Rapid simultaneous clinical monitoring of five oral anti-coagulant drugs in human urine using green microextraction technique coupled with LC- MS/MS

Objectives: There is no analytical method for simultaneous monitoring of warfarin and novel direct oral anticoagulants (NOACs) in human urine samples in a single run in the clinics. Although several studies have reported their measurements in human blood samples, but its sample collection is more invasive and time-consuming, thereby challenging to monitor frequently. Methods: In this work, we developed a fast microextraction technique (ultrasound-assisted salt-induced liquid-liquid microextraction, USA-SI-LLME) coupled with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to rapidly quantify four commonly used NOACs drugs (apix-aban, dabigatran, edoxaban, and rivaroxaban) and warfarin in human urine samples. USA-SI-LLME con-ditions were optimized using a water-miscible organic solvent as an extraction solvent, high salt concentrations, sample pH, and extraction time (-5.5 min). Results and conclusions: The analytical method showed excellent linearities from 0.5 to 500 lg/L for apix-aban, edoxaban, rivaroxaban, warfarin, and 1 ti 500 lg/L for dabigatran. Intra-and inter-day precision values were <9.31% and R2 > 0.99 for all analytes. Limits of detection ranged between 0.07 ti 0.18 lg/L , and relative recoveries ranged between 92.18 and 110.15%. This method was successfully applied to analyze 15 one-spot urine samples from 15 clinical patients who regularly took warfarin or NOACs, and high accuracy was found. We concluded that this method could be used as a non-invasive high-throughput and rapid monitoring of NOACs and warfarin in human urine in clinical settings and could provide timely analysis during emergency care. (c) 2021 The Author(s). Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Automated summary

This study developed a fast microextraction technique coupled with HPLC-MS/MS to simultaneously quantify warfarin and four NOACs in human urine samples. The method showed excellent linearity, precision, detection limits, and relative recoveries, making it a reliable and rapid monitoring tool in clinical settings.