高效液相色谱法测定大鼠血清中高良姜素

HPLC determination of galangin in rat serum

目的:建立测定大鼠血清中高良姜素的HPLC内标法。方法:SD大鼠腹腔注射给药,血清样品用乙酸乙酯萃取,以大黄素为内标;采用Shim-pack C_(18)柱(2.1 mm×250 mm,5μm),以A相:0.1%乙酸-水,B相:乙腈-甲醇-四氢呋喃(15:40:45)为流动相,梯度洗脱[0 min时,A-B(61:39);20 min时,A-B(39:61);30 min时,A-B(0:100);45 min时,A-B(0:100)],流速0.2 mL·min^(-1);检测波长为345 nm,柱温35℃。结果:高良姜素的线性范围为0.34～26.0μg·mL^(-1)(r=0.9998),最低定量限为0.34μg·mL^(-1),方法回收率为90.6%～111.9%,绝对回收率大于85%,日内和日间的RSD均小于6.3%;血清样品的稳定性符合要求。结论:所建立的大鼠血清中高良姜素的HPLC测定方法灵敏、准确、精密、稳定,适用于大鼠体内高良姜的药代动力学研究。

Objective：To develop an HPLC internal standard method for the determination of galangin in rat serum. Methods：The determination was performanced on Shim -pack C18 column（2.1 mm×250 mm,5μm）with the mobile phase consisting of 0.1%acetic acid - water（A） and acetonitrile - methanol - tetrahydrofuran（15：40：45;B） with gradient elution mode at a flow rate of 0.2 mL·min^-1.The following gradient procedure was used：0 - 20 min,39%→61%B;20 - 30 min,61%→100%B;30 -45 min,100%→100%B.Detection wavelength was 345 nm, the column temperature was 35℃.The route of administration of rats was ip,serum samples were extracted with ethyl acetate,methyl parahydroxybenzoate was used as an internal standard.Results：The linear range of galangin was 0.34 to 26.0μg·mL^-1（r = 0.9998 ） with the lower limit of quantification for 0.34μg·mL^-1.The method recovery was 90.6%to 111.9%and the absolute recovery was more than 85%.The intra - day RSD and inter - day RSD were less than 6.3%.Conclusion：The developed method for the quantification of galangin in rat serum is sensitive, accurate,precise,stable,and is suitable for the pharmacokinetic studies of galangal in rats.