Fish Active Packaging with ZnO/Fe-MMT nanoparticles

Fish Active Packaging with ZnO/Fe-MMT 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 Fish Active Packaging with ZnO/Fe-MMT nanoparticles Kaveh Rahmanifarah, Mehdi Mahmoudian, Somayeh Mahmoudi Eskandarabadi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3946040/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 29 Jan, 2025 Read the published version in Scientific Reports → Version 1 posted 12 You are reading this latest preprint version Abstract Today, active packaging has become important to increase food safety and decrease its spoilage. In this study, the aim was to delay spoilage and increase the shelf life of rainbow fish fillets with a new hybrid nanocomposite active packaging. Packaging was fabricated with Ethylene vinyl acetate and active compounds such as rosemary extract, zinc oxide nanoparticles, and modified iron (Fe-MMT). Various experiments such as XRD and FESEM analyses were performed on active films, and for fish fillets, total sulfhydryl, volatile nitrogen, and lipid oxidation tests were conducted to determine the shelf life of fish fillets in packages. The highest increase in volatile nitrogen, which is related to spoilage of fish fillet, was 9.45 mg/100g, although the limit is 25 mg/100g. It can be concluded that films containing active compounds have increased the shelf life of fish fillets until the sixth day of storage without changing the texture, color and odor. Physical sciences/Chemistry/Materials chemistry Physical sciences/Chemistry/Polymer chemistry shelf life rainbow trout antibacterial active packaging nanoparticles Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1. Introduction In recent years, a great demand has aroused for food products with high durability, and this demand has led to a high attention in controlling food and protein products (Leso et al., 2020 ). In this respect, producers and consumers of the food products have paid significant attention to some modern methods in order to enhance the quality of the food and agricultural products to increase their durability (Tiwari, Singh, Negi, Dani, & Rawat, 2021 ). The use of preservative chemicals is a common way to increase the shelf life of food. In spite of the effectiveness of this method in enhancing the durability of food products, it is not so appropriate due to their toxicity and the health risks they pose (Videira-Quintela, Martin, & Montalvo, 2021 ). Packaging technology develops as one of the newest advances in the field of the food industry. It plays a vital role in the storage and preservation of food products, and vast researches have been conducted in this field. Polymers are the most common materials used to make packaging(Hu, Huyan, Ding, Dong, & Yu, 2020 ). These polymers can be natural, like starch or chitosan or, synthetic. Synthetic polymers have advantages over natural polymers, such as high resistance against environmental factors, moisture, and gas permittivity. Various synthetic polymers have been used in this field, such as polyethylene, polypropylene, and polyethylene terephthalate (Tajeddin & Arabkhedri, 2020 ). Although in today's world, packaging of all foods is necessary, meat products need more accurate and effective packaging because of high microbial growth and fast rotting (He, Li, Fei, & Peng, 2021 ). Fish meat and its products are enriched with protein, unsaturated fatty acids, and essential minerals for the human body. However, fast microbial growth and fat oxidation decrease the useful life of the fish meat. Escherichia coli and Staphylococcus are among the factors that cause fast rotting and reduce the fish meat durability(Topuz & Uyar, 2020 ). Therefore, considering the importance of fish meat in people's diets, the use of new technologies in the packaging of this food seems necessary. The use of nanotechnology has provided unique opportunities to develop packaging science, improve its properties, and has a significant impact on increasing the quality and shelf life of packaged food during storage or transportation. Active packaging is a concept that has been developed in recent years with the use of nanotechnology (Abu-Dalo, Jaradat, Albiss, & Al-Rawashdeh, 2019 ). An active package contains some nano-structured metal oxide and, or other additives with antioxidant, antibacterial, and absorbing light or moisture properties that help the durability, health, and quality of food products(Eskandarabadi et al., 2019 ). Oxygen scavenger is one of the standard components for active packaging that is used to reduce the oxygen in the package and thus reduce the growth of microbes. Various materials are used as oxygen scavengers, including metals such as iron and porous materials (Eskandarabadi et al., 2019 ). In addition, substances that have antibacterial properties, inhibit the growth of microbes and are used in packaging. Zinc oxide nanoparticles (X. Li et al., 2021 ) and extracts such as rosemary and anthocyanin (Sharma, Barkauskaite, Jaiswal, & Jaiswal, 2021 ) were studied as substances with antibacterial properties and have been used in the packaging of new foods (Piñeros-Hernandez, Medina-Jaramillo, López-Córdoba, & Goyanes, 2017 ). In the current study, ethylene vinyl acetate (EVA) nanocomposite containing additives such as zinc oxide, and iron stabilized on montmorillonite was produced as transparent films and used to prepare active packaging. This new packaging was used to maintain fish fillet, and the effect of the components on the packaging performance and quality of fish fillet was analyzed. 2. Experimental 2.1. Materials PVA (Mw 89,000–98,000, 99% hydrolyzed), Ethylene vinyl acetate (wt: 33% VA), montmorillonite (MMT), FeSO 4 , Zn(NO 3 ) 2 .6H 2 O, 2,2-diphenyl-1-picrylhydrazyl (DPPH), NaBH 4 , brain heart broth, Muller Hinton agar, chloroform, ethanol, MgO, H 2 SO 4 , Methyl red, H 3 BO 3 , Thiobarbituric acid, Trichloro acetic acid, DTNB (Ellman’s Reagent- 5,5-dithio-bis-(2-nitro benzoic acid)), Tris, glycine, EDTA (Ethylene Diamine Tetraacetic Acid) and HCl were purchased from Merck with high purity and used without further purification. Rosemary essential oil was acquired from Idearaye Pishgam Co (Iran). Fresh fish fillets were purchased from a local market. 2.2. Synthesis of ZnO nanoparticles The method performed by Kundu (Kundu, Karak, Barik, & Saha, 2011 ) was used to synthesize zinc oxide nanoparticles. First, to prepare solution (A), 20 ml of distilled water was used to dissolve 0.6 g of Zn(NO 3 ).6H 2 O under stirring at room temperature. Next, to prepare solution (B), 80 ml of distilled water was used to dissolve 0.3 g of PVA, which was added as a stabilizer at 50°C to obtain a clear solution. Both solutions were mixed, and NH 4 OH (0.1 M) solution was added dropwise until the pH reached 12 and a white precipitate was formed. The precipitated powder was dried in an oven at 90–100 ˚C and finally was calcinated at 500°C. 2.3. Synthesis of Fe-MMT 1.8 g of FeSO 4 was dissolved in a mixture of deionized water and absolute ethanol (6 ml, 1:2 v/v). Then 0.5 g of MMT was added into the above solution and dispersed under ultrasonic conditions. A solution of NaBH 4 (1M) was used to stabilize the FeSO 4 on the surface of MMT (S. Li et al., 2010 ). Finally, the stabilized Fe nanoparticles on MMT sheets were separated by centrifuging and washed three times with ethanol, then dried at 45°C in a vacuum oven. 2.4. Preparation of active films Fabrication of nanocomposite films was conducted using a casting method. Active films were prepared by 10 wt% EVA in chloroform. Fe-MMT and ZnO nanoparticles were added to the above solution according to the values given in Table 1 . To obtain a homogeneous mixture of nanoparticles in the EVA solution, ultrasonic stirring was used for 2 min. Rosemary essential oil was added to some formulations. The final mixtures were spread out on smooth and cleaned glass plates and dried for 12 h at room temperature. The composition of prepared films was tabulated in Table 1 . Table 1 Components of the fabricated films. Sample code ZnO Nano-particles (% wt) Fe-MMT Nano-particles (% wt) Rosemary (% wt) F 1 0 0 0 F 2 0 2 0 F 3 0 0 2 F 4 2 0 0 F 5 2 2 2 2.5. Characterization The Fourier Transform Infrared spectroscopy (FT-IR) analysis was carried out to survey the structural characteristics of active films. The analysis was done using an FT-IR JASCO 4100 instrument, and spectra were recorded from 500 to 4000 cm − 1 . X-ray diffraction (XRD) pattern of the films and crystalline structure of ZnO, Fe-MMT were analyzed with XRD in the 2θ range of 5° to 80° (X’PertPro, Panalitycal.co, Holland). The morphology of nanoparticles and fabricated films were visualized using a field emission scanning electron microscope (FESEM) (Hitachi S4160). UV/VIS Spectrometer (biochrom Ltd. UK) was used to calculate total sulfhydryl groups in fish filets. 2.6. Antioxidation activity The antioxidant activity of the films was estimated by assaying the scavenging ability of DPPH free radicals (Phang, Malek, & Ibrahim, 2013 ). 0.05 g of a film that had been prepared was placed in a glass vial. Then, 5 ml of ethanol was added to the vial. The mixture was left undisturbed for 20 min, allowing the antioxidant to be extracted from the formulation. In another vial, the stock solution was prepared by dissolving 4 mg of DPPH into 100 ml methanol for 30 min. The working solution was obtained by mixing 1 ml film extract solution with 3 ml of stock solution. The resultant mixture was left in a dark environment at room temperature for 30 min. The antioxidation capacity of films was calculated using the following equation: \(I\% =[({A_{blank}} - {A_{sample}})/{A_{blank}}] \times 100\) In this equation, A blank and A sample are the absorbance of the blank and real sample at 515 nm, respectively, and I is the antioxidation activity (in percent). 2.7. Antibacterial test Staphylococcus aureus bacteria were used to evaluate the antibacterial activity of the prepared films. The antibacterial activities of films were tested using the disk diffusion method. For performing the antibacterial test, 50 ml of brain heart broth solution was prepared and located in an autoclave at 121°C for 15 min. Then, the suspensions of homogenized microorganisms (turbidity at 0.5 McFarland standards) were added to the previous solution, and it was kept for 24 h to let bacteria growth. Also, 50 ml of Mueller-Hinton agar was prepared and placed in an autoclave at 121°C for 15 min. The bacteria solution was impregnated onto agar grow medium the bacterial lawn technique; following the solution was poured into plates and then the prepared films with 1×1 dimensions were placed in the culture medium containing bacteria in an incubator for 24–48 h at 37°C in order to culture the bacteria. Finally, after this period of time, the inhibition zone area was measured (Abu-Dalo et al., 2019 ). 2.8. Oxygen absorption To measure oxygen absorption capacity, Fe-MMT nanostructures and the prepared films were placed in a 40 ml glass container sealed with a septum. The relative humidity in the bottles was set at 94% RH with a vessel that contained 1 ml of water. The bottles contain ambient air (20.9% O 2 ). The samples were fixed in the bottles and then, the containers were sealed and kept at room temperature for 24 h. An oxygen sensor instrument (Regensburg, Germany) was used to measure the percentage of oxygen in different periods. 2.9. Sampling and storage conditions for fish fillet Fresh rainbow trout ( Oncorhynchus mykiss ) was purchased from the local market for testing. Fresh trout was thoroughly cleaned, and its fillet was used. The fabricated films were used to evaluate their performance in the shelf life of fish fillets. To perform the experiment, 100 g of fish fillets were weighed and packed in the prepared films, and then these samples were stored in a refrigerator at 4°C. The experiments were performed on packaged fish fillets at intervals of 0, 3, 6, and 9 days of storage. The results are tabulated and discussed. 2.9.1. Moisture The moisture content of fish fillet samples was determined by the AOAC 1990 method (Debnath, Hemavathy, & Bhat, 2002 ). In order to determine the moisture, the samples were placed at 105°C in an oven for 5 h. Moisture content is calculated from the weight difference between fish fillets before and after drying. 2.9.2. pH A homogenizer was used to stir 5 g of the sample (packed meats) in 45 ml distilled water for 1 min. The pH of the resulting mixture was measured at room temperature using a pH meter (Crison Instruments, Barcelona, Spain). 2.9.3. Water binding capacity 5 g of the sample was weighed and placed between filter paper and pressured under a 0.5 kg weight. After 30min, the sample was weighed again and the observed weight difference was reported as the amount of water loss in the sample (Luther, Weber, & Schuster, 1983 ). 2.9.4. Total sulfhydryl group content The content of sulfhydryl groups was measured using DTNB (Ellman, 1959)(Etemadian, Shabanpour, Mahoonak, & Shabani, 2012 ). Briefly, 1 g of fish fillet was dispersed in 20 ml with 1% (p/v) NaCl in tris–glycine buffer (10.4 g tris, 6.9 g glycine, 1.2 g EDTA per liter, pH 8.0). 2.9 ml of 0.5% SDS in tris–glycine buffer was added to 0.1 ml of fish fillet solution and 0.02 ml of Ellman’s reagent (4 mg ml − 1 DTNB in tris–glycine buffer). All samples were stored at 40 ˚C for 15 min. A molar extinction coefficient of 13,600 M − 1 cm − 1 was used to calculate total SH (lmol/g) after measuring absorbance at 412 nm. 2.9.5. Total volatile nitrogen The total volatile nitrogen content of the samples was measured by distillation and Kjeldahl titration. In this method, 10 g of the sample was homogenized with 50 ml of distilled water with a homogenizer and transferred to Kjeldahl distillation balloon with 2 g of magnesium oxide and 250 ml of distilled water. The distillation balloon was connected to the relevant device, and the end hopper of the device was immersed in 25 ml of 2wt% boric acid solution and methyl red reagent (the balloon outlet should be completely immersed in boric acid to prevent vapor escaping). The content of the balloon were heated for 30–40 min. Finally, titration was performed with a 0.1 N sulfuric acid solution. The amount of volatile nitrogen (in mg / 100 g of the sample) was calculated according to the following equation(Mokrani, Oumouna, & Cuesta, 2018 ). $$TVN=14\times the amount of acid consumed \left(ml\right)$$ 2.9.6. Lipid oxidation Thiobarbituric acid (TBARS) as a reactive substances utilized to determination of oxidative activity of the samples based on Buege and Aust methods (Buege & Aust, 1978 ). In this respect, 0.5 g of fish fillet was poured into a 50 ml falcon tube and mixed with 10 ml of thiobarbituric acid solution (including 0.375 g thiobarbituric acid reagent powder, 15% trichloroacetic acid, and hydrochloric acid / 0.25 N). Then, the falcon tube was immersed in a boiling water bath for 10 min to obtain a pink color. Then the falcon tubes were cooled in a cold water bath and the content of tube was centrifuged at 7000 rpm for 10 min immediately, and the adsorption of the supernatant was read at 532 nm. Finally, after drawing the standard curve using 1, 1, 3, and 3-tetratoxy-propane, the amount of thiobarbituric acid was calculated and reported in mg of malondialdehyde per kg of sample(Mokrani et al., 2018 ). 2.9.7. Microbiological analyses For microbiological testing, the plate count method was used. In this method, the number of colonies on a plate can be counted when the colony becomes visible to the naked eye (Mokrani et al., 2018 ). After the growth of gram-negative E. coli. Bacteria, the number of colonies was calculated and reported as follows: Number of colonies = 10 × 10 − 3 × Counted 2.9.8. SDS–PAGE analysis Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) was used to visualize changes in fish fillet protein during the storage time according to the method of Laemmli (Guerrero, Bertrand, & Rochefort, 2011 ). SDS-PAGE analysis was carried out using 5% wt stacking gel and 15% wt separating gel. The protein extract was diluted 1:1 with Laemmli buffer (containing 5% 2-mercaptoethanol) and quickly heated at 100 ˚C for 5 min. Exactly 10 µg of protein was loaded on to each gel. The mobility of the protein bands were calibrated with standards of molecular weight markers. After staining and distaining, the gel was scanned using a gel documentation system (Bio-Rad, USA)(Muhammad, Mahmoud, Fazio, & Sayed, 2018 ). 