An HPLC Method for the Determination of Saxagliptin in Human Plasma with Fluorescence Detection

Demet Dinçel,Mustafa Volkan Kızıltaş,Serife Evrim Kepekci Tekkeli

doi:10.24297/jac.v5i3.2678

Demet Dinçel, Mustafa Volkan Kızıltaş + Show 1 more

Open Access

https://doi.org/10.24297/jac.v5i3.2678

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Abstract

A sensitive and selective HPLC method with fluorometric detection was developed for the determination of saxagliptin (SGX) in human plasma and applied to a pharmacokinetic study. SGX was precolumn derivatized with fluorescamine, and the fluorescent derivative was separated on an RP C18 column using a mobile phase composed of acetonitrile-10 mM orthophosphoric acid by isocratic elution with flow rate of 1.3 ml/min. The method was based on the measurement of the derivative using fluorescence detection at 378 nm, with excitation at 463 nm. The calibration curve was linear over the range of 3.0–100.0 ng/ml. LOD and LOQ were found to be 0.15 and 0.5 ng/ml, respectively. Intraday and interday RSD values were less than 2.84%. The plasma concentration–time profile and pharmacokinetic parameters such as AUC0–t, AUC0–∞, Cmax, tmax, t1/2, were calculated according to the assays.

Highlights

Saxagliptin (SGX), (1S, 3S, 5S) - 2 - [(2S) - 2 - amino - 2 - (3 - hydroxy - 1 - adamantyl) acetyl] - 2 - azabicyclo [3.1.0] hexane - 3 - carbonitrile, is a widely used drug substance for treatment of type 2 diabetes mellitus [1]
To date two liquid chromatography with tandem mass spectrometry (LC–MS/MS) methods were developed and validated for SGX in plasma, one of them describes the assay for human plasma [3] and the other one is for rat plasma [4]
The proposed method is very sensitive with a 0.5 ng/ml limit of quantitation (LOQ) value. This is very important for SGX because the drug has very low plasma concentrations in the pharmacokinetic studies

Summary

Introduction

Saxagliptin (SGX), (1S, 3S, 5S) - 2 - [(2S) - 2 - amino - 2 - (3 - hydroxy - 1 - adamantyl) acetyl] - 2 - azabicyclo [3.1.0] hexane - 3 - carbonitrile, is a widely used drug substance for treatment of type 2 diabetes mellitus [1] It is a reversible dipeptidyl peptidase 4 (DPP4) inhibitor. To date two liquid chromatography with tandem mass spectrometry (LC–MS/MS) methods were developed and validated for SGX in plasma, one of them describes the assay for human plasma [3] and the other one is for rat plasma [4] They are fairly sensitive, accurate and precise methods, a simplier and more cost reduced method is required for the analysis of SGX in human plasma

Methods

Results

Conclusion