Green Extraction and Optimization of Bioactive Compounds from Elderberry (Sambucus nigra L.) by β-Cyclodextrin Assisted Extraction Short title: Optimized Green Extraction of Elderberry Bioactives with β-Cyclodextrin

Green Extraction and Optimization of Bioactive Compounds from Elderberry (Sambucus nigra L.) by β-Cyclodextrin Assisted Extraction Short title: Optimized Green Extraction of Elderberry Bioactives with β-Cyclodextrin | 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 Green Extraction and Optimization of Bioactive Compounds from Elderberry (Sambucus nigra L.) by β-Cyclodextrin Assisted Extraction Short title: Optimized Green Extraction of Elderberry Bioactives with β-Cyclodextrin Vildan EYİZ, Ayşenur ACAR This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7428774/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Phenolics of elderberry ( Sambucus nigra L.) were recovered via a green, β-cyclodextrin (β-CD)–assisted aqueous extraction. Process effects were mapped using response surface methodology (face-centered central composite design) with β-CD 1–3% (w/v), liquid–solid (L/S) ratio 10–30% (w/v), and time 30–90 min. Responses (TPC, TFC, TMAC, TPAC; DPPH, FRAP, CUPRAC) were modeled to identify operating windows. TPC followed a quadratic relationship; TFC a reduced quadratic with an optimum near ~ 2% β-CD, whereas TMAC and TPAC exhibited approximately linear trends across the studied space. DPPH showed clearer parameter sensitivity than FRAP/CUPRAC, and extraction time exerted a secondary effect within 30–90 min. Contour plots consistently indicated low-to-moderate L/S together with ~ 2% β-CD as a region that balances multiple responses while avoiding dilution or mass-transfer limitations. Overall, β-CD–assisted extraction provides an effective, solvent-lean route for recovering elderberry phenolics and delineates a practical window for greener processing. These results motivate follow-up work on inclusion mechanisms, stability during storage and digestion, and scale-up supported by techno-economic and life-cycle assessments. elderberry β-cyclodextrin green extraction response surface methodology Figures Figure 1 Figure 2 Figure 3 1. Introduction Phenolic compounds, a class of secondary metabolites including pigments, have long garnered significant attention due to their diverse functional properties. These compounds play a crucial role in determining the organoleptic characteristics—such as taste, color, and aroma—of plant-based foods and are abundantly present in fruits and vegetables. In particular, phenolics are largely responsible for the characteristic coloration observed in many fruit and vegetable species, contributing both to their visual appeal and potential health benefits 1 . Elderberry ( Sambucus nigra L.) is a rich source of bioactive compounds, particularly polyphenols, and is well known for its strong antioxidant capacity 2 – 5 . Among its notable constituents are anthocyanins, which contribute not only to its health-promoting properties but also to its potential use as a natural colorant. Elderberry powders and extracts have been successfully incorporated into various food products for coloring purposes 6 – 8 . Owing to these functional and technological attributes, elderberry is increasingly utilized as a functional ingredient and natural additive in the development of health-oriented food products 9 (MARTİS). β-CDs are cyclic oligosaccharides produced through the enzymatic degradation of starch. They consist of six, seven, or eight D-(+)-glucopyranose units linked by α-1,4 glycosidic bonds 10 . Among natural cyclodextrins, β-CD is the most accessible, cost-effective, and widely utilized form 11 . Recognized as Generally Regarded As Safe (GRAS), β-CD is extensively used in the food industry due to its ability to form inclusion complexes with various bioactive compounds, thereby enhancing their stability, solubility, and bioavailability 12 . When employed as an extraction medium in aqueous systems, β-CD facilitates the effective recovery of bioactive compounds from plant matrices by interacting with target molecules 13 . Previous studies have demonstrated the successful application of β-CD in improving the extraction efficiency of bioactives from various plant sources, including red beetroot 14 , olive and red grape pomace 15 , 16 , whole pomegranate fruit 17 , and peach pomace 18 . In this study, phytochemicals were extracted from elderberry using β-CD as a green solvent, and the bioactive properties of the resulting extracts were evaluated. The effects of key extraction parameters—namely β-CD concentration, liquid-to-solid ratio, and extraction time—on the recovery of phenolics, antioxidants, flavonoids, anthocyanidins, and proanthocyanidins were systematically examined using a Central Composite Design (CCD). This experimental approach enabled the identification of optimal extraction conditions for maximizing the yield of bioactive compounds from elderberry. 2. Materials and methods 2.1.Materials Approximately 3 kg of elderberries ( Sambucus nigra L.) were procured from a local market in Konya, Turkey. The fruits were thoroughly washed and cleaned to remove any foreign materials. Drying was performed using a vacuum oven (Eksis Makine, Isparta, Turkey) at 50°C. The dried elderberries were then ground into a fine powder using a laboratory blender. The resulting elderberry powder was stored in sealed polyethylene bags at − 20°C until further use. β-CD was obtained from Sigma-Aldrich (St. Louis, MO, USA), while all other analytical-grade chemicals were purchased from Merck (Darmstadt, Germany). 2.2.Extraction Extraction was performed following Kim, Son, Pang, Yang, Lee, Lee, Lee, Park and Yoo 19 , with modifications. Based on the experimental design, predetermined amounts of elderberry powder were accurately weighed into extraction vessels. Corresponding volumes of β-CD solutions were added to the samples according to the design parameters. The mixtures were homogenized at 10,000 rpm for 1 min using an Ultraturrax homogenizer (IKA T25 digital, Staufen im Breisgau, Germany). The samples were placed in a shaking water bath (Nüve, NB-20, Ankara, Türkiye) (60°C, 70 rpm) for 1 h. Following homogenization, the samples were centrifuged at 10,000 × g for 15 min (Hermle Z326K, Germany). The resulting supernatants were filtered (Saha, Turkey) to remove particulates and subsequently stored at + 4°C until further analysis. 2.3.Experimental design In this study, a face-centered CCD was employed to evaluate the effects of three independent variables on the extraction efficiency of bioactive compounds from elderberries. The selected factors and their levels were as follows: β-CD concentration (1%, 2%, 3%) 18 , 20 ,liquid-to-solid ratio (10%, 20%, 30%) 21 , and extraction time (30, 60, and 90 min) 22 , 23 . As presented in Table 1 , the design consisted of 15 experimental runs, including three replicates at the center point to estimate experimental error. 7 response variables were measured for each extract: TPC, antioxidant activity (DPPH, FRAP, and CUPRAC assays), TFC, TMAC, and TPAC. Mathematical models for each response were developed using least squares regression based on the experimental data. The statistical significance of model terms was evaluated at a 90% confidence level using p-values. Analysis of variance (ANOVA), lack-of-fit testing, and determination of coefficients (R²) were performed using Design-Expert Version 10 software (Stat-Ease Inc., Minneapolis, USA). A desirability function approach was applied to determine the optimal extraction conditions by simultaneously maximizing all response variables. No additional experimental validation of the model was conducted. . Table 1 Experimental design and responses of elderberry extracts Std order Run order β-cyclodextrin concentration (%) Liquid-solid ratio (%, w/v) Extraction time (min) TPC (g GAE/kg dm) DPPH (g TE/kg dm) FRAP (g TE/kg dm) CUPRAC (g TE/kg dm) TFC (mg CE/kg dm) TMAC (mg C3G/kg dm) TPAC (g cyanidin/kg dm) 10 1 2 30 30 51.03 98.04 50.89 48.79 18189 9225 17.17 12 2 2 30 90 51.96 98.04 85.86 75.24 20521 8652 26.20 15 3 2 20 60 47.59 74.81 71.01 59.82 19454 8620 22.67 3 4 1 30 60 46.71 85.62 66.89 68.37 16773 7800 23.98 5 5 1 20 30 44.59 83.09 80.80 54.34 16197 7399 23.26 11 6 2 10 90 44.34 84.76 75.10 66.32 20009 8732 25.75 4 7 3 30 60 45.00 78.47 100.19 65.32 14497 7897 23.21 9 8 2 10 30 38.43 78.64 97,30 72.48 19898 8359 24.97 13 9 2 20 60 46.97 68.87 90.72 64.85 18418 9430 24.89 6 10 3 20 30 46.61 73.37 67.53 64.51 19380 8890 19.75 2 11 3 10 60 37.00 54.16 82.04 55.58 18862 9701 24.32 7 12 1 20 90 49.67 74.27 84.03 50.95 17048 8042 25.40 1 13 1 10 60 43.30 76.56 73.61 47.62 18656 9058 20.72 14 14 2 20 60 48.42 85.43 98.66 63.44 18362 8480 27.58 8 15 3 20 90 45.08 59.78 65.67 49.82 17085 8182 24.04 2.4.Extraction analyses 2.4.1.Determination of TPC of extracts The TPC of the elderberry extracts was determined using the Folin–Ciocalteu colorimetric method, as described by Georgiev, Weber, Kneschke, Denev, Bley and Pavlov 24 , with slight modifications. Briefly, each extract was appropriately diluted with distilled water. Then, 0.2 mL of the diluted sample was mixed with 2.0 mL of 0.2 N Folin–Ciocalteu reagent, followed by the addition of 0.8 mL sodium carbonate solution (75 g/L, Na₂CO₃). The mixture was incubated for 30 min at room temperature in the dark. After incubation, the absorbance was measured at 765 nm using a UV-Vis spectrophotometer (Biochrom Libra S22, Cambridge, UK). Results were expressed as grams of gallic acid equivalents per kilogram of dry matter (g GAE/kg dm). 2.4.2.Determination of antiradical activity of extracts The antioxidant capacity of elderberry extracts was evaluated using three widely applied assays: 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Power (FRAP), and Cupric Ion Reducing Antioxidant Capacity (CUPRAC). The DPPH radical scavenging activity was determined with slight modifications based on the method of Tontul and Topuz 25 . Initially, the elderberry extracts were diluted with distilled water. Then, 950 µL of freshly prepared DPPH solution (60 µM in methanol) was added to 50 µL of the diluted extract. The mixture was incubated for 30 min at room temperature in the dark. Absorbance was measured at 516 nm using a UV–Vis spectrophotometer. The FRAP assay was carried out according to the method described by Granato, Karnopp and van Ruth 26 Granato et al. (2015), with minor modifications. The FRAP reagent was freshly prepared by mixing 300 mM acetate buffer (pH 3.6), 10 mM TPTZ solution in 40 mM HCl, and 20 mM ferric chloride hexahydrate in a 10:1:1 (v/v/v) ratio. After pre-incubation of the FRAP reagent at 37°C for 10 min, 100 µL of each extract was added. The mixture was incubated in the dark at 37°C for 30 min, and absorbance was measured at 593 nm. CUPRAC analysis was performed following the procedure described by Apak, Güçlü, Özyürek and Karademir 27 . The reaction mixture was prepared by combining equal volumes (1:1:1, v/v/v) of 7.5 mM neocuproine, 10 mM copper(II) chloride (CuCl₂), and 1 M ammonium acetate buffer (pH 7.0). To this mixture, 0.5 mL of the extract and 0.6 mL of distilled water were added. The final mixture was incubated at room temperature for 30 min in the dark. Absorbance was recorded at 450 nm. A blank was prepared by replacing the sample volume with an equivalent volume of the reaction mixture. All antioxidant activity results were expressed as grams of Trolox equivalents per kilogram of dry matter (g TE/kg dm). 