2.9.9. Sensory evaluation In order to assess the sensory quality of fish fillets, a sensory analysis was conducted by fifty panelists (aged between 25 and 50 years old) selected from the Urmia University, Seafood Quality Control center at the different storage times. The tester panelists were utilized to detect differences in sensory attributes of stored fish fillets in the five different packagings. They were asked to test the samples for the following characteristics: color intensity, acid odor and tissue. Results are reported based on average scores(Cullere et al., 2018 ). Each sensory attribute was rated on a continuous linear scale ranging from 1 (dislike very much) to 5 (like very much). 2.9.10. Statistical analysis Three replications were carried out for each test (n = 3),t and statistical analysis was performed using SPSS16.0 software. SPSS and data mean comparison were estimated by the Duncan test, and values with (P < 0.05) were considered significant. Mean standard error (SE) was presented in Tables and Figures (Cullere et al., 2018 ). 3. Results and Discussion 3.1. Characterization Different applied nanostructures and the prepared nanocomposite films as active packaging’s were characterized by various techniques such as FT-IR, XRD, and FESEM, and then their performance was evaluated in preserving fish fillet. 3.1.1. FT-IR The FT-IR spectra of additives and the fabricated films are shown in Fig. 1 . The FT-IR spectrum of Fe-MMT exhibited characteristic peaks at 3500, 1132, 875, and 630 cm − 1 which corresponded to the hydroxyl groups, Si – O – Si, Si – O – Al and Fe–O bounds, respectively (Y. Li, Xiaojiao, Guo, Zhou, & Na, 2015 ). In the FT-IR spectra of ZnO nanoparticles, peaks that appeared in the area below 1000 cm − 1 were associated with the Zn-O bond, an and the peaks at 3350 and 1560 cm − 1 were related to hydroxyl groups and adsorbed water (Wang, Wu, Fangyuan, & Yang, 2014 ). The characteristic peaks of rosemary were related to O-H, C = O, and C = C functional groups of an aromatic skeleton and appeared at 3400, 1752, and 1515 cm − 1 (El-Badry & Samir, 2015 ). In the EVA films, the peak of the carbonyl group was observed at 1739 cm − 1 , and other characteristic peaks which were related to C-H and C-O bonds appeared at 2923, 1242, and 1020 cm − 1 , respectively (Bessarabov, Тaran, Zagoriy, & Vakhitova, 2019 ). Incorporated additives in the film matrix were not recognizable in this test due to their low percentage (2 wt%) and the complete encapsulation of these compounds by EVA. On the other hand, the technique used in studying the films (FT-IR) was more limited to the surface and reduced the sensitivity of the analysis. 3.1.2. XRD XRD analyses were conducted to study the crystalline structure of nanoparticles and their containing films. XRD pattern of samples are shown in Fig. 2 . The strong diffraction peaks of Fe-MMT are at 2θ = 25, 30, 35 and 43 °(Harun, Almadani, & Radzi, 2016 ). Probably formation of FeOOH leads to appearance of a new peak at 2θ = 35, and therefore, proved successful Fe insertion onto MMT plates. The size of these particles was calculated by about 15 nm. The obvious diffraction peaks of rosemary appeared at 2θ = 20.85, 31, 45.47, 56.51, 66.28, 75.29, and 76.76°. ). Moreover, the XRD pattern of rosemary has a structure that can be described as face-centered cubic. The ZnO nanoparticles showed a crystalline pattern that corresponded to the characteristic hexagonal structure (Lee, Nam, Kim, & Boo, 2012 ). The average size of ZnO nanoparticles was 38 nm, which was obtained from the Scherrer relationship (Adelnia, Cheraghi Bidsorkhi, Ismail, & Matsuura, 2015). As can be seen in the EVA XRD pattern (F 1 ), this polymer has shown a characteristic broaden peak at 2θ = 23° due to its amorphous nature. With the incorporation of Fe-MMT nanoparticles in film F 2 , the characteristic peaks in the Fe-MMT pattern have become wider, which may be due to the polymer insertion between the MMT layers and the dispersion of the monolayers in the EVA matrix. A similar trend has been observed in the rosemary pattern. However, in films F 4 and F 5 , the presence of ZnO nanoparticles can be clearly detected due to the appearance of the relevant peaks. Moreover, rosemary had a significant effect on reducing the characteristic peak intensity of the EVA, which was intensified in the presence of ZnO and Fe-MMT nanostructures. This can be due to the effect of rosemary on the orientation and movement of polymer chains, which has affected the order in the polymer microstructure. 3.1.3. FESEM FESEM images of nanoparticles and the prepared films are shown in Fig. 3 . As can be observed, ZnO nanoparticles had a spherical morphology with a rough surface, and mean particle diameter was about 40 nm. Moreover, FESEM images of MMT and Fe-MMT were taken and compared. Layers of MMT with a smooth surface were visible, while in the FESEM image of Fe-MMT, needle-shaped appendages were seen, which were related to Fe nanoparticles, and proved its successful stabilization. (Almadani, Radzi, & Harun, 2016 ). By comparing the morphology of the film without additives (F 1 ) with the films containing nanoparticles (F 2 , F 3 , F 4 , and F 5 ), it was found that film F 1 had a smooth surface, however, the presence of additives in the films caused thier surface to be uneven and decreased the uniformity of the films. The cracks that can be observed on the surface of F 1 film were due to the high energy of the collision of electrons during the imaging process (Mumin et al., 2015 ). However, these cracks in films containing additives have been drastically reduced, and this was probably due to the reinforcing effect of nanoparticles on film strength. 3.2. Antioxidant activity Antioxidants reduce active materials damaging effects by scavenging free radicals. DPPH is a substance used in antioxidant tests to measure the antioxidant activity of the samples (Zamani, Moradi Delfani, & Jabbari, 2018 ). The results of the antioxidant activity as percentages of free radical scavenging by DPPH are given in Table 2 . According to the results, F 5 , which contained rosemary extract, ZnO, and Fe-MMT nanoparticles inhibited 36.8% DPPH free radical inhibitory and had the greatest effect compared to samples containing other nanoparticles and control film. The same trend was observed during at 3, 6, and 9-day intervals. Over time, with the consumption of antioxidants, inhibitory activity has decreased, and as a result, their effectiveness reduced. The antioxidant properties of rosemary and nanoparticles have been studied by other researches. In a study by Hernandez et al. (Piñeros-Hernandez et al., 2017 ), rosemary was loaded into a starch film to measure the antioxidant effect of the active film. According to the results, with increasing the percentage of rosemary, the free radical inhabitation increased, and the best inhibitory was achieved using 5% of rosemary, (28%). The antioxidant activity of the present film has been significantly improved compared to similar work despite the lower percentage of rosemary. Furthermore, the F 3 film containing rosemary extract has also shown considerable inhibitory activity (32.5%). Table 2 Analysis of the prepared films. Days F 1 F 2 F 3 F 4 F 5 DPPH-free radical 0 7.4 ± 0.01 Ea 28.5 ± 0.01 Ca 32.5 ± 0.01 Ba 27.6 ± 0.05 Da 36.8 ± 0.01 Aa inhibitory(%) 3 6.2 ± 0.07 Eb 25.3 ± 0.1 Cb 31 ± 0.1 Bb 25 ± 0.7 Db 35 ± 0.2 Ab 6 4.8 ± 0.04 Ec 20.1 ± 0.2 Cc 28 ± 0.03 Bc 19.8 ± 0.2 Dc 33.3 ± 0.1 Ac 9 3.1 ± 0.2 Ed 14 ± 0.2 Dd 25 ± 0.5 Bd 15.2 ± 0.1 Cd 30 ± 0.3 Ad Antibacterial 0 0 Ea 5 ± 0.02 Da 17 ± 0.01 Ba 15 ± 0.01 Ca 20 ± 0.05 Aa activity(mm) 3 0 Ea 2 ± 0.2 Db 15.9 ± 0.4 Bb 13.6 ± 0.4 Cb 18.4 ± 0.1 Ab 6 0 Ca 0 Cc 14 ± 0.4 Ac 12.4 ± 0.3 Bc 14.3 ± 0.2 Ac 9 0 Ca 0 Cc 8 ± 0.3 Ad 7 ± 0.2 Bd 8.2 ± 0.2 Ad Different letters (a, b) within each column denote significant differences among same time (P < 0.05). Different letters (A, B) within each raw denote significant differences among same group (P < 0.05). 3.3. Antibacterial activity Staphylococcus aureus was used to evaluate the antibacterial performance of the films. The related tests were performed, and the obtained data are tabulated in Table 2 . Antibacterial activity was recorded as the diameter of the growth inhibition zone and increasing the diameter indicates an improvement in antibacterial properties. Film F 1 , which was free of any additives, did not show antibacterial properties. However, nanocomposite films showed antibacterial activity. The highest activity was observed in films F 3 and F 5 (containing rosemary and hybrid nanoparticles, respectively), and the lowest activity belonged to film F 2 . ZnO and rosemary have been used as antibacterial agents in active packaging (X. Li et al., 2021 ; Piñeros-Hernandez et al., 2017 ), and different mechanisms were proposed for their antibacterial activity(Bajalan, Rouzbahani, Pirbalouti, & Maggi, 2017 ). The antibacterial activity of the films was significantly maintained until the sixth day and then decreased. A review of similar works showed that this stability has improved remarkably. 3.4. Oxygen absorption The oxygen absorption ability of films containing additives was investigated and the results are shown in Fig. 4 . According to the previous studies, the highest oxygen absorption was observed in 100% humidity. When the humidity is high, the reaction between iron and oxygen is increasing, so Fe 2 O 3 .3H 2 O produces and the level of O 2 decreases (Foltynowicz, Bardenshtein, Sängerlaub, Antvorskov, & Kozak, 2017 ). Due to the effect of Fe nanoparticles in oxygen absorption, this ability in the Fe-MMT powder was also evaluated, and its efficiency was proven. Examination of the films showed that F 1 film had little effect on oxygen absorption, while this capability was greatly increased in film F 2 , which contains Fe-MMT nanoparticles. The oxygen absorption in film F 5 , which contained a hybrid of nanoparticles and rosemary extract, has also increased compared to film F 2 , which showed the role of ZnO nanoparticles and rosemary extract in oxygen absorption. As shown in the diagrams, the ability to absorb oxygen was maintained until the fifteenth day and decreased after that. 3.5. Effect of active films on fish fillets shelf life Due to the importance of preserving fish fillets because of their high rate of spoilage, the prepared formulation in the packaging of fish fillets were used and various parameters were evaluated to investigate the performance of the prepared films. 3.5.1. Moisture The moisture content is an important parameter that is affected by fish spoilage. Fish spoilage is a result of lipid oxidation, which leads to the destruction of the protein structure and the ability to hold water in the fish fillet tissue. Therefore, moisture content was measured for fish fillets packed in films. Results are given in Table 3 , and it can be said that all samples had almost constant moisture, and this content has not changed significantly at different times. The moisture content of the fish fillets and films was about 75%, which is also true for these samples. (Hematyar, Rustad, Sampels, & Kastrup Dalsgaard, 2019 ; Seline Glorieux, Olivier Goemaere, Liselot Steen, & Fraeye, 2017). 3.5.2. pH The pH was another factor that was assessed for fish fillets packaged and stored at 4 ˚C for 9 days. The growth of bacteria in the fish fillet leads to the destruction of the protein structure that causes the release of alkaline compounds such as ammonia. This process results in the pH increase. Recorded pHs are listed in Table 3 . The pH value of the control sample (without additives) and films containing nanoparticles increased from 6 to 7 over 9 days. A closer look revealed that the pH increase in the samples containing nanoparticles was less than the control sample, and in fact less ammonia was released which indicated less spoilage of the meat. This protective effect seemed reasonable when considering the antibacterial and antioxidant effect of nanoparticles and rosemary extract (Ashrafi, Jokar, & Mohammadi Nafchi, 2018 ). Considerable differences (P < 0.05) were observed for the control and nanocomposite samples on different days (Van Haute, Raes, Van der Meeren, & Sampers, 2016 ). The lowest pH changes are related to F 5 . The presence of compounds such as rosemary extract, Fe-MMT, and ZnO nanostructures that acting as antioxidant and antibacterial ingredients has led to the observation of this result. The maximum acceptable pH range for meat consumption is 6.8 to 7, and therefore, the meat in all the samples was within the acceptable range in terms of pH during the storage time. Table 3 The efficiency of parameters which affect on rainbow trout fillets kept during storage at 4℃ Experiments Days F 1 F 2 F 3 F 4 F 5 Moisture 0 75.4 ± 0.7 Ac 75.4 ± 0.7 Ac 75.4 ± 0.7 Ac 75.4 ± 0.7 Ac 75.4 ± 0.7 Ac 3 75.8 ± 0.14 Ab 75.5 ± 0.15 Bb 75.6 ± 0.14 Ab 75.8 ± 0.13 Ab 75.2 ± 0.14 Cc 6 75.8 ± 0.28 Cb 76.3 ± 0.14 Aa 75.3 ± 0.7 Bb 75.8 ± 0.14 Db 75.7 ± 0.28 Cb 9 76.6 ± 0.14 Aa 76.3 ± 0.2 BCa 76.2 ± 0.2 CDa 76.4 ± 0.28 Ba 76 ± 0.31 Da pH 0 6.13 ± 0.014 Ac 6.11 ± 0.014 Ac 6.10 ± 0.014 Ac 6.12 ± 0.014 Ad 6.13 ± 0.014 Ac 3 6.6 ± 0.042 Ab 6.33 ± 0.005 Cb 6.26 ± 0.005 Db 6.5 ± 0.021 Bc 6.17 ± 0.042 Ec 6 6.66 ± 0.007 Ab 6.45 ± 0.021 Da 6.45 ± 0.035 Da 6.61 ± 0.005 Bb 6.4 ± 0.042 Cb 9 6.93 ± 0.007 Aa 6.58 ± 0.042 Ca 6.54 ± 0.021 Da 6.81 ± 0.049 Ba 6.47 ± 0.028 Ea Water binding 0 13.5 ± 0.28 Ad 13.6 ± 0.28 Ac 13.5 ± 0.28 Ac 13.6 ± 0.28 Ad 13.4 ± 0.28 Ad capacity 3 16.1 ± 0.2 Cc 17.5 ± 0.2 Ab 16 ± 0.2 Cb 16.9 ± 0.2 Bc 14.7 ± 0.14 Dc 6 17.9 ± 0.21 Bb 18.1 ± 0.3 Aa 16.8 ± 0.2 Da 17.4 ± 0.2 Cb 16.5 ± 0.2 Eb 9 18.2 ± 0.7 Aa 18.2 ± 0.7 Aa 17 ± 0.3 Ca 17.9 ± 0.4 Ba 16.8 ± 0.7 Da Active sulfhydryl 0 7.05 ± 0.02 Aa 7.04 ± 0.02 Aa 7.06 ± 0.02 Aa 7.04 ± 0.02 Aa 7.03 ± 0.02 Aa 3 5.78 ± 0.07 Eb 6.21 ± 0.03 Cb 6.86 ± 0.01 Ab 5.90 ± 0.01 Db 6.82 ± 0.05 Bb 6 4.23 ± 0.02 Dc 5.28 ± 0.05 Cc 6.40 ± 0.05 Bc 5.11 ± 0.03 Ec 6.51 ± 0.04 Ac 9 2.39 ± 0.04 Ed 4.84 ± 0.05 Cd 5.93 ± 0.03 Ad 4.06 ± 0.02 Dd 5.81 ± 0.01 Bd Different letters (a, b) within each column denote significant differences among same time (P < 0.05). Different letters (A, B) within each raw denote significant differences among same group (P < 0.05). 3.5.3. Water binding capacity (WBC) In fish meat spoilage, as a result of lipid oxidation, the amino groups in protein structure is destroyed, and the ability to hold water in the fish fillet tissue is decreased. By degradation of amino acids, the ability to retain interstitial water is decreased, so it affects WBC. (Etemadian et al., 2012 ; Hematyar et al., 2019 ). The water binding capacity was investigated and the results are given in Table 3 . According to the results, it can be said that the amount of water loss of fish fillets due to pressure has not changed significantly over time. However, the observed changes were considered. After the third day of storage, the amount of water loss was measurable in the packed fish fillets. These changes continued on the sixth and ninth storage days. There was a significant difference between the different samples, which was related to the degree of protein degradation. According to the obtained results, the least degradation was observed in samples F 3 and F 5 . 