HPLC法测定柴黄颗粒中柴胡皂苷a、b_1、c、d及黄芩苷含量

Determination of saikosaponin a,saikosaponin b_1,saikosaponin c,saikosaponin d and baicalin in Chaihuang granules by HPLC

目的:建立测定柴黄颗粒中柴胡皂苷a、b_1、C、d及黄芩苷含量的HPLC方法。方法:采用SB-C_(18)色谱柱(250 mm×4.6 mm,5.0μm),以乙腈(A)-0.01%磷酸水溶液(B)为流动相,梯度洗脱(0-30 min,30%A→60%A;30~35 min,60%A),流速1 mL·min^(-1),检测波长为210 nm。结果:样品采用5%的浓氨甲醇溶液超声提取30min,柴胡皂苷a、b_1、c、d及黄芩苷质量浓度分别在3.6~36.0、6.0-60.0、8.4~84.0、9.2~92.0和10.0-100.0μg·mL^(-1)范围内线性关系良好(r≥0.999 2);方法的精密度、稳定性及重复性良好,RSD均小于2.0%;柴胡皂苷a、b_1、c、d及黄芩苷平均回收率为97.5%、95.3%、99.7%、98.4%和95.1%,RSD均小于2.0%;3批样品中柴胡皂苷a、b_1、d及黄芩苷含量范围分别为1.560-1.573、7.501~7.587、9.204~9.356和11.635~11.817 mg·g^(-1),柴胡皂苷c未检出。结论:经方法学验证本方法可用于柴黄颗粒中柴胡皂苷a、b_1、C、d及黄芩苷的含量测定,为柴黄颗粒的全面质量控制提供科学依据。

Objective： To establish an HPLC method for determination of saikosaponin a, saikosaponin b~, saikosaponin c, saikosaponin d and baicalin in Chaihuang granules. Methods： HPLC experiments were performed on a ZORBAX SB-C18 column （250 mm~ 4.6 ram, 5.0 p^m ）. The mobile phase consisted of acetonitrile （A）-0.01% phosphoric acid （ B ）in gradient mode（ 0-30 rain, 30%A -~ 60%A; 30 -35 rain, 60%A ）. The flow rate was 1 mL＂ rain-t The detection wavelength was set at 210 nm. Results： The samples were extracted by ultrasonic with 5% concentrated ammonia solution for 30 rain. Saikosaponin a, saikosaponin b1, saikosaponin c, saikosaponin d and baicalin show a good linear relationship within the ranges of 3.6-36.0, 6.0-60.0, 8.4-84.0, 9.2-92.0 and 10.0- 100.0 μg· mL-1, respectively （ r ≥ 0.999 2 ）. Good precision, stability and reproducibility was achieved with the RSDs below 2.0%. The average recoveries of saikosaponin a, saikosaponin bz, saikosaponin c, saikosaponin d and baicalin were 97.5%, 95.3%, 99.7%, 98.4% and 95.1% with RSDs below 2.0%. The contents of saikosaponins a, bj, d and baiealin in three batches of samples were 1.560-1.573,7.501-7.587,9.204-9.356 and 11.635-11.817 mg·g-1. The saikosaponins c is not detected in three batches of samples. Conclusion： The established method can be used in determination of saikosaponin a, saikosaponin bi, saikosaponin c, saikosaponin d and baiealin, and it provide a scientific basis for the comprehensive quality control of Chaihuang granules.