2.4.3.Determination of TFC of extracts The TFC of the elderberry extracts was determined using the colorimetric method described by Chang, Lin, Chang and Liu 28 , with slight modifications. Briefly, 0.5 mL of the extract was mixed with 2.5 mL of distilled water and 0.15 mL of 5% sodium nitrite (NaNO₂) solution. The mixture was allowed to stand for 5 min at room temperature. Subsequently, 0.3 mL of 10% aluminum chloride (AlCl₃) solution was added, and the reaction was allowed to proceed for an additional 5 min. Then, 1.0 mL of 1 M sodium hydroxide (NaOH) was added to the mixture, which was then vortexed vigorously. The absorbance was measured at 510 nm against a distilled water blank using a UV–Vis spectrophotometer. TFC was expressed as grams of catechin equivalents per kilogram of dry matter (g CE/kg dm). 2.4.4.Determination of Total Monomeric Anthocyanin Content of extracts The TMAC of the elderberry extracts was determined using the pH differential method, as described by 25 , with slight modifications. The extracts were appropriately diluted with buffer solutions at pH 1.0 and pH 4.5. The diluted samples were allowed to equilibrate in the dark for 20 min at room temperature. Absorbance readings were then taken at 510 nm (λmax) and 700 nm against a distilled water blank using a UV–Vis spectrophotometer. The anthocyanin content was calculated and expressed as grams of cyanidin-3-glucoside equivalents per kilogram of dry matter (g C3G/kg dm). 2.4.5.Determination of Total Proanthocyanin Content of extracts The TPAC of the elderberry extracts was determined following the method described by Eyiz, Tontul and Turker 23 , with slight modifications. A solution of FeSO₄·7H₂O (154 ppm) was prepared in a mixture of hydrochloric acid and n -butanol (2:3, v/v). In a stoppered glass tube, 2.5 mL of this reagent was added to 0.25 mL of the extract. The mixture was then incubated in a water bath at 95°C for 15 min. After incubation, the absorbance was measured at 540 nm using a UV–Vis spectrophotometer. Results were expressed as grams of proanthocyanidins per kilogram of dry matter (g TPAC/kg dm). 2.5. Statistical analysis All data were expressed as mean values ± standard deviations based on triplicate measurements. Statistical analyses were performed using one-way analysis of variance (ANOVA) to determine significant differences among experimental groups, and Pearson correlation analysis was applied to assess relationships between the measured variables. ANOVA results were considered statistically significant at p < 0.05, while Pearson correlation coefficients were evaluated at significance levels of p < 0.05 and p < 0.01. In addition, Response Surface Methodology (RSM) based on a face-centered CCD was employed to model and optimize the effects of independent variables—β-CD concentration, liquid-to-solid ratio, and extraction time—on the recovery of bioactive compounds. Design-Expert version 10 software (Stat-Ease Inc., Minneapolis, USA) was used for regression analysis, analysis of variance (ANOVA), lack-of-fit testing, and optimization through desirability function. 3. Results and discussions 3.1.TPC Extracting natural bioactives like polyphenols is often a time-consuming process that uses organic solvents. This can be bad for the environment and limit how useful they are in practice. Furthermore, harsh extraction conditions can result in the degradation of the compound, thereby reducing its bioavailability 29 . TPC values of elderberry extracts ranged from 37.00 to 51.96 g GAE/kg dm, with an average of 45.78 g GAE/kg dm and a coefficient of variation of 8.97% (Table 1 ). Furthermore, TPC values of elderberry extract samples are illustrated in Fig. 1 a. 2% β-CD, 30% liquid-to-solid ratio, 90 min had the highest TPC, while 3% β-CD, 10% liquid-to-solid ratio, 60 min had the lowest (Fig. 1 a). According to the RSM analysis (see Table 2 ), the TPC response was best described by a quadratic model (R² = 0.9330; Adj R² = 0.8124). At a fixed liquid-to-solid ratio of 20 and an extraction time of 30 min, increasing the concentration of β-CD did not notably enhance TPC. Similar trends were observed in phenolic yield studies with mint 30 . Both β-CD concentration (A²) and the liquid-to-solid ratio (B²) exhibited significant negative quadratic effects (p ≈ 0.05). Figure 3a clearly illustrates the combined influence of β-CD concentration and liquid-to-solid ratio on TPC. The orange–yellow regions indicate the highest TPC values (~ 48–50 g GAE/kg dm), corresponding to a moderate β-CD concentration (2%) and a medium–high liquid-to-solid ratio (20–25%). The green and blue zones represent lower TPC levels, with the minimum occurring at a high β-CD concentration (3%) and a low liquid-to-solid ratio (10%). Phenolic compounds are generally highly soluble in water due to their ability to form hydrogen bonds. However, the relatively low water solubility of β-CD 31 and the presence of a secondary hydrogen-bonding network within its structure that limits interactions with water molecules 32 , 33 can reduce mass transfer at high β-CD levels. Additionally, β-CD presents a hydrophobic inner surface. Therefore, it is better for compounds with high polarity instead 34 . Consequently, as the concentration of β-CD increases, the extraction yield of water-soluble phenolic compounds may decrease. In this study, the liquid-to-solid ratio also negatively affected TPC, suggesting that exceeding the optimum level could cause dilution of phenolic compounds in the solvent phase and reduce extraction efficiency. In contrast, studies on whole pomegranates have reported increases in TPC at higher β-CD levels 17 . Such discrepancies are likely due to differences in the plant matrix, extraction methods and liquid-to-solid ratios applied. 3.2.Antioxidant activity analyses (DPPH, FRAP, CUPRAC) Changes in DPPH, FRAP, and CUPRAC antioxidant activities of elderberry extract samples are presented in Table 1 and Figs. 1 b, 1 c, and 1 d. The results showed that antioxidant activities, measured by the DPPH, FRAP, and CUPRAC methods, ranged from 54.16 to 98.04, 50.89 to 100.19, and 47.62 to 75.24 g TE/kg dm, respectively. The highest DPPH activity was observed in the sample, extracted using 2% (w/v) β-CD concentration, 30% liquid-solid ratio, and 90 min extraction time. This same sample exhibited the highest antioxidant activity according to CUPRAC assays (Table 1 ). On the other hand, the maximum FRAP activities were detected in the sample, prepared with 3% (w/v) β-CD concentration, 30% liquid-solid ratio, and 60 min extraction time. According to two-factorial ANOVA, the interaction between β-CD concentration and liquid-solid ratio significantly influenced the antioxidant activities, and Reduced Quadratic was identified as the most suitable. Although the TPC yield was best described by a quadratic model, the DPPH radical scavenging activity was more accurately represented by a reduced quadratic model. A positive correlation between TPC and DPPH values is commonly observed in most plant extracts. Nevertheless, deviations from this pattern have been reported, often attributed to synergistic or antagonistic interactions among the bioactive constituents within the extracts 35 , 36 . However, it is important to note that the FRAP and CUPRAC responses could not be modeled within this experimental design (Table 2 ). Moreover, the liquid-solid ratio exerted an increased decrease effect on antioxidant activities (Figs. 1 b, 1 c, and 1 d). Table 1 shows the relationship between temperature and the decrease in DPPH value when the ß-Cd concentration (1%) and the liquid-solid ratio (20) were kept constant. With regard to the extraction process with β-CD using DES solvent, an initial rise in antiradical activity up to 55°C is observed, which corresponds to an enhancement in binding affinity. This is followed by a subsequent decrease up to 80°C 37 . The observed decline in antioxidant activities with increasing β-CD concentration likely results from the concurrent decrease in TPC extraction, indicating a limitation of higher β-CD concentrations in maximizing antioxidant recovery. Contrary to our findings, previous studies have reported significantly higher antiradical activities in samples extracted with β-CD from red beet and peach pomace compared to those extracted with ethanol 14 , 18 . This discrepancy may be attributed to variations in extraction temperature, time, and method, as these parameters critically affect the efficiency of compound release, solubility, and stability during extraction. Increases were observed in both TPC and DPPH values with the increase in the liquid-solid ratio (Fig. 3a, 3b). The process of extracting bioactive compounds with antioxidant properties, like phenolics and flavonoids, is mainly driven by the difference in concentrations between the solvent and the solid matrix 38 . The gradient is enhanced by an increase in the liquid-to-solid ratio, which facilitates mass transfer by promoting the diffusion of the solvent into the solid matrix. Higher extraction yields, which are typically assessed through antiradical activity, are achieved due to this improved diffusion efficiency 39 , 40 . 3.3.TFC TFC values in elderberry extracts ranged from 14,497 to 20,521 mg CE/kg dm (Fig. 2 a). The highest TFC was observed in Sample 2 (2% β-CD, 30% liquid–solid, 90 min) and Sample 6 (2% β-CD, 10%, 90 min), while the lowest value was found in Sample 7 (1% β-CD, 30%, 60 min) (Table 1 ). RSM showed that a reduced quadratic model best explained the TFC response (Table 2 ). Of the model terms, only the quadratic effect of β-CD concentration (A²) was statistically significant (p = 0.0135), indicating a distinct curvature and an optimal point near approximately 2% β-CD. The effects of the liquid–solid ratio and extraction time (both main and interaction effects) were not significant; therefore, the impact of time within 30–90 minutes was limited, and the influence of the liquid–solid ratio appeared secondary to β-CD concentration. In general, reviews of flavonoid/phenolic extraction emphasise the pivotal role of the liquid-to-solid ratio and extraction time. Deviating from the optimal range can result in diminished yields due to dilution at high liquid-to-solid ratios or mass-transfer limitations at low liquid-to-solid ratios (along with thermal and degradative losses with extended duration), even within green-solvent systems 41 – 43 . The contour plot (Fig. 3c) shows that TFC approaches a maximum at ~ 2% β-CD and moderate liquid–solid ratios (a broad plateau roughly across 50–70%), while the response slightly declines at very low (30%) or very high (90%) ratios. Our experimental maximum (Sample 2) lies within this “~2% β-CD–low/moderate liquid–solid” region, consistent with the model’s trend. Increasing β-CD concentration up to a moderate range and employing moderate liquid–solid (L/S) ratios have been reported across multiple matrices to enhance TFC yield 29 , 30 , 44 , 45 . This pattern has also been reported in comparative β-CD–assisted extraction studies: increasing β-CD to an optimal window and employing moderate liquid–solid ratios enhance TFC yield, whereas excessive β-CD may reduce free water and limit mass transfer 37 . 