3.5.4. Active sulfhydryl To investigate the structural changes of proteins due to oxidation, active sulfhydryl was studied (Etemadian et al., 2012 ). The active sulfhydryl assay is a reliable indicator for checking the quality of fish protein and evaluating its oxidation. During the storage period of fish, protein denaturation leads to thiol group oxidation. As a result, the amount of active sulfhydryl decreased, and disulfide bonds increased. A decrease in the amount of sulfhydryl indicates protein oxidation and tissue destruction. In this study, sulfhydryl content was measured for packaged meat samples. According to the data obtained in Table 3 , changes in the amount of active sulfhydryl were significant from the third day. Changes in the F 1 sample were more obvious. On days 6 and 9, as discussed further, films 3 and 5 were most effective in preventing oxidation, resulting in less oxidized protein and higher levels of active sulfhydryl than other films. There was a significant difference (P < 0.05) between control and samples containing antioxidant and antibacterial active compounds during fish fillet storage. 3.5.5. TVN TVN indicates the quality of meat, and its increase is due to bacterial and enzyme activities (Etemadian et al., 2012 ). Metabolism of amino acids in fish fillet releases some compounds such as ammonia, monoethylamine, diethylamine, and trimethylamine, and causes an unpleasant smell (Ashrafi et al., 2018 ; Echegaray, Domínguez, Franco, Lorenzo, & Carballo, 2018 ). Figure 5 shows the rate of change in the amount of TVN over a period of 9 days. The highest increase in TVN was related to the control sample on the ninth day, which was 9.45 mg / 100g. Of course, the TVN limit is 25 mg / 100g, which even the control sample did not exceed. The smallest change in TVN content was observed in the F 5 sample, which contained all the additives. Also, the F 3 sample, with rosemary extract as an antioxidant, showed good stability. The obtained results showed the role of components, especially rosemary extract in protecting fish fillet. 3.5.6. Lipid oxidation Free radicals are create by the reaction of oxygen with double bonds in lipid oxidation. The oxygen attack on the double bond in the fatty acid triggers the onset of radical reactions. Moreover, the oxidation of polyunsaturated fatty acids (PUFA) can develop an unpleasant off flavor and formation of volatiles. Light, heat, and the presence of metal ions and radicals can cause autoxidation in meat and fish fillets (Echegaray et al., 2018 ). Furthermore, there are various enzymes in fish fillets, which are capable of catalyzing lipid oxidation such as lipoxygenases and myeloperox idases. According to the research, TBA value of 2 mg Kg-1 is the limit (Etemadian et al., 2012 ; Hematyar et al., 2019 ). TBARS-related changes can be seen in Fig. 6 . Although the value of TBA did not exceed the limit during the storage time, the amount of malondialdehyde on the first day of the experiment was less than in subsequent days, which is normal because the amount of this substance in fish fillets increases over time (P < 0.05). The F 5 sample had the lowest increase in TBARS which was related to the antioxidant and antibacterial properties of rosemary and nanoparticles. This film prevented oxidative reactions and reduced the oxidation of fat in meat. Significant differences were seen between different packaged samples. 3.5.7. Microbiological test Bacteria were counted to monitor the shelf life of the meat (Van Haute et al., 2016 ). The results of the microbial test performed on days 0, 3, 6, and 9 for five samples at 4 ° C are shown in Fig. 7 . According to the data from the microbiological test, no bacteria grew on the first day of the test, and the meat was completely healthy. This trend was also observed on the third day in low dilution of bacteria. Over time, the amount of bacteria in the meat has increased, and the largest increase was related to the control sample, and it can be said that the packaging had the ability to control the growth of bacteria The lowest bacterial growth rate was observed in the F 5 film contained all additives (X. Li et al., 2021 ; Piñeros-Hernandez et al., 2017 ), which was consistent with the results of microbial tests. The allowable limit for bacteria is 7 log CFU / g, which by the sixth day no sample has exceeded this limit and meat samples have not been microbial contaminated. 3.5.8. SDS-page pattern Many characteristics of meat tissue depend on myofibrillar proteins. Myosin is one of the most important proteins from the family of myofibrillar proteins. Myosin heavy chains (MHC) are very susceptible to hydrolysis and their size reduction is a measure of meat quality during the storage period. The changes in myosin size in the electrophoresis pattern of SDS-PAGE gel were investigated during the storage period, and the results are shown in Fig S1 -S4. It was observed that MHC was 63 kD at the beginning of the experiment, and decreased to 48 kD on the third day, and remained almost unchanged until the ninth day. As seen in the patterns, all the samples in this test have shown a similar pattern. 3.5.9. Sensory analysis The sensory properties of meat, which are related to color, odor, and texture quality, are given in Table 4 . Color and odor are the main quality characteristics, which can be affected in fish fillets by lipid oxidation and microbial corruption (Hematyar et al., 2019 ). According to the obtained data, all the samples were fresh on the first day and had a pink color. Fish fillet texture also had good elasticity. On the third day of storage, the samples had no odor, were still elastic, and had lost some of their natural color. On the sixth day, changes began to appear. Only on the ninth day, the control sample and sample in the film containing Fe-MMT had a small amount of odor, and the meat was slightly pale. Significant differences were observed between F 3 and F 5 samples with other samples in terms of color, odor, and meat quality(Etemadian et al., 2012 ). Considering microbial, lipid oxidation, etc. sensory analysis of fish fillets in different packages can be fully justified. Table 4 Sensory evaluation in rainbow trout fillets kept during storage at 4 ° C Experiments Days F 1 F 2 F 3 F 4 F 5 Sensory quality 0 4.7 ± 1.1 aA 4.7 ± 1.1 aA 4.7 ± 1.1 aA 4.7 ± 1.1 aA 4.7 ± 1.1 aA Appearance 3 4.2 ± 1.2 aA 4.1 ± 1.4 aA 4.3 ± 1.3 aA 4.2 ± 1.4 aA 4.5 ± 0.7 aA 6 4.0 ± 1.5 aA 3.4 ± 1.2 bAB 3.8 ± 1.3 aAB 4.0 ± 1.1 aA 4.0 ± 0.7 aA 9 3.3 ± 1.5 aB 3.3 ± 1.3 aB 3.4 ± 1.1 aB 3.4 ± 1.1 aB 3.5 ± 0.6 aB Odor 0 4.7 ± 0.7 aA 4.7 ± 0.7 aA 4.7 ± 0.7 aA 4.7 ± 0.7 aA 4.7 ± 0.7 aA 3 4.1 ± 0.9 aA 4.0 ± 1.2 aA 4.1 ± 0.8 aA 4.0 ± 1.0 aA 4.1 ± 1.4 aA 6 3.6 ± 0.6 aAB 3.6 ± 0.9 aA 3.9 ± 0.8 aAB 3.9 ± 1.1 aA 3.9 ± 1.5 aAB 9 2.9 ± 0.8 bB 3.0 ± 0.9 bB 3.6 ± 0.6 aAB 3.2 ± 1.1 abB 3.8 ± 1.5 aB Overall 0 4.6 ± 1.2 aA 4.6 ± 1.2 aA 4.6 ± 1.2 aA 4.6 ± 1.2 aA 4.6 ± 1.2 aA acceptability 3 4.0 ± 1.2 aA 3.9 ± 1.4 aA 4.1 ± 0.9 aA 4.1 ± 1.5 aA 4.1 ± 1.2 aA 6 3.1 ± 0.8 bB 3.6 ± 1.2 abAB 3.9 ± 1.3 aA 3.6 ± 1.0 abAB 4.0 ± 1.4 aA 9 2.4 ± 0.8 bBC 2.7 ± 1.2 bB 3.3 ± 0.9 aB 2.9 ± 1.0 bB 3.7 ± 1.5 aA Different letters (a, b) within each column denote significant differences among same time (P < 0.05). Different letters (A, B) within each raw denote significant differences among same group (P < 0.05). 4. Conclusion Given the concerns about healthy food consumption, it is important to develop a system that will increase food shelf life and ensure its quality. In this study, the aim was to use compounds within the polymer structure to increase the shelf life of fish fillets in the refrigerator at 4 ° C. ZnO and Fe-MMT nanoparticles and rosemary were used agents. Based on the tests performed and the results obtained, the antibacterial and antioxidant activity of the film containing all additives was higher than other films. This result can also be seen in experiments performed on fish fillets. The amount of zone inhibition bacteria grown in meat and the amount of oxidized and degraded proteins in the two films containing rosemary and all additives (F3 and F5, respectively) were much lower and significantly different from other films. Although nanoparticles were also effective against spoilage and increased shelf life, rosemary compound was more effective until the ninth day. Concomitant use of nanoparticles with rosemary extract has much effect on increasing better results due to synergistic effect. Declarations Declaration of competing interest The authors declare no conflict of interests. Competing interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Author Contribution Dr. Rahmani wrote and edited Effect of active films on fish fillets shelf life part in article. Dr. mahmoudian and somayeh mahmoudi eskandarabadi provided pictures and paticipate in writing introduction and all other parts of the article. all authors reviewed the manuscripts. Acknowledgments This work was supported by the fund for Urmia University (Artemia and Aquaculture Research Institute No.77.126 ) The raw/processed data required to reproduce these findings cannot be shared at this time due to legal or ethical reasons. Data availability The datasets used and/or analysed during the current study available from the corresponding author on reasonable request. References Abu-Dalo, M., Jaradat, A., Albiss, B. A., & Al-Rawashdeh, N. A. F. (2019). Green synthesis of TiO2 NPs/pristine pomegranate peel extract nanocomposite and its antimicrobial activity for water disinfection. Journal of Environmental Chemical Engineering, 7 (5), 103370. doi:https://doi.org/10.1016/j.jece.2019.103370 Adelnia, H., Cheraghi Bidsorkhi, H., Ismail, A., & Matsuura, T. (2015). Gas permeability and permselectivity properties of ethylene vinyl acetate/sepiolite mixed matrix membranes. Separation and Purification Technology, 146 , 351–357. doi:10.1016/j.seppur.2015.03.060 Almadani, E., Radzi, S., & Harun, F. (2016). Stearic Acid Esters Synthesized using Iron Exchanged Montmorillonite K10 Clay Catalysts. International Journal of Applied Chemistry, 12 , 62-67. Ashrafi, A., Jokar, M., & Mohammadi Nafchi, A. (2018). Preparation and characterization of biocomposite film based on chitosan and kombucha tea as active food packaging. International Journal of Biological Macromolecules, 108 , 444-454. doi:https://doi.org/10.1016/j.ijbiomac.2017.12.028 Bajalan, I., Rouzbahani, R., Pirbalouti, A. G., & Maggi, F. (2017). Antioxidant and antibacterial activities of the essential oils obtained from seven Iranian populations of Rosmarinus officinalis. Industrial Crops and Products, 107 , 305-311. doi:https://doi.org/10.1016/j.indcrop.2017.05.063 Bessarabov, V., Тaran, N., Zagoriy, G., & Vakhitova, L. (2019). Definition of the thermal and fire-protective properties of ethylene-vinyl acetate copolymer nanocomposites. Eastern-European Journal of Enterprise Technologies, 1 . doi:10.15587/1729-4061.2019.154676 Buege, J. A., & Aust, S. D. (1978). [30] Microsomal lipid peroxidation. In S. Fleischer & L. Packer (Eds.), Methods in Enzymology (Vol. 52, pp. 302-310): Academic Press. Cullere, M., Dalle Zotte, A., Tasoniero, G., Giaccone, V., Szendrő, Z., Szín, M., . . . Matics, Z. (2018). Effect of diet and packaging system on the microbial status, pH, color and sensory traits of rabbit meat evaluated during chilled storage. Meat Science, 141 , 36-43. doi:https://doi.org/10.1016/j.meatsci.2018.03.014 Debnath, S., Hemavathy, J., & Bhat, K. K. (2002). Moisture sorption studies on onion powder. Food Chemistry, 78 (4), 479-482. doi:https://doi.org/10.1016/S0308-8146(02)00161-9 Echegaray, N., Domínguez, R., Franco, D., Lorenzo, J. M., & Carballo, J. (2018). Effect of the use of chestnuts (Castanea sativa Miller) in the finishing diet of Celta pig breed on the shelf-life of meat refrigerated and frozen. Food Research International, 114 , 114-122. doi:https://doi.org/10.1016/j.foodres.2018.07.036 El-Badry, A., & Samir, S. (2015). Essential Oils: A Promising Remedy against Fungal and Bacterial Human Keratitis. Egyptian Journal of Botany and Microbiology, II , 403-431. Eskandarabadi, S. M., Mahmoudian, M., Farah, K. R., Abdali, A., Nozad, E., & Enayati, M. (2019). Active intelligent packaging film based on ethylene vinyl acetate nanocomposite containing extracted anthocyanin, rosemary extract and ZnO/Fe-MMT nanoparticles. Food Packaging and Shelf Life, 22 , 100389. doi:https://doi.org/10.1016/j.fpsl.2019.100389 Etemadian, Y., Shabanpour, B., Mahoonak, A. S., & Shabani, A. (2012). Combination effect of phosphate and vacuum packaging on quality parameters of Rutilus frisii kutum fillets in ice. Food Research International, 45 (1), 9-16. doi:https://doi.org/10.1016/j.foodres.2011.09.026 Foltynowicz, Z., Bardenshtein, A., Sängerlaub, S., Antvorskov, H., & Kozak, W. (2017). Nanoscale, zero valent iron particles for application as oxygen scavenger in food packaging. Food Packaging and Shelf Life, 11 , 74-83. doi:https://doi.org/10.1016/j.fpsl.2017.01.003 Guerrero, M. P., Bertrand, F., & Rochefort, D. (2011). Activity, stability and inhibition of a bioactive paper prepared by large-scale coating of laccase microcapsules. Chemical Engineering Science - CHEM ENG SCI, 66 , 5313-5320. doi:10.1016/j.ces.2011.07.026 Harun, F., Almadani, E., & Radzi, S. (2016). Metal cation exchanged montmorillonite K10 (MMT K10): Surface properties and catalytic activity. 3 , 90-96. He, Y., Li, H., Fei, X., & Peng, L. (2021). Carboxymethyl cellulose/cellulose nanocrystals immobilized silver nanoparticles as an effective coating to improve barrier and antibacterial properties of paper for food packaging applications. Carbohydrate Polymers, 252 , 117156. doi:https://doi.org/10.1016/j.carbpol.2020.117156 Hematyar, N., Rustad, T., Sampels, S., & Kastrup Dalsgaard, T. (2019). Relationship between lipid and protein oxidation in fish. Aquaculture Research, 50 (5), 1393-1403. doi:https://doi.org/10.1111/are.14012 Hu, K., Huyan, Z., Ding, S., Dong, Y., & Yu, X. (2020). Investigation on food packaging polymers: Effects on vegetable oil oxidation. Food Chemistry, 315 , 126299. doi:https://doi.org/10.1016/j.foodchem.2020.126299 Kundu, T. K., Karak, N., Barik, P., & Saha, S. (2011). Optical properties of ZnO nanoparticles prepared by chemical method using poly (vinylalcohol) (PVA) as capping agent. IJSCE, 1 , 19-24. Lee, S., Nam, S.-H., Kim, M.-H., & Boo, J.-H. (2012). Synthesis and Photocatalytic Property of ZnO Nanoparticles Prepared by Spray-Pyrolysis Method. Physics Procedia, 32 , 320-326. doi:10.1016/j.phpro.2012.03.563 Leso, L., Barbari, M., Lopes, M. A., Damasceno, F. A., Galama, P., Taraba, J. L., & Kuipers, A. (2020). Invited review: Compost-bedded pack barns for dairy cows. Journal of Dairy Science, 103 (2), 1072-1099. doi:https://doi.org/10.3168/jds.2019-16864 Li, S., Li, H., Zhu, N., Li, P., Wu, J., Wang, X., & Dang, Z. (2010). Synthesis and characterization of organo-montmorillonite supported iron nanoparticles. Applied Clay Science, 50 , 330–336. doi:10.1016/j.clay.2010.08.021 Li, X., Ren, Z., Wang, R., Liu, L., Zhang, J., Ma, F., . . . Liu, X. (2021). Characterization and antibacterial activity of edible films based on carboxymethyl cellulose, Dioscorea opposita mucilage, glycerol and ZnO nanoparticles. Food Chemistry, 349 , 129208. doi:https://doi.org/10.1016/j.foodchem.2021.129208 Li, Y., Xiaojiao, C., Guo, J., Zhou, S., & Na, P. (2015). Fe/Ti co-pillared clay for enhanced arsenite removal and photo oxidation under UV irradiation. Applied Surface Science, 324 , 179-187. doi:10.1016/j.apsusc.2014.10.111 Luther, H., Weber, E., & Schuster, E. (1983). [Characterization of the water-binding capacity of proteins]. Nahrung, 27 (3), 265-271. doi:10.1002/food.19830270323 Mokrani, D., Oumouna, M., & Cuesta, A. (2018). Fish farming conditions affect to European sea bass (Dicentrarchus labrax L.) quality and shelf life during storage in ice. Aquaculture, 490 , 120-124. doi:https://doi.org/10.1016/j.aquaculture.2018.02.032 Muhammad, O. I., Mahmoud, U. M., Fazio, F., & Sayed, A. E.