ICH-Compliant Development and Validation of RP-HPLC and UV-Visible Spectrophotometric Methods for Quantitative Estimation of Tapentadol in Bulk and Biological Matrices

Abstract

Background: Two novel methods, one an RP-HPLC technique and the other a chromogenic bioanalytical UV-Visible spectrophotometric method were developed and validated for the quantitative estimation of Tapentadol in bulk, pharmaceutical formulations, and human plasma. The bioanalytical method utilized a chromogenic reaction with Gibbs reagent to form a colored complex. Protein precipitation with methanol was used for the plasma extraction process for chromogenic bioanalytical method. Materials and methods: For Chromogenic Bioanalytical method plasma samples were prepared using protein precipitation with methanol. The reaction produced a blue complex measurable at 653 nm, while RPHPLC detection was performed at 270 nm. The RP-HPLC method was validated over a concentration range of 15–105 μg/mL. The mobile phase consisted of 0.1% formic acid (pH 6.8) in water and methanol (80:20, v/v) with a flow rate of 0.7 mL/min at temperature of 25ºC. These methods were rigorously validated to ensure accuracy, precision, and reliability. Results: For HPLC the retention time was recorded at 2.478min, precision was below 2% and the accuracy for HPLC was 99.97%. LLOQ and ULQC samples of bioanalytical method was found to be 20 and 240 μg/mL. Precision, expressed as relative standard deviation (RSD), was consistently below 10%, with in acceptable limits and robustness of the method was within limits. Conclusion: Chromogenic bioanalytical method provides high sensitivity by using Gibbs reagent for quantifying Tapentadol in human plasma. Bioanalytical method offer reliable quantification of TPD in biological matrices. HPLC method demonstrates adequate validation for every parameter, including peak purity assessment as well, ruggedness as well robustness, the range, its specificity along with accuracy, linearity and precision in compliance with ICH criteria. The study's findings demonstrated that this method was suitable for routinely quality control determining tapentadol levels in prescribed dosage forms and bulk.