HPLC-PDA法同时测定茵栀黄注射液中黄芩苷和栀子苷血浆浓度及其在大鼠体内的药动学研究

Simultaneous determination of serum concentration of baicalin and geniposide from Yinzhihuang Injection by HPLC-PDA and pharmacokinetic study in rats

目的建立采用HPLC-PDA法同时测定大鼠血浆中黄芩苷和栀子苷含量的方法,并研究茵栀黄注射液在大鼠血浆中的药物动力学行为。方法色谱柱为DiamonsilC18柱,乙腈-0.3%甲酸水线形梯度洗脱,流速为1mL/min,检测波长分别为240nm和280nm,血浆样品采用甲醇直接沉淀蛋白的方法。结果黄芩苷和栀子苷的线性范围分别是15.25～7625μg/L和6.25～3125μg/L;日内与日间RSD<15%,提取回收率在70.45%和82.54%之间。大鼠单次尾静脉注射茵栀黄注射液4mL/kg后,2个主要药效成分黄芩苷和栀子苷的动力学参数如下:t1/2α为(7.46±4.86)和(3.50±4.26)min,V(c)为(0.0045±0.0024)和(0.0658±0.0806)mL/kg;AUC(0-t)为(11318.75±3550.51)和(683.41±272.90)ng/(min·L)。结论该方法简便、快速、准确、重现性好,适用于黄芩苷和栀子苷血药浓度测定及药代动力学研究。

Objective To set up a determination method of serum concentration of baicalin and geniposide from Yinzhihuang Injection by HPLC-PDA,and study the pharmacokinetics of Yinzhihuang Injection in rat serum.Methods The procedure of HPLC-PDA was performed on a column of Diamonsil C18,taking acetonitrile-0.3%formic acid as mobile phase in aqueous gradient elution.The flow rate was 1 mL/min and detection wavelength was 240 nm and 280 nm.The serum samples were treated with single-step protein precipitation.Results The linear ranges of baicalin and geniposide were,respectively,from 15.25 to 7625 ng/mL,and from 6.25 to 3125 ng/mL.The RSD values of intra-day and inter-day were less than 15%.The extractive recovery was between 70.45%and 82.54%.After the injection of Yinzhihuang Injection（4 mL/kg） in rats tail for one time,the pharmacokinetic parameters of baicalin and geniposide showed as follows：t 1/2α were（7.46±4.86） min and（3.50±4.26） min,V（c） were（0.004 5±0.002 4） mL/kg and（0.065 8±0.080 6） mL/kg;and AUC（0-t） were（11 318.75± 3 550.51） ng/（min·L） and（683.41±272.90） ng/（min·L） .Conclusion The method is simple,quick and accurate with a good repeatability,which can be used in the determination of serum concentration of baicalin and geniposide and pharmacokinetic study.