VALIDATION AND ASSESSMENT OF THE HPLC METHOD PROFILE FOR THE QUANTITATIVE DETERMINATION OF THE API SODIUM 2-((4-AMINO-5-(THIOPHEN-2-YLMETHYL)-4H-1,2,4-TRIAZOL-3-YL)THIO)ACETATE

Abstract

Introduction. Derivatives of 1,2,4-triazole occupy an important place among modern biologically active compounds due to their broad spectrum of pharmacological properties. Of particular interest is sodium 2-((4-amino- 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazol-3-yl)thio)acetate, a promising active pharmaceutical ingredient with proven actoprotective and stress-protective effects. Its implementation into pharmaceutical practice requires reliable analytical methods for quality control. The Aim of the Study. To validate a rapid, accurate, and environmentally justified HPLC method for the quantitative determination of sodium 2-((4-amino-5-(thiophen-2-ylmethyl)-4H-1,2,4-triazol-3-yl)thio)acetate in the substance. Research Methods. The study was performed using high-performance liquid chromatography with diode-array detection (HPLC–DAD) on an Agilent 1260 Infinity system. Analysis was carried out under isocratic conditions on a Zorbax SB-C18 column using a water–acetonitrile mobile phase with the addition of 0,1 % formic acid. Method validation was conducted in accordance with ICH Q2(R1), the State Pharmacopoeia of Ukraine, and the European Pharmacopoeia, including evaluation of sensitivity, linearity, precision, accuracy, and environmental profile (AGREE, GAPI). Results and Discussion. The lower limit of quantification was established at 10 μg/mL with a signal-to-noise ratio of 12,6. The method demonstrated high linearity in the range of 80–120 % of the nominal concentration (R² = 0,9981), satisfactory precision (RSD ≤ 2 %), and accuracy (mean recovery 100,26 %). Assessment based on the principles of green analytical chemistry indicated an acceptable environmental profile of the developed method (AGREE = 0,74). Conclusions. The developed and validated HPLC method is rapid, sensitive, accurate, reproducible, and environmentally justified. It can be recommended for quality control of the substance sodium 2-((4-amino- 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazol-3-yl)thio)acetate and for use during pharmaceutical development and standardization of promising medicinal products based on this API.