Deep-eutectic-solvent-based ultrasound-assisted extraction of polysaccharides from maca: Optimization using Taguchi methodology and comparison with the conventional method

Deep-eutectic-solvent-based ultrasound-assisted extraction of polysaccharides from maca: Optimization using Taguchi methodology and comparison with the conventional method | 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 Research Article Deep-eutectic-solvent-based ultrasound-assisted extraction of polysaccharides from maca: Optimization using Taguchi methodology and comparison with the conventional method Eun Jeong Kim, Choon Young Kim, Kyung Young Yoon This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4340936/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 11 You are reading this latest preprint version Abstract Deep eutectic solvent (DES) was used for the ultrasound-assisted extraction (UAE) of polysaccharides from maca. Extraction parameters affecting the extraction yield were experimentally identified and their significance was further investigated using the Taguchi method. DES prepared from choline chloride and urea afforded the highest yield (20.03%) and was chosen as the solvent for UAE. The optimal extraction parameters were: water content of 30% for DES, ultrasonic power of 300 W, and extraction time of 20. The extraction yield (26.28%) of maca polysaccharides (MPs) obtained using these extraction parameters was more than twice that of MPs obtained by hot-water extraction and UAE with water. Moreover, MPs obtained through DES-based extraction exhibited various biological functions such as inhibiting pancreatic α-amylase and α-glucosidase activities, delaying absorption of glucose and bile acid, and stimulating the probiotic. Therefore, DES can be used to extract polysaccharides from maca with biological action as a highly efficient and non-polluting alternative solvent. Polysaccharide Deep eutectic solvent Ultrasound-assisted extraction Orthogonal experiment design Biological activity Figures Figure 1 Introduction Maca ( Lepidium meyenii ) is a cruciferous plant that grows naturally in the highlands; the roots are mainly used as edible parts. Maca contains bioactive components such as macaene, macamides, glucosinolates, and polysaccharides (Caicai et al., 2018 ; Wang & Zhu, 2019 ) and has traditionally been used as a tonic fertility enhancer for both humans and cattle and to treat diseases such as rheumatism, respiratory disorders, and anemia, among other ailments (Beharry & Heinrich, 2018 ). Maca polysaccharides can reduce alcohol-related liver oxidative damage (Zhang et al., 2017 ) and exhibit antioxidant activity (Zha et al., 2014 ); thus, the use of maca polysaccharides as functional materials is expected. Polysaccharides, which are sugar complexes comprising numerous monosaccharides, are representative natural products that are ubiquitous in animals, plants, and microorganisms. Plant-derived polysaccharides are attractive as pharmaceutical and functional food materials because of their low toxicity and therapeutic properties (Schepetkin & Quinn, 2006 ). Plant polysaccharides prevent blood sugar spikes by delaying glucose absorption, lower blood cholesterol, and exhibit antioxidant and prebiotic effects (Tseng et al., 2008 ; Xu et al., 2013 ). Polysaccharides are highly valuable natural materials used as thickeners, stabilizers, and emulsifiers in the food industry due to their diverse physiochemical properties (Shedden et al., 2001). Extraction is an essential step for obtaining bioactive ingredients contained in plants. Various extraction methods have been used to increase the extraction yield by improving the solubility of these compounds (Oh & Yoon, 2018 ). The extraction method affects the physical and chemical properties of the target ingredients (Shang et al., 2019 ). Notably, the physiological activity of polysaccharides is greatly influenced by their structure (Yi et al., 2019 ); thus, the use of appropriate technologies for extracting polysaccharides is very important. ​ Ultrasound-assisted extraction (UAE) has been widely used for extracting polysaccharides from plants because of its advantages over traditional extraction techniques, including high extraction efficiency, shortened process time, low cost, and solvent and energy consumption (Chavan & Singhal, 2013 ). UAE can be performed by selecting various solvents, such as water, ethanol, and methanol. The yield of the extract and bioactivity in the human body vary depending on the extraction solvent (Rajha et al., 2019 ). Therefore, in extracting the target ingredient, not only the extraction technique but also the solvent selection is very important. Recently, as interest in eco-friendly solvents that can replace existing organic solvents increases, extraction methods using deep eutectic solvents (DESs) are emerging (Pan et al., 2022 ). DESs can be prepared by simply mixing two or more non-toxic substances: a hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD). Bioactive substances can be obtained from plants through hydrogen bonding and electrostatic interactions between the DESs and the target components (Zhang & Wang, 2017 ). DESs also have benefits such as excellent solubilizing power, thermal stability, simple preparation, low cost, and lack of pollution (Pan et al., 2022 ). This study aims to extract maca polysaccharides (MPs) using DES-based UAE and analyze their biological activities. First, a DES solvent with a high extraction yield is selected, and the extraction conditions are optimized to maximize the extraction yield by using an orthogonal experimental design. Finally, the yield and biological activities of the MPs obtained under the optimal DES-based UAE conditions are compared with those obtained using hot-water extraction (HWE) and UAE with water. Materials and Methods Materials The maca ( L. meyenii ) used in this study was purchased from a local farm located in the Gyeongbuk Province of Korea. The maca roots were cleansed, chopped, and subjected to lyophilization (FD8508, IlshinBioBase, Seoul, Korea). Subsequently, the freeze-dried roots were pulverized using a commercial-grade blending apparatus (FM-681C, Hanil, Incheon, Republic of Korea), passed through a 45-mesh screen (Chung Gye Indus, MFG Co., Seoul, Republic of Korea), and stored at − 45°C for further analysis. Preparation and Selection of DES for UAE The preparation of DESs adhered to the thermal protocol described by Pan et al. ( 2022 ). Within this procedure, individual DES, including choline chloride-glycerin (CCG), choline chloride-urea (CCU), and choline chloride-ethylene glycol (CCEG), was composed of two parts at a molar ratio of 1:2. The mixture was then agitated in a shaking water-bath (BS-11, JeioTech, Seoul, Republic of Korea) at 50°C until it achieved a homogeneous, colorless solution. The optimal DES selection was determined by extraction efficacy, setting parameters for water content, ultrasound power, and extraction duration based on established protocols of Chen et al. ( 2012 ) and Zhang and Wang ( 2017 ) using different combination of three types of DES. Briefly, 80 mL of a 3:7 solvent mixture of distilled water (DW) and selected DES was mixed with 2 g of maca powder. The mixture underwent an ultrasonic extraction for 20 min at 240 W power using an ultrasonic probe device (KFS-600N, Korea Process Technology, Seoul, Korea). Following centrifugation, precipitation was completed by adding 95% (v/v) ethanol equivalent to thrice the volume of the supernatant. After freeze-drying, the resulting maca polysaccharides (MPs) were quantified using the phenol-sulfuric acid method (Nielsen, 2017 ), with D-glucose as a standard, and the final yield percentage was calculated using the Eq. (1): Yield (%) = [total sugar content of MPs (g)/weight of sample (g)] × 100 (1) Orthogonal Experiment Design The optimization of the DES-assisted UAE process was facilitated by employing the orthogonal array L 9 according to the Taguchi experimental design (Table 2 ). Variables such as DES water content, ultrasound power, and the extraction time allocated for extraction were categorized into three distinct levels and tested for their influence on the yield of MPs. The quantitative analysis was performed using the Minitab® 18 software (Minitab Inc., State College, PA, USA), and the impact of individual variables was evaluated through Analysis of Variance (ANOVA). Statistical significance between different levels was determined by the least significant difference test, with a p-value threshold of less than 0.05. The optimum condition for each parameter was ascertained by analyzing the signal-to-noise (S/N) ratio, culminating in the determination of the most favorable extraction conditions. Extraction of MPs Hot-water Extraction Method For the extraction of MPs using hot water, a 2.0 g of maca powder was combined with 80 mL of distilled water. The mixture was then agitated in a shaking water bath (JeioTech BS-11, Seoul, Republic of Korea) at a speed of 100 rpm and maintained at a temperature of 90°C for 3 h. Post incubation, supernatant was obtained by a centrifugation step at the force of 4000 ×g for 20 min and then precipitated by adding 95% ethanol corresponding to thrice volume of the supernatant and kept at a cold temperature setting of 4°C for 12 h. The derived precipitates, denoted as MP-H, were separated through another centrifugal step at the same force for 15 min, then cleansed with 80% (v/v) of ethanol, and finally lyophilized. Ultrasound-Assisted Extraction Methods For the UAE process, an aqueous solution of DES was formulated by combining choline chloride-urea (CCU) with water in a 7:3 volume ratio. A 2 g of maca powder was immersed in either 80 mL of distilled water or the DES solution and subjected to ultrasonic waves at 300 W for 20 min using an ultrasonic probe device (KFS-600N, Korprotech). Following the ultrasonic treatment, the denoted MPs (MP-U from water and MP-DU from DES) were collected using the same method used in the hot water extraction to obtain precipitates. Performance of Pancreatic α-Amylase and α-Glucosidase Assays In order to measure the anti-diabetic activity of MPs, the inhibitory effects of MPs on pancreatic α-amylase and α-glucosidase activities were assessed using a modified approach based on Lee and Yoon ( 2022 ), with modifications. For pancreatic α-amylase assay, α-amylase enzyme solution (100 µL of 5 U/mL in 0.02 M sodium phosphate buffer at pH 6.9, type VI-B; Sigma–Aldrich) was reacted with 0.1 mL of MPs at room temperature (RT) for 5 min. Following this, a starch solution (0.1 mL of 1% concentration) was incorporated and allowed to interact for another 5 min at RT. The addition of 0.2 mL DNS reagent, which had been prepared in a 48 mM concentration with 30% sodium potassium tartarate in a 0.5 M of NaOH, followed and the mixture was boiled for 10 min then cooled for an additional 10 min. Afterwards, 1.5 mL of distilled water was added to reaction mixture, and the absorbance was measured at 540 nm for quantification. For α-glucosidase activity assay, 50 µL of MPs were mixed with 0.2 U/mL α-glucosidase enzyme and 200 mM potassium phosphate buffer (pH 6.8) and incubated at 37°C for 15 min, followed by the introduction of 100 µL of 3 mM p -nitrophenyl-α-D-glucopyranoside (Sigma–Aldrich.) to the mixture and incubation for 10 min at 37°C. The reaction was halted by adding 50 µL of 0.1 M NaOH was added. The absorbance was evaluated at 405 nm. The inhibitory effects of MPs on α-amylase and α-glucosidase activities were calculated using the Eq. (2): Inhibition (%) = [( A b ‒ A s )/A b ] ×100 (2) where A b and A s represents the absorbance of the blank (mixture without MPs) and that with MPs, respectively. IC 50 is defined as the concentration of MPs required to inhibit 50% of the enzyme activity. Acarbose (Sigma–Aldrich), an α-glucosidase inhibitor for treating type 2 diabetes mellitus, was used as a positive control. Determination of Glucose and Bile Acid Dialysis Retarding Index The glucose dialysis retardation index (GDRI) serves as an indicator of the ability of MPs to inhibit the absorption of glucose within the gastrointestinal tract. According to the methodology established by Adiotomre et al. ( 1990 ), 13 cm dialysis tube (D7884, Sigma-Aldrich, M w cutoff ≤ 1,200) was prepared by soaking in a 0.1% sodium azide solution (Sigma-Aldrich.) for 24 h, then filled with 6 mL of a 0.1% sodium azide solution containing either 36 mg of glucose alone or combined with 0.2 g of pre-hydrated MPs in an aqueous solution of 0.1% sodium azide for 14 h. The bags were sealed, submerged in 100 mL of 0.1% sodium azide solution and subjected to agitation at 37°C for 6 h. Glucose dispersion from the dialysis bag was monitored and recorded at set intervals at every 0.5, 1, 1.5, 2, and 4 h, by analyzing the glucose content in 1 mL of the dialysate by the DNS method. The GDRI values were subsequently calculated with the following Eq. (3). GDRI value (%) = 100 − [(Total glucose diffused with MPs present/ Total glucose diffused without MPs) ×100] (3) The bile acid dialysis retarding index (BDRI) was determined in the same manner as the GDRI, with one key difference being the inclusion of 15 mM taurocholic acid in a 50 mM phosphate buffer (pH 7.0) instead of glucose. The control setup consisted of the buffer solution without