꾸지뽕나무 잎에 함유된 Chlorogenic acid, Rutin, Isoquercitrin의 분석을 위한 HPLC-DAD를 분석법 검증

강주영*, 노혜령, 김준성, 김봉규

(경상국립대학교 환경산림과학부)

HPLC/DAD Method Validation of Chlorogenic acid, Rutin, and Isoquercitrin Analysis from Cudrania tricuspidata leaves

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

(Division of Environmental and Forest Science, Gyeongsang National University)

요약: 본 연구는 꾸지뽕나무 잎에 함유된 클로로겐산, 루틴, 이소퀘르시트린의 표준정량화를 위해 diode array detector(DAD)가 장착된 고성능 액체 크로마토그래피 (HPLC) 분석법을 확립하고자 실시하였다. 꾸지뽕나무 잎의 주요 화합물은 MS/MS 분석과 표준물질과의 머무름시간(RT) 및 UV 흡광도 비교로 클로로겐산, 루틴, 이소퀘르시트린인 것을 확인하였다. HPLC 분석법에 대한 특이성, 정확성, 정밀도 및 정량 한계 검증을 하였다. HPLC분석으로 세 화합물에 대한 검량선이 각각 0.99% 이상의 상관 계수(r2)로 높은 선형성을 가지는 것을 보여주었다. 클로로겐산의 전체 회수율은 101.63%, 회수율 구간은 101.04-102.68%, 루틴의 전체 회수율은 101.81%, 회수율 구간은 100.28-102.76%, 이소퀘르시트린의 전체 회수율은 102.185%, 회수율 구간은 100.30-104.43%였다. 클로로겐산의 검출한계(LOD)는 0.286µg/ml, 루틴은 0.411µg/ml, 이소퀘르시트린은 0.201µg/ml였다. 클로로겐산의 일간정밀도는 0.236%(RSD%), 루틴은 0.559%(RSD%), 이소퀘르시트린은 0.256%(RSD%)였다. 테스트한 모든 매개변수는 기능성 원료 인정을 위한 분석기준을 충족하였다. 따라서 본 연구에서 확립된 HPLC 분석은 꾸지뽕나무 잎 추출물에서 유래된 건강 관련 기능성 제품 또는 의약품을 개발하는 데 사용될 수 있을 것이다.

Abstract: We attempted to establish a High-performance liquid chromatography (HPLC) analysis method with a diode array detector (DAD) for standard quantification of chlorogenic acid, rutin, and isoquercitrin, which are healthy functional food ingredients in leaves of Cudrania tricuspidata. The major compounds of C. tricuspidata leaves were identified by MS/MS analysis, comparing retention time and UV absorbance with authentic compounds. HPLC method was validated by specificity, accuracy, precision, and limit of quantification. The HPLC method showed that calibration curves for each of the three compounds have high linearity with a correlation coefficient of  (r2)＞ 0.99%. The total recovery rate of chlorogenic acid was 101.63%, the recovery section was 101.04-102.68%, the total recovery rate of rutin was 101.81%, the total recovery section was 100.28-102.76%, the total recovery rate of isoquercitrin was 102.185, and the recovery section was 100.30-104.43%. The detection limits (LOD) for chlorogenic acid are 0.286㎍/ml, rutin 0.411㎍/ml, and isoquercitrin 0.201㎍/ml, respectively. Daily precision of chlorogenic acid was 0.236%(RSD%), rutin was 0.559%(RSD%), and isoquercitrin was 0.256%(RSD%). All tested parameters met the appropriate ranges designated in the guideline. Therefore, the HPLC analysis established in this study can be used to develop health-related functional products or medicines derived from C. tricuspidata leaves extract.

Keyword: Cudrania tricuspidata, chlorogenic acid, rutin, isoquercitrin, HPLC-DAD , Method validation

Acknowledgments: This study was carried out with the support of R&D Program for Forest Science Technology (Project No. "FTIS 2023478B10-2325BC036 ")' and Forest Service's forest industry-customized human resources training project (Short-term Income Forest products governance education center. Project No. RS-2025-00405196), provided by Korea Forest Service (Korea Forestry Promotion Institute).