3.4.TMAC The TMAC of elderberry extracts ranged from 7.399 to 9.701 mg C3G/kg dm (Fig. 2 b). The highest TMAC was recorded for Sample 11 (3% β-CD, 10% liquid-to-solid ratio, 60 min), followed by Sample 6 (2% β-CD, 10%, 90 min). The lowest TMAC was observed in Sample 5 (1% β-CD, 20%, 30 min). In general, higher anthocyanin levels were associated with a low liquid-to-solid ratio (10%) and medium-to-high β-CD concentrations (2–3%), consistent with solid–liquid extraction behavior in which moving outside the optimal L/S window depresses yields via dilution at high L/S or mass-transfer limits at very low L/S 46 , 47 . RSM analysis (Table 2 ) indicated that TMAC was best described by a linear model with limited predictive power; linear and interaction terms were not statistically significant (p > 0.05). Unexplained variance may arise from anthocyanin–protein interactions and degradation during processing/storage 48 . The contour plot (Fig. 3d) shows the combined effect of β-CD concentration and liquid-to-solid ratio on TMAC: TMAC increases as β-CD rises from 1–3%, particularly at 10% L/S, whereas at higher L/S (20–30%) values are generally lower—again consistent with dilution and reduced β-CD–anthocyanin encounter frequency 49 . The enhancement at moderate-to-high β-CD aligns with studies showing that β-CD (or HP-β-CD) forms inclusion complexes with cyanidin-type anthocyanins, improving solubility and protecting against thermal/oxidative or gastrointestinal degradation 12 , 50 . Xie et al. (2019) reported that β-cyclodextrin (β-CD) achieved higher anthocyanin extraction yields than water or aqueous ethanol across all tested mulberry cultivars; molecular docking further showed that cyanidin-3-O-glucoside is encapsulated within the β-CD cavity, thereby enhancing anthocyanin solubility. 3.5.TPAC The TPAC values obtained in this study ranged from 17.98 to 29.72 mg C3G/kg dm (Fig. 2 c). The highest TPAC was recorded in Sample 14 (3% β-CD, 10% liquid–solid ratio, 60 min), followed by Sample 2 (1% β-CD, 10%, 60 min); the lowest was observed in Sample 1 (1% β-CD, 30%, 30 min). In general, higher values were associated with a low liquid–solid ratio (10%) and medium-to-high β-CD levels (2–3%), consistent with recent reports that stepping outside an optimal L/S window depresses yields via dilution at high L/S or mass-transfer limitations at very low L/S 51 , 52 . RSM (Table 2 ) indicated that a linear model provided the best fit for TPAC, in line with measurable but essentially linear effects of β-CD concentration and liquid–solid ratio, while curvature terms were not significant. Similar β-CD assisted extractions have identified β-CD level as a primary driver and limited curvature depending on matrix 18 , 53 . Unexplained variance may arise from anthocyanin/proanthocyanidin–protein interactions and degradation pathways during processing or storage 48 , 52 . The contour plot (Fig. 3e) shows that increasing β-CD raises TPAC at low L/S (10–15%), whereas the benefit weakens above ~ 25% L/S; similar L/S-driven optima and off-optimum declines have been reported in recent RSM studies on anthocyanins 54 . Mechanistically, proanthocyanidins belong to the less-polar fraction of phenolics and interact favorably with the hydrophobic cavity of β-cyclodextrin (β-CD). Formation of β-CD inclusion complexes increases the apparent solubility and dispersibility of PAs, facilitates mass transfer, and can mitigate degradation, thereby improving recovery 33 , 55 , 56 . Consistent with this mechanism, β-/HP-β-CD–assisted extractions from grape- and vine-derived matrices routinely deliver higher polyphenol recoveries—including flavan-3-ols—than conventional solvent systems, with additional gains when β-CD level and process variables are optimized by RSM 45 . Specifically for proanthocyanidins, β-CD has been shown to enhance solubility and extraction yield, and high recoveries have been reported from sources such as grape seeds and cocoa shells 13 . Table 2. Results of RSM analysis of extraction conditions Variables TPC DPPH FRAP CUPRAC TFC TMAC TPAC Regression coefficient P Regression coefficient P Regression coefficient P Regression coefficient P Regression coefficient P Regression coefficient P Regression coefficient P C - - - - - - - - - - -33.13 0.8844 2.33 0.0360 AB 1.15 0.2532 - - - - - - - - 0.44 0.1116 0.52 0.1247 AC -1.65 0.1223 - - - - - - -786.44 0.2331 -0.28 0.2778 -0.36 0.2655 BC -1.24 0.2207 - - - - - - - - -0.07 0.7550 -0.05 0.8741 A 2 -2.31 0.0551 -9.96 0.0164 - - - - -1951.91 0.0135 0.36 0.1905 0.40 0.2313 B 2 -2.36 0.0516 7.29 0.0565 - - - - - - -0.03 0.8890 -0.22 0.4976 C 2 1.13 0.2769 6.21 0.0955 - - - - - - -0.29 0.2690 -0.32 0.3231 Model Quadratic Reduced Quadratic - - Reduced Quadratic Linear Linear Lack of fit 0.0988 0.8252 - - 0.1811 0.4391 0.6129 R 2 0.9330 0.8418 - - 0.6370 0.2375 0.3620 Adj R 2 0.8124 0.7232 - - 0.4354 0.0295 0.1944 Pred R 2 -0.0100 0.4894 - - -0.2299 -0.5011 -0.1734 A: β-cyclodextrin concentration (%), B: liquid-solid ratio (% w/v), C: extraction time (min) 3.6.Pearson correlation coefficient analysis The results of the correlation analysis show a clear link between phenolic compounds and antioxidant capacity at different levels (Table 3 ). As presented in Table 3 , the determined analytes exhibited linearity, with correlation coefficient (r) values well above 0.301. A positive and strong relationship was demonstrated by TPC and DPPH radical scavenging capacity (r = 0.618, p < 0.01). DPPH also showed a significant positive correlation with CUPRAC (r = 0.301, p < 0.05). Antioxidant capacity measured by the FRAP method was found to be significantly correlated with TPAC. The r value was 0.665 and the p value was < 0.01. The same was true of the correlation with CUPRAC. Here the r value was 0.465 and the p value was < 0.01. Despite the interrelatedness and occasional correlation of TPC, antioxidant activity and reducing power 57 , 58 , their optimisation demanded discrete conditions, signifying their independence. The differences can be attributed to the properties of polyphenols themselves 59 , 60 and the impact of the extraction process and coexisting compounds 61 . A significant positive correlation was also exhibited by CUPRAC with TFC (r = 0.325, p < 0.05). In addition, a strong and significant relationship was identified between TFC and TMA levels (r = 0.553, p < 0.01). A strong positive relationship was also identified between TPAC and CUPRAC (r = 0.467, p < 0.01). These results support the decisive role of polyphenolic compounds in the antioxidant properties of the samples, which is an important finding given the potential health benefits. Table 3 Pearson correlation coefficient among TPC, DPPH, FRAP, CUPRAC, TFC, TMAC and TPAC assays of the elderberry extracts TPC DPPH FRAP CUPRAC TFC TMAC TPAC TPC 1 DPPH 0.618** 1 FRAP - - 1 CUPRAC - 0.301* 0.465** 1 TFC - - - 0.325* 1 TMA - - - - 0.553** 1 TPAC - - 0.665** 0.467** - - 1 **Correlation is significant at the 0.01 level *Correlation is significant at the 0.05 level 4. Conclusions The application of β-CD-assisted extraction in the analysis of phenolic subclasses from elderberry exhibited a high degree of efficiency, with the recovery process demonstrating a notable degree of specificity for individual compounds. TPC increased under moderate β-CD conditions, while TFC exhibited a clear optimum at approximately 2% β-CD; extraction time (30–90 min) was secondary. Optimising the concentration of β-CD and the liquid-to-solid ratio can significantly improve the extraction efficiency of phenolic compounds from elderberries. It was found that an optimal β-CD level — particularly around 2% — combined with a moderate liquid-to-solid ratio, was critical for maximising TPC, flavonoid yield and antioxidant activity. Conversely, excessive β-CD concentrations were shown to hinder mass transfer, thereby reducing extraction performance. Antioxidant responses exhibited significant correlation with assay chemistry, manifesting as stronger trends for DPPH than for FRAP/CUPRAC. The models of TMAC and TPAC demonstrated a linear relationship across the range of the experiment; therefore, their application is more suited to providing directional insight than to precise prediction. Synthesizing the model-adequacy assessment: TPC (quadratic), TFC (reduced quadratic, optimum ~ 2% β-CD), and trend-based interpretations for the remaining responses converge on a practical window of ~ 2% β-CD, low-to-moderate L/S (≈ 10–25%), and 60–90 min to balance yields while avoiding dilution and solubility limits. Overall, these findings suggest that β-CD-assisted extraction is an effective, potentially superior alternative to conventional methods for obtaining stable, antioxidant-rich plant extracts. It is recommended that future work should take the following three forms: firstly, the validation of β-CD–phenolic inclusion and complex stability; secondly, the expansion of factor space (temperature, pH, particle size, gentle pre-treatments) and the benchmarking of alternative CDs/green co-solvents; and thirdly, the performance of multi-response optimisation with external validation alongside stability and scale-up assessments (techno-economics/LCA) to support industrial implementation. Declarations Conflict of interest The authors declared no conflict of interest. Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Author Contribution V.E. and A.A. contributed to the conceptualization, methodology, validation, formal analysis, investigation, and visualization. V.E. and A.A. also wrote and reviewed the manuscript. All authors approved the final version of the manuscript. Data Availability Data will be made available on request. 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Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 12 Oct, 2025 Reviews received at journal 06 Oct, 2025 Reviewers agreed at journal 29 Sep, 2025 Reviews received at journal 11 Sep, 2025 Reviewers agreed at journal 04 Sep, 2025 Reviewers invited by journal 02 Sep, 2025 Editor assigned by journal 24 Aug, 2025 Submission checks completed at journal 24 Aug, 2025 First submitted to journal 21 Aug, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-7428774","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":511495357,"identity":"7ba8c42d-3458-4344-80f3-d685f87dc0af","order_by":0,"name":"Vildan EYİZ","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0klEQVRIiWNgGAWjYLCCDwwMCQw8YCYzEcrZGBgYZ6BoYSNCCzMPSVoM7jcfe2xTcS+Pv+fwswcMFdaJDfK9D/BrOcaWbpxzprhY4mybuQHDmfTEBjZ2A7xaJNt4zKRz2xISG84zmEkwth0GaiHgMrAWy38JifPPs3+TYPxHhBZ+NqAWxoaExA1ne4C2NBClJS3dsOdYQrHhmTNlEgnH0o3b2NLwa2FjPnzswY+ahDy5M+nbJD7UWMv2Mx/Dr4UBJR4SGIiISSLVjIJRMApGwYgGANdyPdzOGO3sAAAAAElFTkSuQmCC","orcid":"","institution":"Necmettin Erbakan University","correspondingAuthor":true,"prefix":"","firstName":"Vildan","middleName":"","lastName":"EYİZ","suffix":""},{"id":511495358,"identity":"aaf42597-5dee-4f64-962d-1101032ad988","order_by":1,"name":"Ayşenur ACAR","email":"","orcid":"","institution":"Selçuk University","correspondingAuthor":false,"prefix":"","firstName":"Ayşenur","middleName":"","lastName":"ACAR","suffix":""}],"badges":[],"createdAt":"2025-08-21 18:53:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7428774/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7428774/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90853560,"identity":"f8e792f5-d145-480d-a59e-269a75410a3f","added_by":"auto","created_at":"2025-09-09 03:54:39","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":219761,"visible":true,"origin":"","legend":"\u003cp\u003eResults of total phenolics content and antioxidant activity assays\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7428774/v1/bada2d6aad66769574367f57.png"},{"id":90853954,"identity":"5711ebb6-cdfa-461a-84e2-96aa018e2bec","added_by":"auto","created_at":"2025-09-09 04:02:39","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":208240,"visible":true,"origin":"","legend":"\u003cp\u003eResults of total flavonoids, monomeric anthocyanidins and proanthocyanidins content\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7428774/v1/60e3d9dace24c5a367758cb2.png"},{"id":90854562,"identity":"5ba57507-1b87-436e-a631-2f41e5cf8266","added_by":"auto","created_at":"2025-09-09 04:18:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":371182,"visible":true,"origin":"","legend":"\u003cp\u003eResults modeled by RSM analysis under specified extraction conditions\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7428774/v1/34a512559f94ad0ae17efc2d.png"},{"id":90854915,"identity":"5add0854-ff90-4f79-b1b5-a1ce7a5965d3","added_by":"auto","created_at":"2025-09-09 04:26:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1877865,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7428774/v1/354eb6e4-4b7e-407e-9a2e-d7dbc74ec0d9.