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 A. 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. A. 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 intraday 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 A. 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 A. esculentus L. Moench, is a scientifically reliable and suitable analytical method.

본 연구에서는 표준화된 곰피추출물의 항산화 및 콜레 스테롤 개선에 대한 효능평가를 통해 건강기능식품 소재로서의 가치를 검토하기 위해 총 폴리페놀, 총 플라보노이드 및 dieckol 함량을 측정하였으며 DPPH, ABTS radical 소거능, reducing power 및 FRAP 활성을 통하여 곰피추 출물의 in vitro 항산화 활성을 조사하였고 표준화된 곰피 추출물의 HMG-CoA reductase 저해 활성 및 세포 내 콜레스테롤 생성 억제 효능을 평가하였다. 표준화된 곰피추출물의 총 폴리페놀, 총 플라보노이드 및 dieckol 함량은 각각 9.64±0.04 mg GAE/g, 2.72±0.08 mg RE/g, 27.42±0.66 mg/g 으로 나타났다. 표준화된 곰피추출물의 in vitro 항산화활 성, HMG-CoA reductase 저해활성 및 세포 내 콜레스테롤 생성 억제 효능은 농도의존적으로 증가하는 경향을 보였으며 이는 표준화된 곰피추출물에 함유되어 있는 페놀성 화합물에 기인된 효능으로 사료되며 항산화성분, 항산화 효과, 콜레스테를 개선 효능간의 상관관계가 있음을 확인 하였다. 향후, 표준화된 곰피추출물에 대한 in vivo 모델 에서의 전임상 연구 및 작용기전 입증되면 인체적용시험을 통해 이중기능성을 갖는 건강기능식품의 개발이 가능할 것으로 사료된다.