Determination of Hypericin in Rat Plasma by HPLC-MS/MS / 中国药师

ABSTRACT

Objective: To establish an HPLC-MS/MS method for the determination of hypericin in rat plasma. Methods: The sample was analyzed by HPLC-MS/MS after the addition of internal standard and then protein precipitation using acetonitrile. The sepa-ration was carried out on an Ultimate C18 column (150 mm × 2. 1 mm,5. 0 μm). The mobile phase was composed of acetonitrile∶ 5 mmol·L-1 ammoniumacetate (containing 0. 1% formic acid) (90∶10) at a flow rate of 0. 5 ml·min-1 under 35℃. The detection was performed with multiple teactions monitoring ( MRM) using an electrospray ionization ( ESI) . The precursor/product ion transitions were monitored at m/z 503. 2→m/z 405. 1 for hypericin and m/z 355. 0→m/z 41. 9 for the internal standard pioglitazone ( anion mode). Results:The good linearity of hypericin was obtained within the range of 0. 1-13. 2 ng·ml-1. The correlation coefficient was more than 0. 99 and the lower limit of quantification was 0. 1 ng·ml-1. The extraction recovery was within the range of 84. 19%-98. 71%. The precision of intra-and inter-day was below 18. 47%. Conclusion: The method is fast, sensitive and accurate, which provides research basis for the clinical further study of hypericin.