339, https://doi.org/10.1016/j.foodchem.2016.02.080 (2016). Wangngae, S. et al. Lipid-Based Nanoparticles as Targeted Delivery System in Korat Chicken. ACS Food Sci. Technol. 3, 1913–1919 https://doi.org/10.1021/acsfoodscitech.3c00275 (2023). Cohen, M. et al. Live imaging of GLUT2 glucose-dependent trafficking and its inhibition in polarized epithelial cysts. Open Biol. 4, https://doi.org/10.1098/rsob.140091 (2014). Poulsen, S. B., Fenton, R. A. & Rieg, T. Sodium-glucose cotransport. Curr Opin Nephrol Hypertens. 24, 463–469, https://doi.org/10.1097/MNH.0000000000000152 (2015). Byers, M. S., Howard, C. & Wang, X. Avian and Mammalian Facilitative Glucose Transporters. Microarrays (Basel) . 6, https://doi.org/10.3390/microarrays6020007 (2017). Siu, F. Y., Ye, S., Lin, H. & Li, S. Galactosylated PLGA nanoparticles for the oral delivery of resveratrol: enhanced bioavailability and in vitro anti-inflammatory activity. Int. J. Nanomedicine. 13, 4133–4144, https://doi.org/10.2147/IJN.S164235 (2018). Yeh, Y. C., Kim, S. T., Tang, R., Yan, B. & Rotello, V. M. Insulin-based regulation of glucose-functionalized nanoparticle uptake in muscle cells. J. Mater. Chem. B. 2, 4610–4614, https://doi.org/10.1039/C4TB00608A (2014). Grefner, N. M., Gromova, L. V., Gruzdkov, A. A. & Komissarchik, Y. Y. Interaction of glucose transporters SGLT1 and GLUT2 with cytoskeleton in enterocytes and Caco2 cells during hexose absorption. Cell Tissue Biol. 9, 45–52, https://doi.org/10.1134/s1990519x15010034 (2015). Settembre, C. & Ballabio, A. Lysosome: regulator of lipid degradation pathways. Trends Cell Biol. 24, 743–750, https://doi.org/10.1016/j.tcb.2014.06.006 (2014). Zhang, R., Qin, X., Kong, F., Chen, P. & Pan, G. Improving cellular uptake of therapeutic entities through interaction with components of cell membrane. Drug Deliv. 26, 328–342, https://doi.org/10.1080/10717544.2019.1582730 (2019). Chen, J. et al. Glucosamine derivative modified nanostructured lipid carriers for targeted tumor delivery. J. Mater. Chem. 22, 5770–5783, https://doi.org/10.1039/c2jm15830b (2012). Yostawonkul, J. et al. Nanocarrier-mediated delivery of α-mangostin for non-surgical castration of male animals. Sci. Rep. 7, 16234, https://doi.org/10.1038/s41598-017-16563-3 (2017). Salminen, H., Helgason, T., Kristinsson, B., Kristbergsson, K. & Weiss, J. Formation of solid shell nanoparticles with liquid omega-3 fatty acid core. Food Chem. 141, 2934–2943, https://doi.org/10.1016/j.foodchem.2013.05.120 (2013). Wang, L. J. et al. Fabrication and Characterization of Antioxidant Pickering Emulsions Stabilized by Zein/Chitosan Complex Particles (ZCPs). J. Agric. Food Chem. 63, 2514–2524, https://doi.org/10.1021/jf505227a (2015). Folch, J., Lees, M. & Sloane Stanley, G. H. A simple method for the isolation and purification of total lipides from animal tissues. J. Biol. Chem. 226, 497–509, https://doi.org/10.1016/S0021-9258(18)64849-5 (1957). Metcalfe, L. D., Schmitz, A. A. & Pelka, J. R. Rapid Preparation of Fatty Acid Esters from Lipids for Gas Chromatographic Analysis. Anal. Chem. 38, 514–515, https://doi.org/10.1021/ac60235a044 (1966). Additional Declarations No competing interests reported. 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(NPs).\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-4761693/v1/0f85d54ac6eef472afcbf9cb.png"},{"id":63371603,"identity":"c25b60c8-d116-46de-9609-44257118ce4f","added_by":"auto","created_at":"2024-08-27 11:57:24","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":49143,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectra of nanomaterials (GMS, tuna crude oil, tween 20, poloxamer, alkyl polyglucoside), physical mixture (PM) and lipid-based nanoparticles (NPs and TNPs).\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-4761693/v1/161b1516e9d3952018e4da19.png"},{"id":63370238,"identity":"4693863c-838c-4e4e-8bec-1bdcda00f505","added_by":"auto","created_at":"2024-08-27 11:49:24","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":51955,"visible":true,"origin":"","legend":"\u003cp\u003eNile Red distribution in Pectoralis major muscle of chicken imaging by IVIS of treatment group including non-targeted lipid nanoparticles (NPs) and targeted lipid nanoparticles (TNPs) at 2, 4, 8, 12 and 24 h after oral administration. The data are presented as bar graphs showing means ± SD (n=3). \u003csup\u003e*\u003c/sup\u003e(P\u0026lt;0.05), \u003csup\u003e**\u003c/sup\u003e(P\u0026lt; 0.01) and \u003csup\u003e***\u003c/sup\u003e(P\u0026lt; 0.001).\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-4761693/v1/367ac219296cae49d4343ab2.png"},{"id":63370237,"identity":"3ffd8ab9-68a4-48b1-8cd0-8c1e5ebcf02e","added_by":"auto","created_at":"2024-08-27 11:49:24","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":98249,"visible":true,"origin":"","legend":"\u003cp\u003eLipid nanoparticle preparation\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-4761693/v1/f73e61e1dde4e43872111c44.png"},{"id":98813952,"identity":"5e9beb81-3ce3-44d8-a72f-5ce0de696e18","added_by":"auto","created_at":"2025-12-22 16:08:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2057711,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4761693/v1/0f1c2c80-0d85-4402-a464-9bb261b55fe0.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Enhancement of n-3 PUFAs utilization for functional meat production in slow-growing Korat chicken: evaluation of characteristics of glucose transporter-targeted lipid nanoparticles","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOils containing high amounts of n-3 polyunsaturated fatty acids (n-3 PUFAs) are susceptible to oxidation process [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Many studies have tried to improve the storage stability of these oil sources, and the encapsulation technology was the more efficient [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Another possibility is to use the nanotechnology to improve the utilization of n-3 PUFAs oil source [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. This technology has a high potential, and we investigated its use to enhance the accumulation of n-3 PUFAs in chicken muscles to produce functional meat.\u003c/p\u003e \u003cp\u003eKorat chicken, a slow-growing breed that is very popular in Thailand, has been developed as an alternative option catering to the preferences of Thai chicken producers [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] and could be used to produce functional meat enriched with n-3 PUFAs. Previous studies showed that dietary 4% tuna oil supplementation increased n-3 PUFAs content reaching 19.03% of total fatty acids in Korat chicken meat [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHowever, n-3 PUFAs sources are susceptible to lipid oxidation that can affect the feed quality [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. The protection of these lipids through encapsulation alone is inadequate to enhance the utilization of n-3 PUFAs sources, as a portion of these fatty acids will also be distributed among non-targeted organs such as liver.\u003c/p\u003e \u003cp\u003eDifferent studies showed that lipid nanoparticles could be synthesized by applying the edible oil such as fish oil [\u003cspan additionalcitationids=\"CR9\" citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. However, various nanoparticle size, dispersion ability of particles, zeta-potential (the observation of a robust electric potential at the particle surface boundary demonstrates the existence of a potent repulsive force, which contributes significantly to enhancing the overall physical colloidal stability) and high amount of PUFAs including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) might increase the unstable form of particle size during storage [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] and plain lipid nanoparticles could be degraded by digestive enzymes [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Another important parameter is the ability of nanoparticle surface coating [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Modifying their surface with PEGylating helps to avoid enzyme activity in the digestive tract [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Moreover, generating a surface with a specific substrate recognized with target transporter/receptor could allow the transport of beneficial substance directly to the target organs [\u003cspan additionalcitationids=\"CR15 CR16\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTherefore, we investigated the physicochemical characteristics of lipid nanoparticles and their susceptibility to lipid oxidation during storage and their ability to transfer the bioactive compounds (n-3 PUFAs) into the muscles. For feeding animals, the important point to consider is that lipid nanoparticles need to transfer perfectly to the target organ after the passage through the digestive tract and adsorptive cells that depends on the stability of lipid nanoparticles.\u003c/p\u003e \u003cp\u003eThe aim of our study was to analyze the effects of n-3 PUFAs source, targeting process, and storage temperature and duration on the physicochemical characteristics of lipid nanoparticles, their oxidation susceptibility, and their ability to transfer n-3 PUFAs to chicken meat.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePhysicochemical characteristics of lipid-based nanoparticles containing n-3 PUFAs oils\u003c/h2\u003e \u003cp\u003eThe physical characterization of nanoparticles containing n-3 PUFAs oils (Tuna oil, TO and Algal oil, AO) are presented on Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The mean particle diameter of the NPs groups was higher than that of the TNPs groups (P\u0026thinsp;\u0026le;\u0026thinsp;0.001) whereas tuna oil in both nanoparticle forms had smaller particle diameters than algal oil (P\u0026thinsp;\u0026le;\u0026thinsp;0.001). The TO_TNPs group had the highest PDI value (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and the NPs groups had higher zeta-potential values than the TNPs groups (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and the value was highest in AO_NPs group.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhysical characteristic of n-3 PUFA oils within different type of lipid-nanoparticles\u003csup\u003e1\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAverage diameter (nm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePolydispersity index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eZeta-potential (mV)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e223.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.01\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.315\u0026thinsp;\u0026plusmn;\u0026thinsp;0.014\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-42.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e294.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.83\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.261\u0026thinsp;\u0026plusmn;\u0026thinsp;0.007\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-34.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_TNPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e134.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.365\u0026thinsp;\u0026plusmn;\u0026thinsp;0.008\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-48.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.68\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_TNPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e184.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.21\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.298\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-49.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026le;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026le;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026le;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003e Two different n-3 PUFA oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles.\u003c/p\u003e \u003cp\u003eData are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE (n\u0026thinsp;=\u0026thinsp;3). \u003csup\u003ea\u0026minus;c\u003c/sup\u003e Values in the same column with different superscripts letters are significantly different (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eThermal and chemical properties of lipid-based nanoparticles\u003c/h3\u003e\n\u003cp\u003eThe melting point of glyceryl monostearate (GMS) a raw material used for nanoparticle synthesis was 76.6\u003csup\u003eo\u003c/sup\u003eC, whereas the nanoparticle suspension including TO_NPs, AO_NPs, TO_TNPs and AO_TNPs showed lower endothermic peaks at 46.8, 46.4, 45.3 and 45.0\u003csup\u003eo\u003c/sup\u003eC, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Both the n-3 PUFA sources and nanoparticle types changed the melting point in nanoparticle form by lipid nanoparticles composite of algal oil and the targeting groups had the lowest melting points of lipid nanoparticles.\u003c/p\u003e \u003cp\u003eThe Fourier-transformed infrared spectroscopy (FTIR) analysis of nanomaterials and lipid-based nanoparticles is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The peak intensity of lipid nanoparticles was more flatten than individual spectra of each material. It can be concluded that tuna crude oil was packed tightly inside the nanoparticles. Due to functional group which is composite in the structure of lipid, triglyceride and fatty acid that could be found in tween 20, alkyl polyglucoside, GMS and tuna oil, the peak intensity decreased after the synthesis into lipid-based nanoparticles.\u003c/p\u003e \u003cp\u003eThe FTIR spectra show in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, The bands related to hydroxyl group (3499\u0026thinsp;\u0026minus;\u0026thinsp;3233 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003csub\u003e,\u003c/sub\u003e -OH stretching) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. was found in tween 20, poloxamer, alkyl polyglucoside and GMS. For lipid (2921\u0026thinsp;\u0026minus;\u0026thinsp;2850 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, asymmetric and symmetric stretching vibration of CH\u003csub\u003e2\u003c/sub\u003e groups in lipid alkyl chain, ν\u003csub\u003easym\u003c/sub\u003e CH\u003csub\u003e2\u003c/sub\u003e and ν\u003csub\u003esym\u003c/sub\u003e CH\u003csub\u003e2\u003c/sub\u003e) [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] and fatty acid (1641 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, C\u0026thinsp;=\u0026thinsp;C stretching, 1470\u0026ndash;1460 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, -CH\u003csub\u003e3\u003c/sub\u003e bending and 1240 \u0026minus;\u0026thinsp;1207 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, -CH\u003csub\u003e2\u003c/sub\u003e bending) [\u003cspan additionalcitationids=\"CR21\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] were found in tween 20, alkyl polyglucoside, and tuna oil. The ester linkages (1741\u0026thinsp;\u0026minus;\u0026thinsp;1734 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, ester carbonyl group (-C\u0026thinsp;=\u0026thinsp;O)) [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] was found in GMS, tween 20 and triglyceride in tuna oil but not occurred in alkyl polyglucoside and poloxamer. The cyclic ether (1029 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, cyclic ether (-COC) and pyranose ring (1150 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, -COC glycosidic bond) [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] were found in alkyl polyglucoside. The wavelength at 720 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicated that this long chain carbon [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] was found in GMS, tween 20, alkyl polyglucoside, and tuna oil. In addition, the wavelength ranging at 2921\u0026thinsp;\u0026minus;\u0026thinsp;2850 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-CH stretching) and 1500\u0026thinsp;\u0026minus;\u0026thinsp;500 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-CH bending and long chain carbon) had a reduced intensity in physical mixture and synthesized of lipid nanoparticles.