Determination of 3-Hydroxy pterocarpan, a novel osteogenic compound in rat plasma by liquid chromatography-tandem mass spectrometry: application to pharmacokinetics study

Abstract

A rapid, sensitive and selective LC-MS/MS method for the quantitative analysis of 3-hydroxy pterocarpan (S006-1709) in female rat plasma has been developed and validated. Discovery RP18 column was used for the chromatographic elution using acetonitrile and 0.1% acetic acid in water as mobile phase (80: 20 v/v) at the flow rate of 0.5ml/min. MS/MS analysis was performed in triple quadrupole mass spectrometer with electrospray ionization (ESI) in negative ion mode using biochanin as an internal standard (IS). Extraction of S006-1709 and IS from rat plasma was done by liquid-liquid extraction method using diethyl ether. The LC-MS/MS method was sensitive with 1.95ng/ml as the limit of detection (LOD) and 3.9ng/ml as lower limit of quantification (LLOQ). The method was linear in the concentration range of 3.9ng/ml to 1000ng/ml. The % bias for intraday and interday accuracy was not greater than 4.2 and the % R.S.D for intraday and interday precision was not greater than 13.2. The recovery of S006-1709 and IS were 73.9 to 79.3 % and 85.7% respectively. S006-1709 was found to be stable in various stability studies. The validated LC-MS/MS method was successfully applied for the oral pharmacokinetics study of S006-1709 at 10mg/kg in female S.D. rats.