-D. H. (2018). SDS-PAGE technique as biomarker for fish toxicological studies. Toxicology Reports, 5 , 905-909. doi:https://doi.org/10.1016/j.toxrep.2018.08.020 Mumin, M. A., Akhter, K. F., Dresser, S., van Dinther, S. T., Wu, W., & Charpentier, P. A. (2015). Multifunctional mesoporous silica nanoparticles in poly(ethylene-co-vinyl acetate) for transparent heat retention films. Journal of Polymer Science Part B: Polymer Physics, 53 (12), 851-859. doi:https://doi.org/10.1002/polb.23707 Phang, C.-W., Malek, S. N. A., & Ibrahim, H. (2013). Antioxidant potential, cytotoxic activity and total phenolic content of Alpinia pahangensis rhizomes. BMC Complementary and Alternative Medicine, 13 (1), 243. doi:10.1186/1472-6882-13-243 Piñeros-Hernandez, D., Medina-Jaramillo, C., López-Córdoba, A., & Goyanes, S. (2017). Edible cassava starch films carrying rosemary antioxidant extracts for potential use as active food packaging. Food Hydrocolloids, 63 , 488-495. doi:https://doi.org/10.1016/j.foodhyd.2016.09.034 Seline Glorieux, Olivier Goemaere, Liselot Steen, & Fraeye, I. (2017). Phosphate Reduction in Emulsified Meat Products: Impact of Phosphate Type and Dosage on Quality Characteristics. food technology and biotechnology, 55 , 390–397. Sharma, S., Barkauskaite, S., Jaiswal, A. K., & Jaiswal, S. (2021). Essential oils as additives in active food packaging. Food Chemistry, 343 , 128403. doi:https://doi.org/10.1016/j.foodchem.2020.128403 Tajeddin, B., & Arabkhedri, M. (2020). Chapter 16 - Polymers and food packaging. In M. A. A. AlMaadeed, D. Ponnamma, & M. A. Carignano (Eds.), Polymer Science and Innovative Applications (pp. 525-543): Elsevier. Tiwari, K., Singh, R., Negi, P., Dani, R., & Rawat, A. (2021). Application of nanomaterials in food packaging industry: A review. Materials Today: Proceedings . doi:https://doi.org/10.1016/j.matpr.2021.01.385 Topuz, F., & Uyar, T. (2020). Antioxidant, antibacterial and antifungal electrospun nanofibers for food packaging applications. Food Research International, 130 , 108927. doi:https://doi.org/10.1016/j.foodres.2019.108927 Van Haute, S., Raes, K., Van der Meeren, P., & Sampers, I. (2016). The effect of cinnamon, oregano and thyme essential oils in marinade on the microbial shelf life of fish and meat products. Food Control, 68 , 30-39. doi:https://doi.org/10.1016/j.foodcont.2016.03.025 Videira-Quintela, D., Martin, O., & Montalvo, G. (2021). Recent advances in polymer-metallic composites for food packaging applications. Trends in Food Science & Technology, 109 , 230-244. doi:https://doi.org/10.1016/j.tifs.2021.01.020 Wang, L., Wu, Y., Fangyuan, C., & Yang, X. (2014). Photocatalytic enhancement of Mg-doped ZnO nanocrystals hybridized with reduced graphene oxide sheets. Progress in Natural Science: Materials International, 24 . doi:10.1016/j.pnsc.2014.01.002 Zamani, M., Moradi Delfani, A., & Jabbari, M. (2018). Scavenging performance and antioxidant activity of γ-alumina nanoparticles towards DPPH free radical: Spectroscopic and DFT-D studies. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 201 , 288-299. doi:https://doi.org/10.1016/j.saa.2018.05.004 Additional Declarations No competing interests reported. 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Rahmanifarah","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4ElEQVRIie3RMQrCMBSA4SeBuCS6pgTqFSoFF8GzNAhOQRQXwcGCEM8gDh5CyNySoUsPIDi5u6ijDqYouMW6OeSHhBD4yIMA+Hx/GLMLVQfcTO0evW6zeoRkvxJgSc3BgpSeblM5GLeCc+96mAygvc4aZu4gHJox3+jhDHMZb2U0BFYmkJcOEgIGTjUSyhIkIzvkASBP3QTdqV4KFZQVWULnG+GAsX3FCMVIRQxE30iwwrhPdSEUGc0sKUi3FKmTsEKhI9ULsVubPZKPRRgWxlxc5P0pnwhAwwl8Pp/PV6MnP30/ovNv4WIAAAAASUVORK5CYII=","orcid":"","institution":"Urmia University","correspondingAuthor":true,"prefix":"","firstName":"Kaveh","middleName":"","lastName":"Rahmanifarah","suffix":""},{"id":281297975,"identity":"c8c13d57-f9be-4e36-a5cf-1ce66b0de476","order_by":1,"name":"Mehdi Mahmoudian","email":"","orcid":"","institution":"Urmia University","correspondingAuthor":false,"prefix":"","firstName":"Mehdi","middleName":"","lastName":"Mahmoudian","suffix":""},{"id":281297976,"identity":"2e4f7374-bc8a-47e8-80cd-4506828b0f2a","order_by":2,"name":"Somayeh Mahmoudi Eskandarabadi","email":"","orcid":"","institution":"Urmia University","correspondingAuthor":false,"prefix":"","firstName":"Somayeh","middleName":"Mahmoudi","lastName":"Eskandarabadi","suffix":""}],"badges":[],"createdAt":"2024-02-10 14:04:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3946040/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3946040/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-025-88008-1","type":"published","date":"2025-01-29T15:58:15+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":53193679,"identity":"66bab1f2-89fb-4173-ac75-37cca0488a2d","added_by":"auto","created_at":"2024-03-21 18:03:28","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":219918,"visible":true,"origin":"","legend":"\u003cp\u003eFT-IR spectra of additives and the prepared films\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/861bd0788d590d368b913785.jpeg"},{"id":53193683,"identity":"4309ad3c-7403-4d04-8368-73a5014dc35c","added_by":"auto","created_at":"2024-03-21 18:03:29","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":160787,"visible":true,"origin":"","legend":"\u003cp\u003eXRD pattern of additives prepared films\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/e44e5df3ef95b6f45d147f9b.jpeg"},{"id":53193681,"identity":"b579b31a-a3e4-4378-9b96-be7a42a6ab46","added_by":"auto","created_at":"2024-03-21 18:03:28","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":223455,"visible":true,"origin":"","legend":"\u003cp\u003eFESEM micrographs of ZnO and Fe-MMT nanoparticles and the prepared films\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/458236ca3c46b04dafbea24e.jpeg"},{"id":53193682,"identity":"b5ec44e5-4050-4469-a35b-93cb6104a07d","added_by":"auto","created_at":"2024-03-21 18:03:28","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":151930,"visible":true,"origin":"","legend":"\u003cp\u003eEvolution of oxygen rate of Fe-MMT and prepared films, during shelf life\u003c/p\u003e","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/d15adbca274d384d7b4f3c37.jpeg"},{"id":53193684,"identity":"d8e09147-1228-4973-9921-ef1e1d6f3ebb","added_by":"auto","created_at":"2024-03-21 18:03:29","extension":"jpeg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":96756,"visible":true,"origin":"","legend":"\u003cp\u003eChanges in amount of total volatile nitrogen in fish fillets\u003c/p\u003e","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/0bd16153d89ade964dbd5578.jpeg"},{"id":53193686,"identity":"b2dd31ac-9c0d-4614-8e0c-d62b868a0f8f","added_by":"auto","created_at":"2024-03-21 18:03:29","extension":"jpeg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":114117,"visible":true,"origin":"","legend":"\u003cp\u003eChanges in Oxidative state in fish fillets\u003c/p\u003e","description":"","filename":"floatimage6.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/30cb9e617fe2fefd9fa90ff1.jpeg"},{"id":53193685,"identity":"cb69d493-0dbf-4e0e-9117-cdffa6bd1878","added_by":"auto","created_at":"2024-03-21 18:03:29","extension":"jpeg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":79222,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of packaging on bacteria grows during shelf life\u003c/p\u003e","description":"","filename":"floatimage7.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/f5e3a937f02a668bd307252e.jpeg"},{"id":75351356,"identity":"f58b9d7f-8798-47cd-b958-2d4552d47eeb","added_by":"auto","created_at":"2025-02-03 16:10:04","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2271488,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/3c022337-be92-4b34-87cc-325456d1da96.pdf"},{"id":53193680,"identity":"5972cac7-4dde-4ce1-be3c-d648d8bcf1e9","added_by":"auto","created_at":"2024-03-21 18:03:28","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":210234,"visible":true,"origin":"","legend":"","description":"","filename":"supplementarymaterialR1.docx","url":"https://assets-eu.researchsquare.com/files/rs-3946040/v1/3cdc272b2b282c1a2bd72088.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Fish Active Packaging with ZnO/Fe-MMT nanoparticles","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eIn recent years, a great demand has aroused for food products with high durability, and this demand has led to a high attention in controlling food and protein products (Leso et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). In this respect, producers and consumers of the food products have paid significant attention to some modern methods in order to enhance the quality of the food and agricultural products to increase their durability (Tiwari, Singh, Negi, Dani, \u0026amp; Rawat, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The use of preservative chemicals is a common way to increase the shelf life of food. In spite of the effectiveness of this method in enhancing the durability of food products, it is not so appropriate due to their toxicity and the health risks they pose (Videira-Quintela, Martin, \u0026amp; Montalvo, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePackaging technology develops as one of the newest advances in the field of the food industry. It plays a vital role in the storage and preservation of food products, and vast researches have been conducted in this field. Polymers are the most common materials used to make packaging(Hu, Huyan, Ding, Dong, \u0026amp; Yu, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These polymers can be natural, like starch or chitosan or, synthetic. Synthetic polymers have advantages over natural polymers, such as high resistance against environmental factors, moisture, and gas permittivity. Various synthetic polymers have been used in this field, such as polyethylene, polypropylene, and polyethylene terephthalate (Tajeddin \u0026amp; Arabkhedri, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAlthough in today's world, packaging of all foods is necessary, meat products need more accurate and effective packaging because of high microbial growth and fast rotting (He, Li, Fei, \u0026amp; Peng, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Fish meat and its products are enriched with protein, unsaturated fatty acids, and essential minerals for the human body. However, fast microbial growth and fat oxidation decrease the useful life of the fish meat. Escherichia coli and Staphylococcus are among the factors that cause fast rotting and reduce the fish meat durability(Topuz \u0026amp; Uyar, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Therefore, considering the importance of fish meat in people's diets, the use of new technologies in the packaging of this food seems necessary.\u003c/p\u003e \u003cp\u003eThe use of nanotechnology has provided unique opportunities to develop packaging science, improve its properties, and has a significant impact on increasing the quality and shelf life of packaged food during storage or transportation. Active packaging is a concept that has been developed in recent years with the use of nanotechnology (Abu-Dalo, Jaradat, Albiss, \u0026amp; Al-Rawashdeh, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). An active package contains some nano-structured metal oxide and, or other additives with antioxidant, antibacterial, and absorbing light or moisture properties that help the durability, health, and quality of food products(Eskandarabadi et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Oxygen scavenger is one of the standard components for active packaging that is used to reduce the oxygen in the package and thus reduce the growth of microbes. Various materials are used as oxygen scavengers, including metals such as iron and porous materials (Eskandarabadi et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In addition, substances that have antibacterial properties, inhibit the growth of microbes and are used in packaging. Zinc oxide nanoparticles (X. Li et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and extracts such as rosemary and anthocyanin (Sharma, Barkauskaite, Jaiswal, \u0026amp; Jaiswal, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) were studied as substances with antibacterial properties and have been used in the packaging of new foods (Pi\u0026ntilde;eros-Hernandez, Medina-Jaramillo, L\u0026oacute;pez-C\u0026oacute;rdoba, \u0026amp; Goyanes, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the current study, ethylene vinyl acetate (EVA) nanocomposite containing additives such as zinc oxide, and iron stabilized on montmorillonite was produced as transparent films and used to prepare active packaging. This new packaging was used to maintain fish fillet, and the effect of the components on the packaging performance and quality of fish fillet was analyzed.\u003c/p\u003e"},{"header":"2. Experimental","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003e2.1. Materials\u003c/h2\u003e\n\u003cp\u003ePVA (Mw 89,000\u0026ndash;98,000, 99% hydrolyzed), Ethylene vinyl acetate (wt: 33% VA), montmorillonite (MMT), FeSO\u003csub\u003e4\u003c/sub\u003e, Zn(NO\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003e.6H\u003csub\u003e2\u003c/sub\u003eO, 2,2-diphenyl-1-picrylhydrazyl (DPPH), NaBH\u003csub\u003e4\u003c/sub\u003e, brain heart broth, Muller Hinton agar, chloroform, ethanol, MgO, H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e, Methyl red, H\u003csub\u003e3\u003c/sub\u003eBO\u003csub\u003e3\u003c/sub\u003e, Thiobarbituric acid, Trichloro acetic acid, DTNB (Ellman\u0026rsquo;s Reagent- 5,5-dithio-bis-(2-nitro benzoic acid)), Tris, glycine, EDTA (Ethylene Diamine Tetraacetic Acid) and HCl were purchased from Merck with high purity and used without further purification. Rosemary essential oil was acquired from Idearaye Pishgam Co (Iran). Fresh fish fillets were purchased from a local market.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003e2.2. Synthesis of ZnO nanoparticles\u003c/h2\u003e\n\u003cp\u003eThe method performed by Kundu (Kundu, Karak, Barik, \u0026amp; Saha, \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e) was used to synthesize zinc oxide nanoparticles. First, to prepare solution (A), 20 ml of distilled water was used to dissolve 0.6 g of Zn(NO\u003csub\u003e3\u003c/sub\u003e).6H\u003csub\u003e2\u003c/sub\u003eO under stirring at room temperature. Next, to prepare solution (B), 80 ml of distilled water was used to dissolve 0.3 g of PVA, which was added as a stabilizer at 50\u0026deg;C to obtain a clear solution. Both solutions were mixed, and NH\u003csub\u003e4\u003c/sub\u003eOH (0.1 M) solution was added dropwise until the pH reached 12 and a white precipitate was formed. The precipitated powder was dried in an oven at 90\u0026ndash;100 ˚C and finally was calcinated at 500\u0026deg;C.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.3. Synthesis of Fe-MMT\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1.8 g of FeSO\u003csub\u003e4\u003c/sub\u003e was dissolved in a mixture of deionized water and absolute ethanol (6 ml, 1:2 v/v). Then 0.5 g of MMT was added into the above solution and dispersed under ultrasonic conditions. A solution of NaBH\u003csub\u003e4\u003c/sub\u003e (1M) was used to stabilize the FeSO\u003csub\u003e4\u003c/sub\u003e on the surface of MMT (S. Li et al., \u003cspan class=\"CitationRef\"\u003e2010\u003c/span\u003e). Finally, the stabilized Fe nanoparticles on MMT sheets were separated by centrifuging and washed three times with ethanol, then dried at 45\u0026deg;C in a vacuum oven.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.4. Preparation of active films\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFabrication of nanocomposite films was conducted using a casting method. Active films were prepared by 10 wt% EVA in chloroform. Fe-MMT and ZnO nanoparticles were added to the above solution according to the values given in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. To obtain a homogeneous mixture of nanoparticles in the EVA solution, ultrasonic stirring was used for 2 min. Rosemary essential oil was added to some formulations. The final mixtures were spread out on smooth and cleaned glass plates and dried for 12 h at room temperature. The composition of prepared films was tabulated in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eComponents of the fabricated films.