ABSTRACT

Objective: The aim & objectives of this research work was to develop & validate a simple, selective, linear, precise, and accurate RP-HPLC method for the quantitative determination of Tramadol & Caffeine in bulk. Methods: Optimized liquid chromatographic condition was obtained by performing many trials for the selection of mobile phase and column, for the separation of Tramadol & Caffeine in RPHPLC. The method development was conducted with C18 column (250 X 4.6 mm, 5? particle size) with the flow rate of 1mL/min. the optimized mobile phase conditions were 1% Triflouroacetic acid, Acetonitrile and methanol in the ratio of 70:20:10 (v/v). The method was validated as per ICH guidelines. Results: The method found to be linear, accurate, rugged and robust for validated parameters. The linearity range was determined by external standard calibration method in the concentration range of 1?g/ml to 10?g/ml. The amount of recovery was calculated as 94% – 101% and it wasobserved that all the values are within the limits. The Retention time for Tramadol & Caffeine is observed as 3.01 and 7.39 minutes respectively. Further the precision of the method was confirmed by the repeatable analysis of sample. The results were found to be precise due to low values of the % RSD. It indicated that the method has good precision. Limit of detection for Tramadol & for Caffeine 0.077?g/ml and 0.006?g/ml respectively. Similarly limit of quantification for Tramadol & for Caffeine 0.233?g/ml and 0.017?g/ml respectively. In the robustness study %RSD obtained for change of flow rate and wavelength and ruggedness for change of analyst was found to be below 2, which was within the acceptance criteria. So, simple, sensitive, accurate, precise RP- HPLC methods were developed and validated for the simultaneous estimation of Tramadol & Caffeine. Keywords: RP-HPLC, Method development, Validation, Accuracy, Tramadol & Caffeine
Publication date: 01/09/2021

https://ijbpas.com/pdf/2021/September/MS_IJBPAS_2021_5646.pdf

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https://doi.org/10.31032/IJBPAS/2021/10.9.5646