HPLC-QAMS同时测定芪桑益肝丸中8种成分的含量

Simultaneous Determination of Eight Components in Qisang Yigan Pills by HPLC-QAMS

关键词:  一测多评法  芪桑益肝丸  虎杖苷  白藜芦醇  大黄素  獐牙菜苦苷  龙胆苦苷  獐牙菜苷  毛蕊异黄酮葡萄糖苷  芒柄花素

Key words:  QAMS  Qisang Yigan pills  polydatin  resveratrol  emodin  swertiamarin  gentiopicrin  sweroside  calycosin 7-O-β-D-glucopyranoside  formononetin

摘要:

目的 建立高效液相一测多评法（HPLC-QAMS）同时测定芪桑益肝丸中虎杖苷、白藜芦醇、大黄素、獐牙菜苦苷、龙胆苦苷、獐牙菜苷、毛蕊异黄酮葡萄糖苷和芒柄花素的含量。方法 采用Agilent HC-C₁₈色谱柱（250 mm×4.6 mm，5 μm），柱温30℃；乙腈-0.1%甲酸水溶液为流动相梯度洗脱，流速1.0 mL·min^-1；检测波长分别为290 nm （检测虎杖苷、白藜芦醇和大黄素）、243 nm （检测獐牙菜苦苷、龙胆苦苷、獐牙菜苷）和254 nm （检测毛蕊异黄酮葡萄糖苷、芒柄花素）。以獐牙菜苦苷为内参物，建立其他7个成分的相对校正因子，计算含量。结果 虎杖苷、白藜芦醇、大黄素、獐牙菜苦苷、龙胆苦苷、獐牙菜苷、毛蕊异黄酮葡萄糖苷和芒柄花素质量浓度分别在3.46~69.20，2.09~41.80，2.77~55.40，6.71~134.20，2.88~57.60，1.19~23.80，0.56~11.20，0.89~17.80 μg·mL^-1内线性关系良好（r≥0.999 1），平均加样回收率（RSD）分别为100.05%（0.82%），97.79%（1.07%），98.41%（0.99%），99.75%（0.65%），98.72%（1.21%），99.07%（1.19%），96.99%（1.04%），97.65%（1.38%），一测多评法计算结果与外标法实测值无显著性差异。结论 所建立的HPLC-QAMS方法专属性强、准确度高，为芪桑益肝丸的质量评价提供了参考依据。

Abstract:

OBJECTIVE To establish a HPLC-QAMS method for the determination of polydatin, resveratrol, emodin, swertiamarin, gentiopicrin, sweroside, calycosin 7-O-β-D-glucopyranoside and formononetin in Qisang Yigan pills. METHODS The separation and detection procedure was performed on an Agilent HC-C₁₈(250 mm×4.6 mm, 5 μm) at 30℃. The mobile phase was composed of acetonitrile-0.1% formic acid aqueous solution using gradient elution at a flow rate of 1.0 mL·min^-1. The detecting wavelength was set at 290 nm for polydatin, resveratrol and emodin, 243 nm for swertiamarin, gentiopicrin and sweroside, and 254 nm for calycosin 7-O-β-D-glucopyranoside and formononetin. Using swertiamarin as an internal standard, the relative correction factors of the other seven constituents were calculated, after which the content determination was made. RESULTS Polydatin, resveratrol, emodin, swertiamarin, gentiopicrin, sweroside, calycosin 7-O-β-D-glucopyranoside and formononetin showed good linear relationships within the ranges of 3.46-69.20, 2.09-41.80, 2.77-55.40, 6.71-134.20, 2.88-57.60, 1.19-23.80, 0.56-11.20, 0.89-17.80 μg·mL^-1(r ≥ 0.999 1), whose average recoveries(RSD) were 100.05%(0.82%), 97.79%(1.07%), 98.41%(0.99%), 99.75%(0.65%), 98.72%(1.21%), 99.07%(1.19%), 96.99%(1.04%), 97.65%(1.38%), respectively. There was no significant difference between the calculated values and the measured values of the external standard method. CONCLUSION The established HPLC-QAMS method is specifical, reliable and can be used to provide a reference standard for quality evaluation of Qisang Yigan pills.