Method for determining concentration of sitagliptin in plasma by liquid chromatography-mass spectrometry

Abstract

The invention discloses a method for determining the concentration of sitagliptin in plasma by liquid chromatography-mass spectrometry. The method comprises the following steps: pretreating a plasma sample and determining the sample: taking K2EDTA as an anticoagulant and sitagliptin-d4 as an internal standard; the method comprises the following steps: adding 50 microliters of a sample into pores of a 96-pore plate, adding an internal standard sitagliptin-d4 working solution, uniformly mixing, adding methanol, sealing the plate, uniformly mixing, and centrifuging the sample at 4 DEG C for 10 minutes; adding 100 microliters of centrifuged supernate into another 96-hole collection plate, adding 300 microliters of 50% methanol, and uniformly mixing; centrifuging the sample at 4 DEG C to obtain a test sample; then taking 1 microliter of a test sample, injecting the test sample into a high performance liquid chromatography tandem mass spectrometer, detecting the chromatographic peaks of sitagliptin and internal standard sitagliptin-d4 in the sample, and calculating the sitagliptin concentration in the plasma sample according to the chromatographic peaks of the sitagliptin and the internal standard sitagliptin-d4; according to the detection method, sitagliptin-d4 is taken as an internal standard, a precipitant is added for protein precipitation, supernate is taken, a diluent is added, pretreatment is performed, separation is performed through a chromatographic column, detection is performed through a mass spectrometric detector, and the method is high in detection speed, high in precision and excellent in sensitivity.

Description

technical field [0001] The invention belongs to the technical field of medicine, and more specifically relates to a method for measuring the concentration of sitagliptin in plasma by liquid chromatography-mass spectrometry. Background technique [0002] Sitagliptin (SGT) is a dipeptidyl peptidase 4 (DPP-4) inhibitor that improves glycemic control in patients with type 2 diabetes by increasing the level of active incretin hormones. Incretin hormones, including glucagon-like polypeptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), are released from the gut throughout the day and rise in levels after a meal. Sitagliptin prevents DPP-4 from hydrolyzing incretin hormones, thereby increasing plasma concentrations of the active forms of GLP-1 and GIP. By increasing active incretin levels, sitagliptin increases insulin release and decreases glucagon levels in a glucose-dependent manner. These changes in insulin and glucagon levels reduce glycated hemoglobin A1...

AI Technical Summary

Problems solved by technology

[0003] At present, the analytical methods for the detection of sitagliptin in human plasma are uneven, and most of them use non-isotopic internal standards, which cannot guarantee the consistency of the elution method between the analyte and the internal standard

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