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Electrospray ionization LC‐MS/MS validated method for the determination of the active metabolite (R‐138727) of prasugrel in human plasma and its application to a bioequivalence study
Author(s) -
Lukram Ojikumar,
Zarapkar Mukund,
Kumar Jha Chandan,
Parmar Shivaji,
Tomar Keshav S.,
Hande Amit
Publication year - 2012
Publication title -
drug testing and analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.065
H-Index - 54
eISSN - 1942-7611
pISSN - 1942-7603
DOI - 10.1002/dta.264
Subject(s) - bioequivalence , electrospray ionization , chromatography , active metabolite , metabolite , chemistry , electrospray , human plasma , pharmacology , mass spectrometry , pharmacokinetics , medicine , biochemistry
A rapid and sensitive liquid chromatography tandem mass spectrometry method has been developed and validated for the determination of the active metabolite (R‐138727) of prasugrel in human plasma. Because R‐138727 contains a thiol group, it requires stabilization by derivatizing with N‐ethyl maleimide. Commercially available trandolapril was used as the internal standard (IS). The derivatives of R‐138727 and IS were extracted from human plasma using a liquid‐liquid extraction technique. Chromatography was performed on a Hypurity C18, 5 µ (50 mm × 4.6 mm, i.d.) column, with the mobile phase consisting of acetonitrile and 10 mM ammonium formate (pH 3.0, 50:50 V/V), followed by detection using mass spectrometry. No significant endogenous peaks corresponding to R‐138727 or IS were detected in the blank human plasma samples and no significant matrix effect was observed for R‐138727 and IS in the human plasma samples. The mean recovery for R‐138727 ranged from 90.1 to 104.1%, with the lower limit of quantification set at 1 ng/ml. Linearity was established for concentrations in the range of 1.0–500.12 ng/ml, with a coefficient of determination (r 2 ) of 0.9958. The derivatized R‐138727 was stable in human plasma for 3 months at − 20 °C. This method increased the sensitivity and selectivity, resulting in high‐throughput analysis of R‐138727 using trandolapril as the IS in pharmacokinetic and bioequivalence studies, with a chromatographic run time of 3.7 min. Copyright © 2011 John Wiley & Sons, Ltd.