Development and validation of a liquid chromatography-mass spectrometric method for the determination of DPC 423, an antithrombotic agent, in rat and dog plasma.
A sensitive and selective LC-MS-MS method for the determination of DPC 423 (I), an antithrombotic agent, is described. This method used a solid-phase extraction from 0.1 ml plasma with an Isolute C(2) cartridge. HPLC separation was carried out on a YMC ODS-AQ C(18) column (50x2 mm) at a flow-rate of 300 microliter/min with an analysis time of 5 min. Compounds were eluted using a mobile phase of H(2)O/CH(3)CN/HCOOH: 66:34:0.1 (v/v/v), pH 4.0. A structural analogue of I was used as the internal standard to account for variations in recovery and instrument response. Mass spectrometric detection was carried out with a PE Sciex API III(+) triple quadrupole mass spectrometer equipped with a Turbo IonSpray source as the LC-MS interface. Good intra-day and inter-day assay precision (<10% CV) and accuracy (<10% difference) were observed over a concentration range of 0.005-2.5 microM in plasma. The extraction recoveries were approximately 90% and the method was found to be linear for the assay (r(2)>0.999). The method has been successfully applied to discovery and preclinical pharmacokinetic studies, including a dose range-finding study and toxicokinetic exposure studies in rat and dog.
Chi
Cecilia
C
Bristol-Myers Squibb Company, Metabolism & Pharmacokinetics, PRI, Experimental Station, Rt 141 and Henry Clay Rd., 19803-0353, Wilmington, DE, USA. cecilia.chi@bms.com
Liang
Li
L
Padovani
Patty
P
Unger
Steve
S
D000327:Adsorption; D000818:Animals; D000991:Antithrombins / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004285:Dogs; D013058:Mass Spectrometry / Q000379:methods*; D011720:Pyrazoles / Q000097:blood* / Q000493:pharmacokinetics; D051381:Rats; D015203:Reproducibility of Results; D013450:Sulfones / Q000097:blood* / Q000493:pharmacokinetics
C421624:1-(3-(aminomethyl)phenyl)-N-(3-fluoro-2'-(methylsulfonyl)(1,1'-biphenyl)-4-yl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxamide; D000991:Antithrombins; D011720:Pyrazoles; D013450:Sulfones