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1.0mL/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.
基金项目
自治区创新环境(人才、基地)建设专项科技创新基地建设计划(资源共享平台建设)(PT2018)
中国营养学会全民营养科研基金(CNS-NNSRG2019-96)
国家科技期刊平台
NSTL
万方数据