LC-MS/MS determination of 2-(4-((2-(2S,5R)-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.
As an effective DPP-IV inhibitor, 2-(4-((2-(2S,5R)-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 x 150 mm, 5 microm, 120 A) with a mobile phase of 20 mm 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 microL 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 < or =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.
Kim
Joseph
J
Department of Drug Analysis, Abbott Laboratories, Abbott Park, IL 60064, USA.
Flick
Jeanette
J
Reimer
Michael T
MT
Rodila
Ramona
R
Wang
Perry G
PG
Zhang
Jun
J
Ji
Qin C
QC
El-Shourbagy
Tawakol A
TA
D000284:Administration, Oral; D000818:Animals; D002138:Calibration / Q000592:standards; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004285:Dogs; D004355:Drug Stability; D052216:Glucagon-Like Peptide 1 / Q000097:blood / Q000187:drug effects; D057927:Hydrophobic and Hydrophilic Interactions; D015394:Molecular Structure; D011725:Pyridines / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry; D011759:Pyrrolidines / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
C515139:2-(4-((2-(2-cyano-5-ethynyl-1-pyrrolidinyl)-2-oxoethyl)amino)-4-methyl-1-piperidinyl)-4-pyridinecarboxylic acid; D011725:Pyridines; D011759:Pyrrolidines; D052216:Glucagon-Like Peptide 1