An integrative analytical approach designed for feasible tranexamic acid assay using o‐phthalaldehyde as a fluorogenic probe: applications to tablets, ampoules, and urine

Antifibrinolytic tranexamic acid (TRX) suppresses plasminogen activation to plasmin in a competitive way. TRX is approved for the management of heavy menstrual periods, hereditary angioedema, hemophilia, postpartum hemorrhage, surgery, tooth extraction, and severe blood loss after acute trauma. Here, the practical use of an isoindole derivative was established for a novel, easy‐to‐use, and affordable TRX assay. In the presence of a molecule containing a sulfhydryl group (2‐mercaptoethanol) 0.02% v/v, the primary amine moiety in TRX allows its combination with o‐phthalaldehyde to produce a luminous product. Excitation (338.8 nm) and emission (433.9 nm) wavelengths were used to monitor the isoindole fluorophore yield, and each operational variable was carefully examined and adjusted. The calibration graph was constructed with fluorescence intensity versus TRX concentration, excellent linearity was observed at concentrations between 40 and 950 ng/ml, and limit of detection and limit of quantitation were 41.3 and 13.6 ng/ml, respectively. The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines were used to validate the method. The developed method for TRX assay in various dosage forms and urine was successfully implemented and was shown to be an effective, simple, and quick replacement for the TRX assay in clinical trials and quality control. The current methodology is designed for Tranexamic acid assay in dosage forms and urine. The proposed method depends on isoindole formation between Tranexamic acid and o‐phthalaldehyde. The method lacks extraction and heating steps. The method was validated according to ICH guidelines.