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Green Extraction and Optimization of Bioactive Compounds from Elderberry (Sambucus nigra L.) by β-Cyclodextrin Assisted Extraction Short title: Optimized Green Extraction of Elderberry Bioactives with β-Cyclodextrin","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003ePhenolic compounds, a class of secondary metabolites including pigments, have long garnered significant attention due to their diverse functional properties. These compounds play a crucial role in determining the organoleptic characteristics\u0026mdash;such as taste, color, and aroma\u0026mdash;of plant-based foods and are abundantly present in fruits and vegetables. In particular, phenolics are largely responsible for the characteristic coloration observed in many fruit and vegetable species, contributing both to their visual appeal and potential health benefits \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eElderberry (\u003cem\u003eSambucus nigra\u003c/em\u003e L.) is a rich source of bioactive compounds, particularly polyphenols, and is well known for its strong antioxidant capacity \u003csup\u003e\u003cspan additionalcitationids=\"CR3 CR4\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Among its notable constituents are anthocyanins, which contribute not only to its health-promoting properties but also to its potential use as a natural colorant. Elderberry powders and extracts have been successfully incorporated into various food products for coloring purposes \u003csup\u003e\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Owing to these functional and technological attributes, elderberry is increasingly utilized as a functional ingredient and natural additive in the development of health-oriented food products \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e(MARTİS).\u003c/p\u003e\u003cp\u003eβ-CDs are cyclic oligosaccharides produced through the enzymatic degradation of starch. They consist of six, seven, or eight D-(+)-glucopyranose units linked by α-1,4 glycosidic bonds \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Among natural cyclodextrins, β-CD is the most accessible, cost-effective, and widely utilized form \u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Recognized as Generally Regarded As Safe (GRAS), β-CD is extensively used in the food industry due to its ability to form inclusion complexes with various bioactive compounds, thereby enhancing their stability, solubility, and bioavailability \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. When employed as an extraction medium in aqueous systems, β-CD facilitates the effective recovery of bioactive compounds from plant matrices by interacting with target molecules \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Previous studies have demonstrated the successful application of β-CD in improving the extraction efficiency of bioactives from various plant sources, including red beetroot \u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e, olive and red grape pomace \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e, whole pomegranate fruit \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e, and peach pomace \u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eIn this study, phytochemicals were extracted from elderberry using β-CD as a green solvent, and the bioactive properties of the resulting extracts were evaluated. The effects of key extraction parameters\u0026mdash;namely β-CD concentration, liquid-to-solid ratio, and extraction time\u0026mdash;on the recovery of phenolics, antioxidants, flavonoids, anthocyanidins, and proanthocyanidins were systematically examined using a Central Composite Design (CCD). This experimental approach enabled the identification of optimal extraction conditions for maximizing the yield of bioactive compounds from elderberry.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1.Materials\u003c/h2\u003e\u003cp\u003eApproximately 3 kg of elderberries (\u003cem\u003eSambucus nigra\u003c/em\u003e L.) were procured from a local market in Konya, Turkey. The fruits were thoroughly washed and cleaned to remove any foreign materials. Drying was performed using a vacuum oven (Eksis Makine, Isparta, Turkey) at 50\u0026deg;C. The dried elderberries were then ground into a fine powder using a laboratory blender. The resulting elderberry powder was stored in sealed polyethylene bags at \u0026minus;\u0026thinsp;20\u0026deg;C until further use. β-CD was obtained from Sigma-Aldrich (St. Louis, MO, USA), while all other analytical-grade chemicals were purchased from Merck (Darmstadt, Germany).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2.Extraction\u003c/h2\u003e\u003cp\u003eExtraction was performed following Kim, Son, Pang, Yang, Lee, Lee, Lee, Park and Yoo \u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e, with modifications. Based on the experimental design, predetermined amounts of elderberry powder were accurately weighed into extraction vessels. Corresponding volumes of β-CD solutions were added to the samples according to the design parameters. The mixtures were homogenized at 10,000 rpm for 1 min using an Ultraturrax homogenizer (IKA T25 digital, Staufen im Breisgau, Germany). The samples were placed in a shaking water bath (N\u0026uuml;ve, NB-20, Ankara, T\u0026uuml;rkiye) (60\u0026deg;C, 70 rpm) for 1 h. Following homogenization, the samples were centrifuged at 10,000 \u0026times; g for 15 min (Hermle Z326K, Germany). The resulting supernatants were filtered (Saha, Turkey) to remove particulates and subsequently stored at +\u0026thinsp;4\u0026deg;C until further analysis.\u003c/p\u003e\u003cp\u003e\u003cb\u003e2.3.Experimental design\u003c/b\u003e\u003c/p\u003e\u003cp\u003eIn this study, a face-centered CCD was employed to evaluate the effects of three independent variables on the extraction efficiency of bioactive compounds from elderberries. The selected factors and their levels were as follows: β-CD concentration (1%, 2%, 3%) \u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e ,liquid-to-solid ratio (10%, 20%, 30%) \u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e, and extraction time (30, 60, and 90 min) \u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. As presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the design consisted of 15 experimental runs, including three replicates at the center point to estimate experimental error. 7 response variables were measured for each extract: TPC, antioxidant activity (DPPH, FRAP, and CUPRAC assays), TFC, TMAC, and TPAC.\u003c/p\u003e\u003cp\u003eMathematical models for each response were developed using least squares regression based on the experimental data. The statistical significance of model terms was evaluated at a 90% confidence level using p-values. Analysis of variance (ANOVA), lack-of-fit testing, and determination of coefficients (R\u0026sup2;) were performed using Design-Expert Version 10 software (Stat-Ease Inc., Minneapolis, USA). A desirability function approach was applied to determine the optimal extraction conditions by simultaneously maximizing all response variables. No additional experimental validation of the model was conducted.\u003c/p\u003e\u003cp\u003e.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eExperimental design and responses of elderberry extracts\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"12\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStd order\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRun order\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eβ-cyclodextrin concentration (%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLiquid-solid ratio\u003c/p\u003e\u003cp\u003e(%, w/v)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eExtraction time\u003c/p\u003e\u003cp\u003e(min)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eTPC\u003c/p\u003e\u003cp\u003e(g GAE/kg dm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eDPPH\u003c/p\u003e\u003cp\u003e(g TE/kg dm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eFRAP\u003c/p\u003e\u003cp\u003e(g TE/kg dm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eCUPRAC\u003c/p\u003e\u003cp\u003e(g TE/kg dm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eTFC\u003c/p\u003e\u003cp\u003e(mg CE/kg dm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u003cp\u003eTMAC\u003c/p\u003e\u003cp\u003e(mg C3G/kg dm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c12\"\u003e\u003cp\u003eTPAC\u003c/p\u003e\u003cp\u003e(g cyanidin/kg dm)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e51.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e50.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e48.79\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e18189\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e9225\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e17.17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e12\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e90\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e51.96\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e85.86\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e75.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e20521\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8652\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e26.20\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e15\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e60\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e47.59\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e74.81\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e71.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e59.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e19454\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8620\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e22.67\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e60\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e46.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e85.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e66.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e68.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e16773\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e7800\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e23.98\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e44.59\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e83.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e80.80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e54.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e16197\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e7399\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e23.26\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e11\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e90\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e44.