A sensitive and credible high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established and validated for the determination of isovitexin in rat plasma and various tissues (including heart, liver, lung, kidney, stomach, intestine, muscle, brain and cerebellum). The samples were prepared with methanol by liquid-liquid extraction, and puerarin was used as the internal standard. The chromatographic separation was carried out on an Agilent Poroshell 120 EC-C18 column (4.6mm×50mm, 2.7μm) with a mobile phase consisting of acetonitrile and 0.1% formic acid (21:79, v/v). The MS analysis was performed by multiple reaction monitoring (MRM) with electronic spray ionization source (ESI(-)) for quantitative response of isovitexin (431.0→311.0) and puerarin (415.1→295.0). The linearity of isovitexin in all the biosamples was good, with correlation coefficients greater than 0.9912 within the corresponding concentration range. The intra- and inter-day precisions in plasma and various tissues were less than 11.80%, and the accuracy (RE %) ranged from -4.89% to 4.78%. The extraction recoveries were in the range of 72.70%-90.81%. The present method was successfully applied to pharmacokinetics and tissue distribution of isovitexin in rats after tail vein injection with 2.0mg/kg of the compound. The pharmacokinetic parameters were demonstrated as followed: the half-life (t1/2) was 1.05±0.325h, the apparent volume of mean residual time (MRT) was 1.229±0.429h, and the area under the curve (AUC) was 11.39±5.05μg/mL/h. The results of tissue distribution showed that the main tissue depots for isovitexin in rats were kidney, intestine and liver. The results provided a meaningful insight for the further pharmacological investigation of isovitexin.
Keywords: HPLC-MS/MS; Isovitexin; Pharmacokinetics; Tissue distribution.
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