一测多评法同时测定桑叶中9种绿原酸类和黄酮类成分

刘梦文; 沈静; 阿依努尔·白克热; 贺诗茹; 肖辉; 徐维盛

doi:10.13386/j.issn1002-0306.2023080195

一测多评法同时测定桑叶中9种绿原酸类和黄酮类成分

Simultaneous Determination of Nine Chlorogenic Acids and Flavonoids in Mori Folium by QAMS

摘要

摘要: 目的：建立一测多评法同时测定桑叶中新绿原酸、绿原酸、隐绿原酸、芦丁、异槲皮苷、异绿原酸B、紫云英苷、异绿原酸A和异绿原酸C的含量。方法：采用Agilent TC-C₁₈色谱柱（250 mm×4.6 mm，5 μm），以乙腈（A）-0.1%磷酸水溶液（B）为流动相，梯度洗脱，流速1.0 mL/min，检测波长260、320 nm，柱温30 ℃。以绿原酸为内参物，分别采用外标法和一测多评（多点校正、斜率校正和单点校正）测定桑叶中9个成分的含量并比较4种方法之间的差异，另外采用多点校正和两点校正法进行色谱峰定位，验证该方法的准确性和可行性。结果：新绿原酸、隐绿原酸、芦丁、异槲皮苷、异绿原酸B、紫云英苷、异绿原酸A和异绿原酸C相对于绿原酸的校正因子分别为0.9072、0.8736、0.6207、0.8547、1.1936、0.5501、1.4369和1.2244（多点校正），且在不同条件下相对校正因子耐用性（RSD<1.5%）和重现性（RSD<5%）良好。外标法和一测多评（多点校正、斜率校正和单点校正）测得桑叶中9个成分含量的结果之间无差异（RSD<1.5%）。相较于相对保留时间，多点校正和两点校正可改善色谱峰的准确定位（相对误差|RE|<3%）。结论：建立了桑叶中9种成分的一测多评法，该法准确可行，可用于桑叶的质量控制。

Abstract: Objective: To establish a HPLC method for the determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutin, isoquercitrin, isochlorogenic acid B, astragalin, isochlorogenic acid A and isochlorogenic acid C in Mori Folium using the quantitative analysis of multi-components by single marker (QAMS). Methods: The analysis was performed on an Agilent TC-C₁₈ column (250 mm×4.6 mm, 5 μm), with acetonitrile (A)-0.1% phosphoric acid (B) as mobile phase at the flow rate of 1.0 mL/min for gradient elution, as well as the wavelengths were 260 and 320 nm and the column temperature was 30 ℃. The chlorogenic acid was used as the internal reference. The contents of nine components in Mori Folium were calculated by the external standard method and QAMS (multi-point correction, gradient correction and single-point correction) respectively, and the differences among the four methods were compared. In addition, the accuracy and feasibility of the method were verified by using multi-point correction and two-point correction to locate chromatographic peaks of the components. Results: The relative correction factors of neochlorogenic acid, cryptochlorogenic acid, rutin, isoquercitrin, isochlorogenic acid B, asiaticoside, isochlorogenic acid A and isochlorogenic acid C to chlorogenic acid were 0.9072, 0.8736, 0.6207, 0.8547, 1.1936, 0.5501, 1.4369 and 1.2244, respectively (multi-point correction). The durability (RSD<1.5%) and the reproducibility (RSD<5%) of the relative correction factors were positive under different conditions. The content of nine components in Mori Folium was determined simultaneously by the external standard method and QAMS (multi-point correction, gradient correction and single-point correction), and there was no difference between the results obtained from the four calculation methods (RSD<1.5%). Compared to the relative retention time, multi-point correction and two-point correction could improve the accurate localization of the chromatographic peaks (relative error |RE|<3%). Conclusion: The method of QAMS for nine components determination in Mori Folium was established, which was accurate and feasible and could be applied for the quality control of Mori Folium.