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e84.76\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e75.10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e66.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e20009\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8732\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e25.75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e7\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e60\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e45.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e78.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e100.19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e65.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e14497\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e7897\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e23.21\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e9\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e8\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e38.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e78.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e97,30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e72.48\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e19898\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8359\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e24.97\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e13\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e9\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e60\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e46.97\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e68.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e90.72\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e64.85\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e18418\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e9430\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e24.89\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e30\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e46.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e73.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e67.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e64.51\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e19380\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8890\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e19.75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e11\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e60\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e37.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e54.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e82.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e55.58\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e18862\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e9701\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e24.32\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e7\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e12\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e90\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e49.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e74.27\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e84.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e50.95\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e17048\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8042\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e25.40\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e13\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e60\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e43.30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e76.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e73.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e47.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e18656\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e9058\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e20.72\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e14\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e14\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e60\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e48.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e85.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e98.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e63.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e18362\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8480\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e27.58\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e8\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e15\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e90\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e45.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e59.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e65.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e49.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e17085\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e8182\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e\u003cp\u003e24.04\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.4.Extraction analyses\u003c/h2\u003e\u003cdiv id=\"Sec6\" class=\"Section3\"\u003e\u003ch2\u003e2.4.1.Determination of TPC of extracts\u003c/h2\u003e\u003cp\u003eThe TPC of the elderberry extracts was determined using the Folin\u0026ndash;Ciocalteu colorimetric method, as described by Georgiev, Weber, Kneschke, Denev, Bley and Pavlov \u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e, with slight modifications. Briefly, each extract was appropriately diluted with distilled water. Then, 0.2 mL of the diluted sample was mixed with 2.0 mL of 0.2 N Folin\u0026ndash;Ciocalteu reagent, followed by the addition of 0.8 mL sodium carbonate solution (75 g/L, Na₂CO₃). The mixture was incubated for 30 min at room temperature in the dark. After incubation, the absorbance was measured at 765 nm using a UV-Vis spectrophotometer (Biochrom Libra S22, Cambridge, UK). Results were expressed as grams of gallic acid equivalents per kilogram of dry matter (g GAE/kg dm).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section3\"\u003e\u003ch2\u003e2.4.2.Determination of antiradical activity of extracts\u003c/h2\u003e\u003cp\u003eThe antioxidant capacity of elderberry extracts was evaluated using three widely applied assays: 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Power (FRAP), and Cupric Ion Reducing Antioxidant Capacity (CUPRAC). The DPPH radical scavenging activity was determined with slight modifications based on the method of Tontul and Topuz \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. Initially, the elderberry extracts were diluted with distilled water. Then, 950 \u0026micro;L of freshly prepared DPPH solution (60 \u0026micro;M in methanol) was added to 50 \u0026micro;L of the diluted extract. The mixture was incubated for 30 min at room temperature in the dark. Absorbance was measured at 516 nm using a UV\u0026ndash;Vis spectrophotometer. The FRAP assay was carried out according to the method described by Granato, Karnopp and van Ruth \u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003eGranato et al. (2015), with minor modifications. The FRAP reagent was freshly prepared by mixing 300 mM acetate buffer (pH 3.6), 10 mM TPTZ solution in 40 mM HCl, and 20 mM ferric chloride hexahydrate in a 10:1:1 (v/v/v) ratio. After pre-incubation of the FRAP reagent at 37\u0026deg;C for 10 min, 100 \u0026micro;L of each extract was added. The mixture was incubated in the dark at 37\u0026deg;C for 30 min, and absorbance was measured at 593 nm. CUPRAC analysis was performed following the procedure described by Apak, G\u0026uuml;\u0026ccedil;l\u0026uuml;, \u0026Ouml;zy\u0026uuml;rek and Karademir \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. The reaction mixture was prepared by combining equal volumes (1:1:1, v/v/v) of 7.5 mM neocuproine, 10 mM copper(II) chloride (CuCl₂), and 1 M ammonium acetate buffer (pH 7.0). To this mixture, 0.5 mL of the extract and 0.6 mL of distilled water were added. The final mixture was incubated at room temperature for 30 min in the dark. Absorbance was recorded at 450 nm. A blank was prepared by replacing the sample volume with an equivalent volume of the reaction mixture. All antioxidant activity results were expressed as grams of Trolox equivalents per kilogram of dry matter (g TE/kg dm).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\u003ch2\u003e2.4.3.Determination of TFC of extracts\u003c/h2\u003e\u003cp\u003eThe TFC of the elderberry extracts was determined using the colorimetric method described by Chang, Lin, Chang and Liu \u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e, with slight modifications. Briefly, 0.5 mL of the extract was mixed with 2.5 mL of distilled water and 0.15 mL of 5% sodium nitrite (NaNO₂) solution. The mixture was allowed to stand for 5 min at room temperature. Subsequently, 0.3 mL of 10% aluminum chloride (AlCl₃) solution was added, and the reaction was allowed to proceed for an additional 5 min. Then, 1.0 mL of 1 M sodium hydroxide (NaOH) was added to the mixture, which was then vortexed vigorously. The absorbance was measured at 510 nm against a distilled water blank using a UV\u0026ndash;Vis spectrophotometer. TFC was expressed as grams of catechin equivalents per kilogram of dry matter (g CE/kg dm).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\u003ch2\u003e2.4.4.Determination of Total Monomeric Anthocyanin Content of extracts\u003c/h2\u003e\u003cp\u003eThe TMAC of the elderberry extracts was determined using the pH differential method, as described by \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e, with slight modifications. The extracts were appropriately diluted with buffer solutions at pH 1.0 and pH 4.5. The diluted samples were allowed to equilibrate in the dark for 20 min at room temperature. Absorbance readings were then taken at 510 nm (λmax) and 700 nm against a distilled water blank using a UV\u0026ndash;Vis spectrophotometer. The anthocyanin content was calculated and expressed as grams of cyanidin-3-glucoside equivalents per kilogram of dry matter (g C3G/kg dm).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\u003ch2\u003e2.4.5.Determination of Total Proanthocyanin Content of extracts\u003c/h2\u003e\u003cp\u003eThe TPAC of the elderberry extracts was determined following the method described by Eyiz, Tontul and Turker \u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e, with slight modifications. A solution of FeSO₄\u0026middot;7H₂O (154 ppm) was prepared in a mixture of hydrochloric acid and \u003cem\u003en\u003c/em\u003e-butanol (2:3, v/v). In a stoppered glass tube, 2.5 mL of this reagent was added to 0.25 mL of the extract. The mixture was then incubated in a water bath at 95\u0026deg;C for 15 min. After incubation, the absorbance was measured at 540 nm using a UV\u0026ndash;Vis spectrophotometer. Results were expressed as grams of proanthocyanidins per kilogram of dry matter (g TPAC/kg dm).\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e2.5. Statistical analysis\u003c/h2\u003e\u003cp\u003eAll data were expressed as mean values\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviations based on triplicate measurements. Statistical analyses were performed using one-way analysis of variance (ANOVA) to determine significant differences among experimental groups, and Pearson correlation analysis was applied to assess relationships between the measured variables. ANOVA results were considered statistically significant at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05, while Pearson correlation coefficients were evaluated at significance levels of \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 and \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01.