Determination of tranexamic acid in human plasma by UHPLC coupled with tandem mass spectrometry targeting sub-microgram per milliliter levels

Abstract

Tranexamic acid (TXA) is an antifibrinolytic drug, with the ability to inhibit lysine binding at plasminogen receptors, used in adult trauma patients with on-going or at risk of significant haemorrhage. To understand the pharmacokinetics and pharmacodynamics of this drug in variable age groups undergoing surgeries with high blood loss, effective methods for determination of TXA in biological samples at sub-μg mL −1 are still required. We describe herein the development and validation of a method based on ultra-high performance liquid chromatography coupled to triple quadrupole-tandem mass spectrometry to quantify TXA in human plasma. An inexpensive, simple and efficient sample clean-up was implemented, not requiring matrix-matching calibration. Sample preparation consisted in protein precipitation using acetonitrile containing 0.5% (v/v) formic acid, followed by hydrophilic interaction based chromatographic separation, with elution in isocratic mode using a combination of acetonitrile and water (75:25, v/v), with quantification of TXA based on selected reaction monitoring. Good linearity was achieved (r 2 > 0.997) for TXA concentrations ranging from 30 to 600 ng mL −1 , with LOD of 18 ng mL −1 in plasma. The developed method proved to be selective, sensitive, accurate (96.4-105.7% of nominal values) and precise (RSD ≤ 4.5%). TXA was found to be stable in plasma extracts standing 24 h at room temperature (20°C) or in the autosampler, and after three freeze-thawing cycles. Mean recovery values of TXA spiked plasma samples were ≥91.9%. No significant matrix effects were observed. The proposed methodology was successfully applied to the clinical study of plasma samples recovered during scoliosis surgery of pediatric patients pretreatment with TXA.

Functional Testing for Tranexamic Acid Duration of Action Using Modified Viscoelastometry

“…TXA plasma concentration was assessed by ultra-high-performance liquid chromatography/mass spectrometry (UHPLC-MS/ MS) as described by Barreiros et al [23].…”

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

“…Validation of developed analytical method was done as per mentioned in the FDA and ICH Guidelines for the evaluation parameters such as Specificity/Selectivity, Accuracy, Precision, Linearity/Range, Matrix Factor, Robustness, and Limit of Quantification (LOQ). 13,14…”

“…The data system Mass lynx Version 4.1 software had been used for the quantification purpose. 8,12,13…”

Acetonitrile Adducts of Tranexamic Acid as Sensitive Ions for Quantification at Residue Levels in Human Plasma by UHPLC-MS/MS

“…Despite the highest analytical signal for [M + H] + obtained with 50% (v/v) ACN (Figure 1B), TXA was not retained in the stationary phase in these conditions. Therefore, having in mind the development of an LC-MS/MS method for analysis of plasma samples [30], organic solvent percentages lower than 50% (v/v) were not considered.…”

“…Moreover, the significance of the observed adduct signals for the detection and quantification of TXA in biological samples was evaluated. In the course of our studies related to the development of an LC-MS/MS method to quantify TXA in human plasma, a protonated acetonitrile adduct ([M + ACN + H] + ) was observed as the most abundant ion formed under electrospray ionization in the positive mode (Scheme 1) [30]. Due to the stability displayed by this TXA-ACN protonated adduct ion under the reported conditions, its potential application was recognized for quantification purposes allowing to achieve higher sensitivity.…”