Determination of MK-0431 in human plasma using high turbulence liquid chromatography online extraction and tandem mass spectrometry.
A robust and sensitive method using high turbulence liquid chromatography (HTLC) online extraction with tandem mass spectrometry (MS/MS) for the determination of MK-0431 in human plasma was developed and validated to support the clinical studies. This HTLC online extraction method eliminated the time-consuming offline sample extraction procedures and significantly increased productivity. A narrow bore large particle size reversed-phase column (Cyclone, 50 x 1.0 mm, 60 microm) and a BDS Hypersil C18 column (30 x 2.1 mm, 3 microm) were used as extraction and analytical columns, respectively. The linear dynamic range of the calibration curve was 0.5 to 1000 ng/mL. Intraday validation was conducted using five calibration curves prepared in five lots of human control plasma, and the intraday precision (RSD%) was from 2.4 to 9.0% and the accuracy was from 98.0 to 103% of the nominal value. The intraday precision (RSD%, n = 5) for plasma quality control (QC) samples varied from 2.0 to 5.3% and accuracy from 103 to 105% of the nominal value. The interday precision (RSD%) for 100 sets of plasma QC samples in 29 analytical runs varied from 6.3 to 9.0% and the accuracy from 98.8 to 104% of the nominal value. No significant difference was observed between the interday and intraday precision and accuracy of the QC samples.
Department of Drug Metabolism, Merck Research Laboratories, Merck & Co. Inc., West Point, PA 19486-0004, USA. wei.zeng@merck.com
Wei
W
Zeng
Donald G
DG
Musson
Alison L
AL
Fisher
Amy Qiu
AQ
Wang
D002851:Chromatography, High Pressure Liquid; D005615:Freezing; D006801:Humans; D007004:Hypoglycemic Agents / Q000097:blood*; D009862:Online Systems; D011719:Pyrazines / Q000097:blood*; D011786:Quality Control; D015203:Reproducibility of Results; D000068900:Sitagliptin Phosphate; D012997:Solvents; D053719:Tandem Mass Spectrometry; D014230:Triazoles / Q000097:blood*
D007004:Hypoglycemic Agents; D011719:Pyrazines; D012997:Solvents; D014230:Triazoles; D000068900:Sitagliptin Phosphate