高效液相质谱联用法分析大鼠血浆中异鼠李素-3-O-新橙皮糖苷的浓度及其药代动力学研究
Analysis of isorhamnetin-3-O-neohesperidoside in rat plasma by liquid chromatography/electrospray ionization tandem mass spectrometry and its application to pharmacokinetic studies
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摘要: 目的:建立LC-MS/MS方法测定异鼠李素-3-O-新橙皮糖苷的浓度并研究其在大鼠体内的药代动力学。方法:8只大鼠分别尾静脉注射异鼠李素-3-O-新橙皮糖苷5 mg·kg-1。建立并确证高灵敏度的液质联用法分析大鼠血浆中异鼠李素-3-O-新橙皮糖苷的浓度,采用芦丁作为内标。采用甲醇蛋白沉淀法提取血浆中的异鼠李素-3-O-新橙皮糖苷和芦丁。然后用10 mmol·L-1的乙酸铵/甲醇(20:80,V/V)的流动相在C18色谱柱(150 mm×2.1 mm,I.D.,5 μm)上进行洗脱并采用多反应检测的方式进行质谱分析。结果:异鼠李素-3-O-新橙皮糖苷的在大鼠血浆中的线性范围为0.01-10 μg·mL-1。其定量下限为0.01μg·mL-1。结论:该方法可以用于测定大鼠血浆中异鼠李素-3-O-新橙皮糖苷的浓度。Abstract: AIM:To establish an LC-MS/MS method for determination of isorhamnetin-3-O-neohesperidoside and investigate its application on pharmacokinetic study in rats.METHODS:Eight rats were given 5 mg·kg-1 isorhamnetin-3-O-neohesperidoside after intravenous administration.A highly sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of isorhamnetin-3-O-neohesperidosidein rat plasma using rutin as internal standard.The analytes and rutin(internal standard) were extracted with methanol followed by a rapid isocratic elution with 10 mmol·L-1 ammonium acetate buffer/methanol(20:80,V/V) on a C18 column(150 mm×2.1 mm,I.D.,5 μm) and subsequent analysis by mass spectrometry in the multi-eaction-monitoring mode.RESULTS:The assays were linear over the concentration range of 0.01-10 μg·mL-1 for isorhamnetin-3-O-neohesperidosidein rat plasma.The lower limit of quantifications for isorhamnetin-3-O-neohesperidoside was 0.01 μg·mL-1.CONCLUSION:The validated method is successfully applied to determine the plasma concentrations of isorhamnetin-3-O-neohesperidosidein in rats.
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