title: 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.
abstract: 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.
authors: LifeSan Clinical Research, Division of Centaur Pharmaceuticals Pvt. Ltd., Centaur House, Near Hotel Grand Hyatt, Vakola, Santacruz (E), Mumbai, India 400055. ojitkr@yahoo.co.in.
authors: Ojikumar
authors: O
authors: Lukram
authors: Mukund
authors: M
authors: Zarapkar
authors: Chandan
authors: C
authors: Kumar Jha
authors: Shivaji
authors: S
authors: Parmar
authors: Keshav S
authors: KS
authors: Tomar
authors: Amit
authors: A
authors: Hande
mesh_terms: D000293:Adolescent; D000328:Adult; D002853:Chromatography, Liquid / Q000379:methods; D006801:Humans; D016014:Linear Models; D008297:Male; D010879:Piperazines / Q000097:blood* / Q000378:metabolism*; D000068799:Prasugrel Hydrochloride; D058921:Purinergic P2Y Receptor Antagonists / Q000378:metabolism*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods; D013876:Thiophenes / Q000378:metabolism*; D055815:Young Adult
chemical_list: D010879:Piperazines; D058921:Purinergic P2Y Receptor Antagonists; C510685:R-138727; D013876:Thiophenes; D000068799:Prasugrel Hydrochloride