A Rapid and Sensitive LC-MS/MS Assay for the Determination of Saxagliptin and its Active Metabolite 5-hydroxy Saxagliptin in Human Plasma and its Application to a Pharmacokinetic Study

Abstract

The authors proposed a simple, rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay method for the simultaneous determination of saxagliptin and its active metabolite 5-hydroxy saxagliptin in human plasma. The developed method was fully validated as per the US FDA guidelines. The method utilized stable labeled isotopes saxagliptin-15 N d2 (IS1) and 5-hydroxy saxagliptin-15 N-d2 (IS2) as internal standards for the quantification of saxagliptin and 5-hydroxy saxagliptin, respectively. Analytes and the internal standards were extracted from human plasma by a single step solid-phase extraction technique without drying, evaporation and reconstitution steps. The optimized mobile phase was composed of 0.1% acetic acid in 5 mM ammonium acetate and acetonitrile (30:70, v/v) and delivered at a flow rate of 0.85 mL/min. The method exhibits the linear calibration range of 0.05-100 ng/mL for both the analytes. The precision and accuracy results for both the analytes were well within the acceptance limits. The different stability experiments conducted in aqueous samples and in matrix samples are meeting the acceptance criteria. The chromatographic run time was set at 1.8 min; hence more than 400 samples can be analyzed in a single day.

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

“…In addition, adequate retention of the analytes was a major challenge in the simultaneous analysis of MET, SAXA and 5‐OH SAXA because of their varying polarities. MET, being highly polar, has been shown to have poor retention on conventional C 18 reversed‐phase columns (Michel et al, ; Nielsen et al, ; Zhang et al, ), while SAXA and 5‐OH SAXA can be adequately retained on these columns within acceptable run times (Batta et al, ; Gao et al, ; Xu et al, ).…”

“…There are other reports in which MET has been determined with some DPP‐4 inhibitors, namely, vildagliptin (Pontarolo et al, ) and sitagliptin (Bonde et al, ; Reddy, Ahmed, Ahmad, Mukhopadhyay, & Thangam, ; Swales, Gallagher, Denn, & Peter, ; Scherf‐Clavel & Hogger, ). Likewise, a number of bioanalytical methods are presented for the determination of SAXA (Fura et al, ; Gao, Yuan, Lu, & Yao, ) and with its active metabolite 5‐OH SAXA (Batta et al, ; Xu et al, ) in different biological fluids. However, there are only two reports based on LC–MS/MS which were specifically intended to study the bioequivalence of SAXA/MET extended release (XR) (Gummesson et al, ) and immediate release (IR) (Upreti et al, ) FDC tablet formulation with corresponding monotherapy of these drugs in healthy subjects.…”

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

“…Batta et al (2015) reported an interesting method for the measurement of both saxagliptin and 5-hydroxysaxagliptin in human plasma. Because of the sensitivity issues, a positive ionization mode was preferred over that of negative ionization mode as it gave enhanced signal and permitted multiple reaction monitoring (MRM) detection with better selectivity (Batta et al, 2015). The choice of volatile buffer (i.e.…”

“…ammonium acetate) was crucial as part of the mobile phase as it provided the ionization potential and enabled better sensitivity with symmetrical peak shapes of both saxagliptin and 5-hydroxysaxagliptin. Lastly, the method employed solid-phase extraction (SPE), which not only enabled higher recoveries for both analytes but also gave an alternative option for sample extraction outside of protein precipitation and LLE (Batta et al, 2015).…”

A high‐performance liquid chromatography–tandem mass spectrometry method for simultaneous determination of imigliptin, its five metabolites and alogliptin in human plasma and urine and its application to a multiple‐dose pharmacokinetic study

“…HPLC-MS/MS method has been extensively used to investigate pharmacokinetic profiles of new drug owing to its high specificity, sensitivity and rapid analysis speed, especially for multiple reaction monitoring (MRM) scan mode (Batta et al, 2015;Diao et al, 2013;Shah, Shah, Sanyal, & Shrivastav, 2016;Wang, Jiang, Hu, & Zhao, 2017;Wang, Zheng, Wang, Hu, & Jiang, 2017). Therefore, in view of the previous method, a specific, sensitive and rapid method based on HPLC-MS/MS with MRM scan mode was developed and fully validated to quantify imigliptin, its five metabolites, and alogliptin in human plasma and urine.…”