MPs, and the experimental setup included 0.2 g of MPs prepared under identical pre-hydration for 14 h. Dialysis were performed at 37℃ for 12 h. In order to measure the bile acid diffusion, a 2 mL dialysate sample at various times (0.5, 1, 2, 4, and 8 h) was taken for analysis. The quantification of bile acids in the dialysate was measured based on the taurocholic acid concentration, as previously described by Boyd et al. ( 1996 ). The BRI values were computed with the Eq. (4): BDRI value (%) = 100 − [(Total bile acid diffused with MPs present / Total bile acid diffused without MPs) ×100] The effects of the MPs on the retardation of glucose and bile acid diffusion were compared with those of carboxymethylcellulose (CMC) (Sigma-Aldrich), a commercial dietary fiber. Measurement of Prebiotic Growth Promotion Selection of Probiotic Strains Four types of Lactobacillus strains known for their probiotic benefits within the human gut was utilized: Lactobacillus plantarum (ATCC 8014), L. acidophilus (ATCC 832), L. casei (ATCC 393), and L. rhamnosus (ATCC 7469). Prior to experimental use, each strain was cultivated in Lactobacillus MRS Broth (Becton, Dickinson and Company, Sparks, MD., USA) in an incubator (IB-600M, Jeio Tech, Japan) at 37°C for 24 h. After initial culturing, each bacterium underwent a series of three subcultures to ensure robust and active bacterial colonies for the subsequent assessments. Probiotic Growth Assessment Following the protocols described by Im et al. ( 2016 ), each bacterial strain was first diluted in 0.1% sterilized peptone water (Bacto Pepetone; Becton Dickinson Co.) to standardized the initial bacterial concentration to 4 log Colony-Forming Units per milliliter (CFU/mL). Then, 0.5 mL of each diluted bacterial suspension was inoculated into 8.5 mL of sterile MRS broth. To this environment, 1 mL of the MPs that had been previously dissolved to a concentration of 50 mg/mL in a 30% dimethyl sulfoxide (DMSO from Junsei, Tokyo, Japan) solution or 1 mL of a 30% DMSO solution was added and incubated under controlled conditions at 37°C for 24-h to allow for bacterial growth promotion. Following the growth period, a 1 mL aliquot of each cultured sample was taken, further diluted using 0.1% sterilized peptone water, and subsequently plated to facilitate colony formation. After an additional 24 h of incubation, the resultant colonies were quantified. Statistical Analysis The experimental data are expressed as mean values ± standard deviation from triplicate measurements. Statistical analysis of the experimental results was performed using the SPSS (Ver. 23, IBM Corp., Armonk, NY, USA) software with the significance threshold set at p < 0.05. A one-way analysis of variance ensured the reliability of each set values and Duncan’s multi-range test established the significance versus the mean of the experiments. Results and Discussion Effect of the Type of DES on the Extraction Yield The extraction yields of the MP fraction obtained by UAE using CCG, CCU, and CCEG as solvents are shown in Table 1 . The extraction yields of the MPs obtained using CCG, CCU, and CCEG were 3.72 ± 0.12%, 20.03 ± 0.05%, and 8.73 ± 1.16%, respectively. The highest yield ( p < 0.05) of the MPs was obtained using CCU. Zhang and Wang ( 2017 ) reported that DESs can dissolve various substances such as sugars, polysaccharides, proteins, and amino acids through hydrogen bonding, enabling efficient extraction of polar or non-polar components. CCU afforded a higher extraction yield than the other DESs, which may be due to the higher hydrogen bonding ability and more electrostatic interactions of CCU with the MPs, compared to the other DESs (Zhang & Wang, 2017 ). Therefore, CCU is considered a suitable DES for MP extraction. Table 1 Yield of maca polysaccharide fractions obtained by various DES types DES type Yield (%) CCG 3.72 ± 0.12 c CCU 20.03 ± 0.05 a CCEG 8.73 ± 1.16 b Mean ± S.D ( n = 3) Values with different superscript letters in the same column are significantly different at p < 0.05. CCG, mixture of choline chloride and glycerin CCU, mixture of choline chloride and urea CCEG, mixture of choline chloride and ethylene glycol Orthogonal Optimization of Parameters for DES-based UAE To select the experimental factors, including the water content added to the DES, ultrasonic power, and extraction time, the orthogonal experiment L 9 was carried out to check the effects on the extraction yield. The results are shown in Table 2 . The yield of the MP extract ranged from 5.03 ± 0.16 to 26.28 ± 0.48%, with significant variation depending on the level of each factor. The experimental group with the highest extraction yield was run 9, and the average yield of all MPs was 15.70 ± 0.25%. Based on the average ( \({k}_{i}^{A})\) extraction yield obtained at each factor variable level, as the water content added to the DES increased, the yield of the MPs also increased. The highest yield was obtained with a water content of 30%. Because DESs have high viscosities, it is important to control the viscosity to promote the movement of the target component into the solvent (Wang et al., 2016 ). Zhang and Wang ( 2017 ) reported that the extraction yield increased as the water content added to a DES increased from 0–30%, which is consistent with the results of this study. The yield also increased significantly with increasing ultrasonic power. At extraction times of 0, 20, and 30 min, the yield was 14.69%, 17.33, and 15.07%, respectively; the extraction yield was highest at 20 min. Chen et al. ( 2012 ) reported that as the ultrasonic power increased, the mass transfer rate and extraction efficiency of polysaccharides improved; however, an ultrasonic power that was too high increased the number of bubbles in the solvent, reducing the transfer efficiency of the ultrasonic power. Previous reports also stated that the yield increased at an extraction time of 20 min and then decreased again at 30 min, consistent with the results of this study. Table 2 Factors and results of orthogonal design experiments on different extraction condition of maca polysaccharides Experiment No. Reaction parameters Yield (%) Water content (%) Ultrasonic power (W) Extraction time (min) 1 10 180 10 5.03 ± 0.16 2 10 240 20 14.40 ± 0.54 3 10 300 30 12.22 ± 0.24 4 20 180 20 11.29 ± 0.76 5 20 240 30 18.36 ± 1.68 6 20 300 10 22.84 ± 1.26 7 30 180 30 14.64 ± 0.37 8 30 240 10 16.20 ± 1.25 9 30 300 20 26.28 ± 0.48 K1 31.66 30.96 44.07 K2 52.50 48.97 51.98 K3 57.12 61.35 45.22 k1 10.55 10.32 14.69 k2 17.50 16.32 17.33 k3 19.04 20.45 15.07 R 8.49 10.13 2.67 Optimal level 3 3 2 Mean ± S.D ( n = 3) \({K}_{i}^{A}\) = Ʃ yield at A i \({k}_{i}^{A} = {K}_{i}^{A}/3\) R: Refers to the result of extreme analysis, \({R}_{i}^{A}\) = max { \({k}_{i}^{A}\) } - min{ \({k}_{i}^{A}\) } To examine the degree of influence of the extraction conditions on the yield of each extract, the effect of each factor was calculated based on an orthogonal test. According to the R value ( R = max \({k}_{i}^{A}\) − min \({k}_{i}^{A}\) ), the yield was affected by the parameters in the following order: ultrasonic power (10.13 ± 0.74), moisture content (8.4 ± 0.44), and extraction time (2.67 ± 0.33). In the main effect plot for the S/N ratios, the mean S/N ratios for each factor were also plotted against the test level for each factor parameter (Fig. 1 ). The main effects plot for the extraction yield clearly shows that ultrasonic power had the most significant influence on the extraction yield, followed by the water content (very slight significance). These results are consistent with those of Mudliyar et al. ( 2019 ), who reported that the ultrasound power had the greatest influence on the extraction yield when the conditions for polysaccharide extraction from Tuber aestivum were optimized using the Taguchi method. Because the Taguchi experimental method does not provide a precise estimation of the effect of each factor on the overall process, the percentage contribution determined using ANOVA was employed to compensate for this effect (Abe et al., 2019 ). The Taguchi experimental results (Table 3 ) were processed by ANOVA using Minitab 18, where values were considered significantly different at p < 0.05. The ANOVA of the extraction data indicates that the ultrasound power had the most statistically significant contribution of 48.13% ( p < 0.001), followed by the water content (37.94%; p < 0.001), and the extraction time (3.77%, p value = 0.043). Based on the analysis of the extraction yield using the Taguchi method, the optimal conditions are as follows: water content of 30%, ultrasound power of 300 W, and extraction time of 20 min. Table 3 Analysis of variance results in relation to extraction yield Variance source Degree of freedom (DF) Sum of square (SS) Mean square (MS) F-ratio P-value Contribution (%) Water content 2 367.954 183.977 37.351 < 0.001 37.94 Ultrasonic power 2 466.774 233.387 47.382 < 0.001 48.13 Extraction time 2 36.529 18.265 3.708 0.043 3.77 Error 20 98.513 4.926 10.16 Total 28 969.770 100.00 Extraction Yield of MPs The yield and IC 50 values for the antidiabetic activity of the MPs obtained using HWE and UAE are shown in Table 4 . The yield of the MPs was highest for MP-DU at 26.28 ± 0.48% ( p < 0.05), followed by MP-U (12.1 ± 0.16%); MP-H had the significantly lowest yield (11.67 ± 0.17%) ( p < 0.05). In UAE, the extraction yield was approximately two times higher when the DES was used as the solvent than when distilled water was used. Nam et al. ( 2014 ) reported that in the ultrasound extraction of flavonoids from Flos sophorae , the yield of flavonoids was higher when the DES was used as a solvent than when water was used as the solvent. This indicates that the use of a DES during ultrasonic extraction can lead to an increase in the extraction yield. Table 4 Extraction yield and antidiabetic activity of maca polysaccharides obtained by various extraction methods Sample Yield (%) IC 50 value (µg/mL) α-Amylase inhibitory activity α-Glucosidase inhibitory activity MP-H 11.67 ± 0.17 b 329.62 ± 13.07 a 641.42 ± 2.21 a MP-U 12.10 ± 0.16 b 156.15 ± 18.27 b 119.67 ± 11.21 c MP-DU 26.28 ± 0.48 a 99.21 ± 3.08 c 110.44 ± 4.09 d Acarbose - 89.98 ± 6.60 d 178.15 ± 4.22 b Mean ± S.D ( n = 3) Values with different superscript letters in the same column are significantly different at p < 0.05. MP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively. Inhibitory Effects of MPs on Pancreatic α-Amylase and α-Glucosidase Activities The IC 50 value for the α-amylase inhibitory activity was lowest (89.98 ± 6.60 µg/mL) for acarbose, followed by MP-DU (99.21 ± 3.08 µg/mL), showing an activity equivalent to 90% of acarbose. MP-U and MP-H afforded high IC 50 values of 156.15 ± 1.11 and 359.62 ± 13.07 µg/mL, respectively, indicative of low inhibitory activity. The IC 50 values for the α-glucosidase inhibitory activity were 110.44 ± 4.09, 119.67 ± 11.21, and 641.42 ± 2.21 µg/mL for MP-DU, MP-U, and MP-H, respectively. The IC 50 values of MP-DU and MP-U were significantly lower than that of acarbose (178.15 ± 4.22 µg/mL), indicating that MP-DU and MP-U have high enzyme inhibitory activity. α-Amylase and α-glucosidase inhibitors inhibit the hydrolysis of carbohydrates, thereby reducing glucose release from starch and delaying the absorption of carbohydrates, consequently suppressing the rise in blood sugar (Xu et al., 2018 ). Recently, antidiabetic active substances isolated from natural sources have received considerable attention. The results of this study show that MPs have higher inhibitory activity than acarbose and great potential for use as a natural diabetes treatment. Therefore, MP, especially MP-DU, is expected to play a role as a treatment for diabetes by inhibiting the activity of α-amylase and α-glucosidase to prevent postprandial blood glucose rise. Retarding Effect of MPs on Glucose Absorption The effect of compounds on delaying glucose absorption was reported to have a very high correlation with the in vivo lowering of blood sugar based on an in vitro experiment evaluating the entrapping effect of dietary fiber (Ahmed et al., 2011 ). The glucose content released into the dialysate according to the dialysis time and GDRI, which is a value showing the glucose permeation inhibition effect compared to the control, is shown in Table 5 . At 30 min of dialysis, the glucose content of CMC, the positive control group, was the lowest at 4.51 ± 0.51 mg/100 mL, followed by that of the MP-DU (5.76 ± 0.26 mg/100 mL) and MP-U (6.13 ± 0.19 mg/100 mL) groups, and finally the MP-H (6.18 ± 0.13 mg/100 mL) group, for which the value was significantly lower than that of the control (7.22 ± 0.31 mg/100 mL). After this period, the glucose concentration showed similar trends for all groups up to 4 h of dialysis. Table 5 Retarding effect of maca polysaccharides obtained by various extraction methods on dialysis membrane transport of glucose Dialysis time (h) Sample 0.5 1 1.5 2 4 Glucose content (mg/100 mL) GDRI (%) Glucose content (mg/100 mL) GDRI (%) Glucose content (mg/100 mL) GDRI (%) Glucose content (mg/100 mL) GDRI (%) Glucose content (mg/100 mL) GDRI (%) Control 7.22 ± 0.31 d - 15.38 ± 0.37 d - 22.24 ± 0.31 e - 26.03 ± 0.43 e - 29.11 ± 0.44 f - CMC 4.51 ± 0.51 a 37.53 ± 7.00 a 8.63 ± 0.12 a 43.88 ± 0.79 a 13.11 ± 0.59 a 41.04 ± 2.66 a 15.41 ± 0.39 a 40.79 ± 1.51 a 19.17 ± 0.61 a 34.14 ± 2.10 a MP-H 6.18 ± 0.13 c 14.43 ± 1.73 b 10.69 ± 0.32 c 30.48 ± 2.09 c 16.56 ± 0.41 d 25.54 ± 1.86 d 20.04 ± 0.37 d 23.02 ± 1.40 d 25.81 ± 0.42 d 11.34 ± 1.43 c MP-U 6.13 ± 0.19 c 15.01 ± 2.65 b 9.97 ± 0.12 b 35.21 ± 0.79 b 15.26 ± 0.38 c 31.35 ± 1.70 c 19.20 ± 0.37 c 26.22 ± 1.40 c 25.04 ± 0.35 c 13.98 ± 1.21 c MP-DU 5.76 ± 0.26 bc 20.21 ± 3.61 b 9.40 ± 0.30 b 38.89 ± 2.53 b 14.36 ± 0.35 b 35.41 ± 1.59 b 17.34 ± 0.41 b 33.36 ± 1.58 b 20.60 ± 0.40 b 29.23 ± 1.36 b Mean ± S.D ( n = 3) Values with different superscript letters in the column are significantly different at p < 0.05. GDRI, glucose dialysis retardation index; CMC, carboxymethylcelluose; MP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively. During all dialysis times, the GDRI of CMC was significantly higher than that of the MPs, followed by that of MP-DU, which had a higher GDRI than the other MPs. This shows that MP-DU has a significant effect on retarding glucose absorption. The GDRI increased with the dialysis time up to 1 h of dialysis and then tended to decrease gradually. The highest GDRI values during dialysis were as follows: CMC 43.88 ± 0.79%, MP-DU 38.89 ± 2.53%, MP-U 35.21 ± 0.79%, and MP-H 30.48 ± 2.09%, with MP-DU having the highest GDRI after CMC. It has been reported that polysaccharides form a viscous gel in aqueous solutions, slowing access to small intestine cells, and that the large surface area of polysaccharide particles promotes the trapping effect of glucose, delaying its absorption (Ahmed et al., 2011 ; Abirami et al., 2014 ). Chen et al. ( 2015 ) reported that the GDRI of maca dietary fiber was 12.30‒23.66% after 1 h of dialysis, which is lower than the results of this study. Although the maca samples are the same, these differences are thought to be due to the variety and extraction methods used. Therefore, MP extracted with DES-based UAE in this study effectively lowers the level of serum glucose by delaying the absorption of glucose and is expected to have potential for development as a hypoglycemic agent and low-calorie functional food. Retarding Effect of MPs on Bile Acid Absorption The bile acid dialysis retardation index (BDRI), which shows the effect of the polysaccharides on inhibiting bile acid permeation compared to the control, was used to monitor the effects of fiber on cholesterol metabolism. The amount of bile acid released into the dialysate according to the dialysis time and BDRI is shown in Table 6 . At all dialysis times, the amount of bile acid released into the dialysate with all MPs and CMC was significantly lower than that of the control ( p < 0.05). At 1 h of dialysis, CMC showed the highest BDRI (37.77 ± 2.45%), followed by MP-DU (35.96 ± 2.45%), MP-U (34.06 ± 2.52%), and MP-H (21.68 ± 0.93%). The BDRI values of MP-DU and MP-U were not significantly different from those of CMC, indicating a strong inhibitory effect on the absorption of bile acid. Even at 90 min of dialysis, the BDRI of MP-DU was 35.02 ± 1.66%, which was not significantly different from that of CMC (38.22 ± 1.91%). The BDRI of the MPs increased up to 2 h of dialysis and then decreased. The highest inhibition of bile acid absorption was observed at 2 h of dialysis. Dietary fiber adsorbs free bile acid and inhibits the reabsorption of bile acid, which induces the consumption of cholesterol for the synthesis of bile acid in the body. This process lowers the concentration of cholesterol in the blood, which has a positive effect on cardiovascular diseases such as arteriosclerosis and heart disease (Ioniță-Mîndrican et al., 2022 ). Ma et al. ( 2015 ) reported that the many hydrophobic groups present in dietary fiber increase the adsorption capacity by forming hydrophobic bonds through interaction with bile acid, and that the small size of the dietary fiber particles increases the absorption capacity by increasing the surface area. Therefore, MP-DU and MP-U had a high bile acid absorption inhibitory effect and are expected to have a positive effect on lipid metabolism. Table 6 Retarding effect of maca polysaccharides obtained by various extraction methods on dialysis membrane transport of bile acid Dialysis time (h) Sample 0.5 1 2 4 8 Bile acid content (µmole/L) BRI (%) Bile acid content (µmole/L) BRI (%) Bile acid content (µmole/L) BRI (%) Bile acid content (µmole/L) BRI (%) Bile acid content (µmole/L) BRI (%) Control 98.98 ± 1.63 c - 170.92 ± 5.76 d - 369.25 ± 4.38 d - 497.56 ± 3.39 e - 597.39 ± 5.32 f - CMC 65.54 ± 2.82 a 33.79 ± 2.85 a 106.36 ± 4.18 a 37.77 ± 2.45 a 230.37 ± 3.93 a 37.61 ± 1.06 a 301.78 ± 3.91 a 39.35 ± 0.79 a 370.50 ± 2.92 a 37.98 ± 0.49 a MP-H 80.12 ± 1.63 b 19.05 ± 1.65 b 133.85 ± 1.58 c 21.68 ± 0.93 b 316.63 ± 4.89 c 14.25 ± 1.32 c 425.96 ± 5.23 c 14.39 ± 1.05 d 535.14 ± 3.19 d 10.42 ± 0.53 d MP-U 69.45 ± 1.63 a 29.83 ± 1.65 a 112.71 ± 4.31 ba 34.06 ± 2.52 a 278.55 ± 5.35 b 24.56 ± 1.45 b 372.36 ± 2.49 b 25.16 ± 0.50 b 471.36 ± 2.62 c 21.10 ± 0.44 c MP-DU 75.86 ± 4.44 b 23.36 ± 4.49 b 109.45 ± 4.18 a 35.96 ± 2.45 a 279.53 ± 6.89 b 24.30 ± 1.87 b 383.20 ± 3.02 c 22.99 ± 0.61 c 460.35 ± 3.95 b 22.94 ± 0.66 b Mean ± S.D ( n = 3) Values with different superscript letters in the column are significantly different at p < 0.05. BDRI, bile acid dialysis retardation index; CMC, carboxymethylcelluose; HWE, hot water extraction; UAE, ultrasound-assisted extraction; DES, deep eutectic solvent; MP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively. Probiotic Growth-Stimulation Capacity of MPs To determine the prebiotic effects of the MPs obtained using different extraction methods, the effects of the MPs on the proliferation of lactic acid bacteria ( L. plantarum, L. acidophilus, L. casei, L. rhamnosus ) were measured; the results are summarized in Table 7 . For all strains of lactic acid bacteria, groups treated with MPs exhibited significantly higher growth—approximately 10 times—compared to the control group. MP-H had the greatest effect on the proliferation of L. plantarum and L. acidophilus , with counts of 14.67 ± 2.52 × 10 9 and 11.00 ± 2.00 × 10 9 , respectively, whereas MP-U had the highest effect on the proliferation of L. casei and L. rhamnosus , with counts of 9.00 ± 1.00 × 10 9 and 10.00 ± 1.00 × 10 9 , respectively. The degree of growth for each strain of lactic acid bacterium differed depending on the sample. The control and MP-H groups were most effective for promoting the growth of L. plantarum , whereas the MP-U and MP-DU groups were most effective for promoting the growth of L. rhamnosus and L. acidophilus , respectively. MP-U and MP-DU significantly enhanced the growth of all strains, demonstrating their broad-spectrum effectiveness. Polysaccharides that are not digested in the gastrointestinal tract can stimulate the growth of beneficial intestinal bacteria, such as lactic acid bacteria and bifidobacteria, and have health benefits such as promoting the production of short-chain fatty acids; therefore, they are considered to have a prebiotic effect (Chen et al., 2019 ). The prebiotic activity of polysaccharides varies depending on their physicochemical properties, including molecular weight, monosaccharide composition, glycosidic bonds, and viscosity (Huang et al., 2019 ). Table 7 Effect of maca polysaccharide by various extraction methods on the growth of lactic acid bacteria (CFU/mL) Sample L. plantarum L. acidophilus L. casei L. rhamnosus Control 9.33 ± 1.53☓10 8c 5.00 ± 0.00☓10 8c 3.00 ± 1.00☓10 8c 6.67 ± 1.15☓10 8d MP-H 14.67 ± 2.52☓10 9a 11.00 ± 2.00☓10 9a 4.67 ± 1.53☓10 9b 2.67 ± 0.58☓10 9c MP-U 8.67 ± 1.53☓10 9b 7.00 ± 1.00☓10 9b 9.00 ± 1.00☓10 9a 10.00 ± 1.00☓10 9a MP-DU 7.33 ± 0.58☓10 9b 8.67 ± 0.58☓10 9b 7.67 ± 0.58☓10 9a 7.33 ± 1.53☓10 9b Mean S.D (n = 3) Values with different superscript letters in the column are significantly different at p < 0.05 Control is only DMSO added. MP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively Conclusions The present study investigated the deep eutectic solvent (DES)-based ultrasound-assisted extraction (UAE) for extracting maca polysaccharides (MPs) from maca roots and compared this process with two other extraction techniques (HWE and UAE with water). The highest extraction yield was obtained using choline chloride and urea at a molar ratio of 1:3. As a result of using the Taguchi orthogonal array as a design variable, the extraction yield was 26.28% at 30% water content, 300W ultrasonic power, and 20% extraction time. Under the optimized conditions, the DES-based UAE provided a higher yield of MPs (MP-DU) than the other extraction methods. MP-DU inhibited the α-amylase and β-glucosidase activities and delayed the absorbance of glucose and bile acids. In conclusion, this study not only provides an environmentally friendly, efficient, and economical alternative method to extract MPs, but also provides a scientific basis for the comprehensive utilization of maca as a potential functional food. Abbreviations CCEG Mixture of choline chloride and ethylene glycol CCG Mixture of choline chloride and glycerin CCU Mixture of choline chloride and urea DESs Deep eutectic solvents BDRI Bile acid dialysis retardation index GDRI Glucose dialysis retardation index HWE Hot-water extraction MPs Maca polysaccharides MP-DU MPs prepared by using DES-based UAE MP-U MPs prepared by using UAE with water MP-W MPs prepared by using HWE UAE Ultrasound-assisted extraction Declarations Conflict of Interest The authors declare no conflicts of interest. Funding This research was funded by the Basic Science Research Program through the National Research Foundation of Korea (NRF), which is funded by the Ministry of Education (grant numbers NRF-2021R1A6A1A03040177). Author Contribution Eun Jeong Kim: formal analysis, investigation, data curation, writing-original draft, Choon Young Kim: data curation, writing-review and editing, funding acquisition, Kyung Young Yoon: conceptualization, data curation, writing-review and editing, supervision. References Abe, J. O., Popoola, O. M., Popoola, A. P. I., Ajenifuja, E., & Adebiyi, D. I. (2019). Application of Taguchi design method for optimization of spark plasma sintering process parameters for Ti-6Al-4V/h-BN binary composite. Engineering Research Express , 1 (2), 025043. http://dx.doi.org/10.1088/2631-8695/ab561c . Abirami, A., Nagarani, G., & Siddhuraju, P. (2014). 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4340936","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":299215931,"identity":"98463c9d-cb41-442f-b55f-f500691850fc","order_by":0,"name":"Eun Jeong Kim","email":"","orcid":"","institution":"Yeungnam University","correspondingAuthor":false,"prefix":"","firstName":"Eun","middleName":"Jeong","lastName":"Kim","suffix":""},{"id":299215932,"identity":"593f747c-5ecc-4ef6-9a93-d5d11dc018bc","order_by":1,"name":"Choon Young Kim","email":"","orcid":"","institution":"Yeungnam University","correspondingAuthor":false,"prefix":"","firstName":"Choon","middleName":"Young","lastName":"Kim","suffix":""},{"id":299215933,"identity":"0c6efd6b-be17-4f0d-8ac9-2802bb909366","order_by":2,"name":"Kyung Young Yoon","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYJACxgYGG2bGZuYDUD4bUVrS2Jnb2RJI0nKYn72fx4A4LQbHm489nNnGLM3bzPNNurCNQZ6/gS3tA14tZ46lG25sYzOWbObdJj2zjcFwxgG2wzPwarmRYyb5sI0n2RCkhbeNgXEDA3szfodBtEjU7z/M8wykxZ44LRvbDICBzMMG0pK4gYHtMF4tkmeOpUnOOJcA1MJmbM1zTiJ5xmG2ZLxa+IAhJtlT9p+Zsf/ww9s8ZTa2/e1txni1KBwAEoyQmGCRYGAAIma8GhgY5BtA5B8wmxlvdIyCUTAKRsHIBQC4OkPanZ06IgAAAABJRU5ErkJggg==","orcid":"","institution":"Yeungnam University","correspondingAuthor":true,"prefix":"","firstName":"Kyung","middleName":"Young","lastName":"Yoon","suffix":""}],"badges":[],"createdAt":"2024-04-29 07:20:05","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4340936/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4340936/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":55994288,"identity":"22a382bc-547b-411a-92f9-87f6f1db45b5","added_by":"auto","created_at":"2024-05-07 10:00:51","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":32821,"visible":true,"origin":"","legend":"\u003cp\u003eMain effects plot for signal-to-noise ratios for extraction yield.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4340936/v1/0459e8eb4086950ddbab8265.png"},{"id":55994659,"identity":"07df1aa3-75e2-4c76-9f8e-f5546b2d901e","added_by":"auto","created_at":"2024-05-07 10:08:52","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1009694,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4340936/v1/ec89578d-dc78-4c26-ae7a-89241ae916c4.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Deep-eutectic-solvent-based ultrasound-assisted extraction of polysaccharides from maca: Optimization using Taguchi methodology and comparison with the conventional method","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMaca (\u003cem\u003eLepidium meyenii\u003c/em\u003e) is a cruciferous plant that grows naturally in the highlands; the roots are mainly used as edible parts. Maca contains bioactive components such as macaene, macamides, glucosinolates, and polysaccharides (Caicai et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Wang \u0026amp; Zhu, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) and has traditionally been used as a tonic fertility enhancer for both humans and cattle and to treat diseases such as rheumatism, respiratory disorders, and anemia, among other ailments (Beharry \u0026amp; Heinrich, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Maca polysaccharides can reduce alcohol-related liver oxidative damage (Zhang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and exhibit antioxidant activity (Zha et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2014\u003c/span\u003e); thus, the use of maca polysaccharides as functional materials is expected.