Development and validation of RP HPLC method for determination of thiotriazoline and its impurity in pharmaceutical dosage form
Ivković Branka, Žunić Jelena, Crevar SakaĠMilkica and Zorica Vujić
University of Belgrade, Serbia
: J Diagnos Tech Biomed Anal
Abstract
I n this paper, development of new RP-HPLC method for simultaneous determination of thiotriazoline and its impurity, 3-methyl- 1,2,4-triazolil-5-thione, in the tablets was described. The separation was achieved on the Hypersil GOLD aQ C18 (150x4 mm), 3 µm chromatographic column. The mobile phase consisted of phosphate buffer (pH 3.3) and MeOH (99:1, v/v). Column temperature was set on 30°C, mobile phase flow rate was 1.0 mL/min and detection wavelength was 220 nm. The developed method was validated according to ICH guidelines. Method is linear over the concentrations range 0.139 mg/mL to 0.238 mg/mL (r=0.9992) for thiotriazoline and 0.091 μg/mL to 1.56 μg/mL (r=0.9998) for investigated impurity. Precision was tested at two levels: intraassay precision and intermediate precision. Calculated relative standard deviations were 0.89% and 1.14%, respectively. Accuracy was tested at three concentrations levels (80%, 100% and 120%) and confirmed by calculated recovery values (100.03–101.50% for thiotriazoline; 74.65–88.43% for impurity). Small variations of mobile phase composition did not affect qualitative and quantitative system responses significantly, which proved method’s robustness. Examined impurity 3-methyl-1,2,4-triazolil-5-thione was detected in the tablets but its content was below the limit of quantitation (LOQ) defined by a proposed method (0.091 μg/mL).
Biography
Ivković Branka has completed PhD from University of Belgrade, Faculty of Pharmacy, Department of Pharmaceutical chemistry.
