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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 method for the simultaneous determination of asiaticoside and madecassoside in brain by high performance liquid chromatography (HPLC). Methods: Asiaticoside, madecassoside and centella asiatica were injected into the tail vein of rats. After 30 min, the rats were perfused with saline, and the residual blood was removed from the brain tissue. The brain tissue was removed and precipitated with methanol , The contents of asiaticoside and madecassoside in brain tissue were determined by HPLC. RESULTS: Asiaticoside and madecassoside had a good linearity at 0.5-12 μg / mL with a lower limit of quantitation of 0.5 μg / mL. The intra-and inter-day precision (RSD) was less than 15%, and the accuracy (RE) -5.25% -9.16%, the recovery rate was 70.54% -102.61%. Conclusion: This method is in line with the requirements of biological samples for determination of asiaticoside and madecassoside in rat brain tissue.