\u003c/p\u003e \u003cp\u003ePolysaccharides, which are sugar complexes comprising numerous monosaccharides, are representative natural products that are ubiquitous in animals, plants, and microorganisms. Plant-derived polysaccharides are attractive as pharmaceutical and functional food materials because of their low toxicity and therapeutic properties (Schepetkin \u0026amp; Quinn, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Plant polysaccharides prevent blood sugar spikes by delaying glucose absorption, lower blood cholesterol, and exhibit antioxidant and prebiotic effects (Tseng et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Xu et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Polysaccharides are highly valuable natural materials used as thickeners, stabilizers, and emulsifiers in the food industry due to their diverse physiochemical properties (Shedden et al., 2001).\u003c/p\u003e \u003cp\u003eExtraction is an essential step for obtaining bioactive ingredients contained in plants. Various extraction methods have been used to increase the extraction yield by improving the solubility of these compounds (Oh \u0026amp; Yoon, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The extraction method affects the physical and chemical properties of the target ingredients (Shang et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Notably, the physiological activity of polysaccharides is greatly influenced by their structure (Yi et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2019\u003c/span\u003e); thus, the use of appropriate technologies for extracting polysaccharides is very important. ​\u003c/p\u003e \u003cp\u003eUltrasound-assisted extraction (UAE) has been widely used for extracting polysaccharides from plants because of its advantages over traditional extraction techniques, including high extraction efficiency, shortened process time, low cost, and solvent and energy consumption (Chavan \u0026amp; Singhal, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). UAE can be performed by selecting various solvents, such as water, ethanol, and methanol. The yield of the extract and bioactivity in the human body vary depending on the extraction solvent (Rajha et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Therefore, in extracting the target ingredient, not only the extraction technique but also the solvent selection is very important. Recently, as interest in eco-friendly solvents that can replace existing organic solvents increases, extraction methods using deep eutectic solvents (DESs) are emerging (Pan et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). DESs can be prepared by simply mixing two or more non-toxic substances: a hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD). Bioactive substances can be obtained from plants through hydrogen bonding and electrostatic interactions between the DESs and the target components (Zhang \u0026amp; Wang, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). DESs also have benefits such as excellent solubilizing power, thermal stability, simple preparation, low cost, and lack of pollution (Pan et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThis study aims to extract maca polysaccharides (MPs) using DES-based UAE and analyze their biological activities. First, a DES solvent with a high extraction yield is selected, and the extraction conditions are optimized to maximize the extraction yield by using an orthogonal experimental design. Finally, the yield and biological activities of the MPs obtained under the optimal DES-based UAE conditions are compared with those obtained using hot-water extraction (HWE) and UAE with water.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eMaterials\u003c/h2\u003e\n \u003cp\u003eThe maca (\u003cem\u003eL. meyenii\u003c/em\u003e) used in this study was purchased from a local farm located in the Gyeongbuk Province of Korea. The maca roots were cleansed, chopped, and subjected to lyophilization (FD8508, IlshinBioBase, Seoul, Korea). Subsequently, the freeze-dried roots were pulverized\u003c/p\u003e\n \u003cp\u003eusing a commercial-grade blending apparatus (FM-681C, Hanil, Incheon, Republic of Korea), passed through a 45-mesh screen (Chung Gye Indus, MFG Co., Seoul, Republic of Korea), and stored at \u0026minus;\u0026thinsp;45\u0026deg;C for further analysis.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003ePreparation and Selection of DES for UAE\u003c/h2\u003e\n \u003cp\u003eThe preparation of DESs adhered to the thermal protocol described by Pan et al. (\u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e). Within this procedure, individual DES, including choline chloride-glycerin (CCG), choline chloride-urea (CCU), and choline chloride-ethylene glycol (CCEG), was composed of two parts at a molar ratio of 1:2. The mixture was then agitated in a shaking water-bath (BS-11, JeioTech, Seoul, Republic of Korea) at 50\u0026deg;C until it achieved a homogeneous, colorless solution.\u003c/p\u003e\n \u003cp\u003eThe optimal DES selection was determined by extraction efficacy, setting parameters for water content, ultrasound power, and extraction duration based on established protocols of Chen et al. (\u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e) and Zhang and Wang (\u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e) using different combination of three types of DES. Briefly, 80 mL of a 3:7 solvent mixture of distilled water (DW) and selected DES was mixed with 2 g of maca powder. The mixture underwent an ultrasonic extraction for 20 min at 240 W power using an ultrasonic probe device (KFS-600N, Korea Process Technology, Seoul, Korea). Following centrifugation, precipitation was completed by adding 95% (v/v) ethanol equivalent to thrice the volume of the supernatant. After freeze-drying, the resulting maca polysaccharides (MPs) were quantified using the phenol-sulfuric acid method (Nielsen, \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e), with D-glucose as a standard, and the final yield percentage was calculated using the Eq.\u0026nbsp;(1):\u003c/p\u003e\n \u003cp\u003eYield (%) = [total sugar content of MPs (g)/weight of sample (g)] \u0026times; 100 (1)\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003eOrthogonal Experiment Design\u003c/h2\u003e\n \u003cp\u003eThe optimization of the DES-assisted UAE process was facilitated by employing the orthogonal array L\u003csub\u003e9\u003c/sub\u003e according to the Taguchi experimental design (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). Variables such as DES water content, ultrasound power, and the extraction time allocated for extraction were categorized into three distinct levels and tested for their influence on the yield of MPs. The quantitative analysis was performed using the Minitab\u0026reg; 18 software (Minitab Inc., State College, PA, USA), and the impact of individual variables was evaluated through Analysis of Variance (ANOVA). Statistical significance between different levels was determined by the least significant difference test, with a p-value threshold of less than 0.05. The optimum condition for each parameter was ascertained by analyzing the signal-to-noise (S/N) ratio, culminating in the determination of the most favorable extraction conditions.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003eExtraction of MPs\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eHot-water Extraction Method\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eFor the extraction of MPs using hot water, a 2.0 g of maca powder was combined with 80 mL of distilled water. The mixture was then agitated in a shaking water bath (JeioTech BS-11, Seoul, Republic of Korea) at a speed of 100 rpm and maintained at a temperature of 90\u0026deg;C for 3 h. Post incubation, supernatant was obtained by a centrifugation step at the force of 4000 \u0026times;g for 20 min and then precipitated by adding 95% ethanol corresponding to thrice volume of the supernatant and kept at a cold temperature setting of 4\u0026deg;C for 12 h. The derived precipitates, denoted as MP-H, were separated through another centrifugal step at the same force for 15 min, then cleansed with 80% (v/v) of ethanol, and finally lyophilized.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003eUltrasound-Assisted Extraction Methods\u003c/h2\u003e\n \u003cp\u003eFor the UAE process, an aqueous solution of DES was formulated by combining choline chloride-urea (CCU) with water in a 7:3 volume ratio. A 2 g of maca powder was immersed in either 80 mL of distilled water or the DES solution and subjected to ultrasonic waves at 300 W for 20 min using an ultrasonic probe device (KFS-600N, Korprotech). Following the ultrasonic treatment, the denoted MPs (MP-U from water and MP-DU from DES) were collected using the same method used in the hot water extraction to obtain precipitates.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003ePerformance of Pancreatic \u0026alpha;-Amylase and \u0026alpha;-Glucosidase Assays\u003c/h2\u003e\n \u003cp\u003eIn order to measure the anti-diabetic activity of MPs, the inhibitory effects of MPs on pancreatic \u0026alpha;-amylase and \u0026alpha;-glucosidase activities were assessed using a modified approach based on Lee and Yoon (\u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e), with modifications. For pancreatic \u0026alpha;-amylase assay, \u0026alpha;-amylase enzyme solution (100 \u0026micro;L of 5 U/mL in 0.02 M sodium phosphate buffer at pH 6.9, type VI-B; Sigma\u0026ndash;Aldrich) was reacted with 0.1 mL of MPs at room temperature (RT) for 5 min. Following this, a starch solution (0.1 mL of 1% concentration) was incorporated and allowed to interact for another 5 min at RT. The addition of 0.2 mL DNS reagent, which had been prepared in a 48 mM concentration with 30% sodium potassium tartarate in a 0.5 M of NaOH, followed and the mixture was boiled for 10 min then cooled for an additional 10 min. Afterwards, 1.5 mL of distilled water was added to reaction mixture, and the absorbance was measured at 540 nm for quantification.\u003c/p\u003e\n \u003cp\u003eFor \u0026alpha;-glucosidase activity assay, 50 \u0026micro;L of MPs were mixed with 0.2 U/mL \u0026alpha;-glucosidase enzyme and 200 mM potassium phosphate buffer (pH 6.8) and incubated at 37\u0026deg;C for 15 min, followed by the introduction of 100 \u0026micro;L of 3 mM \u003cem\u003ep\u003c/em\u003e-nitrophenyl-\u0026alpha;-D-glucopyranoside (Sigma\u0026ndash;Aldrich.) to the mixture and incubation for 10 min at 37\u0026deg;C. The reaction was halted by adding 50 \u0026micro;L of 0.1 M NaOH was added. The absorbance was evaluated at 405 nm.\u003c/p\u003e\n \u003cp\u003eThe inhibitory effects of MPs on \u0026alpha;-amylase and \u0026alpha;-glucosidase activities were calculated using the Eq.\u0026nbsp;(2):\u003c/p\u003e\n \u003cp\u003eInhibition (%) = [(\u003cem\u003eA\u003c/em\u003e\u003csub\u003e\u003cem\u003eb\u003c/em\u003e\u003c/sub\u003e \u003cem\u003e‒ A\u003c/em\u003e\u003csub\u003e\u003cem\u003es\u003c/em\u003e\u003c/sub\u003e\u003cem\u003e)/A\u003c/em\u003e\u003csub\u003e\u003cem\u003eb\u003c/em\u003e\u003c/sub\u003e] \u0026times;100 (2)\u003c/p\u003e\n \u003cp\u003ewhere \u003cem\u003eA\u003c/em\u003e\u003csub\u003e\u003cem\u003eb\u003c/em\u003e\u003c/sub\u003e and \u003cem\u003eA\u003c/em\u003e\u003csub\u003e\u003cem\u003es\u003c/em\u003e\u003c/sub\u003e represents the absorbance of the blank (mixture without MPs) and that with MPs, respectively.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eIC\u003c/em\u003e \u003csub\u003e\u0026nbsp;\u003cem\u003e50\u003c/em\u003e\u0026nbsp;\u003c/sub\u003e is defined as the concentration of MPs required to inhibit 50% of the enzyme activity. Acarbose (Sigma\u0026ndash;Aldrich), an \u0026alpha;-glucosidase inhibitor for treating type 2 diabetes mellitus, was used as a positive control.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n \u003ch2\u003eDetermination of Glucose and Bile Acid Dialysis Retarding Index\u003c/h2\u003e\n \u003cp\u003eThe glucose dialysis retardation index (GDRI) serves as an indicator of the ability of MPs to inhibit the absorption of glucose within the gastrointestinal tract. According to the methodology established by Adiotomre et al. (\u003cspan class=\"CitationRef\"\u003e1990\u003c/span\u003e), 13 cm dialysis tube (D7884, Sigma-Aldrich, \u003cem\u003eM\u003c/em\u003e\u003csub\u003e\u003cem\u003ew\u003c/em\u003e\u003c/sub\u003e cutoff\u0026thinsp;\u0026le;\u0026thinsp;1,200) was prepared by soaking in a 0.1% sodium azide solution (Sigma-Aldrich.) for 24 h, then filled with 6 mL of a 0.1% sodium azide solution containing either 36 mg of glucose alone or combined with 0.2 g of pre-hydrated MPs in an aqueous solution of 0.1% sodium azide for 14 h. The bags were sealed, submerged in 100 mL of 0.1% sodium azide solution and subjected to agitation at 37\u0026deg;C for 6 h. Glucose dispersion from the dialysis bag was monitored and recorded at set intervals at every 0.5, 1, 1.5, 2, and 4 h, by analyzing the glucose content in 1 mL of the dialysate by the DNS method. The GDRI values were subsequently calculated with the following Eq. (3).