\u003c/p\u003e\u003cp\u003eIn addition, Response Surface Methodology (RSM) based on a face-centered CCD was employed to model and optimize the effects of independent variables\u0026mdash;β-CD concentration, liquid-to-solid ratio, and extraction time\u0026mdash;on the recovery of bioactive compounds. Design-Expert version 10 software (Stat-Ease Inc., Minneapolis, USA) was used for regression analysis, analysis of variance (ANOVA), lack-of-fit testing, and optimization through desirability function.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results and discussions","content":"\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e3.1.TPC\u003c/h2\u003e\u003cp\u003eExtracting natural bioactives like polyphenols is often a time-consuming process that uses organic solvents. This can be bad for the environment and limit how useful they are in practice. Furthermore, harsh extraction conditions can result in the degradation of the compound, thereby reducing its bioavailability \u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. TPC values of elderberry extracts ranged from 37.00 to 51.96 g GAE/kg dm, with an average of 45.78 g GAE/kg dm and a coefficient of variation of 8.97% (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Furthermore, TPC values of elderberry extract samples are illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea. 2% β-CD, 30% liquid-to-solid ratio, 90 min had the highest TPC, while 3% β-CD, 10% liquid-to-solid ratio, 60 min had the lowest (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea). According to the RSM analysis (see Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), the TPC response was best described by a quadratic model (R\u0026sup2; = 0.9330; Adj R\u0026sup2; = 0.8124). At a fixed liquid-to-solid ratio of 20 and an extraction time of 30 min, increasing the concentration of β-CD did not notably enhance TPC. Similar trends were observed in phenolic yield studies with mint \u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. Both β-CD concentration (A\u0026sup2;) and the liquid-to-solid ratio (B\u0026sup2;) exhibited significant negative quadratic effects (p\u0026thinsp;\u0026asymp;\u0026thinsp;0.05). Figure\u0026nbsp;3a clearly illustrates the combined influence of β-CD concentration and liquid-to-solid ratio on TPC. The orange\u0026ndash;yellow regions indicate the highest TPC values (~\u0026thinsp;48\u0026ndash;50 g GAE/kg dm), corresponding to a moderate β-CD concentration (2%) and a medium\u0026ndash;high liquid-to-solid ratio (20\u0026ndash;25%). The green and blue zones represent lower TPC levels, with the minimum occurring at a high β-CD concentration (3%) and a low liquid-to-solid ratio (10%).\u003c/p\u003e\u003cp\u003ePhenolic compounds are generally highly soluble in water due to their ability to form hydrogen bonds. However, the relatively low water solubility of β-CD \u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e and the presence of a secondary hydrogen-bonding network within its structure that limits interactions with water molecules \u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e can reduce mass transfer at high β-CD levels. Additionally, β-CD presents a hydrophobic inner surface. Therefore, it is better for compounds with high polarity instead \u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e. Consequently, as the concentration of β-CD increases, the extraction yield of water-soluble phenolic compounds may decrease.\u003c/p\u003e\u003cp\u003eIn this study, the liquid-to-solid ratio also negatively affected TPC, suggesting that exceeding the optimum level could cause dilution of phenolic compounds in the solvent phase and reduce extraction efficiency. In contrast, studies on whole pomegranates have reported increases in TPC at higher β-CD levels \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. Such discrepancies are likely due to differences in the plant matrix, extraction methods and liquid-to-solid ratios applied.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e3.2.Antioxidant activity analyses (DPPH, FRAP, CUPRAC)\u003c/h2\u003e\u003cp\u003eChanges in DPPH, FRAP, and CUPRAC antioxidant activities of elderberry extract samples are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed. The results showed that antioxidant activities, measured by the DPPH, FRAP, and CUPRAC methods, ranged from 54.16 to 98.04, 50.89 to 100.19, and 47.62 to 75.24 g TE/kg dm, respectively. The highest DPPH activity was observed in the sample, extracted using 2% (w/v) β-CD concentration, 30% liquid-solid ratio, and 90 min extraction time. This same sample exhibited the highest antioxidant activity according to CUPRAC assays (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). On the other hand, the maximum FRAP activities were detected in the sample, prepared with 3% (w/v) β-CD concentration, 30% liquid-solid ratio, and 60 min extraction time.\u003c/p\u003e\u003cp\u003eAccording to two-factorial ANOVA, the interaction between β-CD concentration and liquid-solid ratio significantly influenced the antioxidant activities, and Reduced Quadratic was identified as the most suitable. Although the TPC yield was best described by a quadratic model, the DPPH radical scavenging activity was more accurately represented by a reduced quadratic model. A positive correlation between TPC and DPPH values is commonly observed in most plant extracts. Nevertheless, deviations from this pattern have been reported, often attributed to synergistic or antagonistic interactions among the bioactive constituents within the extracts \u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e. However, it is important to note that the FRAP and CUPRAC responses could not be modeled within this experimental design (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Moreover, the liquid-solid ratio exerted an increased decrease effect on antioxidant activities (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed). Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows the relationship between temperature and the decrease in DPPH value when the \u0026szlig;-Cd concentration (1%) and the liquid-solid ratio (20) were kept constant. With regard to the extraction process with β-CD using DES solvent, an initial rise in antiradical activity up to 55\u0026deg;C is observed, which corresponds to an enhancement in binding affinity. This is followed by a subsequent decrease up to 80\u0026deg;C \u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e. The observed decline in antioxidant activities with increasing β-CD concentration likely results from the concurrent decrease in TPC extraction, indicating a limitation of higher β-CD concentrations in maximizing antioxidant recovery. Contrary to our findings, previous studies have reported significantly higher antiradical activities in samples extracted with β-CD from red beet and peach pomace compared to those extracted with ethanol \u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. This discrepancy may be attributed to variations in extraction temperature, time, and method, as these parameters critically affect the efficiency of compound release, solubility, and stability during extraction.\u003c/p\u003e\u003cp\u003eIncreases were observed in both TPC and DPPH values with the increase in the liquid-solid ratio (Fig.\u0026nbsp;3a, 3b). The process of extracting bioactive compounds with antioxidant properties, like phenolics and flavonoids, is mainly driven by the difference in concentrations between the solvent and the solid matrix \u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e. The gradient is enhanced by an increase in the liquid-to-solid ratio, which facilitates mass transfer by promoting the diffusion of the solvent into the solid matrix. Higher extraction yields, which are typically assessed through antiradical activity, are achieved due to this improved diffusion efficiency \u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003e3.3.TFC\u003c/h2\u003e\u003cp\u003eTFC values in elderberry extracts ranged from 14,497 to 20,521 mg CE/kg dm (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea). The highest TFC was observed in Sample 2 (2% β-CD, 30% liquid\u0026ndash;solid, 90 min) and Sample 6 (2% β-CD, 10%, 90 min), while the lowest value was found in Sample 7 (1% β-CD, 30%, 60 min) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eRSM showed that a reduced quadratic model best explained the TFC response (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Of the model terms, only the quadratic effect of β-CD concentration (A\u0026sup2;) was statistically significant (p\u0026thinsp;=\u0026thinsp;0.0135), indicating a distinct curvature and an optimal point near approximately 2% β-CD. The effects of the liquid\u0026ndash;solid ratio and extraction time (both main and interaction effects) were not significant; therefore, the impact of time within 30\u0026ndash;90 minutes was limited, and the influence of the liquid\u0026ndash;solid ratio appeared secondary to β-CD concentration.\u003c/p\u003e\u003cp\u003eIn general, reviews of flavonoid/phenolic extraction emphasise the pivotal role of the liquid-to-solid ratio and extraction time. Deviating from the optimal range can result in diminished yields due to dilution at high liquid-to-solid ratios or mass-transfer limitations at low liquid-to-solid ratios (along with thermal and degradative losses with extended duration), even within green-solvent systems \u003csup\u003e\u003cspan additionalcitationids=\"CR42\" citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe contour plot (Fig.\u0026nbsp;3c) shows that TFC approaches a maximum at ~\u0026thinsp;2% β-CD and moderate liquid\u0026ndash;solid ratios (a broad plateau roughly across 50\u0026ndash;70%), while the response slightly declines at very low (30%) or very high (90%) ratios. Our experimental maximum (Sample 2) lies within this \u0026ldquo;~2% β-CD\u0026ndash;low/moderate liquid\u0026ndash;solid\u0026rdquo; region, consistent with the model\u0026rsquo;s trend. Increasing β-CD concentration up to a moderate range and employing moderate liquid\u0026ndash;solid (L/S) ratios have been reported across multiple matrices to enhance TFC yield \u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThis pattern has also been reported in comparative β-CD\u0026ndash;assisted extraction studies: increasing β-CD to an optimal window and employing moderate liquid\u0026ndash;solid ratios enhance TFC yield, whereas excessive β-CD may reduce free water and limit mass transfer \u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003e3.4.TMAC\u003c/h2\u003e\u003cp\u003eThe TMAC of elderberry extracts ranged from 7.399 to 9.701 mg C3G/kg dm (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb). The highest TMAC was recorded for Sample 11 (3% β-CD, 10% liquid-to-solid ratio, 60 min), followed by Sample 6 (2% β-CD, 10%, 90 min). The lowest TMAC was observed in Sample 5 (1% β-CD, 20%, 30 min). In general, higher anthocyanin levels were associated with a low liquid-to-solid ratio (10%) and medium-to-high β-CD concentrations (2\u0026ndash;3%), consistent with solid\u0026ndash;liquid extraction behavior in which moving outside the optimal L/S window depresses yields via dilution at high L/S or mass-transfer limits at very low L/S \u003csup\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eRSM analysis (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) indicated that TMAC was best described by a linear model with limited predictive power; linear and interaction terms were not statistically significant (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Unexplained variance may arise from anthocyanin\u0026ndash;protein interactions and degradation during processing/storage \u003csup\u003e\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe contour plot (Fig.