산초나무(Zanthoxylum schinifolium) 잎의 Chlorogenic Acid, Hesperidin 정량분석을 위한 HPLC-DAD 분석법 개발

김준성*, 강주영, 노혜령, 김봉규

(경상국립대학교 환경산림과학부)

HPLC-DAD method for quantitative analysis of Chlorogenic Acid and Hesperidin in Zanthoxylum schinifolium leaves

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

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

요약: 본 연구는 산초나무 잎에 함유된 Chlorogenic acid, Hesperidin의 표준정량화를 위해 diode array detector(PDA)가 장착된 고성능 액체 크로마토그래피 (HPLC) 분석법을 확립하고자 실시하였다. 산초나무 잎의 주요 화합물은 MS/MS 분석과 표준물질과의 머무름시간(RT) 및 UV 흡광도 비교로 Chlorogenic acid, Hesperidin인 것을 확인하였다. HPLC 분석법에 대한 특이성, 정확성, 정밀도 및 정량 한계 검증을 하였다. HPLC분석으로 두 화합물에 대한 검량선이 각각 0.99% 이상의 상관 계수(R2)로 높은 선형성을 가지는 것을 보여주었다. Chlorogenic acid의 전체 회수율은 106.82%이었고, Hesperidin의 전체 회수율은 107.37%였다. Chlorogenic acid의 검출한계(LOD)는 0.271µg/ml, Hesperidin의 검출한계(LOD)는 0.005µg/ml였다. Chlorogenic acid의 일간정밀도는 0.102%(RSD%), Hesperidin의 일간정밀도는 1.038%(RSD%)였다. 테스트한 모든 매개변수는 기능성 원료 인정을 위한 분석기준을 충족하였다. 따라서 본 연구에서 확립한 HPLC 분석법은 향후 산초나무 잎 유래 건강 관련 기능성 제품 또는 의약품 원료 및 제품 표준화에 적용하고 사용할 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 and Hesperidin, which are healthy functional food ingredients in leaves of Zanthoxylum schinifolium. The major compounds of Z. schinifolium 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 two compounds have high linearity with a correlation coefficient of (r2)＞ 0.99%. The total recovery rate of chlorogenic acid was 106.82%, the total recovery rate of Hesperidin was 107.37%. The detection limits (LOD) for chlorogenic acid are 0.271㎍/ml, Hesperidin are 0.005㎍/ml respectively. Daily precision of chlorogenic acid was 0.102%((RSD%), Hesperidin was 1.038%(RSD%). All tested parameters met the appropriate ranges designated in the guideline. Therefore, the HPLC analysis method established in this study can be applied and used in the future for standardizing health-related functional products or pharmaceutical raw materials and products derived from Z. schinifolium leaves.

Keywords: High-performance liquid chromatography with diode-array detection, Chlorogenic acid, Hesperidin, Zanthoxylum schinifolium

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).