\u003c/p\u003e\n \u003cp\u003eGDRI value (%)\u0026thinsp;=\u0026thinsp;100 \u0026minus; [(Total glucose diffused with MPs present/ Total glucose diffused without MPs) \u0026times;100] (3)\u003c/p\u003e\n \u003cp\u003eThe bile acid dialysis retarding index (BDRI) was determined in the same manner as the GDRI, with one key difference being the inclusion of 15 mM taurocholic acid in a 50 mM phosphate buffer (pH 7.0) instead of glucose. The control setup consisted of the buffer solution without MPs, and the experimental setup included 0.2 g of MPs prepared under identical pre-hydration for 14 h. Dialysis were performed at 37℃ for 12 h. In order to measure the bile acid diffusion, a 2 mL dialysate sample at various times (0.5, 1, 2, 4, and 8 h) was taken for analysis. The quantification of bile acids in the dialysate was measured based on the taurocholic acid concentration, as previously described by Boyd et al. (\u003cspan class=\"CitationRef\"\u003e1996\u003c/span\u003e). The BRI values were computed with the Eq.\u0026nbsp;(4):\u003c/p\u003e\n \u003cp\u003eBDRI value (%)\u0026thinsp;=\u0026thinsp;100 \u0026minus; [(Total bile acid diffused with MPs present / Total bile acid diffused without MPs) \u0026times;100]\u003c/p\u003e\n \u003cp\u003eThe effects of the MPs on the retardation of glucose and bile acid diffusion were compared with those of carboxymethylcellulose (CMC) (Sigma-Aldrich), a commercial dietary fiber.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003eMeasurement of Prebiotic Growth Promotion\u003c/h2\u003e\n \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\n \u003ch2\u003eSelection of Probiotic Strains\u003c/h2\u003e\n \u003cp\u003eFour types of \u003cem\u003eLactobacillus\u003c/em\u003e strains known for their probiotic benefits within the human gut was utilized: \u003cem\u003eLactobacillus plantarum\u003c/em\u003e (ATCC 8014), \u003cem\u003eL. acidophilus\u003c/em\u003e (ATCC 832), \u003cem\u003eL. casei\u003c/em\u003e (ATCC 393), and \u003cem\u003eL. rhamnosus\u003c/em\u003e (ATCC 7469). Prior to experimental use, each strain was cultivated in \u003cem\u003eLactobacillus\u003c/em\u003e MRS Broth (Becton, Dickinson and Company, Sparks, MD., USA) in an incubator (IB-600M, Jeio Tech, Japan) at 37\u0026deg;C for 24 h. After initial culturing, each bacterium underwent a series of three subcultures to ensure robust and active bacterial colonies for the subsequent assessments.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eProbiotic Growth Assessment\u003c/h2\u003e\n \u003cp\u003eFollowing the protocols described by Im et al. (\u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e), each bacterial strain was first diluted in 0.1% sterilized peptone water (Bacto Pepetone; Becton Dickinson Co.) to standardized the initial bacterial concentration to 4 log Colony-Forming Units per milliliter (CFU/mL). Then, 0.5 mL of each diluted bacterial suspension was inoculated into 8.5 mL of sterile MRS broth. To this environment, 1 mL of the MPs that had been previously dissolved to a concentration of 50 mg/mL in a 30% dimethyl sulfoxide (DMSO from Junsei, Tokyo, Japan) solution or 1 mL of a 30% DMSO solution was added and incubated under controlled conditions at 37\u0026deg;C for 24-h to allow for bacterial growth promotion. Following the growth period, a 1 mL aliquot of each cultured sample was taken, further diluted using 0.1% sterilized peptone water, and subsequently plated to facilitate colony formation. After an additional 24 h of incubation, the resultant colonies were quantified.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eStatistical Analysis\u003c/h2\u003e\n \u003cp\u003eThe experimental data are expressed as mean values\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation from triplicate measurements. Statistical analysis of the experimental results was performed using the SPSS (Ver. 23, IBM Corp., Armonk, NY, USA) software with the significance threshold set at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05. A one-way analysis of variance ensured the reliability of each set values and Duncan\u0026rsquo;s multi-range test established the significance versus the mean of the experiments.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003eEffect of the Type of DES on the Extraction Yield\u003c/h2\u003e\n \u003cp\u003eThe extraction yields of the MP fraction obtained by UAE using CCG, CCU, and CCEG as solvents are shown in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. The extraction yields of the MPs obtained using CCG, CCU, and CCEG were 3.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12%, 20.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05%, and 8.73\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16%, respectively. The highest yield (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) of the MPs was obtained using CCU. Zhang and Wang (\u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e) reported that DESs can dissolve various substances such as sugars, polysaccharides, proteins, and amino acids through hydrogen bonding, enabling efficient extraction of polar or non-polar components. CCU afforded a higher extraction yield than the other DESs, which may be due to the higher hydrogen bonding ability and more electrostatic interactions of CCU with the MPs, compared to the other DESs (Zhang \u0026amp; Wang, \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e). Therefore, CCU is considered a suitable DES for MP extraction.\u003c/p\u003e\n \u003cp\u003e\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eYield of maca polysaccharide fractions obtained by various DES types\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDES type\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eYield (%)\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\u003eCCG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCEG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.73\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\"\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;S.D (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\"\u003eValues with different superscript letters in the same column are significantly different at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\"\u003eCCG, mixture of choline chloride and glycerin\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\"\u003eCCU, mixture of choline chloride and urea\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\"\u003eCCEG, mixture of choline chloride and ethylene glycol\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003eOrthogonal Optimization of Parameters for DES-based UAE\u003c/h2\u003e\n \u003cp\u003eTo select the experimental factors, including the water content added to the DES, ultrasonic power, and extraction time, the orthogonal experiment L\u003csub\u003e9\u003c/sub\u003e was carried out to check the effects on the extraction yield. The results are shown in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The yield of the MP extract ranged from 5.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16 to 26.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48%, with significant variation depending on the level of each factor. The experimental group with the highest extraction yield was run 9, and the average yield of all MPs was 15.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25%. Based on the average (\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({k}_{i}^{A})\\)\u003c/span\u003e\u003c/span\u003e extraction yield obtained at each factor variable level, as the water content added to the DES increased, the yield of the MPs also increased. The highest yield was obtained with a water content of 30%. Because DESs have high viscosities, it is important to control the viscosity to promote the movement of the target component into the solvent (Wang et al., \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). Zhang and Wang (\u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e) reported that the extraction yield increased as the water content added to a DES increased from 0\u0026ndash;30%, which is consistent with the results of this study. The yield also increased significantly with increasing ultrasonic power. At extraction times of 0, 20, and 30 min, the yield was 14.69%, 17.33, and 15.07%, respectively; the extraction yield was highest at 20 min. Chen et al. (\u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e) reported that as the ultrasonic power increased, the mass transfer rate and extraction efficiency of polysaccharides improved; however, an ultrasonic power that was too high increased the number of bubbles in the solvent, reducing the transfer efficiency of the ultrasonic power. Previous reports also stated that the yield increased at an extraction time of 20 min and then decreased again at 30 min, consistent with the results of this study.\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eFactors and results of orthogonal design experiments on different extraction condition of maca polysaccharides\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eExperiment No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eReaction parameters\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eYield (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWater content (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eUltrasonic power (W)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eExtraction time (min)\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\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e240\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e300\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e240\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.68\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e300\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e22.84\u0026thinsp;\u0026plusmn;\u0026thinsp;1.26\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e240\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.20\u0026thinsp;\u0026plusmn;\u0026thinsp;1.25\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e300\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e26.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eK1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e31.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eK2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e52.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e51.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eK3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e61.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ek1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ek2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ek3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOptimal level\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;S.D (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({K}_{i}^{A}\\)\u003c/span\u003e\u003c/span\u003e = Ʃ yield at A\u003csub\u003ei\u003c/sub\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({k}_{i}^{A} = {K}_{i}^{A}/3\\)\u003c/span\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eR: Refers to the result of extreme analysis, \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({R}_{i}^{A}\\)\u003c/span\u003e\u003c/span\u003e = max {\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({k}_{i}^{A}\\)\u003c/span\u003e\u003c/span\u003e} - min{\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({k}_{i}^{A}\\)\u003c/span\u003e\u003c/span\u003e}\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003cp\u003eTo examine the degree of influence of the extraction conditions on the yield of each extract, the effect of each factor was calculated based on an orthogonal test. According to the \u003cem\u003eR\u003c/em\u003e value (\u003cem\u003eR\u003c/em\u003e\u0026thinsp;=\u0026thinsp;max\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({k}_{i}^{A}\\)\u003c/span\u003e\u003c/span\u003e\u0026minus; min\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({k}_{i}^{A}\\)\u003c/span\u003e\u003c/span\u003e), the yield was affected by the parameters in the following order: ultrasonic power (10.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.74), moisture content (8.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44), and extraction time (2.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33). In the main effect plot for the S/N ratios, the mean S/N ratios for each factor were also plotted against the test level for each factor parameter (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The main effects plot for the extraction yield clearly shows that ultrasonic power had the most significant influence on the extraction yield, followed by the water content (very slight significance). These results are consistent with those of Mudliyar et al. (\u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e), who reported that the ultrasound power had the greatest influence on the extraction yield when the conditions for polysaccharide extraction from \u003cem\u003eTuber aestivum\u003c/em\u003e were optimized using the Taguchi method.