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e\n\u003ch3\u003eIn vitro storage stability of n-3 PUFA lipid-based nanoparticles\u003c/h3\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eParticle size stability\u003c/h2\u003e \u003cp\u003eThe results of the storage stability on particle size of the n-3 PUFA lipid-based nanoparticles are shown on Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The synthesis of n-3 PUFA source oils in the form of targeted-lipid nanoparticles groups (TO_TNPs and AO_TNPs) can help to maintain the size of the nanoparticles, but the storage conditions were better at low temperature storage (4\u0026deg;C) for 4 weeks, while the particle size of the non-targeted lipid nanoparticles groups changed at a faster rate at all temperature storage (TO_NPs_4\u0026deg;C, TO_NPs_RT, AO_NPs_4\u0026deg;C and AO_NPs_RT).\u003c/p\u003e \u003cp\u003eHowever, during the storage, there were statistically significant changes in nanoparticle size for the AO_NPs_4\u0026deg;C (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), AO_NPs_RT (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01), TO_TNPs_RT (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), TO_TNPs_RT (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and AO_TNPs_RT (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003eDuring the storage, the change in nanoparticle size was faster at room temperature than at 4\u0026deg;C. In addition, the use of algal oil resulted in changes particle size faster in lipid-based nanoparticles (AO_NPs_4\u0026deg;C, AO_NPs_RT, and AO_TNPs_RT).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eStability of n-3 PUFA oils within different type of lipid-nanoparticles\u003csup\u003e1\u003c/sup\u003e on particle diameter during storage at room temperature (RT) or 4\u0026deg;C.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003eParticle diameter (nm)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStart\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eWeek 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eWeek 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWeek 3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eWeek 4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_NPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e225.23\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e215.83\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e233.37\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e197.03\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e230.02\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e4.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_NPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e223.50\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e233.83\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e258.83\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e258.8\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e268.88\u003csup\u003edB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e6.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_NPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e303.53\u003csup\u003edAB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e328.93\u003csup\u003eeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e316.87\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e300.4\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e320.03\u003csup\u003eeA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_NPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e299.60\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e332.17\u003csup\u003eeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e341.83\u003csup\u003edB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e358.53\u003csup\u003eeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e344.78\u003csup\u003efB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e5.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_TNPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e136.50\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e137.9\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e139.67\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e142.73\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e139.4\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.853\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_TNPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e134.00\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e137.47\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e131.23\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e136.5\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e140.82\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_TNPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e176.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e183.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e177.57\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e178.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e183.51\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.312\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_TNPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e176.4\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e187.53\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e194.00\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e192.13\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e195.09\u003csup\u003ebC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e15.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e15.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003e Two different n-3 PUFA oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles.\u003c/p\u003e \u003cp\u003eData are the average\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM) of three independent replicates (n\u0026thinsp;=\u0026thinsp;3). For each attribute, different small letters indicate statistically significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), highly significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) and very highly significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) between different samples at a same time of storage. Different capital letters in a same row indicate very highly significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) over time.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eOxidative stability evaluated with peroxidative value\u003c/h2\u003e \u003cp\u003eThe peroxidative values are presented in the Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The unencapsulated tuna oil (TO_4\u0026deg;C) and tuna oil within lipid nanoparticles were stored at 4\u0026deg;C (TO_NPs_4\u0026deg;C and TO_TNPs_4\u0026deg;C) and had reduced peroxidative values ​​(P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) compared to the groups kept at room temperature except at the start of the experiment where the values ​​were not different. Moreover, no statistically significant differences in peroxide production in unencapsulated algal oil were found between the two storage temperatures (AO_4\u0026deg;C and AO_RT) from week 1 to week 4.\u003c/p\u003e \u003cp\u003eIn addition, algal oil within lipid nanoparticles (AO_NPs_4\u0026deg;C, AO_NPs_RT, AO_TNPs_4\u0026deg;C and AO_TNPs_RT at first day) showed a statistically highly significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) increase in peroxide value after synthesis.\u003c/p\u003e \u003cp\u003eThe synthesis of oil within nanoparticle form and storage at suitable temperature influenced the peroxidative value reaction. Pre-synthetic and post-synthetic oils in nanoparticle form kept at low temperature (4\u0026deg;C) showed a statistically significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) slowdown of peroxidative activity by comparison with the storage at room temperature from week 1 to week 3.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eOxidative stability evaluated with TBARS value\u003c/h2\u003e \u003cp\u003eThe malondialdehyde (MDA) values are presented on the Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Synthesizing tuna oil into the targeted-lipid nanoparticle increased the oxidation stability as the amount of MDA was lower in the non-targeted lipid nanoparticles but for algal oil, the reverse was observed. Moreover, storage at low temperature (4\u0026deg;C) slow down the production rate of MDA. The synthesized algal oil within lipid nanoparticles in the experimental groups (AO_NPs_4\u0026deg;C, AO_NPs_RT, AO_TNPs_4\u0026deg;C and AO_TNPs_RT) had a greater increase in MDA than the unsynthesized algal oil (AO_4\u0026deg;C and AO_RT) (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eStability of n-3 PUFA oil sources within different type of lipid-nanoparticles\u003csup\u003e1\u003c/sup\u003e on peroxidative value during a storage at room temperature (RT) or 4\u0026deg;C.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003ePeroxidative value (meq of oxygen/kg fat)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStart\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eWeek 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eWeek 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWeek 3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eWeek 4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.106\u003csup\u003edB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.110\u003csup\u003ecdB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.113\u003csup\u003ecdB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.079\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.184\u003csup\u003ecC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.009\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.113\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.160\u003csup\u003efB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.213\u003csup\u003eiC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.153\u003csup\u003eeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.306\u003csup\u003edD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.018\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.033\u003csup\u003eaC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.022\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.018\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.026\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.017\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.002\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.032\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.031\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.032\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.041\u003csup\u003eabB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.035\u003csup\u003eaAB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_NPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.094\u003csup\u003ecAB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.108\u003csup\u003ecBC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.129\u003csup\u003edeD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.119\u003csup\u003edCD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.085\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_NPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.095\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.130\u003csup\u003edeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.196\u003csup\u003ehiC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.211\u003csup\u003efC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.112\u003csup\u003ebAB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_NPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.113\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.153\u003csup\u003efB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.161\u003csup\u003efgB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.164\u003csup\u003eeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.161\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_NPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.111\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.210\u003csup\u003egCD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.181\u003csup\u003eghBC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.242\u003csup\u003egD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.159\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_TNPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.059\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.064\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.064\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.062\u003csup\u003ebcB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.039\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_TNPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.063\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.116\u003csup\u003ecdC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.097\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.107\u003csup\u003edBC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.102\u003csup\u003ebBC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_TNPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.107\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.099\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.127\u003csup\u003edB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.108\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.103\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_TNPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.108\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.140\u003csup\u003eefB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.149\u003csup\u003eefBC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.164\u003csup\u003eeC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.114\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.009\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eTwo different n-3 oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles.\u003c/p\u003e \u003cp\u003eData are the average\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM) of three independent replicates (n\u0026thinsp;=\u0026thinsp;3). For each attribute, different small letters indicate very highly significant over time (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) between different samples at a same time of storage. Different capital letters in a same row indicate highly significant differences over time (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eStability of n-3 PUFA oil sources within different type of lipid-nanoparticles\u003csup\u003e1\u003c/sup\u003e on malondialdehyde value during storage at room temperature (RT) or 4\u0026deg;C.