Bioanalytical Tandem Mass Spectrometry Method for Precise Measurement of Tranexamic Acid in Human Plasma With Unveiling Methodological Advancement: Green Assessment With Advanced Metrics

Abstract

For the control of excessive blood loss occurring from major trauma, postpartum bleeding, surgery, or hereditary angioedema, antifibrinolytic compound tranexamic acid (TXM) is highly efficient in controlling blood loss by reversibly binding with on lysine receptor sites. A selective and sensitive method has been developed and validated for the quantitation of TXM in human plasma using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). Tranexamic acid-D2 (TXM-D2) was used as internal standard (ISTD) to minimize the errors in TXM quantification. TXM quantification was performed with positive polarity mode using a Shimadzu high performance liquid chromatography coupled with AB-SCIEX API-4000 tandem mass spectrometer. A Zorbax Eclipse C18 (150 × 4.6 mm, 5 μ) column was used for the TXM quantification, 8-mM ammonium formate buffer with 0.1% formic acid ionization enhancer was used as a mobile phase-A, and acetonitrile was used as a mobile phase-B. (38:62) v/v portion of mobile phases A and B selected to elute the TXM and TXM-D2 with the flow rate of 0.8 mL/min. An electrospray ionization (ESI) technique was selected for detection of TXM in the human plasma. Linearity was assessed in the concentration range from 75 to 15,000 ng/mL by using least squares of weighting factor linear regression (1/X²).