Development and Validation of Tranexamic Acid in Human Plasma by Using Ultra-Performance Liquid Chromatography-Mass Spectrometry

Abstract

Background: In the management of bleeding disorders, a synthetic lysine analogue Tranexamic acid is one of the drugs commonly used. Synthetic lysine analogue used in the management of bleeding disorders. The main purpose of the research aimed at developing and validating the new, simple, and cost–effective solid phase extraction method for evaluating tranexamic acid (TX) in human plasma by Ultra–performance liquid chromatography–Mass spectrometry (UPLC–MS/MS) using tranexamic acid as Internal Standard.Materials and Methods: Excellent separation and elution was fulfilled using acetonitrile: 100mM ammonium formate pH 3.5(60:40 v/v) as the mobile phase, at a flow rate of 0.300mL/min with the injection volume of 5µL. Chromatographic analysis of the analyte and IS was initiated under isocratic conditions with an aim to develop a simple separation process with a short run time.Results: The method showed a calibration curve range from 150.00ng/ml- 15,004.00ng/ml. validation of method was determined by using various supporting data of precision and accuracy with percentage recovery of 76.01% and 78.61% for TX and TXD2 respectively.Conclusion: An easy and sensitive UPLC– MS/MS method has been developed for the evaluation of TX in human plasma. This developed method provides excellent linearity over a wide range of TX, sensitive, and precise. Hence this technique is ideal and suitable for clinical pharmacokinetic studies of tranexamic acid.

Keywords: Tranexamic acid; Retention time; Tandem Mass Spectrometry; Method Validation; Human plasma