\u0026nbsp;3d) shows the combined effect of β-CD concentration and liquid-to-solid ratio on TMAC: TMAC increases as β-CD rises from 1\u0026ndash;3%, particularly at 10% L/S, whereas at higher L/S (20\u0026ndash;30%) values are generally lower\u0026mdash;again consistent with dilution and reduced β-CD\u0026ndash;anthocyanin encounter frequency \u003csup\u003e\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe enhancement at moderate-to-high β-CD aligns with studies showing that β-CD (or HP-β-CD) forms inclusion complexes with cyanidin-type anthocyanins, improving solubility and protecting against thermal/oxidative or gastrointestinal degradation \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e. Xie et al. (2019) reported that β-cyclodextrin (β-CD) achieved higher anthocyanin extraction yields than water or aqueous ethanol across all tested mulberry cultivars; molecular docking further showed that cyanidin-3-O-glucoside is encapsulated within the β-CD cavity, thereby enhancing anthocyanin solubility.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003e3.5.TPAC\u003c/h2\u003e\u003cp\u003eThe TPAC values obtained in this study ranged from 17.98 to 29.72 mg C3G/kg dm (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec). The highest TPAC was recorded in Sample 14 (3% β-CD, 10% liquid\u0026ndash;solid ratio, 60 min), followed by Sample 2 (1% β-CD, 10%, 60 min); the lowest was observed in Sample 1 (1% β-CD, 30%, 30 min). In general, higher values were associated with a low liquid\u0026ndash;solid ratio (10%) and medium-to-high β-CD levels (2\u0026ndash;3%), consistent with recent reports that stepping outside an optimal L/S window depresses yields via dilution at high L/S or mass-transfer limitations at very low L/S \u003csup\u003e\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eRSM (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) indicated that a linear model provided the best fit for TPAC, in line with measurable but essentially linear effects of β-CD concentration and liquid\u0026ndash;solid ratio, while curvature terms were not significant. Similar β-CD assisted extractions have identified β-CD level as a primary driver and limited curvature depending on matrix \u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. Unexplained variance may arise from anthocyanin/proanthocyanidin\u0026ndash;protein interactions and degradation pathways during processing or storage \u003csup\u003e\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe contour plot (Fig.\u0026nbsp;3e) shows that increasing β-CD raises TPAC at low L/S (10\u0026ndash;15%), whereas the benefit weakens above ~\u0026thinsp;25% L/S; similar L/S-driven optima and off-optimum declines have been reported in recent RSM studies on anthocyanins \u003csup\u003e\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eMechanistically, proanthocyanidins belong to the less-polar fraction of phenolics and interact favorably with the hydrophobic cavity of β-cyclodextrin (β-CD). Formation of β-CD inclusion complexes increases the apparent solubility and dispersibility of PAs, facilitates mass transfer, and can mitigate degradation, thereby improving recovery \u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e\u003c/sup\u003e. Consistent with this mechanism, β-/HP-β-CD\u0026ndash;assisted extractions from grape- and vine-derived matrices routinely deliver higher polyphenol recoveries\u0026mdash;including flavan-3-ols\u0026mdash;than conventional solvent systems, with additional gains when β-CD level and process variables are optimized by RSM \u003csup\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e. Specifically for proanthocyanidins, β-CD has been shown to enhance solubility and extraction yield, and high recoveries have been reported from sources such as grape seeds and cocoa shells \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eTable 2.\u0026nbsp;\u003c/strong\u003eResults of RSM analysis of extraction conditions\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\" class=\"fr-table-selection-hover\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVariables\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 13px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTPC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDPPH\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFRAP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCUPRAC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTFC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTMAC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTPAC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegression\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ecoefficient\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegression\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ecoefficient\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegression\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ecoefficient\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegression\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ecoefficient\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegression\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ecoefficient\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegression\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ecoefficient\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegression\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ecoefficient\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-33.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.8844\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e2.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.0360\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2532\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.1116\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e0.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.1247\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-1.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.1223\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-786.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2331\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2778\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2655\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-1.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.7550\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.8741\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003csup\u003e2\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-2.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.0551\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-9.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e0.0164\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-1951.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.0135\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.1905\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e0.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2313\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003csup\u003e2\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-2.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.0516\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e7.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e0.0565\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.8890\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.4976\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003csup\u003e2\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e1.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2769\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e6.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e0.0955\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.2690\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e-0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e0.3231\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eModel\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 13px;\"\u003e\n \u003cp\u003eQuadratic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eReduced Quadratic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003eReduced Quadratic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003eLinear\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003eLinear\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLack of fit\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.0988\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.8252\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.1811\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.4391\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.6129\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eR\u003csup\u003e2\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.9330\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.8418\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.6370\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.2375\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.3620\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAdj R\u003csup\u003e2\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 13px;\"\u003e\n \u003cp\u003e0.8124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.7232\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.4354\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.0295\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.1944\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePred R\u003csup\u003e2\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 13px;\"\u003e\n \u003cp\u003e-0.0100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.4894\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 14px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-0.2299\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-0.5011\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 12px;\"\u003e\n \u003cp\u003e-0.1734\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eA: \u0026beta;-cyclodextrin concentration (%), B: liquid-solid ratio (% w/v), C: extraction time (min)\u003c/p\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003e3.6.Pearson correlation coefficient analysis\u003c/h2\u003e\u003cp\u003eThe results of the correlation analysis show a clear link between phenolic compounds and antioxidant capacity at different levels (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). As presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, the determined analytes exhibited linearity, with correlation coefficient (r) values well above 0.301. A positive and strong relationship was demonstrated by TPC and DPPH radical scavenging capacity (r\u0026thinsp;=\u0026thinsp;0.618, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). DPPH also showed a significant positive correlation with CUPRAC (r\u0026thinsp;=\u0026thinsp;0.301, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Antioxidant capacity measured by the FRAP method was found to be significantly correlated with TPAC. The r value was 0.665 and the p value was \u0026lt;\u0026thinsp;0.01. The same was true of the correlation with CUPRAC. Here the r value was 0.465 and the p value was \u0026lt;\u0026thinsp;0.01. Despite the interrelatedness and occasional correlation of TPC, antioxidant activity and reducing power \u003csup\u003e\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e\u003c/sup\u003e, their optimisation demanded discrete conditions, signifying their independence. The differences can be attributed to the properties of polyphenols themselves \u003csup\u003e\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e\u003c/sup\u003e and the impact of the extraction process and coexisting compounds \u003csup\u003e\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e\u003c/sup\u003e. A significant positive correlation was also exhibited by CUPRAC with TFC (r\u0026thinsp;=\u0026thinsp;0.325, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). In addition, a strong and significant relationship was identified between TFC and TMA levels (r\u0026thinsp;=\u0026thinsp;0.553, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). A strong positive relationship was also identified between TPAC and CUPRAC (r\u0026thinsp;=\u0026thinsp;0.467, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). These results support the decisive role of polyphenolic compounds in the antioxidant properties of the samples, which is an important finding given the potential health benefits.