高效液相法同时测定大鼠脑组织中积雪草苷和羟基积雪草苷浓度

Simultaneous Determination of Asiaticoside and Madecassoside in Rat Brain by HPLC

目的:建立高效液相色谱(HPLC)同时测定脑组织中积雪草苷和羟基积雪草苷浓度的方法。方法:大鼠尾静脉注射积雪草苷、羟基积雪草苷和积雪草总苷,30 min后用生理盐水进行心脏灌注,去除脑组织中残留血液,取脑组织,经甲醇沉淀蛋白后,利用HPLC测定脑组织中积雪草苷和羟基积雪草苷浓度。结果:积雪草苷和羟基积雪草苷在0.5-12μg/mL线性关系良好,定量下限为0.5μg/mL,日内和日间精密度(RSD)均小于15%,准确度(RE)在-5.25%-9.16%,提取回收率在70.54%-102.61%。结论:该方法符合生物样品测定要求,可以同时测定大鼠脑组织中积雪草苷和羟基积雪草苷的浓度。

Objective： To establish a determination method of asiaticoside and madecassoside concentration in rat brain by HPLC. Methods： After the rat caudal vein injection of total triterpene saponin of Centella asiatica, asiaticoside and madecassoside for 30 minutes, brain tissue was obtained by conducting the heart perfusion with physiological saline to remove residual blood in the brain. The brain tissue was extracted by methanol, then detected by HPLC for both asisticoside and madecassoside concentration. Results： The linear range was 0.5-12 μg/mL for asiaticoside and madecassoside. The lower limit of quantification was 0.5 μg/mL; The intra-day and inter-day variations of RSD were less than 15%, with relative error （ RE ） ranging from -5.25% to 9.16% and the extraction recovery ranging from 70.54% to 102.61%. Conclusion： This method can satisfy the requirement of biological determination of drugs and could be applied to concentration determination ofasiaticoside and madecassoside.