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003eMalondialdehyde value (\u0026micro;mol MDA/kg oil)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStart\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eWeek 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eWeek 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWeek 3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eWeek 4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.01\u003csup\u003eeC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.81\u003csup\u003edB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.95\u003csup\u003ebcA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.89\u003csup\u003edC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.32\u003csup\u003edeC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e13.76\u003csup\u003eeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.08\u003csup\u003eefB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.97\u003csup\u003efC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.53\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17.9\u003csup\u003efC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.23\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.53\u003csup\u003eaC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.25\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.70\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.15\u003csup\u003eaD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.23\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.36\u003csup\u003eaC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.60\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.40\u003csup\u003eabAB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.82\u003csup\u003eaD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_NPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.61\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.88\u003csup\u003egC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.41\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.92\u003csup\u003edB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e15.65\u003csup\u003eefB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_NPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.61\u003csup\u003edA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22.42\u003csup\u003eiD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.38\u003csup\u003edB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e15.72\u003csup\u003edC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e21.73\u003csup\u003egD\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_NPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.21\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.38\u003csup\u003efC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.10\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.75\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.51\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_NPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.88\u003csup\u003ecA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21.06\u003csup\u003ehC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.65\u003csup\u003egC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.75\u003csup\u003eeC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.55\u003csup\u003edeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_TNPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e13.70\u003csup\u003eeC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.03\u003csup\u003eeC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.42\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.58\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.07\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTO_TNPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e13.64\u003csup\u003eeA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.58\u003csup\u003efA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.70\u003csup\u003eeA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.74\u003csup\u003eeB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e21.68\u003csup\u003egB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_TNPs_4\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.54\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.31\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.92\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.00\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.64\u003csup\u003ebC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAO_TNPs_RT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.54\u003csup\u003ebA\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.89\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.15\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.61\u003csup\u003ecB\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11.4c\u003csup\u003edC\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003e Two different n-3 oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles.\u003c/p\u003e \u003cp\u003eData are the average\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM) of three independent replicates (n\u0026thinsp;=\u0026thinsp;3). For each attribute, different small letters indicate very highly significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) between different samples at a same time of storage. Different capital letters in a same row indicate very highly significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) over time.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eTransfer of lipid nanoparticles to the target organ\u003c/h2\u003e \u003cp\u003eThe Nile red distribution in Pectoralis major sample is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The Nile red distribution was increased within targeted-lipid nanoparticles (TNPs) at 8 and 12 h after oral gavage by comparison with non-targeted lipid nanoparticles (NPs). Then it decreased to the initial content at 24 h. The results demonstrated that the targeted lipid nanoparticles had the potential to be absorbed into the bloodstream and transferred to the target organ faster than other forms of nanoparticles.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eFatty acid composition of breast meat of Korat chicken\u003c/h2\u003e \u003cp\u003eThe fatty acid profile of breast meat of the chickens after oral gavage with the lipid-based nanoparticles is shown on Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. C18:3n3 content in breast meat of TNPs treatment at 4 h was decreased (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) than that in the NPs treatment. NPs treatment at 4, 8 and 12 equal to control (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) whereas TNPs treatment at all time point lower than control (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The EPA content in TNPs group at 4 h was higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) than that in the NPs treatment. The DHA content in TNPs treatment at 4 h was higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) than that in the NPs. By the way, the highest of DHA content was showed in the TNPs group at 24 h. No significant different in EPA contents at 2, 8, 12 and 24 h, and DHA content at 2, 8 and 12 h compared between NPs and TNPs treatment. C18:2n6 contents no different compared between NPs and TNPs at each time. C20:4n6 contents in TNPs treatment at 24 h higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) than that in NPs treatment. But at 2, 4, 8 and 12 h showed no difference (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). C18:1n9 contents in TNPs treatment at 24 h were lowest (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) compared to that in NPs and control treatment.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eCoefficient of efficacy of lipid-nanoparticles\u003c/h2\u003e \u003cp\u003eThe coefficient of efficacy of targeted and non-targeted lipid nanoparticles are showed on Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e. TNPs had higher potential to transfer EPA and DHA into the chicken skeletal muscle than the NPs groups. The coefficient of efficacy of TNPs was higher than that of the NPs treatment in each time. At 2 h both NPs and TNPs treatments were able to transfer the n-3 PUFA but after that the TNPs treatment showed higher potential to transfer the n-3 PUFA at 4 and 24 h in TNPs treatment. The n-3 PUFA amount at 8 and 12 h after oral administration was decreased. In contrast, the n-3 PUFA content in NPs treatment was reduced at 4 h then increased slightly until 24 h.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFatty acid composition of TO oil within different type of lipid-nanoparticles\u003csup\u003e1\u003c/sup\u003e at 2, 4, 8, 12 and 24 h after oral administration.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003eFatty acid composition (% of total fatty acids)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC16:0\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC18:0\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC18:1n9\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC18:2n6\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eC18:3n3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eC20:4n6\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eEPA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eDHA\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.33\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.94\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.35\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e23.21\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.21\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.71\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 2h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.23\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.01\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003csup\u003eabc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e26.11\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.20\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.63\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 2h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19.52\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.69\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.24\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e24.65\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.27\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.85\u003csup\u003eabc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 4h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.75\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.09\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.32\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e22.47\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.16\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.57\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 4h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18.60\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.18\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.20\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e26.60\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.28\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.46\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 8h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.64\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.35\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.29\u003csup\u003ebcd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e22.02\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.21\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.82\u003csup\u003eabc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 8h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.16\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.83\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003csup\u003eabc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25.23\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.25\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.40\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 12h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25.89\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.27\u003csup\u003ebcd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e18.75\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.20\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.78\u003csup\u003eabc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 12h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e23.09\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.85\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.23\u003csup\u003eabc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e22.38\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.23\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.22\u003csup\u003ebcd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 24h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19.75\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.21\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.20\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e27.77\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.23\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.55\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 24h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.49\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.50\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.12\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e35.62\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.29\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.43\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.203\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.058\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003e Tuna oil (TO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles.\u003c/p\u003e \u003cp\u003eStatistical analysis is based on F-test. Data are the average\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM) of three independent replicates (n\u0026thinsp;=\u0026thinsp;3). For each attribute, different small letters indicate statistically significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), highly significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) and very highly significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCoefficient of transfer efficacy of TO within different type of lipid-nanoparticles\u003csup\u003e1\u003c/sup\u003e at 2, 4, 8, 12 and 24 h after oral administration relative to control.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eCoefficient of efficacy (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEPA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDHA\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.002\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.013\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 2h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.76\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 2h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 4h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-1.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e-1.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 4h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 8h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.64\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 8h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.93\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 12h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 12h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6.52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNPs at 24h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNPs at 24h\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16.98\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003e Tuna oil (TO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles.\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eBy comparison with AO nanoparticles, TO nanoparticles had lower diameter, higher PD index and lower ZP. AO contains a high amount of long-chain unsaturated fatty acids such as DHA. The fatty acid can move to the surface of the nanoparticles, resulting in an enlarged particle size [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. It seems to be indicated the n-3 PUFA oil can change the uniformity of the nanoparticle diameter. Moreover, the PD index involved their particle size population. As is obviously in AO nanoparticles lower PD index than TO nanoparticles. A larger PD index value indicated a broad particle size distribution of the formulation whereas a low value of the PD index indicated monodisperse samples [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. This implies that AO can maintain the uniformity of particle size better than TO. In reference to the quality of TO contains phospholipids [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], this compound had potential to lesser particle size [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] meanwhile can increase zeta-potential value with their negative charged [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTO and AO was applied in this present study is unrefined oil and refined oil, respectively. Unrefined TO contains impurities such as phospholipid, free fatty acid, aldehyde, ketones and pigment [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] that rapidly activated by pro-oxidant effect more than refined AO result to higher oxidized of TO at the beginning of experiment whereas algal oil could be better potent lipid peroxidation by greater quality, lower oxidative rate and might be prevented lipid oxidation by the carotenoids activity [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Thus, the lipid oxidative products were higher for TO than the AO at start of experiment.