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePearson correlation coefficient among TPC, DPPH, FRAP, CUPRAC, TFC, TMAC and TPAC assays of the elderberry extracts\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"8\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTPC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eDPPH\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFRAP\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eCUPRAC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eTFC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eTMAC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eTPAC\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTPC\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDPPH\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.618**\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eFRAP\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCUPRAC\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.301*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.465**\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTFC\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.325*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTMA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.553**\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTPAC\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.665**\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.467**\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"8\"\u003e**Correlation is significant at the 0.01 level\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"8\"\u003e*Correlation is significant at the 0.05 level\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"4. Conclusions","content":"\u003cp\u003eThe application of β-CD-assisted extraction in the analysis of phenolic subclasses from elderberry exhibited a high degree of efficiency, with the recovery process demonstrating a notable degree of specificity for individual compounds. TPC increased under moderate β-CD conditions, while TFC exhibited a clear optimum at approximately 2% β-CD; extraction time (30\u0026ndash;90 min) was secondary. Optimising the concentration of β-CD and the liquid-to-solid ratio can significantly improve the extraction efficiency of phenolic compounds from elderberries. It was found that an optimal β-CD level \u0026mdash; particularly around 2% \u0026mdash; combined with a moderate liquid-to-solid ratio, was critical for maximising TPC, flavonoid yield and antioxidant activity. Conversely, excessive β-CD concentrations were shown to hinder mass transfer, thereby reducing extraction performance. Antioxidant responses exhibited significant correlation with assay chemistry, manifesting as stronger trends for DPPH than for FRAP/CUPRAC. The models of TMAC and TPAC demonstrated a linear relationship across the range of the experiment; therefore, their application is more suited to providing directional insight than to precise prediction. Synthesizing the model-adequacy assessment: TPC (quadratic), TFC (reduced quadratic, optimum\u0026thinsp;~\u0026thinsp;2% β-CD), and trend-based interpretations for the remaining responses converge on a practical window of ~\u0026thinsp;2% β-CD, low-to-moderate L/S (\u0026asymp;\u0026thinsp;10\u0026ndash;25%), and 60\u0026ndash;90 min to balance yields while avoiding dilution and solubility limits. Overall, these findings suggest that β-CD-assisted extraction is an effective, potentially superior alternative to conventional methods for obtaining stable, antioxidant-rich plant extracts. It is recommended that future work should take the following three forms: firstly, the validation of β-CD\u0026ndash;phenolic inclusion and complex stability; secondly, the expansion of factor space (temperature, pH, particle size, gentle pre-treatments) and the benchmarking of alternative CDs/green co-solvents; and thirdly, the performance of multi-response optimisation with external validation alongside stability and scale-up assessments (techno-economics/LCA) to support industrial implementation.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eConflict of interest\u003c/h2\u003e\u003cp\u003eThe authors declared no conflict of interest.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eV.E. and A.A. contributed to the conceptualization, methodology, validation, formal analysis, investigation, and visualization. V.E. and A.A. also wrote and reviewed the manuscript. All authors approved the final version of the manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eData will be made available on request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAlara OR, Abdurahman NH, Ukaegbu CI (2021) Curr Res Food Sci 4:200\u0026ndash;214\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eErturk Y, Ercisli S, Tosun M (2009)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEgea I, S\u0026aacute;nchez-Bel P, Romojaro F, Pretel MT (2010) Plant Foods Hum Nutr 65(2):121\u0026ndash;129\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRop O, Ercişli S, Mlcek J, Jurikova T, Hoza I (2014) Turk J Agric For 38(2):224\u0026ndash;232\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVeberic R, Slatnar A, Bizjak J, Stampar F, Mikulic-Petkovsek M (2015) LWT 60(1):509\u0026ndash;517\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNile SH, Park SW (2014) Nutr 30(2):134\u0026ndash;144\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBaeza R, S\u0026aacute;nchez V, Salierno G, Molinari F, L\u0026oacute;pez P, Chirife J (2021) Food Sci Technol Int 27(2):135\u0026ndash;144\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNajgebauer-Lejko D, Liszka K, Tabaszewska M, Domagała J (2021) Molecules 26(8):2345\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMarțiș GS, Mureșan V, Marc RM et al (2021) Antioxidants 10(7):1093\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBiwer A, Antranikian G, Heinzle E (2002) Appl Microbiol Biotechnol 59(6):609\u0026ndash;617\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDel Valle EM (2004) Process Biochem 39(9):1033\u0026ndash;1046\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFernandes A, Rocha MA, Santos LM et al (2018) Food Chem 245:426\u0026ndash;431\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCai R, Yuan Y, Cui L, Wang Z, Yue T (2018) Trends Food Sci Technol 79:19\u0026ndash;27\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTutunchi P, Roufegarinejad L, Hamishehkar H, Alizadeh A (2019) Food Chem 297:124994\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eL\u0026oacute;pez-Miranda S, Serrano-Mart\u0026iacute;nez A, Hern\u0026aacute;ndez-S\u0026aacute;nchez P et al (2016) Food Chem 203:379\u0026ndash;385\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlbahari P, Jug M, Radić K et al (2018) LWT 92, 22\u0026ndash;31\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDiamanti AC, Igoumenidis PE, Mourtzinos I, Yannakopoulou K, Karathanos VT (2017) Food Chem 214:61\u0026ndash;66\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDarra NE, Rajha HN, Debs E et al (2018) (1) Int. 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Process effects were mapped using response surface methodology (face-centered central composite design) with β-CD 1\u0026ndash;3% (w/v), liquid\u0026ndash;solid (L/S) ratio 10\u0026ndash;30% (w/v), and time 30\u0026ndash;90 min. Responses (TPC, TFC, TMAC, TPAC; DPPH, FRAP, CUPRAC) were modeled to identify operating windows. TPC followed a quadratic relationship; TFC a reduced quadratic with an optimum near ~\u0026thinsp;2% β-CD, whereas TMAC and TPAC exhibited approximately linear trends across the studied space. DPPH showed clearer parameter sensitivity than FRAP/CUPRAC, and extraction time exerted a secondary effect within 30\u0026ndash;90 min. Contour plots consistently indicated low-to-moderate L/S together with ~\u0026thinsp;2% β-CD as a region that balances multiple responses while avoiding dilution or mass-transfer limitations. Overall, β-CD\u0026ndash;assisted extraction provides an effective, solvent-lean route for recovering elderberry phenolics and delineates a practical window for greener processing. These results motivate follow-up work on inclusion mechanisms, stability during storage and digestion, and scale-up supported by techno-economic and life-cycle assessments.\u003c/p\u003e","manuscriptTitle":"Green Extraction and Optimization of Bioactive Compounds from Elderberry (Sambucus nigra L.) by β-Cyclodextrin Assisted Extraction Short title: Optimized Green Extraction of Elderberry Bioactives with β-Cyclodextrin","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-09 03:54:34","doi":"10.21203/rs.3.rs-7428774/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-12T17:21:20+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-06T18:34:00+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"144713438230473446096799463050672466636","date":"2025-09-29T20:48:17+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-12T00:14:27+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"183687346449482216619746837476485174937","date":"2025-09-04T18:30:37+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-02T15:16:22+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-08-25T02:59:14+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-08-25T02:57:58+00:00","index":"","fulltext":""},{"type":"submitted","content":"Food Analytical Methods","date":"2025-08-21T18:50:47+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"food-analytical-methods","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Food Analytical Methods](https://www.springer.com/journal/12161)","snPcode":"12161","submissionUrl":"https://submission.nature.com/new-submission/12161/3","title":"Food Analytical Methods","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"9daf1256-d944-4174-b04d-a13bfac0ef7c","owner":[],"postedDate":"September 9th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-02-11T21:39:17+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-09 03:54:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7428774","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7428774","identity":"rs-7428774","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}