title: LC-MS/MS analysis of metformin, saxagliptin and 5-hydroxy saxagliptin in human plasma and its pharmacokinetic study with a fixed-dose formulation in healthy Indian subjects.
abstract: A specific and rapid liquid chromatography-tandem mass spectrometry method is proposed for the simultaneous determination of metformin (MET), saxagliptin (SAXA) and its active metabolite, 5-hydroxy saxagliptin (5-OH SAXA) in human plasma. Sample preparation was accomplished from 50 μL plasma sample by solid-phase extraction using sodium dodecyl sulfate as an ion-pair reagent. Reversed-phase chromatographic resolution of analytes was possible within 3.5 min on ACE 5CN (150 × 4.6 mm, 5 μm) column using acetonitrile and10.0 mm ammonium formate buffer, pH 5.0 (80:20, v/v) as the mobile phase. Triple quadrupole mass spectrometric detection was performed using electrospray ionization in the positive ionization mode. The calibration curves showed good linearity (r2≥ 0.9992) over the established concentration range with limit of quantification of 1.50, 0.10 and 0.20 ng/mL for MET, SAXA and 5-OH SAXA respectively. The extraction recoveries obtained from spiked plasma samples were highly consistent for MET (75.12-77.84%), SAXA (85.90-87.84%) and 5-OH SAXA (80.32-82.69%) across quality controls. The validated method was successfully applied to a bioequivalence study with a fixed-dose formulation consisting of 5 mg SAXA and 500 mg MET in 18 healthy subjects. The reproducibility of the assay was demonstrated by reanalysis of 87 incurred samples.
authors: Department of Chemistry School of Sciences, Gujarat University, Ahmedabad, India.
authors: Priyanka A
authors: PA
authors: Shah
authors: Department of Chemistry School of Sciences, Gujarat University, Ahmedabad, India.
authors: Jaivik V
authors: JV
authors: Shah
authors: Department of Chemistry, St Xavier's College, Ahmedabad, India.
authors: Mallika
authors: M
authors: Sanyal
authors: Department of Chemistry School of Sciences, Gujarat University, Ahmedabad, India.
authors: Pranav S
authors: PS
authors: Shrivastav
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives* / Q000097:blood; D002853:Chromatography, Liquid / Q000379:methods*; D004151:Dipeptides / Q000097:blood*; D006801:Humans; D008687:Metformin / Q000097:blood*; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C558749:5-hydroxysaxagliptin; D004151:Dipeptides; D008687:Metformin; C502994:saxagliptin; D000218:Adamantane
keywords: 5-hydroxy saxagliptin; LC-MS/MS; human plasma; ion-pair solid phase extraction; metformin; saxagliptin