\u003c/p\u003e \u003cp\u003eOn the reverse, TO_NPs had lower peroxidative values than AO_NPs and TO_TNPs had lower peroxidative values than AO_TNPs. In contrast, the MDA content of TO_NPs and TO_TNPs was higher than that of AO_NPs and AO_TNPs. The possibility of the increase of peroxide value (PV) due to the sources had high free fatty acid induce lipid peroxidation stage [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. AO had higher PV after lipid nanoparticle synthesis indicated that the hydroperoxides was produced continuously with higher rate from these free fatty acids, especially the oxidation of n-3 PUFA [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] whereas TO had lower oxidized rate which change into the MDA slowly than that AO. By the way, the lower of the MDA content in AO because the changed of the n-3 PUFAs oxidized into the secondary product [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Moreover, the products higher in oxidation may have a low PV when the primary oxidation metabolites are largely metabolized to secondary oxidation metabolites [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Therefore, the MDA for AO in lipid nanoparticles may rapidly shifted to another compound quicker than TO in lipid nanoparticles. By the way, the further study needs to invest the volatile compound of n-3 PUFAs oxidized to confirm the phenomenon. Therefore, tuna oil loaded in lipid nanoparticles was selected to further investigated the potential of the targeting process on the in vivo biodistribution, fatty acid profile, and coefficient of efficacy.\u003c/p\u003e \u003cp\u003eThe lower melting point of lipid nanoparticles mixtures than that of GMS due to the lipid matrix within the nanoparticles. Oils containing n-3 PUFAs (TO and AO) have liquid lipid properties leading to convert crystalline state to an amorphous state due to the disordered arrangement of molecules [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMore than that, TO in lipid nanoparticles has higher endothermic peak than that algal oil due to undesirable compounds of tuna oil. Unrefined oil or triglyceride are more complex than those of the fatty acids and required more thermal energy [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Moreover, the melting point was affected by the high content in unsaturated fatty acids in the formulation [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. The polyunsaturated fatty acids implying a larger space to be accommodated into the polymeric matrix indicate induced a more amorphous structure of the lipid carrier [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Thus, there are reasonable that the endothermic peak in TO treatment was shifted at higher temperature than that AO treatment. The DSC is an effective tool to investigate the melting behavior and crystalline state of nanocarriers and their impact on some properties such as stability of lipid nanoparticles, which is useful for further use in animal feed processing.\u003c/p\u003e \u003cp\u003eDuring the storage at room temperature (RT), the diameter of nanoparticles was higher than that of nanoparticles stored at 4\u0026deg;C particularly after 3 and 4 weeks of storage particularly for NPs treatment whereas there was no effect of temperature storage for TNPs treatment. The diameter of nanoparticles increased with the storage duration. Similarly with literature researches indicated the diameter of lipid nanoparticle was enlarged by storage for long time particularly unmodified-surface nanoparticles [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. The reason is the aggregation of surfactants between nanoparticle [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e] or the interaction of reactive oxygen species (ROS) with surface of nanoparticle [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. In contrast, the modified-surface nanoparticles can resolve the conflict and enhance oxidative stability by preventing pre-oxidation on the surface of the nanoparticle boundary [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Furthermore, the reduce particle size during storage due to the major compounds such as n-3 PUFAs in the matrix were descended by high temperature [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe peroxidative value and the MDA level were higher during the storage at RT by comparison with the treatments stored at 4\u0026deg;C. The high temperature can increased the kinetic energy contact of oil with oxygen [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. The higher of temperature had increase the rate of hydroperoxide decomposition. The reactivity of redox reactions had greater rates [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The amphiphilic lipid was reacted with the pro-oxidation at the interface and hydroperoxide was produced. The free radicals penetrate through the surface layer and stimulated the oxidation with n-3 PUFAs [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. Thus, the study of storage stability with different temperature indicated that the lipid oxidation continuously occurred during of storage particularly under high temperature for unmodified-surface nanoparticles. This should be noted that the storage of n-3 PUFAs in the lipid nanoparticle should be given priority for improving the sustainable quality prolonged enough for further procedures such as the drying process.\u003c/p\u003e \u003cp\u003eThe smaller nanoparticles formability in TNPs treatments might caused by the alkyl polyglucoside that was not present in the NPs groups. The nanoparticles synthesis with targeting had greater ability to perform lipid nanoparticles [\u003cspan additionalcitationids=\"CR51 CR52\" citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe stable of particles size in TNPs groups were indicated that the alkyl polyglucoside which use as the targeting lipid nanoparticles, have protective property to maintain the physical structure during storage. The change of particle size was faster in algal oil group possibly due to high amount of free fatty acids that have a carboxylic group with anionic, and are able to transfer into the surface of nanoparticles [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. However, the treatment AO_TNPs_4\u0026deg;C was different because the low temperature condition slowdown the breakdown of triglycerides to free fatty acids, particularly, n-3 PUFAs such as DHA that can affect the particle size [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. It is possible that the algal oil group had higher amount of free fatty acids inducing an increase in the nanoparticle size.\u003c/p\u003e \u003cp\u003eAlthough in the previous studies demonstrated that the storage conditions no effect on particle size of nanoparticles [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan additionalcitationids=\"CR56\" citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e] contrast with present study shown kept at 4\u0026deg;C and RT are effect diameter of the unmodified surface nanoparticle. This indicated that NPs is unstable form due to may interact with ROS present at the interface or surrounding aqueous phase [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e] such as impurities in crude oil which act as pro-oxidants [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn addition, the PDI values describe the width or spread of the particles size distribution of the nanoparticles ranged between 0.1 and 0.3 indicating a good stability of the emulsion [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. However, the TO_NPs group had the highest PDI value, possibly due to attractive hydrophobic interactions between the particles [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. The use of liquid oils such as fish oil can make the nanoparticles more physically stable by reducing the crystalline structure. Therefore, it prevents the incorporation of nanoparticles [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e] and increases PDI value. Moreover, the lower PDI value in AO nanoparticles indicated that is more dispersion in suspension than that TO nanoparticles. The PD Index is based on size of nanoparticles [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. The higher of the multiple particle size population show more PDI value while higher uniformity is lower polydisperse result to lower the PD Index [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMoreover, zeta-potential found in all groups were lower than \u0026minus;\u0026thinsp;30 mV. The NPs groups had lower values than the TNPs groups, (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001). The synthesis including tween 20, poloxamer and alkyl polyglucoside, resulted in a decrease in the electric potential difference of the nanoparticles [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFor the NPs groups, the n-3 PUFA oils source influenced the zeta-potential value. This is due to the movement of carboxylic fatty acids in the region near the outer layer of the nanoparticles. This resulted in more negative charge conversion [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. The TNPs groups contained alkyl polyglucoside. When this raw material is used in combination with a poloxamer, it results in a lower voltage difference than that observed for the NPs groups. Moreover, unrefined tuna oil contained phospholipids which are anionic groups [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e] and may have contributed to the high zeta-potential in TO_NPs group.\u003c/p\u003e \u003cp\u003eThe melting temperature was higher for NPs nanoparticles than for TNPs nanoparticles because of targeting process effect on decreasing particle size. The size of particle became smaller resulting in decreased melting point [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e]. By the way, the surface-modification of lipid nanoparticles could be effect on increase the particle size of nanoparticles [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] whereas the particle size was decreased in the present study due to the ratio of targeting surfactant in formulation [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIt has been shown that the active substances and raw materials have similar properties, namely the lipid molecules, which affect the form of nanoparticles. The hydroxyl group (3499\u0026thinsp;\u0026minus;\u0026thinsp;3233 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, -OH stretching) was found in raw materials with properties as an emulsifier, including tween 20 [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], poloxamer [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e], alkyl polyglucoside [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e] and GMS [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. However, after the synthesis, the nanoparticles containing tuna oil had no effect on triglyceride degradation as it did not convert to free fatty acids, which should appear at wavelength of \u003cem\u003e1711\u003c/em\u003e cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe wavelength at 1095 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-OCC, ether bond) provided by tween 20 and poloxamer implicated in the properties of emulsifier hydrophilic part of the molecule. The parts of raw materials, alkyl polyglucoside occur the wavelength at 1029 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicated the vibration of functional group of cyclic ether (-COC) and Pyranose ring and 1150 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-COC glycosidic bond) [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe peak wavelength was changed towards a lower intensity in the physical mixture and lipid nanoparticles. It indicated that the raw material was integrated into the system before the nanoparticles would be formed. Moreover, the peak that disappeared at wavelength below 1095 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in both mixtures was due to the structure of the molecules (including CH-stretching, long chain carbon) that bind together very densely. The nanomaterial changed their position from its own specific properties. The groups that had the hydrophobic properties occurred together inside the particles, while the hydrophilic groups occurred around the periphery of the particles. This phenomenon presented the similar wavelength peak of -OH stretching a functional group found in the surfactant family of tween 20, alkyl polyglucoside, and poloxamer. Moreover, the apparent wavelength in the spectrum at 1741\u0026thinsp;\u0026minus;\u0026thinsp;1734 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 1641 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1150 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e showed that the nanoparticles contained oil, such as tuna oil, inside the mixtures.\u003c/p\u003e \u003cp\u003eThe wavelength ranging at 2921\u0026thinsp;\u0026minus;\u0026thinsp;2850 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-CH stretching) and 1500\u0026thinsp;\u0026minus;\u0026thinsp;500 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-CH bending and long chain carbon) disappeared or had a reduced intensity, indicating that the molecules in the mixture are moving according to the molecular properties mentioned above. These functional groups can be found in fat-containing raw materials, which have nonpolar molecules. This property is more prone to interactions together by noncovalent bonds such as van der Waals binding within particles [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. Furthermore, the absence of new peaks in both mixtures showed that during the synthesis, there was no reaction that formed intermolecular covalent bonds [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e, \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe targeted-lipid nanoparticles were able to slow down the peroxidative reaction by comparison with the oil within non-targeted lipid nanoparticles because the surfactants act as protectors avoiding the inside oil to react with outside oxidants. The peroxidative value was significantly different in weekly storage of each treatment. This means that the initial lipid oxidation reaction was dynamic over the time storage and this constantly occurring leads to increase rate of the reaction [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e]. In addition, the amount of peroxide in some week was found to be low. This could mean that the substance has already converted to another molecule, or it may not have happened yet because of the product of initial stage was quickly oxidized into the next stage.\u003c/p\u003e \u003cp\u003eHowever, the industry recommended peroxidative value for fish oil should not exceed 18 meq active O\u003csub\u003e2\u003c/sub\u003e/kg after 1 month storage [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e]. So, the n-3 PUFAs lipid nanoparticles can be kept more than one month because their peroxidative values are lower than the recommended value. By the ways, further study should carry out the long-term storage period to confirm our data on storage stability for commercial products.\u003c/p\u003e \u003cp\u003eThe MDA content was higher for NPs nanoparticles than for TNPs nanoparticles suggesting a higher susceptibility to oxidation for NPs nanoparticles. May be, the triglycerides were hydrolyzed to free fatty acids with negatively charged carboxylic groups causing deposition in the outer layer of the nanoparticles. The resulting negative ions induced lipid oxidation [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. During the synthesis, lipids were heated may be favoring fatty acid oxidation.