Bioanalytical method development and validation of novel antithrombotic agent S002-333 by LC-MS/MS and its application to pharmacokinetic studies.
A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using an electrospray ionization (ECI) source for the quantification of novel anti-thrombotic agent S002-333 [2-(4-methoxy-benzenesulfonyl)-2,3,4,9-tetrahydro-1H-β-carboxylic acid amide] in rabbit plasma was developed and validated. The extraction from plasma was carried out by simple protein precipitation extraction method. The chromatographic separation was performed on an Ultramex Cyno, (150 x 4.6 mm, 5 µm) with a guard column, using acetonitrile-water (75:25,v/v) with flow rate of 0.6 mL/min as the mobile phase. The tandem mass spectrometer was tuned in the multiple reaction monitoring mode to monitor the m/z transitions 386.4/215.4 for S002-333 and m/z 393.4/171 for the internal standard dexamethasone, using positive ion mode. The MS/MS response was linear over the concentration range from 1.56 to 200 ng/mL, with a lower limit of detection of 0.78 ng/mL. The accuracy and precision of the method were within the acceptable limit of ±20% at the lower limit of quantitation and ±15% at other concentrations and showed no significant matrix effect. The validated method can be used in most or all stages of the screening and optimizing process for future method validation of pharmacokinetic studies.
Pharmacokinetics and Metabolism Division, Central Drug Research Institute, CSIR, Post Box 173, Lucknow 226001UP, India. rabi.cdri@gmail.com
R S
RS
Bhatta
D
D
Kumar
Y S
YS
Chhonker
A K
AK
Saxena
G K
GK
Jain
D000818:Animals; D002243:Carbolines / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005343:Fibrinolytic Agents / Q000097:blood* / Q000493:pharmacokinetics; D008297:Male; D011817:Rabbits; D013449:Sulfonamides / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000379:methods*
C000592767:2-(4'-methoxybenzenesulfonyl)-2,3,4,9-tetrahydro-1H-pyrido(3,4-b)indole-3-carboxylic acid amide; D002243:Carbolines; D005343:Fibrinolytic Agents; D013449:Sulfonamides