산초나무(Zanthoxylum schinifolium) 잎의 Chlorogenic Acid와 Hesperidin 정량분석을 위한 HPLC-DAD 분석법 개발 및 평가
Validation of HPLC-DAD Method for Quantitative Analysis of Chlorogenic Acid and Hesperidin in Zanthoxylum schinifolium Leaves Extract
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초록
본 연구는 Diode Array Detector (DAD)가 장착된 고성능 액체 크로마토그래피(HPLC)를 이용하여 산초나무 잎에 함유된 chlorogenic acid와hesperidin을 동시에 분석할 수 있는 표준 정량법 개발을 확립하고자 실시하였다. 산초나무 잎의 chlorogenic acid와 hesperidin은 MS/MS 분석및 표준물질과의 유지시간(RT), UV 흡광도 비교를 통해 확인하였고, 산초나무 잎의 지표물질로 선정하였다. 개발한 분석방법은 지표물질을 대상으로특이성, 정확성, 정밀도, 검출한계, 정량한계, 선형성에 대해 최적화하고 검증하였다. HPLC 분석으로 두 화합물에 대한 검량선은 각각 0.99% 이상의높은 상관계수(r2)를 보였다. Chlorogenic acid의 전체 회수율은 106.82%이었고, hesperidin의 전체 회수율은 107.37%였다. Chlorogenic acid의검출한계(LOD)는 0.271μg/ml, Hesperidin의 검출한계(LOD)는 0.005μg/ml였다. Chlorogenic acid의 일간정밀도는 0.102% (RSD%), Hesperidin의일간정밀도는 1.038% (RSD%)였다. 본 연구에서 개발한 HPLC-DAD 분석방법은 기능성 원료 인정을 위한 분석기준을 충족하였다. 따라서 본연구에서 확립한 HPLC 분석법은 향후 산초나무 잎에서 유래한 생리활성물질의 대량추출 및 관련제품을 개발할 때 원료 표준화에 적용가능할 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 Zanthoxylumschinifolium. The chlorogenic acid and hesperidin from Z. schinifolium leaves were identified by MS/MS analysis, comparing retentiontime and UV absorbance with authentic compounds. HPLC method was validated by specificity, accuracy, precision, and limit ofquantification. The HPLC method showed that calibration curves for each compounds have high linearity with a correlation coefficient(r 2) of ＞ 0.99%. The total recovery rate of chlorogenic acid and hesperidin was 106.82% and 107.37%, respectively. The detectionlimits (LOD) for chlorogenic acid and hesperidin are 0.271㎍/ml and 0.005㎍/ml, respectively. Daily precision of chlorogenic acid was0.102% (RSD%), and 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 to standardize health-related functional productsor pharmaceutical raw materials and products derived from Z. schinifolium leaves.
참고문헌 (21)
- Aly SH, "Comparative metabolic study of the chloroform fraction of three Cystoseira species based on UPLC", ESI, J. of Enzyme Inhibition and Medicinal Chemistry, pp.2024, 39(1)[DOI]
- Bae SM, "Studies on proximate composition, fatty acids and volatile compounds of Zanthoxylum schinifolium fruit according to harvesting time", Korean J. of Medicinal Crop Science, 19(1), pp.1~8 (2011)
- Choi HJ, "Antiviral activity of Zanthoxylum species against influenza virus", Korean J. of Medicinal Crop Science, 16(4), pp.273~278 (2008)
- Dinda B, "Dietary plant flavonoids in prevention of obesity and diabetes", Advances in Protein Chemistry and Structural Biology, 120, pp.159~235 (2019)
- Guideline IHT, "Validation of analytical procedures: Text and methodology", Q2 (R1), 1(20), pp.05~ (2005)
- Hong JH, "Analytical method validation of cynaroside in domestic Anthriscus sylvestris (L.) Hoffm, leaves extract for standardization as a functional ingredient using RP-HPLC", J. of the Korean Society of Food Science and Nutrition, 50(4), pp.395~402 (2021)
- Jang MJ, "Effects of antioxidative, DPPH radical scavenging activity and antithrombogenic by the extract of sancho(Zanthoxylum schinifolium)", The Korean J. of Nutrition, 38(5), pp.386~394 (2005)
- Jin KS, "Anti-oxidant, anti-melanogenic, and anti-inflammatory activities of Zanthoxylum schinifolium extract and its solvent fractions", Microbiology and Biotechnology Letters, 40(4), pp.371~379 (2012)
- Kim GD, "Covid-19 pandemic and immune regulatory functional foods", Food Science and Industry, 55(3), pp.244~263 (2022)
- Kim J, "Antimicrobial activities of Zanthoxylum schinifolium and Zanthoxylum piperitum leaves", Korean J. of Food Preservation, 7(2), pp.195~200 (2000)
- Kim JS, "Chemical composition and antitumor apoptogenic activity of methylene chloride extracts from the leaves of Zanthoxylum schinifolium", J. of Life Science, 16(3), pp.546~554 (2006)
- Lee JS, "Optimization of extraction conditions of continentalic and kaurenoic acids from Aralia continetalis by HPLC", UV and their validation, 2020, pp.58, 672~677
- Lee JW, "Volatile flavor components of Korean sancho fruit and tree(Zanthoxylum schinifolium)", Korean J. of Food and Nutrition, 11, pp.493~498 (1998)
- Min K., "The status analysis of using forest bio-resource in the Korean bioindustry", Korean J. of Forest Economics, 29(1), pp.37~49 (2022)
- Pyrzynska K, "Hesperidin: A review on extraction methods, stability and biological activities", Nutrients, 14(12), pp.2387~ (2022)
- Raffa D, "Recent discoveries of anticancer flavonoids", Europena J. of Medicinal Chemistry, 142, pp.213~228 (2017)
- Rodriguez JP, "HPLC-UV analysis of sample preparation influence on flavonoid yield from Cirsium japonicum var. maackii", Applied Biological Chemistry, 60, pp.519~525 (2017)
- Ryu GH, "HPLC analytical method validation for xanthohumol in Hop(Humulus lupulus)extract for its standardization as a functional ingredient", J. of the Korean Society of Food Science and Nutritionr, 53(2), pp.157~163 (2024)
- Ryu JH, "Morphological characteristics and genetic diversity analysis of cultivated Sancho(Zanthoxylum schinifolium)and Chopi(Zanthoxylum piperitum)in Korea", Korean J. of Plant Resources, 29(5), pp.555~563 (2016)
- Wang L, "The biological activity mechanism of chlorogenic acid and its applications in food industry : A review", Frontiers in Nutrition, 9, pp.943911~ (2022)
- Zhang R, "Flavonoid-rich sesame leaf extract-mediated synthesis of nanozymes : Extraction optimization, chemical composition identification and bioactivity evaluation", Food Chemistry, 456, pp.140021~ (2024)