\u003c/p\u003e \u003cp\u003eThe measurement of lipid-based nanoparticles by in vivo study indicated that the targeted lipid-based nanoparticles showed the potential to transfer the fatty acids directly into skeletal muscle cells of Korat chickens.\u003c/p\u003e \u003cp\u003eNile red staining showed that FA deposition in muscles was more efficient by using TNPs compared to NPs nanoparticles at 8 h and 12 h after oral gavage with lipid-based nanoparticles. The lipid nanoparticles seem to act as a target carrier specific to the receptors at the microvilli. This phenomenon is confirmed by our previous research [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e]. Specific-transporters such as Sodium-glucose transporter 1 (SGLT1) and glucose transporter (GLUT) 2, 5, 7, etc., serve to accept single molecules such as glucose and galactose into enterocytes [\u003cspan additionalcitationids=\"CR75\" citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e]. For EPA deposition, the amount deposited 4 h after oral gavage was higher with TNPs nanoparticles compared with NPs nanoparticles. There was no difference between treatments for the other time points for EPA deposition. For DHA deposition, the amount deposited between 4 h and 24 h post-gavage was higher with TNPs nanoparticles compared with NPs nanoparticles. TNPs treatment was more efficient than NPs treatment and the efficacy of deposition was higher for DHA than for EPA that could be explained by a higher amount of DHA in nanoparticles compared to the EPA content.\u003c/p\u003e \u003cp\u003eThe results revealed the potential for both the receptors to function as individual molecules within the cell and to bind to those same molecules when attached to the nanoparticles and introduced into the cell. For example, nanoparticle uptake by microvilli receptor-specific monomolecular such as galactose [\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e], L-carnitine [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e] and taurocholic acid [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e] has been studied in digestive tract and muscle cells [\u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e78\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe utilization of receptor-specific molecules enhances the uptake of essential substances into the bloodstream and subsequent delivery to the targeted organs [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. There are indications that intestinal cells possess mechanisms capable of responding to the absorption of active substances in the nanoparticle form, wherein these nanoparticles are transported into the bloodstream through penetration into intestinal cells and subsequently directed towards target cells with specific transporter including GLUT via the endocytosis pathways [\u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e79\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHowever, intracellular transport is divided into several pathways that affects the delivery of nanoparticles into the bloodstream differently [\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e80\u003c/span\u003e]. Consequently, the comprehension and elucidation of the nanoparticle delivery pathway in functional intestinal cells at the molecular level will enable the identification of the underlying processes involved in nanoparticle transmission to the target organs. This understanding can further facilitate the development of appropriate models utilizing lipid nanoparticles.\u003c/p\u003e \u003cp\u003eThe coefficient of efficacy in TNPs groups indicated that the targeting process had potential to transfer the n-3 PUFA into target organ. By the way, the slightly decrease of n-3 PUFA amount in this group might due to the entering process of targeting-lipid nanoparticles in the cell via receptor-mediated endocytosis and sorting into lysosome, endocytosis with low pH in endosome make the ligand dissociated from their receptor [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. Then, the endosome vesical combined with lysosome for degradation and the cargos were degraded by hydrolases to be single molecules [\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e80\u003c/span\u003e]. Moreover, plain lipid nanoparticles are discriminated and weakened and could be recognized by the reticuloendothelial system [\u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e81\u003c/span\u003e]. On the other hand, targeting lipid nanoparticles escape the phagocytosis, resulting in a longer circulation time [\u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e82\u003c/span\u003e] and might enhanced the n-3 PUFA accumulation. Further study should be conducted in haematology and serum biochemistry to determine the response of lipid nanoparticles in chicken.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThis study provides conclusive evidence that nanotechnology has the potential to enhance the utilization of n-3 PUFA oil resources through nanoparticle synthesis. However, it is crucial to consider the source of the oil used for synthesis, as it can significantly affects parameters such as nanoparticle size, nanoparticle dispersion, and zeta-potential. Based on the outcomes of thermal and chemical analysis, it is established that n-3 PUFA oils were successfully encapsulated within lipid nanoparticles along with solid lipid, glyceryl monostearate. This was evidenced by the observed reduction in melting points as indicated by DSC analysis. Furthermore, the utilization of n-3 PUFA oil in nanoparticle form exhibited the ability to hinder lipid oxidation, with the rate of oxidation being contingent upon the storage temperature. Finally, the in vivo distribution findings substantiated the efficacy of targeted lipid-based nanoparticles to deposit long-chain n-3 PUFA in the chicken muscles.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eExperimental design for in vitro study\u003c/h2\u003e \u003cp\u003eFor the in vitro study, four preparations were considered, each with three replications. These were compared with free oils (crude tuna oil, TO; feed grade, T. C. Union Agrotech Co. Ltd., Bangkok, Thailand and algal oil, AO (Schizochytrium sp.); Shanghai Wellboost Health Food Co., Ltd, Shanghai, China), and TO and AO loaded non-targeted nanoparticles (NP) and targeted nanoparticles (TNPs), respectively, including TO_NPs, AO_NPs, TO_TNPs and AO_TNPs. The physicochemical characteristics and in vitro storage stability of lipid nanoparticles were investigated.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eLipid nanoparticles synthesis\u003c/h2\u003e \u003cp\u003eThe n-3 PUFA loaded lipid-based nanoparticles were prepared by hot and high-pressure homogenization approach (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e) as previously described [\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e83\u003c/span\u003e]. In this method, the lipid phase consisting of GMS (5 g), span 80 (3 g) (Croda, Bangkok, Thailand) and n-3 PUFAs (20 g) (tuna oil or algal oil) was blended and melted above 70\u0026deg;C to form an organic phase. The fatty acid composition of crude tuna oil and algal oil is indicated on Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e. Meanwhile, the aqueous phase was prepared by dispersing emulsifying agent, glycerol (2.5 g) Tween 20 (3 g), Poloxamer 188 (2 g) (Croda, Bangkok, Thailand) in distilled water. The aqueous phase was added drop wise to lipid phase at above 70\u0026deg;C with continuous agitation at 300 rpm for 3 min to get a primary emulsion. The pre-emulsion was then homogenized by using high-speed homogenizer (IKA, Altra-Turrac T25, Germany) at 8,000 rpm for 3 min. Then, it was sonicated (Qsonica sonicator, USA) at 30 A pulse on 30 sec and off 10 sec interval for 5 min to obtain n-3 PUFA loaded non-targeted lipid-based nanoparticles (NPs). For targeted lipid-based nanoparticles (TNPs), 2 g of alkyl polyglucoside was blended with organic phase preparation. Subsequently the dispersion was cooled in ice water bath. The prepared dispersion was stored in airtight container at 4\u0026deg;C.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFatty acid composition of n-3 PUFA oils (% of total fatty acids)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFatty acid composition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTO\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAO\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTO loaded in nanoparticles\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePalmitic Acid (C16:0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e38.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e29.43\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStearic Acid (C18:0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.46\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOleic Acid (C18:1n9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e22.64\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLinoleic Acid (C18:2n6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.69\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eα-Linolenic acid (C18:3n3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eArachidonic Acid (C20:4n6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEicosapentaenoic acid (C20:5n3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDocosahexaenoic acid (C22:6n3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e37.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e9.24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSFA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e39.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e52.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e43.95\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMUFA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e28.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePUFA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e36.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e27.23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003en-3 PUFA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e15.24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003en-6 PUFA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.99\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003en-6/n-3 ratio\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.79\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cem\u003eSFA\u003c/em\u003e Saturated fatty acid, \u003cem\u003eMUFA\u003c/em\u003e Monounsaturated fatty acid, \u003cem\u003ePUFA\u003c/em\u003e Polyunsaturated fatty acid, \u003cem\u003en-3 PUFA\u003c/em\u003e Omega-3 polyunsaturated fatty acids, \u003cem\u003en-6 PUFA\u003c/em\u003e Omega-6 polyunsaturated fatty acids.\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003e Two different n-3 PUFA oil sources (tuna or algal, TO or AO) within different type of lipid targeted (TNPs) or not (NPs) nanoparticles\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003ePhysicochemical characteristic analysis\u003c/h2\u003e \u003cp\u003eA Zetasizer 3000HS (Malvern Instruments, Malvern, UK) was employed to characterize NPs and TNPs for particle diameter, polydispersity index (PDI), and zeta-potential.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eThermal and chemical properties of lipid nanoparticles\u003c/h2\u003e \u003cdiv id=\"Sec19\" class=\"Section3\"\u003e \u003ch2\u003eDifferential scanning calorimetry (DSC)\u003c/h2\u003e \u003cp\u003eDSC is a thermo-analytical technique in which the difference in the amount for heat required to increase the temperature of a sample and reference is measured as a function of temperature. Usually, both the sample and reference are maintained at nearly the same temperature throughout the experiment. Generally, the temperature program for a DSC analysis is designed such that the sample holder temperature increases linearly as a function of time. The reference sample should have a well-defined heat capacity over the range of temperatures to be scanned. DSC (Pyris I DSC, Perkin Elmer, USA) was performed on samples of nanoparticles with and without feed or oil ingredient components to determine whether there were any interactions with the core-shell components.\u003c/p\u003e \u003cp\u003eTo determine the phase transition temperatures of the nanoparticles content n-3 PUFA and GMS, dried samples were placed in open aluminum pans. An empty pan was used as a reference pan compared with sample pan. Baseline correction and data treatment were conducted with OriginPro 9.85 (OriginLab, Northampton, Massachusetts, USA).\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eFourier-transformed infrared spectroscopy (FTIR)\u003c/h2\u003e \u003cp\u003eIn order to evaluate potential interactions between functionalized ligands and excipients within the lipid nanoparticles, a FTIR spectrum was acquired. The samples, consisting of solid lipid, liquid lipid, glucose-functionalized ligand, surfactant, physical mixture, non-target lipid nanoparticles loaded with tuna oil and algal oil, as well as targeting lipid nanoparticles loaded with tuna oil and algal oil, were meticulously ground with potassium bromide (KBr) to generate an infrared transparent matrix. Subsequently, FTIR scanning was performed using a Shimadzu Corporation FTIR-8400S spectrometer, covering the spectral range of 4,000 to 500 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e with a resolution of 1.0 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. Baseline correction and data treatment were conducted with OriginPro 9.85.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eIn vitro storage stability\u003c/h2\u003e \u003cp\u003eThe assessment of storage stability was conducted following the methodology outlined as previously described [\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e83\u003c/span\u003e] with some modifications. Free oil and oil synthesized within nanoparticles (n\u0026thinsp;=\u0026thinsp;3) were stored at 4\u0026deg;C in stability cabinets during four weeks. The hydrodynamic diameter (HD) was measured at different time points by dynamic light scattering (DLS) using a Malvern Zetasizer at an angle of 90\u0026deg; at 25\u0026deg;C and the oxidative stability was investigated by measuring PV and TBARS value.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eIn vitro oxidative stability and peroxide value\u003c/h2\u003e \u003cp\u003eThe determination of the primary oxidation products was carried out using the procedure outlined as previously described [\u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e84\u003c/span\u003e]. In order to measure the PV, the sample was added to 9.8 mL of a chloroform: methanol mixture (2:1, v/v) and mixed for 5 seconds on a vortex mixer. Subsequently, a solution of ammonium thiocyanate (50 \u0026micro;L) was introduced into the mixture, followed by the addition of iron (II) solution (50 \u0026micro;L). The iron (II) solution was prepared by combining BaCl\u003csub\u003e2\u003c/sub\u003e and FeSO\u003csub\u003e4\u003c/sub\u003e at a final concentration of 0.144 mol L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, resulting in individual concentrations of 0.132, respectively. After 5 min incubation period at room temperature, the absorbance of the samples was measured at 500 nm against a blank (solution without any sample) by a Shimadzu-1800 UV\u0026ndash;visible spectrophotometer (Shimadzu, Tokyo, Japan). The quantities of lipid hydroperoxides were determined by employing an external standard curve constructed with cumene hydroperoxide (purity 80%, sigma-Aldrich Co, USA) with concentrations ranging between 0 and 16 \u0026micro;M.