\u003c/p\u003e\n \u003cp\u003eBecause the Taguchi experimental method does not provide a precise estimation of the effect of each factor on the overall process, the percentage contribution determined using ANOVA was employed to compensate for this effect (Abe et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). The Taguchi experimental results (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e) were processed by ANOVA using Minitab 18, where values were considered significantly different at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05. The ANOVA of the extraction data indicates that the ultrasound power had the most statistically significant contribution of 48.13% (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), followed by the water content (37.94%; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), and the extraction time (3.77%, \u003cem\u003ep\u003c/em\u003e value\u0026thinsp;=\u0026thinsp;0.043). Based on the analysis of the extraction yield using the Taguchi method, the optimal conditions are as follows: water content of 30%, ultrasound power of 300 W, and extraction time of 20 min.\u003c/p\u003e\n \u003cp\u003e\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAnalysis of variance results in relation to extraction yield\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eVariance source\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDegree of freedom (DF)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSum of square (SS)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean square (MS)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eF-ratio\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eContribution (%)\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\u003eWater content\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e367.954\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e183.977\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e37.351\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e37.94\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUltrasonic power\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e466.774\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e233.387\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e47.382\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e48.13\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eExtraction time\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e36.529\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.265\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.708\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.043\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.77\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eError\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e98.513\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.926\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e969.770\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e100.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003eExtraction Yield of MPs\u003c/h2\u003e\n \u003cp\u003eThe yield and \u003cem\u003eIC\u003c/em\u003e\u003csub\u003e\u003cem\u003e50\u003c/em\u003e\u003c/sub\u003e values for the antidiabetic activity of the MPs obtained using HWE and UAE are shown in Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e. The yield of the MPs was highest for MP-DU at 26.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48% (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), followed by MP-U (12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16%); MP-H had the significantly lowest yield (11.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17%) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). In UAE, the extraction yield was approximately two times higher when the DES was used as the solvent than when distilled water was used. Nam et al. (\u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e) reported that in the ultrasound extraction of flavonoids from \u003cem\u003eFlos sophorae\u003c/em\u003e, the yield of flavonoids was higher when the DES was used as a solvent than when water was used as the solvent. This indicates that the use of a DES during ultrasonic extraction can lead to an increase in the extraction yield.\u003c/p\u003e\n \u003cp\u003e\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eExtraction yield and antidiabetic activity of maca polysaccharides obtained by various extraction methods\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eYield (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e value (\u0026micro;g/mL)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026alpha;-Amylase inhibitory activity\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026alpha;-Glucosidase inhibitory activity\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\u003eMP-H\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e329.62\u0026thinsp;\u0026plusmn;\u0026thinsp;13.07\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e641.42\u0026thinsp;\u0026plusmn;\u0026thinsp;2.21\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMP-U\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e156.15\u0026thinsp;\u0026plusmn;\u0026thinsp;18.27\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e119.67\u0026thinsp;\u0026plusmn;\u0026thinsp;11.21\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMP-DU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e99.21\u0026thinsp;\u0026plusmn;\u0026thinsp;3.08\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e110.44\u0026thinsp;\u0026plusmn;\u0026thinsp;4.09\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAcarbose\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e89.98\u0026thinsp;\u0026plusmn;\u0026thinsp;6.60\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e178.15\u0026thinsp;\u0026plusmn;\u0026thinsp;4.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"4\"\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;S.D (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"4\"\u003eValues with different superscript letters in the same column are significantly different at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"4\"\u003eMP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n \u003ch2\u003eInhibitory Effects of MPs on Pancreatic \u0026alpha;-Amylase and \u0026alpha;-Glucosidase Activities\u003c/h2\u003e\n \u003cp\u003eThe \u003cem\u003eIC\u003c/em\u003e\u003csub\u003e\u003cem\u003e50\u003c/em\u003e\u003c/sub\u003e value for the \u0026alpha;-amylase inhibitory activity was lowest (89.98\u0026thinsp;\u0026plusmn;\u0026thinsp;6.60 \u0026micro;g/mL) for acarbose, followed by MP-DU (99.21\u0026thinsp;\u0026plusmn;\u0026thinsp;3.08 \u0026micro;g/mL), showing an activity equivalent to 90% of acarbose. MP-U and MP-H afforded high \u003cem\u003eIC\u003c/em\u003e\u003csub\u003e\u003cem\u003e50\u003c/em\u003e\u003c/sub\u003e values of 156.15\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11 and 359.62\u0026thinsp;\u0026plusmn;\u0026thinsp;13.07 \u0026micro;g/mL, respectively, indicative of low inhibitory activity. The \u003cem\u003eIC\u003c/em\u003e\u003csub\u003e\u003cem\u003e50\u003c/em\u003e\u003c/sub\u003e values for the \u0026alpha;-glucosidase inhibitory activity were 110.44\u0026thinsp;\u0026plusmn;\u0026thinsp;4.09, 119.67\u0026thinsp;\u0026plusmn;\u0026thinsp;11.21, and 641.42\u0026thinsp;\u0026plusmn;\u0026thinsp;2.21 \u0026micro;g/mL for MP-DU, MP-U, and MP-H, respectively. The \u003cem\u003eIC\u003c/em\u003e\u003csub\u003e\u003cem\u003e50\u003c/em\u003e\u003c/sub\u003e values of MP-DU and MP-U were significantly lower than that of acarbose (178.15\u0026thinsp;\u0026plusmn;\u0026thinsp;4.22 \u0026micro;g/mL), indicating that MP-DU and MP-U have high enzyme inhibitory activity. \u0026alpha;-Amylase and \u0026alpha;-glucosidase inhibitors inhibit the hydrolysis of carbohydrates, thereby reducing glucose release from starch and delaying the absorption of carbohydrates, consequently suppressing the rise in blood sugar (Xu et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). Recently, antidiabetic active substances isolated from natural sources have received considerable attention. The results of this study show that MPs have higher inhibitory activity than acarbose and great potential for use as a natural diabetes treatment. Therefore, MP, especially MP-DU, is expected to play a role as a treatment for diabetes by inhibiting the activity of \u0026alpha;-amylase and \u0026alpha;-glucosidase to prevent postprandial blood glucose rise.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003eRetarding Effect of MPs on Glucose Absorption\u003c/h2\u003e\n \u003cp\u003eThe effect of compounds on delaying glucose absorption was reported to have a very high correlation with the in vivo lowering of blood sugar based on an in vitro experiment evaluating the entrapping effect of dietary fiber (Ahmed et al., \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e). The glucose content released into the dialysate according to the dialysis time and GDRI, which is a value showing the glucose permeation inhibition effect compared to the control, is shown in Table \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e. At 30 min of dialysis, the glucose content of CMC, the positive control group, was the lowest at 4.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51 mg/100 mL, followed by that of the MP-DU (5.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26 mg/100 mL) and MP-U (6.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 mg/100 mL) groups, and finally the MP-H (6.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13 mg/100 mL) group, for which the value was significantly lower than that of the control (7.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31 mg/100 mL). After this period, the glucose concentration showed similar trends for all groups up to 4 h of dialysis.\u003c/p\u003e\n \u003cp\u003e\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eRetarding effect of maca polysaccharides obtained by various extraction methods on dialysis membrane transport of glucose\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" style=\"width: 1.6608%;\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"10\" style=\"width: 14.8362%;\"\u003e\n \u003cp\u003eDialysis time (h)\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\" rowspan=\"2\" style=\"width: 1.6608%;\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.203%;\"\u003e\n \u003cp\u003e0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.203%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.4106%;\"\u003e\n \u003cp\u003e1.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.203%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 6.6136%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003eGlucose content\u003c/p\u003e\n \u003cp\u003e(mg/100 mL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003eGDRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003eGlucose content\u003c/p\u003e\n \u003cp\u003e(mg/100 mL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003eGDRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003eGlucose content\u003c/p\u003e\n \u003cp\u003e(mg/100 mL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.5719%;\"\u003e\n \u003cp\u003eGDRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003eGlucose content\u003c/p\u003e\n \u003cp\u003e(mg/100 mL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003eGDRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.2494%;\"\u003e\n \u003cp\u003eGlucose content\u003c/p\u003e\n \u003cp\u003e(mg/100 mL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003eGDRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.6608%;\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e7.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e15.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e22.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.5719%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e26.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.2494%;\"\u003e\n \u003cp\u003e29.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.6608%;\"\u003e\n \u003cp\u003eCMC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e4.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e37.53\u0026thinsp;\u0026plusmn;\u0026thinsp;7.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e8.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e43.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e13.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.5719%;\"\u003e\n \u003cp\u003e41.04\u0026thinsp;\u0026plusmn;\u0026thinsp;2.66\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e15.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e40.79\u0026thinsp;\u0026plusmn;\u0026thinsp;1.51\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.2494%;\"\u003e\n \u003cp\u003e19.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.61\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e34.14\u0026thinsp;\u0026plusmn;\u0026thinsp;2.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.6608%;\"\u003e\n \u003cp\u003eMP-H\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e6.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e14.43\u0026thinsp;\u0026plusmn;\u0026thinsp;1.73\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e10.