GC/MS法检测大鼠血浆中积雪酸

Determination of Asiatic Acid in Rat Plasma by GC/MS

建立大鼠血浆积雪甙代谢产物积雪酸的气相色谱-质谱(GC/MS)检测方法。在碱性环境中提取血浆样品中积雪酸,高氯酸四丁基铵(TBAP)与之形成离子对缔合物,经固相萃取、净化、衍生,采用内标法进行GC/MS检测。积雪酸和内标化合物乌苏酸的保留时间分别为15.2 min和10.4 min,最低定量限为1.0 ng/mL,药物浓度2.5、10、50 ng/mL测定日内RSD分别为5.48%、1.86%、1.32%(n=6),日间RSD分别为9.36%、5.11%、1.91%(n=90),相对回收率分别为101%、102%、99.1%。用GC/MS检测积雪甙在动物体内代谢产物积雪酸,有灵敏度高、专一性强的优点,能满足药代动力学研究要求。

Method for the determination of the asiaticoside metabolite asiatic acid （target compound） and ursolic acid （internal standard） was developed. The analytes were extracted from alkalized plasma, ion pair extracted with TBAP, further purified and concentrated by SPE method, derived and detected finally by GC/MS. The retention time for asiatic acid and ursolic acid were 15.2 rain and 10.4 rain. LLOQ of the method was 1.0 ng/mL. Relative recoveries for three concentrations （2.5,10,50 ng/mL） were 101%, 102% and 99.1% respectively, and intra-day RSD were 5.48%, 1.86%, and 1.32% （n=6）, inter-day RSD were 9.36%, 5.11% and 1.91% （n=90）. The method was sensitive, specific, reliable and suitable for pharmacokinetic study.