Simultaneous determination of sitagliptin and simvastatin in human plasma by LC‐MS/MS and its application to a human pharmacokinetic study

Abstract

A simple, rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay method has been developed and validated for simultaneous quantification of sitagliptin and simvastatin in human plasma. Carbamazepine was used as an internal standard (IS). The analytes and IS were extracted from the human plasma by liquid-liquid extraction technique. The reconstituted samples were chromatographed on an Alltima HP C(18) column using an isocratic solvent mixture [acetonitrile-5 mm ammonium acetate (pH 4.5), 85:15 (v/v)] at a flow rate of 1.0 mL/min. Method validation was performed as per Food and Drug Administration guidelines and the results met the acceptance criteria. The calibration curves obtained were linear (r(2) ≥ 0.99) over the concentration range of 0.10-501 and 0.05-105 ng/mL for sitagliptin and simvastatin, respectively. The results of the intra- and inter-day precision and accuracy studies were well within the acceptable limits. Both the analytes were found to be stable in a battery of stability studies. The method is precise and sensitive enough for its intended purpose. A run time of 3.0 min for each sample made it possible to analyze more than 300 plasma samples per day. The developed assay was successfully applied to a pharmacokinetic study in human volunteers.

A concise review of the bioanalytical methods for the quantitation of sitagliptin, an important dipeptidyl peptidase‐4 (DPP4) inhibitor, utilized for the characterization of the drug

“…Several LC-MS/MS method have been reported for the estimation of sitagliptin in biological matrices, such as plasma, urine and hemodialysate. The majority of the reported methods are used in clinical applications (Zeng et al, , 2010Nirogi et al, 2008;Hess et al, 2011;Bonde et al, 2013;Burugula et al, 2013;Reddy et al, 2015;Scherf-Clavel and Högger, 2015) compared with preclinical applications (Yao et al, 2011;Gananadhamu et al, 2012;Patel et al, 2014). Shantikumar et al (2015) reported an LC-QTOF/MS method for the determination of five gliptins (vildagliptin, saxagliptins, sitagliptins, linagliptins and teneligliptin) in clinical application.…”

“…Patel et al (2014) reported a simple protein precipitation using MeOH which was free from matrix effect and gave high recovery and simple extraction. Nirogi et al (2008) and Burugula et al (2013) described various methods to quantitatively determine the plasma concentrations in human plasma using LLE using a combination of TBME, dichloromethane and n-hexane as extraction solvent. A C 18 column was used to measure sitagliptin concentrations from human plasma; it gives good peak shape, resolution and reproducibility.…”

A sensitive and selective liquid chromatography mass spectrometry method for simultaneous estimation of anti-diabetic drugs inhibiting DPP-4 enzyme in human plasma: overcoming challenges associated with low recovery and sensitivity

“…Literature reports highlights that few methods for quantification of gliptins (except teneligliptin) [8][9][10] are available. It also demonstrates poor recovery and sensitivity of vildagliptin and saxagliptin because of non-specific binding 11 .…”

Development and Validation of LC–MS/MS Method for Simultaneous Determination of Metformin and Four Gliptins in Human Plasma

“…Furthermore, different methods based on spectrophotometry [20,21]; spectrofluorometry [21,22]; capillary electrophoresis [23]; HPLC [24][25][26]; UPLC [27] and LC-MS/MS [28][29][30][31] were also developed for the quantification of sitagliptin in single as well as in multidrug component formulations including metformin. Linagliptin was reported to be quantitatively analyzed using HPLC [32] and LC-MS/MS [33,34] techniques.…”