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSample\u003c/p\u003e\n\u003cp\u003ecode\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eZnO Nano-particles\u003c/p\u003e\n\u003cp\u003e(% wt)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eFe-MMT Nano-particles (% wt)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eRosemary\u003c/p\u003e\n\u003cp\u003e(% wt)\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n\u003ch2\u003e2.5. Characterization\u003c/h2\u003e\n\u003cp\u003eThe Fourier Transform Infrared spectroscopy (FT-IR) analysis was carried out to survey the structural characteristics of active films. The analysis was done using an FT-IR JASCO 4100 instrument, and spectra were recorded from 500 to 4000 cm \u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eX-ray diffraction (XRD) pattern of the films and crystalline structure of ZnO, Fe-MMT were analyzed with XRD in the 2\u0026theta; range of 5\u0026deg; to 80\u0026deg; (X\u0026rsquo;PertPro, Panalitycal.co, Holland).\u003c/p\u003e\n\u003cp\u003eThe morphology of nanoparticles and fabricated films were visualized using a field emission scanning electron microscope (FESEM) (Hitachi S4160). UV/VIS Spectrometer (biochrom Ltd. UK) was used to calculate total sulfhydryl groups in fish filets.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n\u003ch2\u003e2.6. Antioxidation activity\u003c/h2\u003e\n\u003cp\u003eThe antioxidant activity of the films was estimated by assaying the scavenging ability of DPPH free radicals (Phang, Malek, \u0026amp; Ibrahim, \u003cspan class=\"CitationRef\"\u003e2013\u003c/span\u003e). 0.05 g of a film that had been prepared was placed in a glass vial. Then, 5 ml of ethanol was added to the vial. The mixture was left undisturbed for 20 min, allowing the antioxidant to be extracted from the formulation. In another vial, the stock solution was prepared by dissolving 4 mg of DPPH into 100 ml methanol for 30 min. The working solution was obtained by mixing 1 ml film extract solution with 3 ml of stock solution. The resultant mixture was left in a dark environment at room temperature for 30 min. The antioxidation capacity of films was calculated using the following equation:\u003c/p\u003e\n\u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(I\\% =[({A_{blank}} - {A_{sample}})/{A_{blank}}] \\times 100\\)\u003c/span\u003e \u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eIn this equation, A \u003csub\u003eblank\u003c/sub\u003e and A \u003csub\u003esample\u003c/sub\u003e are the absorbance of the blank and real sample at 515 nm, respectively, and \u003cem\u003eI\u003c/em\u003e is the antioxidation activity (in percent).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003e2.7. Antibacterial test\u003c/h2\u003e\n\u003cp\u003eStaphylococcus aureus bacteria were used to evaluate the antibacterial activity of the prepared films. The antibacterial activities of films were tested using the disk diffusion method. For performing the antibacterial test, 50 ml of brain heart broth solution was prepared and located in an autoclave at 121\u0026deg;C for 15 min. Then, the suspensions of homogenized microorganisms (turbidity at 0.5 McFarland standards) were added to the previous solution, and it was kept for 24 h to let bacteria growth. Also, 50 ml of Mueller-Hinton agar was prepared and placed in an autoclave at 121\u0026deg;C for 15 min. The bacteria solution was impregnated onto agar grow medium the bacterial lawn technique; following the solution was poured into plates and then the prepared films with 1\u0026times;1 dimensions were placed in the culture medium containing bacteria in an incubator for 24\u0026ndash;48 h at 37\u0026deg;C in order to culture the bacteria. Finally, after this period of time, the inhibition zone area was measured (Abu-Dalo et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n\u003ch2\u003e2.8. Oxygen absorption\u003c/h2\u003e\n\u003cp\u003eTo measure oxygen absorption capacity, Fe-MMT nanostructures and the prepared films were placed in a 40 ml glass container sealed with a septum. The relative humidity in the bottles was set at 94% RH with a vessel that contained 1 ml of water. The bottles contain ambient air (20.9% O\u003csub\u003e2\u003c/sub\u003e). The samples were fixed in the bottles and then, the containers were sealed and kept at room temperature for 24 h. An oxygen sensor instrument (Regensburg, Germany) was used to measure the percentage of oxygen in different periods.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n\u003ch2\u003e2.9. Sampling and storage conditions for fish fillet\u003c/h2\u003e\n\u003cp\u003eFresh rainbow trout (\u003cem\u003eOncorhynchus mykiss\u003c/em\u003e) was purchased from the local market for testing. Fresh trout was thoroughly cleaned, and its fillet was used. The fabricated films were used to evaluate their performance in the shelf life of fish fillets. To perform the experiment, 100 g of fish fillets were weighed and packed in the prepared films, and then these samples were stored in a refrigerator at 4\u0026deg;C. The experiments were performed on packaged fish fillets at intervals of 0, 3, 6, and 9 days of storage. The results are tabulated and discussed.\u003c/p\u003e\n\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.1. Moisture\u003c/h2\u003e\n\u003cp\u003eThe moisture content of fish fillet samples was determined by the AOAC 1990 method (Debnath, Hemavathy, \u0026amp; Bhat, \u003cspan class=\"CitationRef\"\u003e2002\u003c/span\u003e). In order to determine the moisture, the samples were placed at 105\u0026deg;C in an oven for 5 h. Moisture content is calculated from the weight difference between fish fillets before and after drying.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.2. pH\u003c/h2\u003e\n\u003cp\u003eA homogenizer was used to stir 5 g of the sample (packed meats) in 45 ml distilled water for 1 min. The pH of the resulting mixture was measured at room temperature using a pH meter (Crison Instruments, Barcelona, Spain).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.3. Water binding capacity\u003c/h2\u003e\n\u003cp\u003e5 g of the sample was weighed and placed between filter paper and pressured under a 0.5 kg weight. After 30min, the sample was weighed again and the observed weight difference was reported as the amount of water loss in the sample (Luther, Weber, \u0026amp; Schuster, \u003cspan class=\"CitationRef\"\u003e1983\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.4. Total sulfhydryl group content\u003c/h2\u003e\n\u003cp\u003eThe content of sulfhydryl groups was measured using DTNB (Ellman, 1959)(Etemadian, Shabanpour, Mahoonak, \u0026amp; Shabani, \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e). Briefly, 1 g of fish fillet was dispersed in 20 ml with 1% (p/v) NaCl in tris\u0026ndash;glycine buffer (10.4 g tris, 6.9 g glycine, 1.2 g EDTA per liter, pH 8.0). 2.9 ml of 0.5% SDS in tris\u0026ndash;glycine buffer was added to 0.1 ml of fish fillet solution and 0.02 ml of Ellman\u0026rsquo;s reagent (4 mg ml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e DTNB in tris\u0026ndash;glycine buffer). All samples were stored at 40 ˚C for 15 min. A molar extinction coefficient of 13,600 M\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003ecm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e was used to calculate total SH (lmol/g) after measuring absorbance at 412 nm.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.5. Total volatile nitrogen\u003c/h2\u003e\n\u003cp\u003eThe total volatile nitrogen content of the samples was measured by distillation and Kjeldahl titration. In this method, 10 g of the sample was homogenized with 50 ml of distilled water with a homogenizer and transferred to Kjeldahl distillation balloon with 2 g of magnesium oxide and 250 ml of distilled water. The distillation balloon was connected to the relevant device, and the end hopper of the device was immersed in 25 ml of 2wt% boric acid solution and methyl red reagent (the balloon outlet should be completely immersed in boric acid to prevent vapor escaping). The content of the balloon were heated for 30\u0026ndash;40 min. Finally, titration was performed with a 0.1 N sulfuric acid solution. The amount of volatile nitrogen (in mg / 100 g of the sample) was calculated according to the following equation(Mokrani, Oumouna, \u0026amp; Cuesta, \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equa\" class=\"mathdisplay\"\u003e$$TVN=14\\times the amount of acid consumed \\left(ml\\right)$$\u003c/div\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.6. Lipid oxidation\u003c/h2\u003e\n\u003cp\u003eThiobarbituric acid (TBARS) as a reactive substances utilized to determination of oxidative activity of the samples based on Buege and Aust methods (Buege \u0026amp; Aust, \u003cspan class=\"CitationRef\"\u003e1978\u003c/span\u003e). In this respect, 0.5 g of fish fillet was poured into a 50 ml falcon tube and mixed with 10 ml of thiobarbituric acid solution (including 0.375 g thiobarbituric acid reagent powder, 15% trichloroacetic acid, and hydrochloric acid / 0.25 N). Then, the falcon tube was immersed in a boiling water bath for 10 min to obtain a pink color. Then the falcon tubes were cooled in a cold water bath and the content of tube was centrifuged at 7000 rpm for 10 min immediately, and the adsorption of the supernatant was read at 532 nm. Finally, after drawing the standard curve using 1, 1, 3, and 3-tetratoxy-propane, the amount of thiobarbituric acid was calculated and reported in mg of malondialdehyde per kg of sample(Mokrani et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.7. Microbiological analyses\u003c/h2\u003e\n\u003cp\u003eFor microbiological testing, the plate count method was used. In this method, the number of colonies on a plate can be counted when the colony becomes visible to the naked eye (Mokrani et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). After the growth of gram-negative E. coli. Bacteria, the number of colonies was calculated and reported as follows:\u003c/p\u003e\n\u003cp\u003eNumber of colonies\u0026thinsp;=\u0026thinsp;10 \u0026times; 10\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e \u0026times; Counted\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.8. SDS\u0026ndash;PAGE analysis\u003c/h2\u003e\n\u003cp\u003eSodium dodecyl sulfate\u0026ndash;polyacrylamide gel electrophoresis (SDS\u0026ndash;PAGE) was used to visualize changes in fish fillet protein during the storage time according to the method of Laemmli (Guerrero, Bertrand, \u0026amp; Rochefort, \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e). SDS-PAGE analysis was carried out using 5% wt stacking gel and 15% wt separating gel. The protein extract was diluted 1:1 with Laemmli buffer (containing 5% 2-mercaptoethanol) and quickly heated at 100 ˚C for 5 min. Exactly 10 \u0026micro;g of protein was loaded on to each gel. The mobility of the protein bands were calibrated with standards of molecular weight markers. After staining and distaining, the gel was scanned using a gel documentation system (Bio-Rad, USA)(Muhammad, Mahmoud, Fazio, \u0026amp; Sayed, \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.9. Sensory evaluation\u003c/h2\u003e\n\u003cp\u003eIn order to assess the sensory quality of fish fillets, a sensory analysis was conducted by fifty panelists (aged between 25 and 50 years old) selected from the Urmia University, Seafood Quality Control center at the different storage times. The tester panelists were utilized to detect differences in sensory attributes of stored fish fillets in the five different packagings. They were asked to test the samples for the following characteristics: color intensity, acid odor and tissue. Results are reported based on average scores(Cullere et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). Each sensory attribute was rated on a continuous linear scale ranging from 1 (dislike very much) to 5 (like very much).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\n\u003ch2\u003e2.9.10. Statistical analysis\u003c/h2\u003e\n\u003cp\u003eThree replications were carried out for each test (n\u0026thinsp;=\u0026thinsp;3),t and statistical analysis was performed using SPSS16.0 software. SPSS and data mean comparison were estimated by the Duncan test, and values with (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) were considered significant. Mean standard error (SE) was presented in Tables and Figures (Cullere et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1. Characterization\u003c/h2\u003e\n\u003cp\u003eDifferent applied nanostructures and the prepared nanocomposite films as active packaging\u0026rsquo;s were characterized by various techniques such as FT-IR, XRD, and FESEM, and then their performance was evaluated in preserving fish fillet.\u003c/p\u003e\n\u003cdiv id=\"Sec22\" class=\"Section3\"\u003e\n\u003ch2\u003e3.1.1. FT-IR\u003c/h2\u003e\n\u003cp\u003eThe FT-IR spectra of additives and the fabricated films are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. The FT-IR spectrum of Fe-MMT exhibited characteristic peaks at 3500, 1132, 875, and 630 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e which corresponded to the hydroxyl groups, Si \u0026ndash; O \u0026ndash; Si, Si \u0026ndash; O \u0026ndash; Al and Fe\u0026ndash;O bounds, respectively (Y. Li, Xiaojiao, Guo, Zhou, \u0026amp; Na, \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e). In the FT-IR spectra of ZnO nanoparticles, peaks that appeared in the area below 1000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e were associated with the Zn-O bond, an and the peaks at 3350 and 1560 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e were related to hydroxyl groups and adsorbed water (Wang, Wu, Fangyuan, \u0026amp; Yang, \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e). The characteristic peaks of rosemary were related to O-H, C\u0026thinsp;=\u0026thinsp;O, and C\u0026thinsp;=\u0026thinsp;C functional groups of an aromatic skeleton and appeared at 3400, 1752, and 1515 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (El-Badry \u0026amp; Samir, \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e). In the EVA films, the peak of the carbonyl group was observed at 1739 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, and other characteristic peaks which were related to C-H and C-O bonds appeared at 2923, 1242, and 1020 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, respectively (Bessarabov, Тaran, Zagoriy, \u0026amp; Vakhitova, \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). Incorporated additives in the film matrix were not recognizable in this test due to their low percentage (2 wt%) and the complete encapsulation of these compounds by EVA. On the other hand, the technique used in studying the films (FT-IR) was more limited to the surface and reduced the sensitivity of the analysis.