Development and Validation of Ultra Performance Liquid Chromatographic Method Simultaneous Estimation of Aspirin and Ticlopidine Hydrochloride in Pharmaceutical Formulation

Abstract

Objective: The combined dose tablet formulation containing aspirin and ticlopidine hydrochloride is used to reduce the formation of harmful blood clots in blood vessels which helps in preventing a heart attack or stroke in people with heart disease. The objective of the present work was to develop an accurate, precise, economical and less time consuming UPLC method and subsequently validate the method as per ICH guidelines, for routine quality control of combined dose tablet formulation containing above mentioned drug. Methods: The present work describes development and validation of ultra performance liquid chromatographic method for simultaneous quantitation of aspirin and ticlopidine hydrochloride in tablets. The chromatographic separation was carried out using Supelcosil LC-18 (3.3 cm × 4.6 mm × 3 µm) column using mixture of Potassium Phosphate buffer (0.01M) and Acetonitrile (20:80 v/v) as mobile phase with a flow rate of 0.6 ml per minute, total run time was 3 minutes. The detection and quantitation of chromatographic peaks was carried out at 230 nm. Results: The retention time for aspirin (ASP) and ticlopidine hydrochloride (TPH) was 0.529 minutes and 1.530 minutes, respectively. The linearity of detector response was observed in the concentration range of 20 – 60 µg/ml and 50-150 µg/ml for ASP and TPH, respectively. The LOD and LOQ for ASP and TPH was found to be 0.399 & 1.209 and 3.385 & 10.260, respectively. The accuracy of the method was found in the range of 99.92 – 100.58 % and 98.91 – 100.52 for ASP and TPH, respectively. The relative standard deviation for intra-day and inter-day precision studies was below 2 % indicating that the method is precise. The robustness of the proposed method was evaluated by varying the chromatographic parameters and the effect of these changes on retention time and tailing factor was studied. The % RSD for retention time and tailing factor was below 2 % indicating that the method is robust. The forced degradation studies were carried out to evaluate the specificity of the method. As the method was able to quantitate ASP and TPH selectively in presence of degradation products, the method was found to be specific. As the results of validation studies were within standard limits, the method was successfully applied for simultaneous estimation of ASP and TPH in marketed tablet formulation. Conclusion: The method has distinct advantages over reported UV and HPLC methods and hence the developed UPLC method can be used for routine quality control of Aspirin and Ticlopidine hydrochloride combined tablet formulation.