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e30.48\u0026thinsp;\u0026plusmn;\u0026thinsp;2.09\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e16.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.5719%;\"\u003e\n \u003cp\u003e25.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1.86\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e20.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e23.02\u0026thinsp;\u0026plusmn;\u0026thinsp;1.40\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.2494%;\"\u003e\n \u003cp\u003e25.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e11.34\u0026thinsp;\u0026plusmn;\u0026thinsp;1.43\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.6608%;\"\u003e\n \u003cp\u003eMP-U\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e6.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e15.01\u0026thinsp;\u0026plusmn;\u0026thinsp;2.65\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e9.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e35.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e15.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.5719%;\"\u003e\n \u003cp\u003e31.35\u0026thinsp;\u0026plusmn;\u0026thinsp;1.70\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e19.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e26.22\u0026thinsp;\u0026plusmn;\u0026thinsp;1.40\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.2494%;\"\u003e\n \u003cp\u003e25.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e13.98\u0026thinsp;\u0026plusmn;\u0026thinsp;1.21\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.6608%;\"\u003e\n \u003cp\u003eMP-DU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e5.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e20.21\u0026thinsp;\u0026plusmn;\u0026thinsp;3.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e9.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e38.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2.53\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e14.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.5719%;\"\u003e\n \u003cp\u003e35.41\u0026thinsp;\u0026plusmn;\u0026thinsp;1.59\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.8388%;\"\u003e\n \u003cp\u003e17.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e33.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.58\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 5.2494%;\"\u003e\n \u003cp\u003e20.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.3642%;\"\u003e\n \u003cp\u003e29.23\u0026thinsp;\u0026plusmn;\u0026thinsp;1.36\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"11\" style=\"width: 22.0059%;\"\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;S.D (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"11\" style=\"width: 22.0059%;\"\u003eValues with different superscript letters in the column are significantly different at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"11\" style=\"width: 22.0059%;\"\u003eGDRI, glucose dialysis retardation index; CMC, carboxymethylcelluose; MP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n \u003cp\u003eDuring all dialysis times, the GDRI of CMC was significantly higher than that of the MPs, followed by that of MP-DU, which had a higher GDRI than the other MPs. This shows that MP-DU has a significant effect on retarding glucose absorption. The GDRI increased with the dialysis time up to 1 h of dialysis and then tended to decrease gradually. The highest GDRI values during dialysis were as follows: CMC 43.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79%, MP-DU 38.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2.53%, MP-U 35.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79%, and MP-H 30.48\u0026thinsp;\u0026plusmn;\u0026thinsp;2.09%, with MP-DU having the highest GDRI after CMC. It has been reported that polysaccharides form a viscous gel in aqueous solutions, slowing access to small intestine cells, and that the large surface area of polysaccharide particles promotes the trapping effect of glucose, delaying its absorption (Ahmed et al., \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e; Abirami et al., \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e). Chen et al. (\u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e) reported that the GDRI of maca dietary fiber was 12.30‒23.66% after 1 h of dialysis, which is lower than the results of this study. Although the maca samples are the same, these differences are thought to be due to the variety and extraction methods used. Therefore, MP extracted with DES-based UAE in this study effectively lowers the level of serum glucose by delaying the absorption of glucose and is expected to have potential for development as a hypoglycemic agent and low-calorie functional food.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n \u003ch2\u003eRetarding Effect of MPs on Bile Acid Absorption\u003c/h2\u003e\n \u003cp\u003eThe bile acid dialysis retardation index (BDRI), which shows the effect of the polysaccharides on inhibiting bile acid permeation compared to the control, was used to monitor the effects of fiber on cholesterol metabolism. The amount of bile acid released into the dialysate according to the dialysis time and BDRI is shown in Table \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e. At all dialysis times, the amount of bile acid released into the dialysate with all MPs and CMC was significantly lower than that of the control (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). At 1 h of dialysis, CMC showed the highest BDRI (37.77\u0026thinsp;\u0026plusmn;\u0026thinsp;2.45%), followed by MP-DU (35.96\u0026thinsp;\u0026plusmn;\u0026thinsp;2.45%), MP-U (34.06\u0026thinsp;\u0026plusmn;\u0026thinsp;2.52%), and MP-H (21.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93%). The BDRI values of MP-DU and MP-U were not significantly different from those of CMC, indicating a strong inhibitory effect on the absorption of bile acid. Even at 90 min of dialysis, the BDRI of MP-DU was 35.02\u0026thinsp;\u0026plusmn;\u0026thinsp;1.66%, which was not significantly different from that of CMC (38.22\u0026thinsp;\u0026plusmn;\u0026thinsp;1.91%). The BDRI of the MPs increased up to 2 h of dialysis and then decreased. The highest inhibition of bile acid absorption was observed at 2 h of dialysis. Dietary fiber adsorbs free bile acid and inhibits the reabsorption of bile acid, which induces the consumption of cholesterol for the synthesis of bile acid in the body. This process lowers the concentration of cholesterol in the blood, which has a positive effect on cardiovascular diseases such as arteriosclerosis and heart disease (Ioniță-M\u0026icirc;ndrican et al., \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e). Ma et al. (\u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e) reported that the many hydrophobic groups present in dietary fiber increase the adsorption capacity by forming hydrophobic bonds through interaction with bile acid, and that the small size of the dietary fiber particles increases the absorption capacity by increasing the surface area. Therefore, MP-DU and MP-U had a high bile acid absorption inhibitory effect and are expected to have a positive effect on lipid metabolism.\u003c/p\u003e\n \u003cp\u003e\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab6\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eRetarding effect of maca polysaccharides obtained by various extraction methods on dialysis membrane transport of bile acid\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" style=\"width: 1.478%;\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"11\" style=\"width: 14.6682%;\"\u003e\n \u003cp\u003eDialysis time (h)\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\" rowspan=\"2\" style=\"width: 1.478%;\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.1442%;\"\u003e\n \u003cp\u003e0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.1442%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.1979%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.1442%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.2161%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 3.2785%;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003eBile acid content (\u0026micro;mole/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003eBRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003eBile acid content (\u0026micro;mole/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003eBRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003eBile acid content (\u0026micro;mole/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.2362%;\"\u003e\n \u003cp\u003eBRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003eBile acid content (\u0026micro;mole/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003eBRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 4.1653%;\"\u003e\n \u003cp\u003eBile acid content (\u0026micro;mole/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003eBRI\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.478%;\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e98.98\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e170.92\u0026thinsp;\u0026plusmn;\u0026thinsp;5.76\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e369.25\u0026thinsp;\u0026plusmn;\u0026thinsp;4.38\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.2362%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e497.56\u0026thinsp;\u0026plusmn;\u0026thinsp;3.39\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 4.1653%;\"\u003e\n \u003cp\u003e597.39\u0026thinsp;\u0026plusmn;\u0026thinsp;5.32\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.478%;\"\u003e\n \u003cp\u003eCMC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e65.54\u0026thinsp;\u0026plusmn;\u0026thinsp;2.82\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e33.79\u0026thinsp;\u0026plusmn;\u0026thinsp;2.85\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e106.36\u0026thinsp;\u0026plusmn;\u0026thinsp;4.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e37.77\u0026thinsp;\u0026plusmn;\u0026thinsp;2.45\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e230.37\u0026thinsp;\u0026plusmn;\u0026thinsp;3.93\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.2362%;\"\u003e\n \u003cp\u003e37.61\u0026thinsp;\u0026plusmn;\u0026thinsp;1.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e301.78\u0026thinsp;\u0026plusmn;\u0026thinsp;3.91\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e39.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 4.1653%;\"\u003e\n \u003cp\u003e370.50\u0026thinsp;\u0026plusmn;\u0026thinsp;2.92\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e37.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.478%;\"\u003e\n \u003cp\u003eMP-H\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e80.12\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e19.05\u0026thinsp;\u0026plusmn;\u0026thinsp;1.65\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e133.85\u0026thinsp;\u0026plusmn;\u0026thinsp;1.58\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e21.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e316.63\u0026thinsp;\u0026plusmn;\u0026thinsp;4.89\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.2362%;\"\u003e\n \u003cp\u003e14.25\u0026thinsp;\u0026plusmn;\u0026thinsp;1.32\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e425.96\u0026thinsp;\u0026plusmn;\u0026thinsp;5.23\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e14.39\u0026thinsp;\u0026plusmn;\u0026thinsp;1.05\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 4.1653%;\"\u003e\n \u003cp\u003e535.14\u0026thinsp;\u0026plusmn;\u0026thinsp;3.19\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e10.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.478%;\"\u003e\n \u003cp\u003eMP-U\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e69.45\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e29.83\u0026thinsp;\u0026plusmn;\u0026thinsp;1.65\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e112.71\u0026thinsp;\u0026plusmn;\u0026thinsp;4.31\u003csup\u003eba\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e34.06\u0026thinsp;\u0026plusmn;\u0026thinsp;2.52\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e278.55\u0026thinsp;\u0026plusmn;\u0026thinsp;5.35\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.2362%;\"\u003e\n \u003cp\u003e24.56\u0026thinsp;\u0026plusmn;\u0026thinsp;1.45\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e372.36\u0026thinsp;\u0026plusmn;\u0026thinsp;2.49\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e25.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 4.1653%;\"\u003e\n \u003cp\u003e471.36\u0026thinsp;\u0026plusmn;\u0026thinsp;2.62\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e21.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 1.478%;\"\u003e\n \u003cp\u003eMP-DU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e75.86\u0026thinsp;\u0026plusmn;\u0026thinsp;4.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e23.36\u0026thinsp;\u0026plusmn;\u0026thinsp;4.49\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e109.45\u0026thinsp;\u0026plusmn;\u0026thinsp;4.