[논문]Optimization of Ascorbic Acid Extraction from Rugosa Rose (Rosa rugosa Thunb.) Fruit Using Response Surface Methodology and Validation of the Analytical Method

UM, Min; KIM, Ji-Woo; LEE, Jae-Won

doi:10.5658/wood.2020.48.3.364

Optimization of Ascorbic Acid Extraction from Rugosa Rose (Rosa rugosa Thunb.) Fruit Using Response Surface Methodology and Validation of the Analytical Method 원문보기

목재공학 = Journal of the Korean wood science and technology, v.48 no.3, 2020년, pp.364 - 375

UM, Min (Department of Wood Science and Engineering, College of Agriculture and Life Sciences, Chonnam National University) , KIM, Ji-Woo (Department of Wood Science and Engineering, College of Agriculture and Life Sciences, Chonnam National University) , LEE, Jae-Won (Department of Wood Science and Engineering, College of Agriculture and Life Sciences, Chonnam National University)

Abstract ▼ AI-Helper

In this study, ultrasound-assisted extraction was performed to extract ascorbic acid from rugosa rose (Rosa rugosa Thunb.) fruit. The optimal conditions were investigated by response surface methodology, using two variable including reaction time (16-44 min) and temperature (16-44℃). The ascorbic acid extraction was sensitive to the reaction time rather than the reaction temperature, and the optimal conditions for ascorbic acid extraction were 25℃ and 30 min. Ascorbic acid and gallic acid in the rugosa rose fruit extract were completely separated by HPLC, with a resolution factor of over 1.5 between the two. The correlation coefficient of the ascorbic acid was 0.999 in a linearity test for 50-150 ㎍/mL concentration of extract. The limit of detection and limit of quantification values were 0.16 ㎍/mL and 29.89 ㎍/mL, respectively. The relative standard deviations (RSD) for repeatability and reproducibility were determined, and each RSD showed good precision at less than 5% (N=6).

주제어

표/그림 (9)

표 Table 1. Ascorbic acid extraction from rugosa rose fruit by central composite design
그림 Fig. 1. Response surface plot presenting effect of temperature and time for ascorbic acid extraction from rugosa rose fruit.
표 Table 2. ANOVA analysis of central composite design for optimizing ascorbic acid extraction from rugosa rose fruit
그림 Fig. 2. Specificity for ascorbic acid using HPLC at 100 μg/mL extract; (A) methanol, (B) rugosa rose fruit extract, (C) ascorbic acid and gallic acid standard, (D) ascorbic acid, (E) gallic acid.
표 Table 3. Linearity range, limit of detection and limit of quantification for ascorbic acid from rugose rose fruit
표 Table 4. Repeatability relative standard deviation and reproducibility relative standard deviation for ascorbic acid from rugosa rose fruit
표 Table 5. Validation of robustness depending on solvent manufacturer, flow rate, and column temperature
표 Table 5. (Continued)
그림 Fig. 3. Absorbed power density (APD) and extract yield of ascorbic acid depending on ultrasound-assisted extraction scale.

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제안 방법

, 2018). Based on these parameters, the optimal UAE conditions were investigated by response surface methodology, using the two independent variables of reaction time and temperature. In addition, we validated the ascorbic acid extract to ensure the reliability of our results.
The specificity, linearity, limit of detection, limit of quantification, precision, and robustness were assessed according to the guidelines of the International Conference on Harmonization (ICH) were conducted for method validation to determine ascorbic acid extraction. Finally, we analyzed the extraction efficiency by taking into account the absorbed power density (W/mL) and absorbed energy density (J/mL) for UAE at the laboratory scale.
, 2012). In this study, ascorbic acid was extracted from rugosa rose fruit using a central composite design that considered two other independent variables: temperature and time. The experimental results according to the extraction conditions are shown in Table 1.
, 2001). In this study, experimental robustness determinations were carried out for different solvent manufacturers (A and B), flow rates (1.0 and 0.6 mL/min), and column temperatures (30 and 35℃) (Table 5). The flow rate had the strongest influence on the ascorbic acid analysis.
The precision for ascorbic acid was determined by HPLC for three different extract concentration (50 µg/mL, 100 µg/mL, and 150 µg/mL), by different analysts (analyst 1 and analyst 2), and at different dates (day 1 and day 2). The precision experiments were carried out both intra-day (within a day) and inter-day (between days).

대상 데이터

All experiments in this study were performed in more than triplicate (RSM N = 3, Validation N = 6). Two way ANOVA was performed to determine the significant differences between mean values.
Ascorbic acid (No 23024, 99.7-100.5%) and gallic acid (No G7384, 97.5-100.5%) were purchased from Sigma Aldrich (St. Louis, MO, USA). Methanol for HPLC analysis was purchased from Duksan Pure Chemicals Co.
The rugosa rose was grown at Chonnam National University (35°10'25.6"N, 126°54'00.8"E, South Korea) and harvested in November 2016.

데이터처리

The Pearson correlation coefficient was used to determine the correlation between the variables used, and the differences were considered significant at a p-value < 0.05.
All experiments in this study were performed in more than triplicate (RSM N = 3, Validation N = 6). Two way ANOVA was performed to determine the significant differences between mean values. The Pearson correlation coefficient was used to determine the correlation between the variables used, and the differences were considered significant at a p-value < 0.

이론/모형

The specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and robustness were assessed according to the guidelines of the International Conference on Harmonization (ICH) for method validation in order to determine the presence of ascorbic acid in the extract obtained by RSM at optimal conditions (Shabir, 2003; Sahu et al., 2018).

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