A Sensitive and Rapid LC-MS/MS Method for Quantification of Aminocaproic Acid in Human Plasma and its Pharmacokinetic Application

Abstract

This paper describes the determination of aminocaproic acid in human plasma by simple, sensitive, and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using aminocaproic D6 acid as the internal standard (ISD). Extraction of aminocaproic acid and ISD was achieved from 200 μL plasma through protein precipitation with acetonitrile. Acetonitrile and 5 mM ammonium acetate buffer in the ratio 60:40 was used as the mobile phase at the flow rate of 1.0 mL/min. The method was developed with a short chromatographic run time of 3.0 min using Ace phenyl column. The retention times of both analyte and ISD were found to be 1.50 ± 0.1 min. The quantification of the separated components was done in positive ion mode by Multiple Reaction Monitoring (MRM) with mass transitions from m/z 132.2 (parent ion) to m/z 79.2 (product ion) for aminocaproic acid and from m/z 138.2 (parent ion) to m/z 74.3 (product ion) for the ISD. The method was validated as per the USFDA guidelines. The linearity was established over the concentration range of 0.3-80 μg/ mL with r2>0.98. The % recovery was 86.69 % and 98.29 %, respectively for aminocaproic acid and ISD. The inter- and intra- day accuracy and precision (%CV) were in the range of 100.91 to 103.80 % and 0.91 to 3.63 %, respectively. The developed method was successfully applied for pharmacokinetic study in twelve healthy adult male subjects under fasting conditions and the results were authenticated by performing incurred sample reanalysis.