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e35.96\u0026thinsp;\u0026plusmn;\u0026thinsp;2.45\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e279.53\u0026thinsp;\u0026plusmn;\u0026thinsp;6.89\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.2362%;\"\u003e\n \u003cp\u003e24.30\u0026thinsp;\u0026plusmn;\u0026thinsp;1.87\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.9617%;\"\u003e\n \u003cp\u003e383.20\u0026thinsp;\u0026plusmn;\u0026thinsp;3.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e22.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.61\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" style=\"width: 4.1653%;\"\u003e\n \u003cp\u003e460.35\u0026thinsp;\u0026plusmn;\u0026thinsp;3.95\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 1.1824%;\"\u003e\n \u003cp\u003e22.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.66\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"12\" style=\"width: 19.9399%;\"\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;S.D (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"12\" style=\"width: 19.9399%;\"\u003eValues with different superscript letters in the column are significantly different at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"12\" style=\"width: 19.9399%;\"\u003eBDRI, bile acid dialysis retardation index; CMC, carboxymethylcelluose; HWE, hot water extraction; UAE, ultrasound-assisted extraction; DES, deep eutectic solvent; MP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\n \u003ch2\u003eProbiotic Growth-Stimulation Capacity of MPs\u003c/h2\u003e\n \u003cp\u003eTo determine the prebiotic effects of the MPs obtained using different extraction methods, the effects of the MPs on the proliferation of lactic acid bacteria (\u003cem\u003eL. plantarum, L. acidophilus, L. casei, L. rhamnosus\u003c/em\u003e) were measured; the results are summarized in Table \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e. For all strains of lactic acid bacteria, groups treated with MPs exhibited significantly higher growth\u0026mdash;approximately 10 times\u0026mdash;compared to the control group. MP-H had the greatest effect on the proliferation of \u003cem\u003eL. plantarum\u003c/em\u003e and \u003cem\u003eL. acidophilus\u003c/em\u003e, with counts of 14.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.52 \u0026times; 10\u003csup\u003e9\u003c/sup\u003e and 11.00\u0026thinsp;\u0026plusmn;\u0026thinsp;2.00 \u0026times; 10\u003csup\u003e9\u003c/sup\u003e, respectively, whereas MP-U had the highest effect on the proliferation of \u003cem\u003eL. casei\u003c/em\u003e and \u003cem\u003eL. rhamnosus\u003c/em\u003e, with counts of 9.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 \u0026times; 10\u003csup\u003e9\u003c/sup\u003e and 10.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 \u0026times; 10\u003csup\u003e9\u003c/sup\u003e, respectively. The degree of growth for each strain of lactic acid bacterium differed depending on the sample. The control and MP-H groups were most effective for promoting the growth of \u003cem\u003eL. plantarum\u003c/em\u003e, whereas the MP-U and MP-DU groups were most effective for promoting the growth of \u003cem\u003eL. rhamnosus\u003c/em\u003e and \u003cem\u003eL. acidophilus\u003c/em\u003e, respectively. MP-U and MP-DU significantly enhanced the growth of all strains, demonstrating their broad-spectrum effectiveness. Polysaccharides that are not digested in the gastrointestinal tract can stimulate the growth of beneficial intestinal bacteria, such as lactic acid bacteria and bifidobacteria, and have health benefits such as promoting the production of short-chain fatty acids; therefore, they are considered to have a prebiotic effect (Chen et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). The prebiotic activity of polysaccharides varies depending on their physicochemical properties, including molecular weight, monosaccharide composition, glycosidic bonds, and viscosity (Huang et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003e\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab7\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eEffect of maca polysaccharide by various extraction methods on the growth of lactic acid bacteria (CFU/mL)\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 \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eL. plantarum\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eL. acidophilus\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eL. casei\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eL. rhamnosus\u003c/em\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\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53☓10\u003csup\u003e8c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00☓10\u003csup\u003e8c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00☓10\u003csup\u003e8c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15☓10\u003csup\u003e8d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMP-H\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.52☓10\u003csup\u003e9a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.00\u0026thinsp;\u0026plusmn;\u0026thinsp;2.00☓10\u003csup\u003e9a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53☓10\u003csup\u003e9b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58☓10\u003csup\u003e9c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMP-U\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53☓10\u003csup\u003e9b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00☓10\u003csup\u003e9b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00☓10\u003csup\u003e9a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00☓10\u003csup\u003e9a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMP-DU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58☓10\u003csup\u003e9b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58☓10\u003csup\u003e9b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58☓10\u003csup\u003e9a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53☓10\u003csup\u003e9b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eMean S.D (n\u0026thinsp;=\u0026thinsp;3)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eValues with different superscript letters in the column are significantly different at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eControl is only DMSO added.\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eMP-H, MP-U, and MP-DU, maca polysaccharides obtained by using hot-water extraction, ultrasound-assisted extraction with water, and DES-based ultrasound-assisted extraction, respectively\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe present study investigated the deep eutectic solvent (DES)-based ultrasound-assisted extraction (UAE) for extracting maca polysaccharides (MPs) from maca roots and compared this process with two other extraction techniques (HWE and UAE with water). The highest extraction yield was obtained using choline chloride and urea at a molar ratio of 1:3. As a result of using the Taguchi orthogonal array as a design variable, the extraction yield was 26.28% at 30% water content, 300W ultrasonic power, and 20% extraction time. Under the optimized conditions, the DES-based UAE provided a higher yield of MPs (MP-DU) than the other extraction methods. MP-DU inhibited the α-amylase and β-glucosidase activities and delayed the absorbance of glucose and bile acids. In conclusion, this study not only provides an environmentally friendly, efficient, and economical alternative method to extract MPs, but also provides a scientific basis for the comprehensive utilization of maca as a potential functional food.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eCCEG Mixture of choline chloride and ethylene glycol\u003c/p\u003e\n\u003cp\u003eCCG Mixture of choline chloride and glycerin\u003c/p\u003e\n\u003cp\u003eCCU Mixture of choline chloride and urea \u003c/p\u003e\n\u003cp\u003eDESs Deep eutectic solvents\u003c/p\u003e\n\u003cp\u003eBDRI Bile acid dialysis retardation index\u003c/p\u003e\n\u003cp\u003eGDRI Glucose dialysis retardation index\u003c/p\u003e\n\u003cp\u003eHWE Hot-water extraction\u003c/p\u003e\n\u003cp\u003eMPs Maca polysaccharides\u003c/p\u003e\n\u003cp\u003eMP-DU MPs prepared by using DES-based UAE\u003c/p\u003e\n\u003cp\u003eMP-U MPs prepared by using UAE with water\u003c/p\u003e\n\u003cp\u003eMP-W MPs prepared by using HWE\u003c/p\u003e\n\u003cp\u003eUAE Ultrasound-assisted extraction \u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eConflict of Interest\u003c/h2\u003e \u003cp\u003eThe authors declare no conflicts of interest.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis research was funded by the Basic Science Research Program through the National Research Foundation of Korea (NRF), which is funded by the Ministry of Education (grant numbers NRF-2021R1A6A1A03040177).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eEun Jeong Kim: formal analysis, investigation, data curation, writing-original draft, Choon Young Kim: data curation, writing-review and editing, funding acquisition, Kyung Young Yoon: conceptualization, data curation, writing-review and editing, supervision.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbe, J. 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Protective effect of polysaccharide from maca (\u003cem\u003eLepidium meyenii\u003c/em\u003e) on Hep-G2 cells and alcoholic liver oxidative injury in mice. \u003cem\u003eInternational Journal of Biological Macromolecules\u003c/em\u003e, \u003cem\u003e99\u003c/em\u003e, 63\u0026ndash;70. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ijbiomac.2017.01.125\u003c/span\u003e\u003cspan address=\"10.1016/j.ijbiomac.2017.01.125\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"food-and-bioprocess-technology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Food and Bioprocess Technology](https://www.springer.com/journal/11947)","snPcode":"11947","submissionUrl":"https://submission.nature.com/new-submission/11947/3","title":"Food and Bioprocess Technology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Polysaccharide, Deep eutectic solvent, Ultrasound-assisted extraction, Orthogonal experiment design, Biological activity","lastPublishedDoi":"10.21203/rs.3.rs-4340936/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4340936/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eDeep eutectic solvent (DES) was used for the ultrasound-assisted extraction (UAE) of polysaccharides from maca. Extraction parameters affecting the extraction yield were experimentally identified and their significance was further investigated using the Taguchi method. DES prepared from choline chloride and urea afforded the highest yield (20.03%) and was chosen as the solvent for UAE. The optimal extraction parameters were: water content of 30% for DES, ultrasonic power of 300 W, and extraction time of 20. The extraction yield (26.28%) of maca polysaccharides (MPs) obtained using these extraction parameters was more than twice that of MPs obtained by hot-water extraction and UAE with water. Moreover, MPs obtained through DES-based extraction exhibited various biological functions such as inhibiting pancreatic α-amylase and α-glucosidase activities, delaying absorption of glucose and bile acid, and stimulating the probiotic. Therefore, DES can be used to extract polysaccharides from maca with biological action as a highly efficient and non-polluting alternative solvent.\u003c/p\u003e","manuscriptTitle":"Deep-eutectic-solvent-based ultrasound-assisted extraction of polysaccharides from maca: Optimization using Taguchi methodology and comparison with the conventional method","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-05-07 10:00:46","doi":"10.21203/rs.3.rs-4340936/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-05-22T05:31:03+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-05-19T03:29:58+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-05-15T16:51:21+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"66126786364602880107007373974884845147","date":"2024-05-15T05:01:23+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-05-13T09:49:40+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"140363743235556137362805202140611868862","date":"2024-05-12T09:55:17+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"168768974538483579167750670033370934980","date":"2024-05-10T04:31:04+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-05-10T03:58:32+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-05-01T11:23:30+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-05-01T02:13:17+00:00","index":"","fulltext":""},{"type":"submitted","content":"Food and Bioprocess Technology","date":"2024-04-29T07:12:33+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"food-and-bioprocess-technology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Food and Bioprocess Technology](https://www.springer.com/journal/11947)","snPcode":"11947","submissionUrl":"https://submission.nature.com/new-submission/11947/3","title":"Food and Bioprocess Technology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"f439321e-6a92-47f9-8220-7b6a622286f7","owner":[],"postedDate":"May 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2024-10-03T11:23:15+00:00","versionOfRecord":[],"versionCreatedAt":"2024-05-07 10:00:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4340936","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4340936","identity":"rs-4340936","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}