Authors: Hua Wei Jun Wen Rui Xie Houwen Lin Guorong Fan Yutian Wu
Publish Date: 2009/09/17
Volume: 395, Issue: 5, Pages: 1461-
Abstract
A sensitive liquid chromatographytandem mass spectrometry LCMS/MS method followed by a 96well protein precipitation has been developed and fully validated for the determination of Phakellistatin 13 PK13 a new cyclic heptapeptide isolated from the sponge Phakellia fusca Thiele in rat plasma After protein precipitation of the plasma samples 50 μL in a 96well plate by methanol 200 μL containing the internal standard Pseudostellarin B 20 ng/mL the plate was vortex mixed for 3 min Following filtration for 5 min the filtrate was directly injected into the LCMS/MS system The analytes were separated on an XBC18 analytical column 5 µm 50 mm × 46 mm id using an eluent of methanol–water 8515 v/v and detected by electrospray ionization mass spectrometry in the negative multiple reaction monitoring mode with a chromatographic run time of 50 min The method was sensitive with a lower limit of quantification LLOQ of 01 ng/mL with good linearity r 0999 over the quantitation range of 01–5 ng/mL The validation results demonstrated that this method was significantly specific accurate precise and was successfully applied in measuring levels of PK13 in rat plasma following intravenous administration of 20 50 and 100 µg/kg of peptide in rats respectively which was suitable for the preclinical pharmacokinetic studies on PK13This work was supported by Shanghai Key Laboratory for Pharmaceutical Metabolites Research Grant NO 08DZ2270900 Pharmacokinetics Platform of the Innovation Drug Research and National HighTech Research and Development Project founded by Ministry of Science and Technology of PR China Grant NO 2006AA09Z423
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