Development of assay method for analysis of a novel Kardiazol substance by HPLC

Abstract

Introduction Kardiazol ([3-allil-4-(41-metoxi fenil)-3N-thiazol-2-iliden]-(32-trifluoromethylphenyl) amin hydrobromide) is a novel substance that was synthesized in Danylo Halytsky Lviv National Medical University. Quality of medicines depends from many factors. One of the main is quality of its components and first of all active pharmaceutical ingredients (API). Therefore, developing assay method method for analysis of Kardiazol is quite necessary as far as it is promising substance that has cardioprotective [1], inta-inflammatory, analgetic, hypolipidemic and antioxidant effects. Materials and Methods HPLC analysis has been carried out with usage of Shimadzu Nexera X2 LC-30 AD (Japan). Separation was performed on ACE 5 C8 (250 4.6 mm, particle size μm). The binary solvent system of the mobile phase was used: the solvent A (water) and the solvent B (acetonitrile). The following linear gradient elution profile was used: 55% A/45% B–0 min, 55% A/45% B–3 min, 20% A/80% B–7 min, 20% A/80% B–14 min, 55% A/45% B–16 min and 55% A/45% B–20 min. The flow rate was 1mL/min and injection volume was 10 L. The column temperature was constant 35 ˚C. The chromatograms were recorded at 300 nm. The weights of substance was diluted in acetonitrile with final concentration 400μg/ml. Results and discussion In suggested conditions retention time of the investigated component was about 13.9 min. The column performance was determinate for its main indexes such as theoretical plate number (more then 65000) and symmetry factor (about 1.00). The method was validated according to Ukrainian Pharmacopoeia [2] and ICH guidelines [3] in terms of specificity, linearity, precision and accuracy. The validation data of proposed method met all requirements. The linearity was made and expressed by the following quadratic equation: R2=0.9998 (y=0.9956x+0.05031), the linearity range 80-120% of standard solution concertation.[...].