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec23\" class=\"Section3\"\u003e\n\u003ch2\u003e3.1.2. XRD\u003c/h2\u003e\n\u003cp\u003eXRD analyses were conducted to study the crystalline structure of nanoparticles and their containing films. XRD pattern of samples are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The strong diffraction peaks of Fe-MMT are at 2\u0026theta;\u0026thinsp;=\u0026thinsp;25, 30, 35 and 43 \u0026deg;(Harun, Almadani, \u0026amp; Radzi, \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). Probably formation of FeOOH leads to appearance of a new peak at 2\u0026theta;\u0026thinsp;=\u0026thinsp;35, and therefore, proved successful Fe insertion onto MMT plates. The size of these particles was calculated by about 15 nm. The obvious diffraction peaks of rosemary appeared at 2\u0026theta;\u0026thinsp;=\u0026thinsp;20.85, 31, 45.47, 56.51, 66.28, 75.29, and 76.76\u0026deg;. ). Moreover, the XRD pattern of rosemary has a structure that can be described as face-centered cubic. The ZnO nanoparticles showed a crystalline pattern that corresponded to the characteristic hexagonal structure (Lee, Nam, Kim, \u0026amp; Boo, \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e). The average size of ZnO nanoparticles was 38 nm, which was obtained from the Scherrer relationship (Adelnia, Cheraghi Bidsorkhi, Ismail, \u0026amp; Matsuura, 2015). As can be seen in the EVA XRD pattern (F\u003csub\u003e1\u003c/sub\u003e), this polymer has shown a characteristic broaden peak at 2\u0026theta;\u0026thinsp;=\u0026thinsp;23\u0026deg; due to its amorphous nature. With the incorporation of Fe-MMT nanoparticles in film F\u003csub\u003e2\u003c/sub\u003e, the characteristic peaks in the Fe-MMT pattern have become wider, which may be due to the polymer insertion between the MMT layers and the dispersion of the monolayers in the EVA matrix. A similar trend has been observed in the rosemary pattern. However, in films F\u003csub\u003e4\u003c/sub\u003e and F\u003csub\u003e5\u003c/sub\u003e, the presence of ZnO nanoparticles can be clearly detected due to the appearance of the relevant peaks. Moreover, rosemary had a significant effect on reducing the characteristic peak intensity of the EVA, which was intensified in the presence of ZnO and Fe-MMT nanostructures. This can be due to the effect of rosemary on the orientation and movement of polymer chains, which has affected the order in the polymer microstructure.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec24\" class=\"Section3\"\u003e\n\u003ch2\u003e3.1.3. FESEM\u003c/h2\u003e\n\u003cp\u003eFESEM images of nanoparticles and the prepared films are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. As can be observed, ZnO nanoparticles had a spherical morphology with a rough surface, and mean particle diameter was about 40 nm. Moreover, FESEM images of MMT and Fe-MMT were taken and compared. Layers of MMT with a smooth surface were visible, while in the FESEM image of Fe-MMT, needle-shaped appendages were seen, which were related to Fe nanoparticles, and proved its successful stabilization. (Almadani, Radzi, \u0026amp; Harun, \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). By comparing the morphology of the film without additives (F\u003csub\u003e1\u003c/sub\u003e) with the films containing nanoparticles (F\u003csub\u003e2\u003c/sub\u003e, F\u003csub\u003e3\u003c/sub\u003e, F\u003csub\u003e4\u003c/sub\u003e, and F\u003csub\u003e5\u003c/sub\u003e), it was found that film F\u003csub\u003e1\u003c/sub\u003e had a smooth surface, however, the presence of additives in the films caused thier surface to be uneven and decreased the uniformity of the films. The cracks that can be observed on the surface of F\u003csub\u003e1\u003c/sub\u003e film were due to the high energy of the collision of electrons during the imaging process (Mumin et al., \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e). However, these cracks in films containing additives have been drastically reduced, and this was probably due to the reinforcing effect of nanoparticles on film strength.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec25\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2. Antioxidant activity\u003c/h2\u003e\n\u003cp\u003eAntioxidants reduce active materials damaging effects by scavenging free radicals. DPPH is a substance used in antioxidant tests to measure the antioxidant activity of the samples (Zamani, Moradi Delfani, \u0026amp; Jabbari, \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). The results of the antioxidant activity as percentages of free radical scavenging by DPPH are given in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. According to the results, F\u003csub\u003e5\u003c/sub\u003e, which contained rosemary extract, ZnO, and Fe-MMT nanoparticles inhibited 36.8% DPPH free radical inhibitory and had the greatest effect compared to samples containing other nanoparticles and control film. The same trend was observed during at 3, 6, and 9-day intervals. Over time, with the consumption of antioxidants, inhibitory activity has decreased, and as a result, their effectiveness reduced. The antioxidant properties of rosemary and nanoparticles have been studied by other researches. In a study by Hernandez et al. (Pi\u0026ntilde;eros-Hernandez et al., \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e), rosemary was loaded into a starch film to measure the antioxidant effect of the active film. According to the results, with increasing the percentage of rosemary, the free radical inhabitation increased, and the best inhibitory was achieved using 5% of rosemary, (28%). The antioxidant activity of the present film has been significantly improved compared to similar work despite the lower percentage of rosemary. Furthermore, the F\u003csub\u003e3\u003c/sub\u003e film containing rosemary extract has also shown considerable inhibitory activity (32.5%).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eAnalysis of the prepared films.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eDays\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eDPPH-free radical\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eEa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e28.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eCa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e32.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eBa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e27.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e36.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003einhibitory(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003eEb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e25.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003eBb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eDb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003eEc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eCc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eBc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e19.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eDc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e33.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eEd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eDd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003csup\u003eBd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e15.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003eCd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAntibacterial\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003csup\u003eEa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eBa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eCa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eactivity(mm)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003csup\u003eEa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eDb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e15.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003csup\u003eBb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e18.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003csup\u003eCa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003csup\u003eCc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e12.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003csup\u003eBc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003csup\u003eCa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003csup\u003eCc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eBd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eDifferent letters (a, b) within each column denote significant differences among same time (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003cp\u003eDifferent letters (A, B) within each raw denote significant differences among same group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec26\" class=\"Section2\"\u003e\n\u003ch2\u003e3.3. Antibacterial activity\u003c/h2\u003e\n\u003cp\u003eStaphylococcus aureus was used to evaluate the antibacterial performance of the films. The related tests were performed, and the obtained data are tabulated in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. Antibacterial activity was recorded as the diameter of the growth inhibition zone and increasing the diameter indicates an improvement in antibacterial properties. Film F\u003csub\u003e1\u003c/sub\u003e, which was free of any additives, did not show antibacterial properties. However, nanocomposite films showed antibacterial activity. The highest activity was observed in films F\u003csub\u003e3\u003c/sub\u003e and F\u003csub\u003e5\u003c/sub\u003e (containing rosemary and hybrid nanoparticles, respectively), and the lowest activity belonged to film F\u003csub\u003e2\u003c/sub\u003e. ZnO and rosemary have been used as antibacterial agents in active packaging (X. Li et al., \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e; Pi\u0026ntilde;eros-Hernandez et al., \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e), and different mechanisms were proposed for their antibacterial activity(Bajalan, Rouzbahani, Pirbalouti, \u0026amp; Maggi, \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe antibacterial activity of the films was significantly maintained until the sixth day and then decreased. A review of similar works showed that this stability has improved remarkably.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec27\" class=\"Section2\"\u003e\n\u003ch2\u003e3.4. Oxygen absorption\u003c/h2\u003e\n\u003cp\u003eThe oxygen absorption ability of films containing additives was investigated and the results are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e. According to the previous studies, the highest oxygen absorption was observed in 100% humidity. When the humidity is high, the reaction between iron and oxygen is increasing, so Fe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e.3H\u003csub\u003e2\u003c/sub\u003eO produces and the level of O\u003csub\u003e2\u003c/sub\u003e decreases (Foltynowicz, Bardenshtein, S\u0026auml;ngerlaub, Antvorskov, \u0026amp; Kozak, \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e). Due to the effect of Fe nanoparticles in oxygen absorption, this ability in the Fe-MMT powder was also evaluated, and its efficiency was proven. Examination of the films showed that F\u003csub\u003e1\u003c/sub\u003e film had little effect on oxygen absorption, while this capability was greatly increased in film F\u003csub\u003e2\u003c/sub\u003e, which contains Fe-MMT nanoparticles. The oxygen absorption in film F\u003csub\u003e5\u003c/sub\u003e, which contained a hybrid of nanoparticles and rosemary extract, has also increased compared to film F\u003csub\u003e2\u003c/sub\u003e, which showed the role of ZnO nanoparticles and rosemary extract in oxygen absorption. As shown in the diagrams, the ability to absorb oxygen was maintained until the fifteenth day and decreased after that.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec28\" class=\"Section2\"\u003e\n\u003ch2\u003e3.5. Effect of active films on fish fillets shelf life\u003c/h2\u003e\n\u003cp\u003eDue to the importance of preserving fish fillets because of their high rate of spoilage, the prepared formulation in the packaging of fish fillets were used and various parameters were evaluated to investigate the performance of the prepared films.\u003c/p\u003e\n\u003cdiv id=\"Sec29\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.1. Moisture\u003c/h2\u003e\n\u003cp\u003eThe moisture content is an important parameter that is affected by fish spoilage. Fish spoilage is a result of lipid oxidation, which leads to the destruction of the protein structure and the ability to hold water in the fish fillet tissue. Therefore, moisture content was measured for fish fillets packed in films. Results are given in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, and it can be said that all samples had almost constant moisture, and this content has not changed significantly at different times. The moisture content of the fish fillets and films was about 75%, which is also true for these samples. (Hematyar, Rustad, Sampels, \u0026amp; Kastrup Dalsgaard, \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e; Seline Glorieux, Olivier Goemaere, Liselot Steen, \u0026amp; Fraeye, 2017).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec30\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.2. pH\u003c/h2\u003e\n\u003cp\u003eThe pH was another factor that was assessed for fish fillets packaged and stored at 4 ˚C for 9 days. The growth of bacteria in the fish fillet leads to the destruction of the protein structure that causes the release of alkaline compounds such as ammonia. This process results in the pH increase. Recorded pHs are listed in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. The pH value of the control sample (without additives) and films containing nanoparticles increased from 6 to 7 over 9 days. A closer look revealed that the pH increase in the samples containing nanoparticles was less than the control sample, and in fact less ammonia was released which indicated less spoilage of the meat. This protective effect seemed reasonable when considering the antibacterial and antioxidant effect of nanoparticles and rosemary extract (Ashrafi, Jokar, \u0026amp; Mohammadi Nafchi, \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). Considerable differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) were observed for the control and nanocomposite samples on different days (Van Haute, Raes, Van der Meeren, \u0026amp; Sampers, \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). The lowest pH changes are related to F\u003csub\u003e5\u003c/sub\u003e. The presence of compounds such as rosemary extract, Fe-MMT, and ZnO nanostructures that acting as antioxidant and antibacterial ingredients has led to the observation of this result. The maximum acceptable pH range for meat consumption is 6.8 to 7, and therefore, the meat in all the samples was within the acceptable range in terms of pH during the storage time.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe efficiency of parameters which affect on rainbow trout fillets kept during storage at 4℃\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eExperiments\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eDays\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eMoisture\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003csup\u003eBb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003eCc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eBb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003eDb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eBCa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eCDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eBa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003epH\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.014\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.014\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.014\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.014\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.014\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003csup\u003eDb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.021\u003csup\u003eBc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042\u003csup\u003eEc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.007\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.021\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.035\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003csup\u003eBb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.007\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042\u003csup\u003eCa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.021\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.049\u003csup\u003eBa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.028\u003csup\u003eEa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eWater binding\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ecapacity\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eCc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eBc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003eDc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003eBb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e18.