\u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eIn vitro oxidative stability and thiobarbituric acid reactive substances\u003c/h2\u003e \u003cp\u003eThe secondary oxidation products were monitored by quantification of the thiobarbituric acid (TBA) reactive substances (TBARS) as previously described [\u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e85\u003c/span\u003e]. In brief, the samples were mixed with 1.8 mL of deionized water and 4.0 mL of TBA solution. TBA solution was prepared by dissolving 15 g of trichloroacetic acid (15% w/v) and 0.375 g of TBA (0.375%) in 100 mL of 0.25 mol L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e HCl. In the following, the mixtures were heated in a boiling water bath for 15 min and then cooled to room temperature. Finally, the mixtures were centrifuged (2000 g for 15 min). The intensity of the color created as a result of the reaction between TBA with malondialdehyde (MDA), an important by-product of lipid peroxidation, was measured at 532 nm. The standard curve of 1,1,3,3-tetraethoxypropane was used to determine the MDA concentrations.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eExperimental design for in vivo study\u003c/h2\u003e \u003cp\u003e All procedures in the present study were approved by the Ethics Committee on Animal Use of the Suranaree University of technology, Nakhon Ratchasima, Thailand (SUT-IACUC-009/2021). The experiment was conducted at Suranaree University of Technology (SUT) farm. The experimental model employed a completely randomized design, comprising three treatments and three replicates. The treatments consisted of control, tuna oil-loaded non-targeted lipid nanoparticles and tuna oil-loaded targeted lipid nanoparticles. Thirty-three female chickens, aged 56 days with an average weight ranging from 750 to 800 g, were individually housed in cages for a period of 7 days as an adaptation phase prior beginning the experiment. The chickens received commercial diet and water ad libitum for the whole period and withdrawal feeding 12 h before investigation. At 63 days of age, the lipid nanoparticles (2 ml for each chicken) were administered through oral administration at once, utilizing a syringe affixed to a designated feeding tube. After administration, the chickens were euthanized by cervical dislocation at distinct time intervals after gavage, 2, 4, 8, 12, and 24 h (3 chickens per treatment and per time interval). At each time point, all the breast muscle of the chickens was harvested and stored at -25\u0026deg;C for imaging purposes, as well as for the subsequent analysis of fatty acid composition.\u003c/p\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003eIn vivo imaging of tissue biodistribution\u003c/h2\u003e \u003cp\u003eThe investigation of nanoparticle distribution in muscle samples involved assessing the quantity of nanoparticles deposited in the target organ by utilizing the optical in vivo imaging system (IVIS), which captured the signal emitted by the fluorescent molecules. The fluorescence was observed within the appropriate wavelength range during the absorption (excitation) and emission processes. This methodology bears resemblance to a confocal laser-scanning microscope. However, the IVIS system is specifically designed for detecting signals from larger specimens. In this study, this instrument was employed to validate the presence of lipid nanoparticles within the target organ, the muscle.\u003c/p\u003e \u003cp\u003eThe In vivo imaging of nanoparticle accumulation in birds was assessed following the methodology as previously described [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. The birds were administered NR-NPs or NR-Glu-NPs orally at a dose of 1 mg/kg (equivalent to 3.14 \u0026micro;M, 2 mL) via oral gavage. Samples were eliminate unwanted fluid before quantify the near-infrared fluorescence signal intensities by using an In Vivo Imaging System MS FX PRO (Carestream Health Inc., Rochester, NY, USA) with an excitation band pass filter at 540 nm and an emission at 600 nm. Images were processed using Bruker Molecular Imaging software version 7.1.3.20550 (Bruker, Billerica, MA, USA). Mean intensity was performed to analyze statistics. The Nile red concentration was equivalent in all experiments (n\u0026thinsp;=\u0026thinsp;3 birds per treatment).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003eFatty acid composition\u003c/h2\u003e \u003cp\u003eThe lipids were extracted from approximately 5 g of each muscle sample using 90 ml of chloroform: methanol (2:1, v/v) [\u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e86\u003c/span\u003e]. After that, the methylation was conducted with around 20 to 25 mg of extracted fat [\u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e]. The fatty acid methyl esters (FAME) were analyzed using gas chromatography (Hewlett-Packard 7890A; Agilent Technologies, Santa Clara, CA, USA) with a capillary column (SP 2560, Supelco Inc., Bellefonte, PA, USA, 100 m \u0026times; 0.25 mm i.d., 0.20-\u0026micro;m film thickness) and a flame ionization detector. The carrier gas was helium at a flow rate of 0.95 ml/min. The temperatures of the injector and detector were 260\u0026deg;C. The initial column temperature was 70\u0026deg;C. It raised to 175\u0026deg;C at a rate of 13\u0026deg;C/min, and then to 240\u0026deg;C at a rate of 4\u0026deg;C/min. Compound were identified and quantified by using Masshunter software (v10.0.707.0, Agilent, USA). Coefficient of efficacy (%) was performed to evaluate the potential of lipid nanoparticles using the following formula:\u003c/p\u003e \u003cp\u003eCoefficient of efficacy (%) = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\left(\\frac{\\text{F}\\text{A}\\text{n}\\:\\text{i}\\text{n}\\:\\text{s}\\text{a}\\text{m}\\text{p}\\text{l}\\text{e}\\:\\text{t}\\text{r}\\text{e}\\text{a}\\text{t}\\text{e}\\text{d}\\:-\\:\\text{F}\\text{A}\\text{n}\\:\\text{i}\\text{n}\\:\\text{u}\\text{n}\\text{t}\\text{r}\\text{e}\\text{a}\\text{t}\\text{e}\\text{d}}{\\text{F}\\text{A}\\text{n}\\:\\text{a}\\text{m}\\text{o}\\text{u}\\text{n}\\text{t}\\:\\text{i}\\text{n}\\:\\text{s}\\text{u}\\text{s}\\text{p}\\text{e}\\text{n}\\text{s}\\text{i}\\text{o}\\text{n}\\text{s}\\:\\text{u}\\text{s}\\text{e}\\text{d}\\:}\\right)\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eAnalysis of variance will be performed by GLM procedure for a completely randomized design using SPSS Version 18.0 (SPSS Inc., Chicago, I11., USA). University Edition with the following statistical model:\u003c/p\u003e \u003cp\u003eYij\u0026thinsp;=\u0026thinsp;\u0026micro;\u0026thinsp;+\u0026thinsp;τi\u0026thinsp;+\u0026thinsp;εij\u003c/p\u003e \u003cp\u003eWhere: Yij\u0026thinsp;=\u0026thinsp;the dependent variable, \u0026micro;\u0026thinsp;=\u0026thinsp;the overall mean, τi\u0026thinsp;=\u0026thinsp;the treatment effect, and εij\u0026thinsp;=\u0026thinsp;the random residual error. Significant differences between treatment means were assessed by Tukey's multiple comparison tests after a significant F-test. The level of statistically significance will establish at P\u0026thinsp;\u0026lt;\u0026thinsp;0.05. The values for physical characteristic of lipid nanoparticles were expressed as means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error (SE) and the value for storage stability and fatty acid composition were expressed as means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of mean (SEM), which represents the pooled SEM for the model.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003eData availability\u003c/h2\u003e \u003cp\u003eThe datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eEthical statement\u003c/h2\u003e \u003cp\u003e The study was approved by the Ethics Committee on Animal Use of the Suranaree University of technology, Nakhon Ratchasima, Thailand (SUT-IACUC-009/2021). All methods were performed in accordance with the relevant guidelines and regulations.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eARRIVE Guidelines statement\u003c/strong\u003e \u003cp\u003eThis study is reported in accordance with ARRIVE guidelines (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://arriveguidelines.org\u003c/span\u003e\u003cspan address=\"https://arriveguidelines.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e \u003c/p\u003e\u003cp\u003e \u003cstrong\u003eCompeting interests:\u003c/strong\u003e \u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis research project was supported by the National Research Council of Thailand (NRCT) : NRCT5-RGJ63007-093, as well as by Suranaree University of Technology (SUT), Thailand Science Research and Innovation (TSRI), and National Science, Research and Innovation Fund (NSRF) (NRIIS number 160352).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eW.M. and A.M. conducted conceptualization, funding acquisition, and project administration. P.H., T.Y., A.K., A.M. and W.M. conceived and designed the methodology and investigation. T.Y. performed lipid nanoparticles synthesis. P.H. and W.M. performed and investigated the physicochemical characteristic, in vitro studies and fatty acid composition. P.H., W.T. and A.A. performed and investigated the samples imaging. P.H., T.Y., A.K., E.B., C.B. and W.M. performed data curation and formal analysis. P.H., E.B. and W.M. performed writing \u0026ndash; original draft. P.H., E.B., C.B. and W.M. performed writing \u0026ndash; review \u0026amp; editing the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eThis research project was supported by the National Research Council of Thailand (NRCT) : NRCT5-RGJ63007-093, the Royal Golden Jubilee Ph.D. (RGJ-PHD) Program as well as by Suranaree University of Technology (SUT). The authors also highly appreciate the Center of Excellence on Technology and Innovation for Korat chicken Business Development, SUT and French National Institute for Agriculture, Food, and Environment (INRAE), UMR Biologie des Oiseaux et Aviculture for partly supporting this research.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003ePacker, L., Weber, S. U. \u0026amp; Rimbach, G. Molecular aspects of alpha-tocotrienol antioxidant action and cell signalling. \u003cem\u003eJ. Nutr\u003c/em\u003e. 131, 369s-373s, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/jn/131.2.369S\u003c/span\u003e\u003cspan address=\"10.1093/jn/131.2.369S\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2001).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChang, M. et al. Optimization of cultivation conditions for efficient production of carotenoid-rich DHA oil by Schizochytrium sp. S31. Process Biochemistry. 94, 190\u0026ndash;197, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.1016/j.procbio.2020.04.007\u003c/span\u003e\u003cspan address=\"10.1016/j.procbio.2020.04.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, Y., Ghosh, S, \u0026amp; Nickerson M. T. Microencapsulation of flaxseed oil by lentil protein isolate-κ-carrageenan and -ι-carrageenan based wall materials through spray and freeze drying. Molecules. 27, 3195. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/molecules27103195\u003c/span\u003e\u003cspan address=\"10.3390/molecules27103195\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKharat, M \u0026amp; McClements D. J. Fabrication and characterization of nanostructured lipid carriers (NLC) using a plant-based emulsifier: Quillaja saponin. Food Res. Int. 126, 108601. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.foodres.2019.108601\u003c/span\u003e\u003cspan address=\"10.1016/j.foodres.2019.108601\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKaewsatuan, P. et al. Comparative proteomics revealed duodenal metabolic function associated with feed efficiency in slow-growing chicken. Poult. Sci. 101, 101824, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.psj.2022.101824\u003c/span\u003e\u003cspan address=\"10.1016/j.psj.2022.101824\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHang, T. T. T., Molee, W. \u0026amp; Khempaka, S. Linseed oil or tuna oil supplementation in slow-growing chicken diets: Can their meat reach the threshold of a \u0026ldquo;high in n-3 polyunsaturated fatty acids\u0026rdquo; product? J. Appl. Poult. Res. 27, 389\u0026ndash;400, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.3382/japr/pfy010\u003c/span\u003e\u003cspan address=\"10.3382/japr/pfy010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSanguansri, L. \u003cem\u003eet al.\u003c/em\u003e Encapsulation of mixtures of tuna oil, tributyrin and resveratrol in a spray dried powder formulation. Food Funct. 4, 1794\u0026ndash;1802, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/c3fo60197h\u003c/span\u003e\u003cspan address=\"10.1039/c3fo60197h\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhu, Q. Y. \u003cem\u003eet al.\u003c/em\u003e Preparation of Deep Sea Fish Oil-Based Nanostructured Lipid Carriers with Enhanced Cellular Uptake. J. Nanosci. Nanotechnol. 15, 9539\u0026ndash;9547, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1166/jnn.2015.10880\u003c/span\u003e\u003cspan address=\"10.1166/jnn.2015.10880\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShahparast, Y., Eskandani, M., Rajaei, A. \u0026amp; Yari Khosroushahi, A. Preparation, Physicochemical Characterization and Oxidative Stability of Omega-3 Fish Oil/alpha-Tocopherol-co-Loaded Nanostructured Lipidic Carriers. Adv. Pharm. Bull. 9, 393\u0026ndash;400, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.15171/apb.2019.046\u003c/span\u003e\u003cspan address=\"10.15171/apb.2019.046\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTaha, E. \u003cem\u003eet al.\u003c/em\u003e Cod liver oil nano-structured lipid carriers (Cod-NLCs) as a promising platform for nose to brain delivery: Preparation, in vitro optimization, ex vivo cytotoxicity \u0026amp; in vivo biodistribution utilizing radioiodinated zopiclone. Int. J. Pharm. X. 5, 100160, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.1016/j.ijpx.2023.100160\u003c/span\u003e\u003cspan address=\"10.1016/j.ijpx.2023.100160\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, T. \u0026amp; Luo, Y. Biological fate of ingested lipid-based nanoparticles: current understanding and future directions. Nanoscale. 11, 11048\u0026ndash;11063, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/c9nr03025e\u003c/span\u003e\u003cspan address=\"10.1039/c9nr03025e\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang, X., Chen, G., Zhang, T., Ma, Z. \u0026amp; Wu, B. Effects of PEGylated lipid nanoparticles on the oral absorption of one BCS II drug: a mechanistic investigation. Int. J. Nanomedicine. 9, 5503\u0026ndash;5514, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2147/ijn.S73340\u003c/span\u003e\u003cspan address=\"10.2147/ijn.S73340\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, H., Guissi, N. E. I., Su, Z., Ping, Q. \u0026amp; Sun, M. Effects of surface hydrophilic properties of PEG-based mucus-penetrating nanostructured lipid carriers on oral drug delivery. RSC Advances. 6, 84164\u0026ndash;84176, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/c6ra18724b\u003c/span\u003e\u003cspan address=\"10.1039/c6ra18724b\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQin, J. J., Wang, W., Sarkar, S. \u0026amp; Zhang, R. Oral delivery of anti-MDM2 inhibitor SP141-loaded FcRn-targeted nanoparticles to treat breast cancer and metastasis. J. Control Release. 237, 101\u0026ndash;114, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jconrel.2016.07.008\u003c/span\u003e\u003cspan address=\"10.1016/j.jconrel.2016.07.008\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKou, L. \u003cem\u003eet al.\u003c/em\u003e Transporter-Guided Delivery of Nanoparticles to Improve Drug Permeation across Cellular Barriers and Drug Exposure to Selective Cell Types. Front. Pharmacol. 9, 27, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fphar.2018.00027\u003c/span\u003e\u003cspan address=\"10.3389/fphar.2018.00027\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKrishna, K. V. \u003cem\u003eet al.\u003c/em\u003e Design and Biological Evaluation of Lipoprotein-Based Donepezil Nanocarrier for Enhanced Brain Uptake through Oral Delivery. ACS Chem. Neurosci. 10, 4124\u0026ndash;4135, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/acschemneuro.9b00343\u003c/span\u003e\u003cspan address=\"10.1021/acschemneuro.9b00343\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTunki, L. \u003cem\u003eet al.