까치밥나무 잎 주요 플라보놀의 HPLC 기반 동시 정량분석법 검증

노혜령*, 강주영, 김봉규

(경상국립대학교 환경산림과학부)

Simultaneous Quantification and Method Validation of Major Flavonols in Ribes mandshuricum Leaves by HPLC-DAD

Hye-Ryeong Noh*, Ju-Yeong Kang, Bong-Gyu Kim

Division of Environmental and Forest Science, Gyeongsang National University)

요약: 본 연구에서는 까치밥나무(Ribes mandshuricum) 잎에 존재하는 quercetin 3-O-sophoroside 및 kaempferol 3-O-sophoroside의 동시 정량을 위한 HPLC-DAD 분석법을 확립하고, 그 타당성을 평가하였다. 대상 화합물들은 표준물질과의 retention time 및 UV 스펙트럼 비교를 통해 확인되었으며, MS/MS 분석을 통해 추가 검증하였다. 이들 화합물은 품질 평가용 지표물질(marker compound) 로 선정되었다. 분석법 검증은 국제의약품규제조화위원회(ICH, International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use) 지침에 따라 특이성, 정확도, 정밀도, 직선성, 검출한계(LOD), 정량한계(LOQ) 항목에 대해 수행되었다. 검량곡선은 우수한 직선성(r² > 0.999) 을 나타냈다. 평균 회수율은 quercetin 3-O-sophoroside 에서 99.04%, kaempferol 3-O-sophoroside 에서 98.39%로 확인되었다. 검출한계는 각각 0.012μg/mL, 0.038μg/mL, 일내 정밀도(RSD%)는 각각 0.09–0.14%, 0.13–0.74% 수준이었다. 확립된 HPLC-DAD 분석법은 까치밥나무 잎을 기능성식품 또는 약용 소재로 활용하기 위한 표준화 및 품질관리에 신뢰성 있게 적용될 수 있으며, 생리활성 물질의 산업적 추출 및 품질관리 체계에도 응용할 수 있을 것으로 기대된다.

Abstract: This study established and validated an HPLC-DAD analytical method for the simultaneous quantification of quercetin 3-O-sophoroside and kaempferol 3-O-sophoroside in leaves of Ribes mandshuricum. The target compounds were identified by comparing retention times and UV spectra with authentic standards, and were further confirmed by MS/MS analysis. These compounds were selected as marker compounds for quality assessment. The analytical method was validated following the guidelines of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH), including evaluations of specificity, accuracy, precision, linearity, limit of detection (LOD), and limit of quantification (LOQ). The calibration curves exhibited excellent linearity (r² > 0.999). The average recovery rates were 99.04% for quercetin 3-O-sophoroside and 98.39% for kaempferol 3-O-sophoroside. The LODs were 0.012 μg/mL and 0.038 μg/mL, respectively. Intra-day precision (RSD%) ranged from 0.09–0.14% for quercetin 3-O-sophoroside and 0.13–0.74% for kaempferol 3-O-sophoroside. The validated HPLC-DAD method can be reliably applied for the standardization of R. mandshuricum leaves as functional food or medicinal materials, and shows potential for use in industrial-scale extraction and quality control of bioactive compounds.

Keyword: High-performance liquid chromatography with diode-array detection, quercetin 3-O-sophoroside, kaempferol 3-O-sophoroside, Ribes mandshuricum leaves

Acknowledgments: This study was supported by the Korea Forest Service through the Forest Industry–Customized Human Resources Training Project (Short-term Income Forest Products Governance Education Center; Project No. RS-2025-00405196), administered by the Korea Forestry Promotion Institute.