一测多评法和指纹图谱在维血宁合剂质量控制中的应用  

Application of the quantitative analysis of multi-components by single marker and fingerprint atlas in the quality control of Weixuening mixture

作　　者：董泰玮 杨焕 邹俊波 张小飞[1] 问思华 史亚军[1] 张益萌 孙静[1] DONG Tai-wei;YANG Huan;ZOU Jun-bo;ZHANG Xiao-fei;ZHANG Yi-meng;WEN Si-hua;SHI Ya-jun;SUN Jing(College of Pharmacy,Shaanxi University of Traditional Chinese Medicine,Xi’an 712046,China)

机构地区：[1]陕西中医药大学药学院,西安712046

出　　处：《药物分析杂志》2022年第11期1989-1999,共11页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金(82104412);维血宁合剂的二次开发研究(2019K01-19)。

摘　　要：目的:采用一测多评(QAMS)法测定维血宁合剂中8个有效成分的含量,建立“中间体-成品”的HPLC指纹图谱,为其质量控制提供参考和依据。方法:采用高效液相色谱法,使用Acclaim120 C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇(A)-0.2%磷酸水溶液(B)-0.1%磷酸(C)为流动相,进行梯度洗脱,流速为1.0 mL·min-1,检测波长为210 nm,柱温为30℃。以虎杖苷为内参物,分别考察儿茶素、表儿茶素、白藜芦醇、槲皮素、木犀草素、染料木素和芒柄花素7个成分的相对校正因子。采用相对校正因子计算3批维血宁合剂提取液、清膏和成品中7个成分的含量和指纹图谱,与外标法(ESM)进行比较,验证QAMS法的准确性和可行性。结果:儿茶素、表儿茶素、虎杖苷、白藜芦醇、槲皮素、木犀草素、染料木素和芒柄花素的线性范围分别为1.066~53.300、1.062~53.100、4.384~219.200、1.118~55.900、1.036~51.800、1.110~55.400、1.040~52.000、0.530~53.000μg·mL^(-1)。以虎杖苷为参照物,其他7个成分的相对校正因子分别为0.3754、0.3486、0.5853、0.5972、0.5176、0.5887、0.6599,在不同的实验条件下重复性较好。9个批次提取液、清膏、成品中8个成分的含量范围:虎杖苷为45.26~90.45μg·mL^(-1),儿茶素为21.29~38.18μg·mL^(-1),表儿茶素为15.06~21.46μg·mL^(-1),白藜芦醇为6.26~8.57μg·mL^(-1),槲皮素为1.22~2.13μg·mL^(-1),木犀草素为3.35~4.44μg·mL^(-1),染料木素为1.97~3.17μg·mL^(-1),芒柄花素为0.65~1.13μg·mL^(-1)。3批维血宁合剂的指纹图谱有27个共有峰,并且指认了以上8个成分峰,绝大部分相似度在0.9以上。结论:以虎杖苷为内参物建立的QAMS法操作简单,省时高效,可为维血宁合剂的质量控制提供参考。Objective:To establish a quantitative analysis of multi-components by single marker(QAMS)method for the determination of 8 isoflavones in Weixuening mixture,and establish the HPLC fingerprint of“intermediate finished product”,so as to provide reference and basis for its quality control.Methods:The Acclaim 120-C18column(250 mm×4.6 mm,5μm)was used with methanol(A)-0.2%phosphoric acid aqueous solution(B)-0.1%phosphoric acid(C)as mobile phase.Gradient elution was performed at a flow rate of 1.0 mL·min-1,with the detection wavelength of 210 nm and the column temperature of 30℃.Using polydatin as the reference substance,relative correction factors of catechin,epicatechin,resveratrol,quercetin,luteolin,genistein and formononetin were calculated,respectively.The relative correction factors were used to calculate the contents and fingerprints of 7 substances to be tested in 3 batches of Weixuening mixture,and the differences between the two methods were compared to verify the accuracy and feasibility.Results:The linear ranges of catechin,epicatechin,polydatin,resveratrol,quercetin,luteolin,genistein and formononetin were 1.066-53.300,1.062-53.100,4.384-219.200,1.118-55.900,1.036-51.800,1.110-55.400,1.040-52.000,0.530-53.000μg·mL^(-1),respectively.The relative correction factors of catechin,epicatechin,resveratrol,quercetin,luteolin,genistein and formononetin were 0.3754,0.3486,0.5853,0.5972,0.5176,0.5887,0.6599,respectively.The repeatability was good under different experimental conditions.At the same time,the content range of 8 components in the extracts,clear extracts and mixture was polydatin(45.26-90.45μg·mL^(-1)),catechin(21.29-38.18μg·mL^(-1)),epicatechin(15.06-21.46μg·mL^(-1)),resveratrol(6.26-8.57μg·mL^(-1)),quercetin(1.22-2.13μg·mL^(-1)),luteolin(3.35-4.44μg·mL^(-1)),genistein(1.97-3.17μg·mL^(-1)),and anthocyanin(0.65-1.13μg·mL^(-1)).There were 27 common peaks in the fingerprint of three batches of Weixuening mixture,and 8 peaks of polydatin,quercetin,catechin,epicatechin,luteolin,antho

关 键 词：一测多评法 指纹图谱 高效液相色谱 相对校正因子 相似度分析 维血宁合剂 虎杖苷 儿茶素 槲皮素 白藜芦醇 

分 类 号：R917[医药卫生—药物分析学]