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KMID : 0881720220370020039
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Journal of Food Hygiene and Safety
2022 Volume.37 No. 2 p.39 ~ p.45
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Improvement and Validation of an Analytical Method for Quercetin-3-O-gentiobioside and Isoquercitrin in Abelmoschus esculentus L. Moench
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Han Xionggao
Choi Sun-Il
Men Xiao
Lee Se-Jeong
Jin Hee-Gu
Oh Hyun-Ji
Cho Se-Haeng
Lee Boo-Yong
Lee Ok-Hwan
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Abstract
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This study aimed to investigate the validation and modify the analytical method to determine quercetin-3-O-gentiobioside and isoquercitrin in Abelmoschus esculentus L. Moench for the standardization of ingredients in development of functional health products. The analytical method was validated based on the ICH (International Conference for Harmonization) guidelines to verify the reliability and validity there of on the specificity, linearity, accuracy, precision, detection limit and quantification limit. For the HPLC analysis method, the peak retention time of the index component of the standard solution and the peak retention time of the index component of Abelmoschus esculentus L. Moench powder sample were consistent with the spectra thereof, confirming the specificity. The calibration curves of quercetin-3-O-gentiobioside and isoquercitrin showed a linearity with a near-one correlation coefficient (0.9999 and 0.9999), indicating the high suitability thereof for the analysis. Abelmoschus esculentus L. Moench powder sample of a known concentration were prepared with low, medium, and high concentrations of standard substances and were calculated for the precision and accuracy. The precision of quercetin-3-O-gentiobioside and isoquercitrin was confirmed for intra-day and daily. As a result, the intra-day precision was found to be 0.50-1.48% and 0.77- 2.87%, and the daily precision to be 0.07-3.37% and 0.58-1.37%, implying an excellent precision at level below 5%.
As a result of accuracy measurement, the intra-day accuracy of quercetin-3-O-gentiobioside and isoquercitrin was found to be 104.87-109.64% and the daily accuracy thereof was found to be 106.85-109.06%, reflecting high level of accuracy. The detection limits of quercetin-3-O-gentiobioside and isoquercitrin were 0.24 μg/mL and 0.16 μg/mL, respectively, whereas the quantitation limits were 0.71 μg/mL and 0.49 μg/mL, confirming that detection was valid at the low concentrations as well. From the analysis, the established analytical method was proven to be excellent with high level of results from the verification on the specificity, linearity, precision, accuracy, detection limit and quantitation limit thereof. In addition, as a result of analyzing the content of Abelmoschus esculentus L. Moench powder samples using a validated analytical method, quercetin-3-O-gentiobioside was analyzed to contain 1.49±0.01 mg/dry weight g, while isoquercitrin contained 1.39±0.01 mg/dry weight g. The study was conducted to verify that the simultaneous analysis on quercetin-3-O-gentiobioside and isoquercitrin, the indicators of Abelmoschus esculentus L. Moench, is a scientifically reliable and suitable analytical method.
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KEYWORD
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Abelmoschus esculentus L. Moench, Quercetin-3-O-gentiobioside, Isoquercitrin, HPLC-PDA, Method validation
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