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003eEb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e18.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e18.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003csup\u003eCa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003csup\u003eBa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eDa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eActive sulfhydryl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eAa\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003eEb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eCb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eAb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eDb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eBb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eDc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eCc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eBc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eEc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003eAc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003eEd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eCd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eAd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eDd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eBd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eDifferent letters (a, b) within each column denote significant differences among same time (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003cp\u003eDifferent letters (A, B) within each raw denote significant differences among same group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec31\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.3. Water binding capacity (WBC)\u003c/h2\u003e\n\u003cp\u003eIn fish meat spoilage, as a result of lipid oxidation, the amino groups in protein structure is destroyed, and the ability to hold water in the fish fillet tissue is decreased. By degradation of amino acids, the ability to retain interstitial water is decreased, so it affects WBC. (Etemadian et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e; Hematyar et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). The water binding capacity was investigated and the results are given in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. According to the results, it can be said that the amount of water loss of fish fillets due to pressure has not changed significantly over time. However, the observed changes were considered. After the third day of storage, the amount of water loss was measurable in the packed fish fillets. These changes continued on the sixth and ninth storage days. There was a significant difference between the different samples, which was related to the degree of protein degradation. According to the obtained results, the least degradation was observed in samples F\u003csub\u003e3\u003c/sub\u003e and F\u003csub\u003e5\u003c/sub\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec32\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.4. Active sulfhydryl\u003c/h2\u003e\n\u003cp\u003eTo investigate the structural changes of proteins due to oxidation, active sulfhydryl was studied (Etemadian et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e). The active sulfhydryl assay is a reliable indicator for checking the quality of fish protein and evaluating its oxidation. During the storage period of fish, protein denaturation leads to thiol group oxidation. As a result, the amount of active sulfhydryl decreased, and disulfide bonds increased. A decrease in the amount of sulfhydryl indicates protein oxidation and tissue destruction. In this study, sulfhydryl content was measured for packaged meat samples. According to the data obtained in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, changes in the amount of active sulfhydryl were significant from the third day. Changes in the F\u003csub\u003e1\u003c/sub\u003e sample were more obvious. On days 6 and 9, as discussed further, films 3 and 5 were most effective in preventing oxidation, resulting in less oxidized protein and higher levels of active sulfhydryl than other films. There was a significant difference (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) between control and samples containing antioxidant and antibacterial active compounds during fish fillet storage.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec33\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.5. TVN\u003c/h2\u003e\n\u003cp\u003eTVN indicates the quality of meat, and its increase is due to bacterial and enzyme activities (Etemadian et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e). Metabolism of amino acids in fish fillet releases some compounds such as ammonia, monoethylamine, diethylamine, and trimethylamine, and causes an unpleasant smell (Ashrafi et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e; Echegaray, Dom\u0026iacute;nguez, Franco, Lorenzo, \u0026amp; Carballo, \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). Figure\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e shows the rate of change in the amount of TVN over a period of 9 days. The highest increase in TVN was related to the control sample on the ninth day, which was 9.45 mg / 100g. Of course, the TVN limit is 25 mg / 100g, which even the control sample did not exceed. The smallest change in TVN content was observed in the F\u003csub\u003e5\u003c/sub\u003e sample, which contained all the additives. Also, the F\u003csub\u003e3\u003c/sub\u003e sample, with rosemary extract as an antioxidant, showed good stability. The obtained results showed the role of components, especially rosemary extract in protecting fish fillet.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec34\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.6. Lipid oxidation\u003c/h2\u003e\n\u003cp\u003eFree radicals are create by the reaction of oxygen with double bonds in lipid oxidation. The oxygen attack on the double bond in the fatty acid triggers the onset of radical reactions. Moreover, the oxidation of polyunsaturated fatty acids (PUFA) can develop an unpleasant off flavor and formation of volatiles. Light, heat, and the presence of metal ions and radicals can cause autoxidation in meat and fish fillets (Echegaray et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). Furthermore, there are various enzymes in fish fillets, which are capable of catalyzing lipid oxidation such as lipoxygenases and myeloperox idases. According to the research, TBA value of 2 mg Kg-1 is the limit (Etemadian et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e; Hematyar et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). TBARS-related changes can be seen in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e. Although the value of TBA did not exceed the limit during the storage time, the amount of malondialdehyde on the first day of the experiment was less than in subsequent days, which is normal because the amount of this substance in fish fillets increases over time (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The F\u003csub\u003e5\u003c/sub\u003e sample had the lowest increase in TBARS which was related to the antioxidant and antibacterial properties of rosemary and nanoparticles. This film prevented oxidative reactions and reduced the oxidation of fat in meat. Significant differences were seen between different packaged samples.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec35\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.7. Microbiological test\u003c/h2\u003e\n\u003cp\u003eBacteria were counted to monitor the shelf life of the meat (Van Haute et al., \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). The results of the microbial test performed on days 0, 3, 6, and 9 for five samples at 4 \u0026deg; C are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e. According to the data from the microbiological test, no bacteria grew on the first day of the test, and the meat was completely healthy. This trend was also observed on the third day in low dilution of bacteria. Over time, the amount of bacteria in the meat has increased, and the largest increase was related to the control sample, and it can be said that the packaging had the ability to control the growth of bacteria The lowest bacterial growth rate was observed in the F\u003csub\u003e5\u003c/sub\u003e film contained all additives (X. Li et al., \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e; Pi\u0026ntilde;eros-Hernandez et al., \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e), which was consistent with the results of microbial tests. The allowable limit for bacteria is 7 log CFU / g, which by the sixth day no sample has exceeded this limit and meat samples have not been microbial contaminated.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec36\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.8. SDS-page pattern\u003c/h2\u003e\n\u003cp\u003eMany characteristics of meat tissue depend on myofibrillar proteins. Myosin is one of the most important proteins from the family of myofibrillar proteins. Myosin heavy chains (MHC) are very susceptible to hydrolysis and their size reduction is a measure of meat quality during the storage period. The changes in myosin size in the electrophoresis pattern of SDS-PAGE gel were investigated during the storage period, and the results are shown in Fig \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003e-S4. It was observed that MHC was 63 kD at the beginning of the experiment, and decreased to 48 kD on the third day, and remained almost unchanged until the ninth day. As seen in the patterns, all the samples in this test have shown a similar pattern.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec37\" class=\"Section3\"\u003e\n\u003ch2\u003e3.5.9. Sensory analysis\u003c/h2\u003e\n\u003cp\u003eThe sensory properties of meat, which are related to color, odor, and texture quality, are given in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e. Color and odor are the main quality characteristics, which can be affected in fish fillets by lipid oxidation and microbial corruption (Hematyar et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). According to the obtained data, all the samples were fresh on the first day and had a pink color. Fish fillet texture also had good elasticity. On the third day of storage, the samples had no odor, were still elastic, and had lost some of their natural color. On the sixth day, changes began to appear. Only on the ninth day, the control sample and sample in the film containing Fe-MMT had a small amount of odor, and the meat was slightly pale. Significant differences were observed between F\u003csub\u003e3\u003c/sub\u003e and F\u003csub\u003e5\u003c/sub\u003e samples with other samples in terms of color, odor, and meat quality(Etemadian et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e). Considering microbial, lipid oxidation, etc. sensory analysis of fish fillets in different packages can be fully justified.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab4\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eSensory evaluation in rainbow trout fillets kept during storage at 4 \u0026deg; C\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eExperiments\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eDays\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSensory quality\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAppearance\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003ebAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003csup\u003eaAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eOdor\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003csup\u003eaAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003csup\u003eaAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003csup\u003eaAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003csup\u003eaAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003csup\u003eabB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eOverall\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eacceptability\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003eabAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0\u003csup\u003eabAB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003csup\u003ebBC\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003csup\u003eaB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0\u003csup\u003ebB\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003csup\u003eaA\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eDifferent letters (a, b) within each column denote significant differences among same time (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003cp\u003eDifferent letters (A, B) within each raw denote significant differences among same group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003eGiven the concerns about healthy food consumption, it is important to develop a system that will increase food shelf life and ensure its quality. In this study, the aim was to use compounds within the polymer structure to increase the shelf life of fish fillets in the refrigerator at 4 \u0026deg; C. ZnO and Fe-MMT nanoparticles and rosemary were used agents. Based on the tests performed and the results obtained, the antibacterial and antioxidant activity of the film containing all additives was higher than other films. This result can also be seen in experiments performed on fish fillets. The amount of zone inhibition bacteria grown in meat and the amount of oxidized and degraded proteins in the two films containing rosemary and all additives (F3 and F5, respectively) were much lower and significantly different from other films. Although nanoparticles were also effective against spoilage and increased shelf life, rosemary compound was more effective until the ninth day. Concomitant use of nanoparticles with rosemary extract has much effect on increasing better results due to synergistic effect.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eDeclaration of competing interest\u003c/h2\u003e\n\u003cp\u003eThe authors declare no conflict of interests.\u003c/p\u003e\n\u003ch2\u003eCompeting interests\u003c/h2\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eDr. Rahmani wrote and edited Effect of active films on fish fillets shelf life part in article. Dr. mahmoudian and somayeh mahmoudi eskandarabadi provided pictures and paticipate in writing introduction and all other parts of the article. all authors reviewed the manuscripts.\u003c/p\u003e\n\u003ch2\u003eAcknowledgments\u003c/h2\u003e\n\u003cp\u003eThis work was supported by the fund for Urmia University (Artemia and Aquaculture Research Institute No.77.126 )\u003c/p\u003e\n\u003cp\u003eThe raw/processed data required to reproduce these findings cannot be shared at this time due to legal or ethical reasons.