HPLC-DAD双波长法测定护肝片中5个柴胡皂苷的含量  

Simultaneous determination of five saikosaponins in Hugan tablets by HPLC-DAD dual wavelength method

作　　者：邹惠亮 陈柯 陈敏 陈褚建 ZOU Hui-liang;CHEN Ke;CHEN Min;CHEN Chu-jian(Huzhou Institute for Food and Drug Control,Huzhou 313000,China)

机构地区：[1]湖州市食品药品检验研究院

出　　处：《药物分析杂志》2019年第12期2248-2253,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立测定护肝片中柴胡皂苷a、柴胡皂苷b1、柴胡皂苷b2、柴胡皂苷c、柴胡皂苷d的定量分析方法。方法:采用Agilent Eclipse Plus C18(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈(A)-水(B),梯度洗脱(0~40 min,30%A→53%A;40~48 min,53%A→98%A),检测波长210 nm(柴胡皂苷a、柴胡皂苷c、柴胡皂苷d)、254 nm(柴胡皂苷b1、b2),流速1 mL·min-1,柱温25℃。结果:5个柴胡皂苷分离度良好;柴胡皂苷a、柴胡皂苷b1、柴胡皂苷b2、柴胡皂苷c、柴胡皂苷d进样量分别在0.013 5~0.674、0.0156~0.781、0.015 2~0.762、0.015 1~0.757和0.013 1~0.653μg范围内线性关系良好(r≥0.999 5);稳定性及重复性试验RSD均低于2.0%;平均回收率在99.7%~102.9%,RSD均小于2.0%(n=6)。12个生产厂家50批样品中均未检测到柴胡皂苷a、柴胡皂苷c、柴胡皂苷d,除2个厂家外的其余厂家样品中均检测到柴胡皂苷b1、柴胡皂苷b2。结论:所建立的方法简便、快捷、重复性好,可用于护肝片中5个柴胡皂苷的含量测定,为护肝片的全面质量控制提供科学依据。Objective:To establish a quantitative method for determination of saikosaponin a,saikosaponin b1,saikosaponin b2,saikosaponin c and saikosaponin d in Hugan tablets. Methods:HPLC experiment was performed on an Agilent Eclipse Plus C18 column(250 mm×4.6 mm,5.0 μm). The mobile phase consisted of acetonitrile(A)-water(B)in gradient mode(0-40 min,30%A → 53%A;40-48 min,53%A → 98%A). The detection wavelength was set at 210 nm for saikosaponin a,saikosaponin c,saikosaponin d and 254 nm for saikosaponin b1,saikosaponin b2. The flow rate was 1 mL·min-1. The column temperature was maintained at 25 ℃. Results:The results showed that the separation of the five saikosaponins was good. Saikosaponin a,saikosaponin b1,saikosaponin b2,saikosaponinc and saikosaponin d showed good linear relationship within the range of 0.0135-0.674,0.0156-0.781,0.0152-0.762,0.0151-0.757 and 0.0131-0.653 μg,respectively(r ≥0.9995). Good precision,stability and reproducibility was achieved with the RSDs below 2.0%. The average recoveries of saikosaponin a,saikosaponin b1,saikosaponin b2,saikosaponinc and saikosaponin d were among 99.7%-102.9% with RSDs below 2.0%. Saikosaponin a,saikosaponin c and saikosaponin d was not detected in fifty batches of samples from twelve companies. Saikosaponin b1 and saikosaponin b2 were detected in most samples except samples from two companies. Conclusion:The method is simple,rapid and reproducible. It can be used for the determination of the five saikosaponins in Hugan Tables,providing a scientific basis for the comprehensive quality control of Hugan Tables.

关 键 词：护肝片 柴胡 柴胡皂苷 含量测定 高效液相色谱-二极管阵列检测器 

分 类 号：R917[医药卫生—药物分析学]