\u003c/em\u003e Modulating the site-specific oral delivery of sorafenib using sugar-grafted nanoparticles for hepatocellular carcinoma treatment. Eur. J. Pharm. Sci. 137, 104978, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ejps.2019.104978\u003c/span\u003e\u003cspan address=\"10.1016/j.ejps.2019.104978\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBanu, V. \u0026amp; Rajendran, S. Corrosion inhibition of carbon steel by tween 20 self-assembling monolayers. J. Chem. Pharm. Res. 7, 127\u0026ndash;132, (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKiefer, J. \u003cem\u003eet al.\u003c/em\u003e Vibrational structure of the polyunsaturated fatty acids eicosapentaenoic acid and arachidonic acid studied by infrared spectroscopy. J. Mol. Struct. 965, 121\u0026ndash;124, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.molstruc.2009.11.052\u003c/span\u003e\u003cspan address=\"10.1016/j.molstruc.2009.11.052\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGholami, H., Yeganeh, H., Burujeny, S. B., Sorayya, M. \u0026amp; Shams, E. Vegetable Oil Based Polyurethane Containing 1,2,3-Triazolium Functional Groups as Antimicrobial Wound Dressing. J. Polym. Environ. 26, 462\u0026ndash;473, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10924-017-0964-y\u003c/span\u003e\u003cspan address=\"10.1007/s10924-017-0964-y\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRaeisi, S., Ojagh, S. M., Quek, S. Y., Pourashouri, P. \u0026amp; Sala\u0026uuml;n, F. Nano-encapsulation of fish oil and garlic essential oil by a novel composition of wall material: Persian gum-chitosan. LWT. 116, 108494, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.1016/j.lwt.2019.108494\u003c/span\u003e\u003cspan address=\"10.1016/j.lwt.2019.108494\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTakeungwongtrakul, S., Karnjanapratum, S., Kaewthong, P., Nalinanon, S. Change in fatty acid profile, volatile compounds and FTIR spectra of samrong seed oil during storage. Int. J. Agric. Technol. 16, 475\u0026ndash;484, (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMd Salim, R., Asik, J. \u0026amp; Sarjadi, M. S. Chemical functional groups of extractives, cellulose and lignin extracted from native Leucaena leucocephala bark. Wood Sci. Technol. 55, 295\u0026ndash;313, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00226-020-01258-2\u003c/span\u003e\u003cspan address=\"10.1007/s00226-020-01258-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNg W, S., Lee, C. S., Cheng, S. F., Chuah, C. H. \u0026amp; Wong, S. F. Biocompatible Polyurethane Scaffolds Prepared from Glycerol Monostearate-Derived Polyester Polyol. J. Polym. Environ. 26, 2881\u0026ndash;2900, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10924-017-1175-2\u003c/span\u003e\u003cspan address=\"10.1007/s10924-017-1175-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCaldas, A. R. \u003cem\u003eet al.\u003c/em\u003e Omega-3- and Resveratrol-Loaded Lipid Nanosystems for Potential Use as Topical Formulations in Autoimmune, Inflammatory, and Cancerous Skin Diseases. Pharmaceutics. 13, 1202, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/pharmaceutics13081202\u003c/span\u003e\u003cspan address=\"10.3390/pharmaceutics13081202\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMudalige, T. \u003cem\u003eet al.\u003c/em\u003e in \u003cem\u003eNanomaterials for Food Applications\u003c/em\u003e (eds Amparo L\u0026oacute;pez Rubio, Maria Jos\u0026eacute; Fabra Rovira, Marta mart\u0026iacute;nez Sanz, \u0026amp; Laura G\u0026oacute;mez G\u0026oacute;mez-Mascaraque) 313\u0026ndash;353 (Elsevier, 2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eŠimat, V. \u003cem\u003eet al.\u003c/em\u003e Production and Refinement of Omega-3 Rich Oils from Processing By-Products of Farmed Fish Species. Foods. 8, 125, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/foods8040125\u003c/span\u003e\u003cspan address=\"10.3390/foods8040125\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSreedhar, R., Kumar, V. S., Bhaskaran Pillai, A. K. \u0026amp; Mangalathillam, S. Omega-3 Fatty Acid Based Nanolipid Formulation of Atorvastatin for Treating Hyperlipidemia. Adv. Pharm. Bull. 9, 271\u0026ndash;280, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.15171/apb.2019.031\u003c/span\u003e\u003cspan address=\"10.15171/apb.2019.031\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u0026Aacute;lvarez-Benedicto, E. \u003cem\u003eet al.\u003c/em\u003e Optimization of phospholipid chemistry for improved lipid nanoparticle (LNP) delivery of messenger RNA (mRNA). Biomater Sci. 10, 549\u0026ndash;559, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/D1BM01454D\u003c/span\u003e\u003cspan address=\"10.1039/D1BM01454D\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSathivel, S. Thermal and flow properties of oils from salmon head. J. Am. Oil Chem. Soc. 82, 147\u0026ndash;152, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11746-005-1057-6\u003c/span\u003e\u003cspan address=\"10.1007/s11746-005-1057-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShi, T. Q., Wang, L. R., Zhang, Z. X., Sun, X. M. \u0026amp; Huang, H. Stresses as First-Line Tools for Enhancing Lipid and Carotenoid Production in Microalgae. Front. Bioeng. Biotechnol. 8, 610, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fbioe.2020.00610\u003c/span\u003e\u003cspan address=\"10.3389/fbioe.2020.00610\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSehl, A. et al. How do algae oils change the omega-3 polyunsaturated fatty acids market?. OCL. 29, 20. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.orFg/10.1051/ocl/2022018\u003c/span\u003e\u003cspan address=\"https://doi.orFg/10.1051/ocl/2022018\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTengku Mohamad, T. R. \u0026amp; Birch, E. Physicochemical characterisation and oxidative stability of refined hoki oil, unrefined hoki oil and unrefined tuna oil. Int. J. Food Sci. Technol. 48, 2331\u0026ndash;2339, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/ijfs.12222\u003c/span\u003e\u003cspan address=\"10.1111/ijfs.12222\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGaweł, S., Wardas, M., Niedworok, E., Wardas, P., 2004. Dialdehyd malonowy (MDA) jako wskaźnik proces\u0026oacute;w peroksydacji lipid\u0026oacute;w w organizmie [Malondialdehyde (MDA) as a lipid peroxidation marker]. Wiad. Lek. 57, 453\u0026ndash;455.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNogueira, M., Scolaro, B., Milne, G. \u0026amp; Castro, I. Oxidation products from omega-3 and omega-6 fatty acids during a simulated shelf life of edible oils. LWT. 101, 113\u0026ndash;122, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.lwt.2018.11.044\u003c/span\u003e\u003cspan address=\"10.1016/j.lwt.2018.11.044\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIsmail, A., Bannenberg, G., Rice, H. B., Schutt, E. \u0026amp; MacKay, D. Oxidation in EPA- and DHA-rich oils: an overview. Lipid Technol. 28, 55\u0026ndash;59, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.1002/lite.201600013\u003c/span\u003e\u003cspan address=\"10.1002/lite.201600013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYin H, Purification of fish oils and production of protein powders with EPA and DHA enriched fish oils [Doctoral Dissertations]. [Louisiana, U.S.]. Louisiana State University; 2011.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBarros, D., Reed, P., Alves, M., Santos, R. \u0026amp; Oliva, A. Biocompatibility and Antimicrobial Activity of Nanostructured Lipid Carriers for Topical Applications Are Affected by Type of Oils Used in Their Composition. \u003cem\u003ePharmaceutics\u003c/em\u003e. 13, 1950, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/pharmaceutics13111950\u003c/span\u003e\u003cspan address=\"10.3390/pharmaceutics13111950\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLacatusu, I. \u003cem\u003eet al.\u003c/em\u003e Ivy leaves extract based \u0026ndash; lipid nanocarriers and their bioefficacy on antioxidant and antitumor activities. RSC Advances. 6, 77243\u0026ndash;77255, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/c6ra12016d\u003c/span\u003e\u003cspan address=\"10.1039/c6ra12016d\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNejadmansouri, M., Hosseini, S. M. H., Niakousari, M. \u0026amp; Yousefi, G. Physicochemical properties and storage stability of ultrasound-mediated WPI-stabilized fish oil nanoemulsions. Food Hydrocoll. 61, 801\u0026ndash;811, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.foodhyd.2016.07.011\u003c/span\u003e\u003cspan address=\"10.1016/j.foodhyd.2016.07.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZaky, M. F. \u003cem\u003eet al.\u003c/em\u003e Influence of Surface-Modification via PEGylation or Chitosanization of Lipidic Nanocarriers on In Vivo Pharmacokinetic/Pharmacodynamic Profiles of Apixaban. Pharmaceutics. 15, 1668, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/pharmaceutics15061668\u003c/span\u003e\u003cspan address=\"10.3390/pharmaceutics15061668\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhosa, A., Reddi, S. \u0026amp; Saha, R. N. Nanostructured lipid carriers for site-specific drug delivery. Biomed. Pharmacother. 103, 598\u0026ndash;613, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.biopha.2018.04.055\u003c/span\u003e\u003cspan address=\"10.1016/j.biopha.2018.04.055\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTerada, T., Kulkarni, J. A., Huynh, A., Tam, Y. Y. C. \u0026amp; Cullis, P. Protective Effect of Edaravone against Cationic Lipid-Mediated Oxidative Stress and Apoptosis. Biol. Pharm. Bull. 44, 144\u0026ndash;149, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1248/bpb.b20-00679\u003c/span\u003e\u003cspan address=\"10.1248/bpb.b20-00679\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeulewaeter, S. \u003cem\u003eet al.\u003c/em\u003e Continuous freeze-drying of messenger RNA lipid nanoparticles enables storage at higher temperatures. J. Control. Release. 357, 149\u0026ndash;160, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jconrel.2023.03.039\u003c/span\u003e\u003cspan address=\"10.1016/j.jconrel.2023.03.039\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTang, C. H., Chen, H. L. \u0026amp; Dong, J. R. Solid Lipid Nanoparticles (SLNs) and Nanostructured Lipid Carriers (NLCs) as Food-Grade Nanovehicles for Hydrophobic Nutraceuticals or Bioactives. Appl. Sci. 13, 1726, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/app13031726\u003c/span\u003e\u003cspan address=\"10.3390/app13031726\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShah, N. V. \u003cem\u003eet al.\u003c/em\u003e Nanostructured lipid carriers for oral bioavailability enhancement of raloxifene: Design and in vivo study. J. Adv. Res. 7, 423\u0026ndash;434, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jare.2016.03.002\u003c/span\u003e\u003cspan address=\"10.1016/j.jare.2016.03.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFadda, A. \u003cem\u003eet al.\u003c/em\u003e Innovative and Sustainable Technologies to Enhance the Oxidative Stability of Vegetable Oils. Sustainability. 14, 849, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/su14020849\u003c/span\u003e\u003cspan address=\"10.3390/su14020849\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMusakhanian, J., Rodier, J. D. \u0026amp; Dave, M. Oxidative Stability in Lipid Formulations: a Review of the Mechanisms, Drivers, and Inhibitors of Oxidation. AAPS PharmSciTech. 23, 151, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1208/s12249-022-02282-0\u003c/span\u003e\u003cspan address=\"10.1208/s12249-022-02282-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, C. \u003cem\u003eet al.\u003c/em\u003e Emulsion structure design for improving the oxidative stability of polyunsaturated fatty acids. Compr. Rev. Food Sci. Food Saf. 19, 2955\u0026ndash;2971, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/1541-4337.12621\u003c/span\u003e\u003cspan address=\"10.1111/1541-4337.12621\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLing, S. S., Yuen, K. H., Magosso, E. \u0026amp; Barker, S. A. Oral bioavailability enhancement of a hydrophilic drug delivered via folic acid-coupled liposomes in rats. J. Pharm. Pharmacol. 61, 445\u0026ndash;449, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1211/jpp/61.04.0005\u003c/span\u003e\u003cspan address=\"10.1211/jpp/61.04.0005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKou, L. \u003cem\u003eet al.\u003c/em\u003e Cotransporting Ion is a Trigger for Cellular Endocytosis of Transporter-Targeting Nanoparticles: A Case Study of High-Efficiency SLC22A5 (OCTN2)-Mediated Carnitine-Conjugated Nanoparticles for Oral Delivery of Therapeutic Drugs. Adv. Healthc. Mater. 6, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/adhm.201700165\u003c/span\u003e\u003cspan address=\"10.1002/adhm.201700165\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTian, C. \u003cem\u003eet al.\u003c/em\u003e Improving intestinal absorption and oral bioavailability of curcumin via taurocholic acid-modified nanostructured lipid carriers. Int. J. Nanomedicine. 12, 7897\u0026ndash;7911, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2147/IJN.S145988\u003c/span\u003e\u003cspan address=\"10.2147/IJN.S145988\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTian, C. \u003cem\u003eet al.\u003c/em\u003e N-acetyl-L-cysteine functionalized nanostructured lipid carrier for improving oral bioavailability of curcumin: preparation, in vitro and in vivo evaluations. Drug Deliv. 24, 1605\u0026ndash;1616, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/10717544.2017.1391890\u003c/span\u003e\u003cspan address=\"10.1080/10717544.2017.1391890\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuill\u0026eacute;n, M. D. \u0026amp; Cabo, N. Infrared spectroscopy in the study of edible oils and fats. \u003cem\u003eJ. Sci. Food Agric\u003c/em\u003e. 75, 1\u0026ndash;11, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.1002\u003c/span\u003e\u003cspan address=\"https://doi.org/https://doi.org/10.1002\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e/(SICI)1097-0010(199709)75:1\u0026lt;1::AID-JSFA842\u0026gt;3.0.CO;2-R (1997).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAverina, E., M\u0026uuml;ller, R., Popov, D. \u0026amp; Radnaeva, L. Physical and chemical stability of nanostructured lipid drug carriers (NLC) based on natural lipids from Baikal region (Siberia, Russia). \u003cem\u003ePharmazie\u003c/em\u003e. 66, 348\u0026ndash;356, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1691/ph.2011.0326\u003c/span\u003e\u003cspan address=\"10.1691/ph.2011.0326\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePornputtapitak, W. \u003cem\u003eet al.\u003c/em\u003e Effect of Oil Content on Physiochemical Characteristics of gamma-Oryzanol-Loaded Nanostructured Lipid Carriers. J. Oleo. Sci. 68, 699\u0026ndash;707, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5650/jos.ess18127\u003c/span\u003e\u003cspan address=\"10.5650/jos.ess18127\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaedi, A., Rostamizadeh, K., Parsa, M., Dalali, N. \u0026amp; Ahmadi, N. Preparation and characterization of nanostructured lipid carriers as drug delivery system: Influence of liquid lipid types on loading and cytotoxicity. Chem. Phys. Lipids. 216, 65\u0026ndash;72, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.chemphyslip.2018.09.007\u003c/span\u003e\u003cspan address=\"10.1016/j.chemphyslip.2018.09.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eL\u0026uacute;cio, M. \u003cem\u003eet al.