\u003c/p\u003e\n\u003ch2\u003eData availability\u003c/h2\u003e\n\u003cp\u003eThe datasets used and/or analysed during the current study available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbu-Dalo, M., Jaradat, A., Albiss, B. A., \u0026amp; Al-Rawashdeh, N. A. F. (2019). Green synthesis of TiO2 NPs/pristine pomegranate peel extract nanocomposite and its antimicrobial activity for water disinfection. \u003cem\u003eJournal of Environmental Chemical Engineering, 7\u003c/em\u003e(5), 103370. doi:https://doi.org/10.1016/j.jece.2019.103370\u003c/li\u003e\n\u003cli\u003eAdelnia, H., Cheraghi Bidsorkhi, H., Ismail, A., \u0026amp; Matsuura, T. (2015). Gas permeability and permselectivity properties of ethylene vinyl acetate/sepiolite mixed matrix membranes. \u003cem\u003eSeparation and Purification Technology, 146\u003c/em\u003e, 351\u0026ndash;357. doi:10.1016/j.seppur.2015.03.060\u003c/li\u003e\n\u003cli\u003eAlmadani, E., Radzi, S., \u0026amp; Harun, F. (2016). Stearic Acid Esters Synthesized using Iron Exchanged Montmorillonite K10 Clay Catalysts. \u003cem\u003eInternational Journal of Applied Chemistry, 12\u003c/em\u003e, 62-67. \u003c/li\u003e\n\u003cli\u003eAshrafi, A., Jokar, M., \u0026amp; Mohammadi Nafchi, A. (2018). Preparation and characterization of biocomposite film based on chitosan and kombucha tea as active food packaging. \u003cem\u003eInternational Journal of Biological Macromolecules, 108\u003c/em\u003e, 444-454. doi:https://doi.org/10.1016/j.ijbiomac.2017.12.028\u003c/li\u003e\n\u003cli\u003eBajalan, I., Rouzbahani, R., Pirbalouti, A. G., \u0026amp; Maggi, F. (2017). Antioxidant and antibacterial activities of the essential oils obtained from seven Iranian populations of Rosmarinus officinalis. \u003cem\u003eIndustrial Crops and Products, 107\u003c/em\u003e, 305-311. doi:https://doi.org/10.1016/j.indcrop.2017.05.063\u003c/li\u003e\n\u003cli\u003eBessarabov, V., Тaran, N., Zagoriy, G., \u0026amp; Vakhitova, L. (2019). Definition of the thermal and fire-protective properties of ethylene-vinyl acetate copolymer nanocomposites. \u003cem\u003eEastern-European Journal of Enterprise Technologies, 1\u003c/em\u003e. doi:10.15587/1729-4061.2019.154676\u003c/li\u003e\n\u003cli\u003eBuege, J. A., \u0026amp; Aust, S. D. (1978). [30] Microsomal lipid peroxidation. In S. Fleischer \u0026amp; L. Packer (Eds.), \u003cem\u003eMethods in Enzymology\u003c/em\u003e (Vol. 52, pp. 302-310): Academic Press.\u003c/li\u003e\n\u003cli\u003eCullere, M., Dalle Zotte, A., Tasoniero, G., Giaccone, V., Szendrő, Z., Sz\u0026iacute;n, M., . . . Matics, Z. (2018). Effect of diet and packaging system on the microbial status, pH, color and sensory traits of rabbit meat evaluated during chilled storage. \u003cem\u003eMeat Science, 141\u003c/em\u003e, 36-43. doi:https://doi.org/10.1016/j.meatsci.2018.03.014\u003c/li\u003e\n\u003cli\u003eDebnath, S., Hemavathy, J., \u0026amp; Bhat, K. K. (2002). Moisture sorption studies on onion powder. \u003cem\u003eFood Chemistry, 78\u003c/em\u003e(4), 479-482. doi:https://doi.org/10.1016/S0308-8146(02)00161-9\u003c/li\u003e\n\u003cli\u003eEchegaray, N., Dom\u0026iacute;nguez, R., Franco, D., Lorenzo, J. M., \u0026amp; Carballo, J. (2018). Effect of the use of chestnuts (Castanea sativa Miller) in the finishing diet of Celta pig breed on the shelf-life of meat refrigerated and frozen. \u003cem\u003eFood Research International, 114\u003c/em\u003e, 114-122. doi:https://doi.org/10.1016/j.foodres.2018.07.036\u003c/li\u003e\n\u003cli\u003eEl-Badry, A., \u0026amp; Samir, S. (2015). Essential Oils: A Promising Remedy against Fungal and Bacterial Human Keratitis. \u003cem\u003eEgyptian Journal of Botany and Microbiology, II\u003c/em\u003e, 403-431. \u003c/li\u003e\n\u003cli\u003eEskandarabadi, S. M., Mahmoudian, M., Farah, K. R., Abdali, A., Nozad, E., \u0026amp; Enayati, M. (2019). Active intelligent packaging film based on ethylene vinyl acetate nanocomposite containing extracted anthocyanin, rosemary extract and ZnO/Fe-MMT nanoparticles. \u003cem\u003eFood Packaging and Shelf Life, 22\u003c/em\u003e, 100389. doi:https://doi.org/10.1016/j.fpsl.2019.100389\u003c/li\u003e\n\u003cli\u003eEtemadian, Y., Shabanpour, B., Mahoonak, A. S., \u0026amp; Shabani, A. (2012). Combination effect of phosphate and vacuum packaging on quality parameters of Rutilus frisii kutum fillets in ice. \u003cem\u003eFood Research International, 45\u003c/em\u003e(1), 9-16. doi:https://doi.org/10.1016/j.foodres.2011.09.026\u003c/li\u003e\n\u003cli\u003eFoltynowicz, Z., Bardenshtein, A., S\u0026auml;ngerlaub, S., Antvorskov, H., \u0026amp; Kozak, W. (2017). Nanoscale, zero valent iron particles for application as oxygen scavenger in food packaging. \u003cem\u003eFood Packaging and Shelf Life, 11\u003c/em\u003e, 74-83. doi:https://doi.org/10.1016/j.fpsl.2017.01.003\u003c/li\u003e\n\u003cli\u003eGuerrero, M. P., Bertrand, F., \u0026amp; Rochefort, D. (2011). Activity, stability and inhibition of a bioactive paper prepared by large-scale coating of laccase microcapsules. \u003cem\u003eChemical Engineering Science - CHEM ENG SCI, 66\u003c/em\u003e, 5313-5320. doi:10.1016/j.ces.2011.07.026\u003c/li\u003e\n\u003cli\u003eHarun, F., Almadani, E., \u0026amp; Radzi, S. (2016). Metal cation exchanged montmorillonite K10 (MMT K10): Surface properties and catalytic activity.\u003cem\u003e 3\u003c/em\u003e, 90-96. \u003c/li\u003e\n\u003cli\u003eHe, Y., Li, H., Fei, X., \u0026amp; Peng, L. (2021). Carboxymethyl cellulose/cellulose nanocrystals immobilized silver nanoparticles as an effective coating to improve barrier and antibacterial properties of paper for food packaging applications. \u003cem\u003eCarbohydrate Polymers, 252\u003c/em\u003e, 117156. doi:https://doi.org/10.1016/j.carbpol.2020.117156\u003c/li\u003e\n\u003cli\u003eHematyar, N., Rustad, T., Sampels, S., \u0026amp; Kastrup Dalsgaard, T. (2019). Relationship between lipid and protein oxidation in fish. \u003cem\u003eAquaculture Research, 50\u003c/em\u003e(5), 1393-1403. doi:https://doi.org/10.1111/are.14012\u003c/li\u003e\n\u003cli\u003eHu, K., Huyan, Z., Ding, S., Dong, Y., \u0026amp; Yu, X. (2020). Investigation on food packaging polymers: Effects on vegetable oil oxidation. \u003cem\u003eFood Chemistry, 315\u003c/em\u003e, 126299. doi:https://doi.org/10.1016/j.foodchem.2020.126299\u003c/li\u003e\n\u003cli\u003eKundu, T. K., Karak, N., Barik, P., \u0026amp; Saha, S. (2011). Optical properties of ZnO nanoparticles prepared by chemical method using poly (vinylalcohol) (PVA) as capping agent. \u003cem\u003eIJSCE, 1\u003c/em\u003e, 19-24. \u003c/li\u003e\n\u003cli\u003eLee, S., Nam, S.-H., Kim, M.-H., \u0026amp; Boo, J.-H. (2012). Synthesis and Photocatalytic Property of ZnO Nanoparticles Prepared by Spray-Pyrolysis Method. \u003cem\u003ePhysics Procedia, 32\u003c/em\u003e, 320-326. doi:10.1016/j.phpro.2012.03.563\u003c/li\u003e\n\u003cli\u003eLeso, L., Barbari, M., Lopes, M. A., Damasceno, F. A., Galama, P., Taraba, J. L., \u0026amp; Kuipers, A. (2020). Invited review: Compost-bedded pack barns for dairy cows. \u003cem\u003eJournal of Dairy Science, 103\u003c/em\u003e(2), 1072-1099. doi:https://doi.org/10.3168/jds.2019-16864\u003c/li\u003e\n\u003cli\u003eLi, S., Li, H., Zhu, N., Li, P., Wu, J., Wang, X., \u0026amp; Dang, Z. (2010). Synthesis and characterization of organo-montmorillonite supported iron nanoparticles. \u003cem\u003eApplied Clay Science, 50\u003c/em\u003e, 330\u0026ndash;336. doi:10.1016/j.clay.2010.08.021\u003c/li\u003e\n\u003cli\u003eLi, X., Ren, Z., Wang, R., Liu, L., Zhang, J., Ma, F., . . . Liu, X. (2021). Characterization and antibacterial activity of edible films based on carboxymethyl cellulose, Dioscorea opposita mucilage, glycerol and ZnO nanoparticles. \u003cem\u003eFood Chemistry, 349\u003c/em\u003e, 129208. doi:https://doi.org/10.1016/j.foodchem.2021.129208\u003c/li\u003e\n\u003cli\u003eLi, Y., Xiaojiao, C., Guo, J., Zhou, S., \u0026amp; Na, P. (2015). Fe/Ti co-pillared clay for enhanced arsenite removal and photo oxidation under UV irradiation. \u003cem\u003eApplied Surface Science, 324\u003c/em\u003e, 179-187. doi:10.1016/j.apsusc.2014.10.111\u003c/li\u003e\n\u003cli\u003eLuther, H., Weber, E., \u0026amp; Schuster, E. (1983). [Characterization of the water-binding capacity of proteins]. \u003cem\u003eNahrung, 27\u003c/em\u003e(3), 265-271. doi:10.1002/food.19830270323\u003c/li\u003e\n\u003cli\u003eMokrani, D., Oumouna, M., \u0026amp; Cuesta, A. (2018). Fish farming conditions affect to European sea bass (Dicentrarchus labrax L.) quality and shelf life during storage in ice. \u003cem\u003eAquaculture, 490\u003c/em\u003e, 120-124. doi:https://doi.org/10.1016/j.aquaculture.2018.02.032\u003c/li\u003e\n\u003cli\u003eMuhammad, O. I., Mahmoud, U. M., Fazio, F., \u0026amp; Sayed, A. E.-D. H. (2018). SDS-PAGE technique as biomarker for fish toxicological studies. \u003cem\u003eToxicology Reports, 5\u003c/em\u003e, 905-909. doi:https://doi.org/10.1016/j.toxrep.2018.08.020\u003c/li\u003e\n\u003cli\u003eMumin, M. A., Akhter, K. F., Dresser, S., van Dinther, S. T., Wu, W., \u0026amp; Charpentier, P. A. (2015). Multifunctional mesoporous silica nanoparticles in poly(ethylene-co-vinyl acetate) for transparent heat retention films. \u003cem\u003eJournal of Polymer Science Part B: Polymer Physics, 53\u003c/em\u003e(12), 851-859. doi:https://doi.org/10.1002/polb.23707\u003c/li\u003e\n\u003cli\u003ePhang, C.-W., Malek, S. N. A., \u0026amp; Ibrahim, H. (2013). Antioxidant potential, cytotoxic activity and total phenolic content of Alpinia pahangensis rhizomes. \u003cem\u003eBMC Complementary and Alternative Medicine, 13\u003c/em\u003e(1), 243. doi:10.1186/1472-6882-13-243\u003c/li\u003e\n\u003cli\u003ePi\u0026ntilde;eros-Hernandez, D., Medina-Jaramillo, C., L\u0026oacute;pez-C\u0026oacute;rdoba, A., \u0026amp; Goyanes, S. (2017). Edible cassava starch films carrying rosemary antioxidant extracts for potential use as active food packaging. \u003cem\u003eFood Hydrocolloids, 63\u003c/em\u003e, 488-495. doi:https://doi.org/10.1016/j.foodhyd.2016.09.034\u003c/li\u003e\n\u003cli\u003eSeline Glorieux, Olivier Goemaere, Liselot Steen, \u0026amp; Fraeye, I. (2017). Phosphate Reduction in Emulsified Meat Products: Impact of Phosphate Type and Dosage on Quality Characteristics. \u003cem\u003efood technology and biotechnology, 55\u003c/em\u003e, 390\u0026ndash;397. \u003c/li\u003e\n\u003cli\u003eSharma, S., Barkauskaite, S., Jaiswal, A. K., \u0026amp; Jaiswal, S. (2021). Essential oils as additives in active food packaging. \u003cem\u003eFood Chemistry, 343\u003c/em\u003e, 128403. doi:https://doi.org/10.1016/j.foodchem.2020.128403\u003c/li\u003e\n\u003cli\u003eTajeddin, B., \u0026amp; Arabkhedri, M. (2020). Chapter 16 - Polymers and food packaging. In M. A. A. AlMaadeed, D. Ponnamma, \u0026amp; M. A. Carignano (Eds.), \u003cem\u003ePolymer Science and Innovative Applications\u003c/em\u003e (pp. 525-543): Elsevier.\u003c/li\u003e\n\u003cli\u003eTiwari, K., Singh, R., Negi, P., Dani, R., \u0026amp; Rawat, A. (2021). Application of nanomaterials in food packaging industry: A review. \u003cem\u003eMaterials Today: Proceedings\u003c/em\u003e. doi:https://doi.org/10.1016/j.matpr.2021.01.385\u003c/li\u003e\n\u003cli\u003eTopuz, F., \u0026amp; Uyar, T. (2020). Antioxidant, antibacterial and antifungal electrospun nanofibers for food packaging applications. \u003cem\u003eFood Research International, 130\u003c/em\u003e, 108927. doi:https://doi.org/10.1016/j.foodres.2019.108927\u003c/li\u003e\n\u003cli\u003eVan Haute, S., Raes, K., Van der Meeren, P., \u0026amp; Sampers, I. (2016). The effect of cinnamon, oregano and thyme essential oils in marinade on the microbial shelf life of fish and meat products. \u003cem\u003eFood Control, 68\u003c/em\u003e, 30-39. doi:https://doi.org/10.1016/j.foodcont.2016.03.025\u003c/li\u003e\n\u003cli\u003eVideira-Quintela, D., Martin, O., \u0026amp; Montalvo, G. (2021). Recent advances in polymer-metallic composites for food packaging applications. \u003cem\u003eTrends in Food Science \u0026amp; Technology, 109\u003c/em\u003e, 230-244. doi:https://doi.org/10.1016/j.tifs.2021.01.020\u003c/li\u003e\n\u003cli\u003eWang, L., Wu, Y., Fangyuan, C., \u0026amp; Yang, X. (2014). Photocatalytic enhancement of Mg-doped ZnO nanocrystals hybridized with reduced graphene oxide sheets. \u003cem\u003eProgress in Natural Science: Materials International, 24\u003c/em\u003e. doi:10.1016/j.pnsc.2014.01.002\u003c/li\u003e\n\u003cli\u003eZamani, M., Moradi Delfani, A., \u0026amp; Jabbari, M. (2018). Scavenging performance and antioxidant activity of \u0026gamma;-alumina nanoparticles towards DPPH free radical: Spectroscopic and DFT-D studies. \u003cem\u003eSpectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 201\u003c/em\u003e, 288-299. doi:https://doi.org/10.1016/j.saa.2018.05.004\u003c/li\u003e\n\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":"shelf life, rainbow trout, antibacterial, active packaging, nanoparticles","lastPublishedDoi":"10.21203/rs.3.rs-3946040/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3946040/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eToday, active packaging has become important to increase food safety and decrease its spoilage. In this study, the aim was to delay spoilage and increase the shelf life of rainbow fish fillets with a new hybrid nanocomposite active packaging. Packaging was fabricated with Ethylene vinyl acetate and active compounds such as rosemary extract, zinc oxide nanoparticles, and modified iron (Fe-MMT). Various experiments such as XRD and FESEM analyses were performed on active films, and for fish fillets, total sulfhydryl, volatile nitrogen, and lipid oxidation tests were conducted to determine the shelf life of fish fillets in packages. The highest increase in volatile nitrogen, which is related to spoilage of fish fillet, was 9.45 mg/100g, although the limit is 25 mg/100g. It can be concluded that films containing active compounds have increased the shelf life of fish fillets until the sixth day of storage without changing the texture, color and odor.\u003c/p\u003e","manuscriptTitle":"Fish Active Packaging with ZnO/Fe-MMT nanoparticles","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-21 18:03:23","doi":"10.21203/rs.3.rs-3946040/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-08-07T03:32:10+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-07-26T08:30:11+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"222033108270454168284231953997974097674","date":"2024-07-09T21:30:50+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"27538574458503631667768020861383438413","date":"2024-05-22T12:28:12+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"1bd85132-7781-4479-a111-47ffd37df9c1","date":"2024-03-30T13:30:44+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-03-29T08:00:17+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"5b85e5e9-5fd8-43f0-8fb5-f6093fae29ae","date":"2024-03-29T06:26:28+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-03-28T13:03:30+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-03-23T11:13:28+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-03-18T17:40:35+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-03-18T17:39:11+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-02-10T13:44:23+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":"1f138766-60c3-42d4-a5ae-74a24f0b31c4","owner":[],"postedDate":"March 21st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":29619460,"name":"Physical sciences/Chemistry/Materials chemistry"},{"id":29619461,"name":"Physical sciences/Chemistry/Polymer chemistry"}],"tags":[],"updatedAt":"2025-02-03T16:04:34+00:00","versionOfRecord":{"articleIdentity":"rs-3946040","link":"https://doi.org/10.1038/s41598-025-88008-1","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-01-29 15:58:15","publishedOnDateReadable":"January 29th, 2025"},"versionCreatedAt":"2024-03-21 18:03:23","video":"","vorDoi":"10.1038/s41598-025-88008-1","vorDoiUrl":"https://doi.org/10.1038/s41598-025-88008-1","workflowStages":[]},"version":"v1","identity":"rs-3946040","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3946040","identity":"rs-3946040","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}