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LC‐MS/MS determination of 2‐(4‐((2‐(2 S ,5 R )‐2‐Cyano‐5‐ethynyl‐1‐pyrrolidinyl)‐2‐oxoethylamino)‐4‐methyl‐1‐piperidinyl)‐4‐pyridinecarboxylic acid (ABT‐279) in dog plasma with high‐throughput protein precipitation sample preparation
Author(s) -
Kim Joseph,
Flick Jeanette,
Reimer Michael T.,
Rodila Ramona,
Wang Perry G.,
Zhang Jun,
Ji Qin C.,
ElShourbagy Tawakol A.
Publication year - 2007
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.855
Subject(s) - chemistry , chromatography , plasma , quantum mechanics , physics
As an effective DPP‐IV inhibitor, 2‐(4‐((2‐(2 S ,5 R )‐2‐Cyano‐5‐ethynyl‐1‐pyrrolidinyl)‐2‐oxoethylamino)‐4‐methyl‐1‐piperidinyl)‐4‐pyridinecarboxylic acid (ABT‐279), is an investigational drug candidate under development at Abbott Laboratories for potential treatment of type 2 diabetes. In order to support the development of ABT‐279, multiple analytical methods for an accurate, precise and selective concentration determination of ABT‐279 in different matrices were developed and validated in accordance with the US Food and Drug Administration Guidance on Bioanalytical Method Validation. The analytical method for ABT‐279 in dog plasma was validated in parallel to other validations for ABT‐279 determination in different matrices. In order to shorten the sample preparation time and increase method precision, an automated multi‐channel liquid handler was used to perform high‐throughput protein precipitation and all other liquid transfers. The separation was performed through a Waters YMC ODS‐AQ column (2.0 × 150 mm, 5 µm, 120 Å) with a mobile phase of 20 m m ammonium acetate in 20% acetonitrile at a flow rate of 0.3 mL/min. Data collection started at 2.2 min and continued for 2.0 min. The validated linear dynamic range in dog plasma was between 3.05 and 2033.64 ng/mL using a 50 µL sample volume. The achieved r 2 coefficient of determination from three consecutive runs was between 0.998625 and 0.999085. The mean bias was between −4.1 and 4.3% for all calibration standards including lower limit of quantitation. The mean bias was between −8.0 and 0.4% for the quality control samples. The precision, expressed as a coefficient of variation (CV), was ≤4.1% for all levels of quality control samples. The validation results demonstrated that the high‐throughput method was accurate, precise and selective for the determination of ABT‐279 in dog plasma. The validated method was also employed to support two toxicology studies. The passing rate was 100% for all 49 runs from one validation study and two toxicology studies. Copyright © 2007 John Wiley & Sons, Ltd.