Determination of sitagliptin in human urine and hemodialysate using turbulent flow online extraction and tandem mass spectrometry.
High turbulence liquid chromatography (HTLC, or turbulent flow online extraction) and tandem mass spectrometry (MS/MS) methods for the determination of sitagliptin in human urine and hemodialysate were developed and validated to support clinical studies. A narrow bore large particle size reversed-phase column (Cyclone, 50 mm x 1.0 mm, 60 microm) and a BDS Hypersil C18 column (30 mm x 2.1 mm, 3 microm) were used as extraction and analytical columns, respectively. For the urine assay, the LLOQ was 0.1 microg/ml, the linear calibration range was 0.1 to 50 microg/ml, the interday precision (R.S.D.%, n=5) was 2.3-6.5%, and the accuracy was 96.9-106% of the nominal value. For the urine quality control samples (QCs), the intraday precision (R.S.D.%, n=5) and accuracy were 1.8-2.6% and 96.2-106% of the nominal value, respectively. The interday precision (R.S.D.%) for 56 sets of urine QCs over a 6-month period varied from 3.8% to 5.5% and the accuracy from 102% to 105% of the nominal value. For the hemodialysate assay, the LLOQ was 0.01 ng/ml, the linear dynamic range was 0.01-5.0 ng/ml, the interday precision was 1.6-4.1%, and the accuracy was 89.8-104% of the nominal value. For hemodialysate QCs, the intraday precision and accuracy varied from 2.3% to 8.9% and from 99.8% to 111% of the nominal value, respectively. These results demonstrated that both methods are selective, accurate, precise, reproducible, and suitable for quantifying sitagliptin in hemodialysate and human urine samples.
Zeng
Wei
W
Department of Drug Metabolism, Merck Research Laboratories, Merck & Co. Inc., West Point, PA 19486-0004, USA. wei_zeng@merck.com
Musson
Donald G
DG
Fisher
Alison L
AL
Chen
Li
L
Schwartz
Michael S
MS
Woolf
Eric J
EJ
Wang
Amy Qiu
AQ
D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D015312:Hemodialysis Solutions / Q000032:analysis*; D006801:Humans; D011719:Pyrazines / Q000032:analysis* / Q000652:urine; D012680:Sensitivity and Specificity; D000068900:Sitagliptin Phosphate; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014230:Triazoles / Q000032:analysis* / Q000652:urine
D015312:Hemodialysis Solutions; D011719:Pyrazines; D014230:Triazoles; D000068900:Sitagliptin Phosphate