HPLC法测定柴黄颗粒中柴胡皂苷a、b₁、c、d及黄芩苷含量

目的：建立测定柴黄颗粒中柴胡皂苷a、b₁、c、d及黄芩苷含量的HPLC方法。方法：采用SB-C₁₈色谱柱（250 mm×4.6 mm，5.0 μm），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30 min，柴胡皂苷a、b₁、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10.0~100.0 μg·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、b₁、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、b₁、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、b₁、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 b₁,saikosaponin c,saikosaponin d and baicalin in Chaihuang granules.Methods: HPLC experiments were performed on a ZORBAX SB-C₁₈ column(250 mm×4.6 mm,5.0 μm).The mobile phase consisted of acetonitrile(A)-0.01% phosphoric acid(B)in gradient mode(0-30 min,30%A→60%A;30 -35 min,60%A).The flow rate was 1 mL·min^-1. The detection wavelength was set at 210 nm.Results: The samples were extracted by ultrasonic with 5% concentrated ammonia solution for 30 min.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 b₁,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,b₁,d and baicalin 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 b₁,saikosaponin c,saikosaponin d and baicalin,and it provide a scientific basis for the comprehensive quality control of Chaihuang granules.

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