酶解法结合HPLC测定鸢尾苷在小鼠体内的组织分布  

Tissue distribution of tectorigenin after tectoridin gavage in mice by enzymatic hydrolysis and HPLC analysis

作　　者：华晶[1] 李雪[1] 唐仕炜 王凌[1] 蒋学华[1] 

机构地区：[1]四川大学华西药学院

出　　处：《华西药学杂志》2018年第1期66-69,共4页West China Journal of Pharmaceutical Sciences

摘　　要：目的 采用酶解与HPLC相结合的方法测定灌胃鸢尾苷后,小鼠组织及血浆中的鸢尾黄素及其Ⅱ相代谢物葡萄糖醛酸结合物的浓度,并研究其组织分布。方法 生物样品经β-葡萄糖醛酸苷酶水解,用乙酸乙酯萃取后进样分析;色谱柱为Shimadzu VP-ODS（250 mm×4.6 mm,5μm）,流动相为甲醇-乙腈-2%乙酸溶液（44∶10∶46）,流速1.0 m L·min^（-1）,柱温25℃,检测波长266 nm。予小鼠灌胃鸢尾苷,检测酶解前、后组织中的药物浓度。结果 鸢尾黄素在组织匀浆样品中,在标准曲线浓度范围内的线性关系良好,精密度及准确度均符合生物样品的测定要求。结论 所用的酶解与HPLC相结合的分析方法,适用于研究鸢尾黄素在小鼠体内的组织分布,给予小鼠灌胃鸢尾苷后,游离态和结合态鸢尾黄素在各组织中均有分布,且AUC0→∞大小的顺序为肾〉肝〉肺≈脾〉心。OBJECTIVE To determine tectorigenin and glucuronide - conjugated tectorigenin in mice tissue and plasma, then investi- gate their tissue distribution by a method combining β - glucuronidase hyrolysis with HPLC analysis. METHODS Bio - samples were hydrolyzed by β- glucuronidase and extracted with ethyl acetate. Chromatographic separation was performed on a Shimadzu VP - ODS column（250 mm ×4.6 mm,5 μm） with UV detection at 266 rim. A mixture of methanol - aeetonitrile -2% HAc aqueous solution （44 ： 10 ： 46） was used as the mobile phase at a flow rate of 1.0 mL min-1. After method validation,this method was applied to determine tectoridin metabolite in the mice tissue and plasma after tectoridin administered intragastrically to mice. RESULTS Calibra- tion curves for tectorigenin had good linearity in every kind of tissue homogenate sample and plasma over a certain range. The accuracies and precisions met the requirements of biological sample＇ s determination. CONCLUSION The present method is successfully applied to study the tissue distribution of tectoridin metabolite in mice. Glucuroxaide and free tectorigenin are widely distributed in mice and the levels of AUC0→∞ were kidney 〉 liver 〉 lung≈-spleed 〉 heart. Tectoridin is metabolized to tectorigenin and glucuronide - conju- gated tectorigenin and wide spread in mice after intragastric administration.

关 键 词：鸢尾苷 鸢尾黄素 Ⅱ相代谢物 Β-葡萄糖醛酸苷酶 酶解法 组织分布 高效液相色谱法 内标法 

分 类 号：R917[医药卫生—药物分析学]