Analysis of isorhamnetin-3-O-neohesperidoside in rat plasma by liquid chromatography/electrospray ionization tandem mass spectrometry and its application to pharmacokinetic studies

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 C-18 column (150 mm x 2.1 mm, I.D., 5 mu m) 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 mu g.mL(-1) for isorhamnetin-3-O-neohesperidosidein rat plasma. The lower limit of quantifications for isorhamnetin-3-O-neohesperidoside was 0.01 mu g-mL(-1). CONCLUSION: The validated method is successfully applied to determine the plasma concentrations of isorhamnetin-3-O-neohesperidosidein in rats.