\u003c/em\u003e Nanostructured Lipid Carriers Enriched Hydrogels for Skin Topical Administration of Quercetin and Omega-3 Fatty Acid. \u003cem\u003ePharmaceutics\u003c/em\u003e. 15, 2078, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/pharmaceutics15082078\u003c/span\u003e\u003cspan address=\"10.3390/pharmaceutics15082078\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCameron-Smith, D., Albert, B. B. \u0026amp; Cutfield, W. S. Fishing for answers: is oxidation of fish oil supplements a problem? J. Nutr. Sci. 4, e36, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/jns.2015.26\u003c/span\u003e\u003cspan address=\"10.1017/jns.2015.26\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAwad, T. S., Helgason, T., Weiss, J., Decker, E. A. \u0026amp; McClements, D. J. Effect of Omega-3 Fatty Acids on Crystallization, Polymorphic Transformation and Stability of Tripalmitin Solid Lipid Nanoparticle Suspensions. Cryst. Growth Des. 9, 3405\u0026ndash;3411, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/cg8011684\u003c/span\u003e\u003cspan address=\"10.1021/cg8011684\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHelgason, T., Awad, T. S., Kristbergsson, K., McClements, D. J. \u0026amp; Weiss, J. Influence of Polymorphic Transformations on Gelation of Tripalmitin Solid Lipid Nanoparticle Suspensions. J. Am. Oil Chem. Soc. 85, 501\u0026ndash;511, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.1007/s11746-008-1219-9\u003c/span\u003e\u003cspan address=\"10.1007/s11746-008-1219-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSurassmo, S., Ruktanonchai, U., Srinuanchai, W. \u0026amp; Yostawonkul, J. Formulation development of plain nanoemulsion based on the influence of surfactant combinations. Chiang Mai J. Sci. 40, 994\u0026ndash;999, (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOomah, B. D. Flaxseed as a functional food source. J. Sci. Food Agric. 81, 889\u0026ndash;894, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/jsfa.898\u003c/span\u003e\u003cspan address=\"10.1002/jsfa.898\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2001).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkhoond Zardini, A., Mohebbi, M., Farhoosh, R. \u0026amp; Bolurian, S. Production and characterization of nanostructured lipid carriers and solid lipid nanoparticles containing lycopene for food fortification. J. Food Sci. Technol. 55, 287\u0026ndash;298, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s13197-017-2937-5\u003c/span\u003e\u003cspan address=\"10.1007/s13197-017-2937-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLuo, Q. \u003cem\u003eet al.\u003c/em\u003e Ascorbate-conjugated nanoparticles for promoted oral delivery of therapeutic drugs via sodium-dependent vitamin C transporter 1 (SVCT1). Artif. Cells Nanomed. Biotechnol. 46, 198\u0026ndash;208, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/21691401.2017.1417864\u003c/span\u003e\u003cspan address=\"10.1080/21691401.2017.1417864\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKannan, K. Compatibility studies of camptothecin with various pharmaceutical excipients used in the development of nanoparticle formulation. Int. J. Pharm. Pharm. Sci. 5, 315\u0026ndash;321, (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePajić, N. Z. B. \u003cem\u003eet al.\u003c/em\u003e Alkyl polyglucoside vs. ethoxylated surfactant-based microemulsions as vehicles for two poorly water-soluble drugs: physicochemical characterization and in vivo skin performance. Acta Pharm. 67, 415\u0026ndash;439, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/doi:10.1515/acph-2017-0036\u003c/span\u003e\u003cspan address=\"doi:10.1515/acph-2017-0036\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAryani, D., Siswodihardjo, S., Soeratri, W., Rahmasari, F. \u0026amp; Sari, D. Experimental development and molecular docking: nanostructured lipid carriers (nlcs) of coenzyme q10 using stearic acid and different liquid lipids as lipid matrix. Int. J. Appl. Pharm. 13, 108\u0026ndash;116, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.22159/ijap.2021v13i1.39890\u003c/span\u003e\u003cspan address=\"10.22159/ijap.2021v13i1.39890\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePezeshki, A., Ghanbarzadeh, B., Mohammadi, M., Fathollahi, I. \u0026amp; Hamishehkar, H. Encapsulation of Vitamin A Palmitate in Nanostructured Lipid Carrier (NLC)-Effect of Surfactant Concentration on the Formulation Properties. Adv. Pharm. Bull. 4, 563\u0026ndash;568, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5681/apb.2014.083\u003c/span\u003e\u003cspan address=\"10.5681/apb.2014.083\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePalmquist, D. L. \u0026amp; Jenkins, T. C. Challenges with fats and fatty acid methods. J. Anim. Sci. 81, 3250\u0026ndash;3254, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2527/2003.81123250x\u003c/span\u003e\u003cspan address=\"10.2527/2003.81123250x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2003).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChang, C. \u0026amp; Nickerson, M. Stability and in vitro release behaviour of encapsulated omega fatty acid-rich oils in lentil protein isolate-based microcapsules. Int. J. Food Sci. Nutr. 69, 1\u0026ndash;12, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/09637486.2017.1336513\u003c/span\u003e\u003cspan address=\"10.1080/09637486.2017.1336513\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKomaiko, J., Sastrosubroto, A. \u0026amp; McClements, D. Encapsulation of ω-3 fatty acids in nanoemulsion-based delivery systems fabricated from natural emulsifiers: Sunflower phospholipids. Food Chem. 203, 331\u0026ndash;339, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.foodchem.2016.02.080\u003c/span\u003e\u003cspan address=\"10.1016/j.foodchem.2016.02.080\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWangngae, S. \u003cem\u003eet al.\u003c/em\u003e Lipid-Based Nanoparticles as Targeted Delivery System in Korat Chicken. ACS Food Sci. Technol. 3, 1913\u0026ndash;1919 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/acsfoodscitech.3c00275\u003c/span\u003e\u003cspan address=\"10.1021/acsfoodscitech.3c00275\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCohen, M. \u003cem\u003eet al.\u003c/em\u003e Live imaging of GLUT2 glucose-dependent trafficking and its inhibition in polarized epithelial cysts. Open Biol. 4, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1098/rsob.140091\u003c/span\u003e\u003cspan address=\"10.1098/rsob.140091\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePoulsen, S. B., Fenton, R. A. \u0026amp; Rieg, T. Sodium-glucose cotransport. Curr Opin Nephrol Hypertens. 24, 463\u0026ndash;469, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1097/MNH.0000000000000152\u003c/span\u003e\u003cspan address=\"10.1097/MNH.0000000000000152\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eByers, M. S., Howard, C. \u0026amp; Wang, X. Avian and Mammalian Facilitative Glucose Transporters. \u003cem\u003eMicroarrays (Basel)\u003c/em\u003e. 6, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/microarrays6020007\u003c/span\u003e\u003cspan address=\"10.3390/microarrays6020007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSiu, F. Y., Ye, S., Lin, H. \u0026amp; Li, S. Galactosylated PLGA nanoparticles for the oral delivery of resveratrol: enhanced bioavailability and in vitro anti-inflammatory activity. Int. J. Nanomedicine. 13, 4133\u0026ndash;4144, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2147/IJN.S164235\u003c/span\u003e\u003cspan address=\"10.2147/IJN.S164235\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYeh, Y. C., Kim, S. T., Tang, R., Yan, B. \u0026amp; Rotello, V. M. Insulin-based regulation of glucose-functionalized nanoparticle uptake in muscle cells. J. Mater. Chem. B. 2, 4610\u0026ndash;4614, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/C4TB00608A\u003c/span\u003e\u003cspan address=\"10.1039/C4TB00608A\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGrefner, N. M., Gromova, L. V., Gruzdkov, A. A. \u0026amp; Komissarchik, Y. Y. Interaction of glucose transporters SGLT1 and GLUT2 with cytoskeleton in enterocytes and Caco2 cells during hexose absorption. Cell Tissue Biol. 9, 45\u0026ndash;52, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1134/s1990519x15010034\u003c/span\u003e\u003cspan address=\"10.1134/s1990519x15010034\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSettembre, C. \u0026amp; Ballabio, A. Lysosome: regulator of lipid degradation pathways. Trends Cell Biol. 24, 743\u0026ndash;750, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.tcb.2014.06.006\u003c/span\u003e\u003cspan address=\"10.1016/j.tcb.2014.06.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang, R., Qin, X., Kong, F., Chen, P. \u0026amp; Pan, G. Improving cellular uptake of therapeutic entities through interaction with components of cell membrane. Drug Deliv. 26, 328\u0026ndash;342, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/10717544.2019.1582730\u003c/span\u003e\u003cspan address=\"10.1080/10717544.2019.1582730\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen, J. \u003cem\u003eet al.\u003c/em\u003e Glucosamine derivative modified nanostructured lipid carriers for targeted tumor delivery. J. Mater. Chem. 22, 5770\u0026ndash;5783, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/c2jm15830b\u003c/span\u003e\u003cspan address=\"10.1039/c2jm15830b\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYostawonkul, J. \u003cem\u003eet al.\u003c/em\u003e Nanocarrier-mediated delivery of α-mangostin for non-surgical castration of male animals. Sci. Rep. 7, 16234, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-017-16563-3\u003c/span\u003e\u003cspan address=\"10.1038/s41598-017-16563-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSalminen, H., Helgason, T., Kristinsson, B., Kristbergsson, K. \u0026amp; Weiss, J. Formation of solid shell nanoparticles with liquid omega-3 fatty acid core. Food Chem. 141, 2934\u0026ndash;2943, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.foodchem.2013.05.120\u003c/span\u003e\u003cspan address=\"10.1016/j.foodchem.2013.05.120\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, L. J. \u003cem\u003eet al.\u003c/em\u003e Fabrication and Characterization of Antioxidant Pickering Emulsions Stabilized by Zein/Chitosan Complex Particles (ZCPs). J. Agric. Food Chem. 63, 2514\u0026ndash;2524, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/jf505227a\u003c/span\u003e\u003cspan address=\"10.1021/jf505227a\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFolch, J., Lees, M. \u0026amp; Sloane Stanley, G. H. A simple method for the isolation and purification of total lipides from animal tissues. J. Biol. Chem. 226, 497\u0026ndash;509, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0021-9258(18)64849-5\u003c/span\u003e\u003cspan address=\"10.1016/S0021-9258(18)64849-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1957).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMetcalfe, L. D., Schmitz, A. A. \u0026amp; Pelka, J. R. Rapid Preparation of Fatty Acid Esters from Lipids for Gas Chromatographic Analysis. Anal. Chem. 38, 514\u0026ndash;515, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/ac60235a044\u003c/span\u003e\u003cspan address=\"10.1021/ac60235a044\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1966).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"n-3 polyunsaturated fatty acids, Lipid-based nanoparticles, Physiochemical characteristics, In vitro storage stability, In vivo biodistribution, Slow-growing chickens","lastPublishedDoi":"10.21203/rs.3.rs-4761693/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4761693/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe aim of this research was to investigate the synthesis of suitable carrier of nanoparticles for improving the utilization of n-3 polyunsaturated fatty acids (n-3 PUFAs) source in chicken diets. Lipid nanoparticles were successfully prepared with two different n-3 oil sources, tuna and algal oils using hot and high-pressure homogenization method. Four preparations were defined as followed: non-targeting lipid nanoparticles containing tuna oil (TO_NPs), non-targeting lipid nanoparticles containing algal oil (AO_NPs), targeting lipid nanoparticles containing tuna oil (TO_TNPs) and targeting lipid nanoparticles containing algal oil (AO_TNPs). A second study was conducted for the targeting procedure, the treatments as followed: Control, TO_NPs and TO_TNPs. Thirty-three slow-growing chickens were examined during the post-administration kinetic at 2, 4, 8, 12 and 24 h. The physicochemical characteristics of lipid nanoparticles, storage stability and in vivo biodistribution were evaluated. The results showed that the particle diameters of TO_NPs and AO_NPs were 223.7 and 294.4 nm, whereas the particle diameters of TO_TNPs and AO_TNPs were 134.7 and 184.0 nm, respectively. The polydispersity index (PDI) and zeta-potential of nanoparticles showed a good distribution and stability in colloid dispersions, respectively. Moreover, the nanoparticles of the TNPs groups were less susceptible to lipid oxidation than that of the NPs groups during a storage at 4\u0026deg;C. The study of the biodistribution based on the Nile red intensity and the determination of n-3 PUFAs composition in chicken meat confirmed the effectiveness of targeted lipid-based nanoparticles to transport directly fatty acids into the skeletal muscle cells of chicken.\u003c/p\u003e","manuscriptTitle":"Enhancement of n-3 PUFAs utilization for functional meat production in slow-growing Korat chicken: evaluation of characteristics of glucose transporter-targeted lipid nanoparticles","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-27 11:49:19","doi":"10.21203/rs.3.rs-4761693/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-05-02T06:50:50+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-03T14:17:15+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"136589161813917781892068261750400762957","date":"2025-03-27T07:15:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"60690223664776056281669337317804029401","date":"2025-03-27T04:25:49+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"178557333385235561658436844131890387093","date":"2025-03-27T02:34:08+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"144471420881666447801074490474387639354","date":"2024-12-27T05:51:01+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"292804912926967036583053808416592130672","date":"2024-11-08T13:38:54+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-08T11:25:50+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"269264644822528785516336175277280646364","date":"2024-08-28T09:58:42+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-08-14T09:50:51+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-08-14T09:50:12+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-07-30T17:09:29+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-07-30T17:06:02+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-07-18T10:01:55+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9fe1f370-ef31-4c73-891c-52abb6a44482","owner":[],"postedDate":"August 27th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":36577677,"name":"Biological sciences/Biochemistry/Lipids/Fats"},{"id":36577679,"name":"Biological sciences/Biochemistry/Lipids/Fatty acids"},{"id":36577681,"name":"Biological sciences/Biochemistry/Lipids/Lipid peroxides"},{"id":36577682,"name":"Biological sciences/Biochemistry/Lipids/Oils"},{"id":36577683,"name":"Physical sciences/Materials science/Nanoscale materials/Nanoparticles"},{"id":36577684,"name":"Physical sciences/Nanoscience and technology/Nanobiotechnology/Nanoparticles"},{"id":36577685,"name":"Biological sciences/Chemical biology/Transporters"},{"id":36577687,"name":"Physical sciences/Chemistry/Biochemistry/Lipids/Fatty acids"},{"id":36577689,"name":"Physical sciences/Chemistry/Biochemistry/Lipids/Lipid peroxides"},{"id":36577690,"name":"Physical sciences/Chemistry/Biochemistry/Lipids/Oils"},{"id":36577692,"name":"Physical sciences/Nanoscience and technology/Nanomedicine/Imaging techniques and agents"}],"tags":[],"updatedAt":"2025-12-22T16:02:00+00:00","versionOfRecord":{"articleIdentity":"rs-4761693","link":"https://doi.org/10.1038/s41598-025-33149-6","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-12-21 15:57:58","publishedOnDateReadable":"December 21st, 2025"},"versionCreatedAt":"2024-08-27 11:49:19","video":"","vorDoi":"10.1038/s41598-025-33149-6","vorDoiUrl":"https://doi.org/10.1038/s41598-025-33149-6","workflowStages":[]},"version":"v1","identity":"rs-4761693","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4761693","identity":"rs-4761693","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}