UV SPECTROPHOTOMETRIC AND RP- HPLC METHODS FOR SIMULTANEOUS ESTIMATION OF ISONIAZID, RIFAMPICIN AND PIPERINE IN PHARMACEUTICAL DOSAGE FORM
Keywords:Isoniazid, Rifampicin, Piperine, UV Spectrophotometry, RP-HPLC, Validation
Objective: Simple, reliable, sensitive and accurate spectrophotometric and RP-HPLC methods for the estimation of INH, RIFA and PIPE in pure and pharmaceutical dosage form.
Methods: In the first Absorption correction method, methanol and distilled water were used as diluent. The wavelengths selected for the analysis were 262 nm, 338 nm and 477 nm for INH, PIPE and RIFA respectively. The Second RP â€“ HPLC method has been developed using Acetonitrile as diluent. Successful separation of drugs was achieved on LC18 100 Aâ° column (250 x 4.6 mm, 5 Î¼) using 0.01M Sodium Dihydrogen Orthophosphate, pH 6.5 and acetonitrile (40:60, % v/v) as mobile phase with flow rate of 0.9 mL/min. The wavelength of detection was 282 nm. Validation of developed methods was done according to ICH Q2 (R1) guideline.
Results: Calibration curve was linear over the concentration range of 12-34.5 Î¼g/mL (INH), 8-23 Î¼g/mL (RIFA) and 0.4-1.15 Î¼g/mL (PIPE) respectively for absorption correction method and 30- 330 Î¼g/mL (INH), 20-220 Î¼g/mL (RIFA) and 1-11 Î¼g/mL (PIPE) for RP â€“ HPLC method with r2 value greater than 0.995. Accuracy of methods was determined by recovery studies and it was found to be 98 to 102 %. The % RSD values for all the validation parameters were less than 2.0 % for both the methods.
Conclusion: The developed RP-HPLC and UV spectrophotometric method were successfully applied for the quantitative determination of cited drugs in pharmaceutical dosage form.
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