DEVELOPMENT AND VALIDATION OF THREE NEW SPECTROPHOTOMETRIC METHODS FOR THE DETERMINATION OF LAMIVUDINE IN PURE AND PHARMACEUTICAL DOSAGE FORMS
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ABSTRACT
In this study three rapid, simple, accurate, economical and reproducible spectrophotometric methods for the quantitative determination of lamivudine in pure form and tablet formulations were developed and validated. The first method is based on dissolution of standard lamivudine powder or extraction of the drug from tablet formulation using methanol as solvent. The resulting extract was filtered and scanned using Helios Zeta Model 164617 UV/Vis spectrophotometer; having a λmax of 273 nm. The second method is based on the formation of a coloured hydrazone by reacting the hydrazine group in 2,4- dinitrophenyl hydrazine with the carbonyl carbon in lamivudine under acidic condition (85 % H2SO4) for 10 minutes. The orange-red coloured hydrazone formed was allowed to stand for 2 hours for complete colour development, then scanned using spectrophotometer and was observed to have a λmax of 438 nm. The third method is based on the diazotization reaction of lamivudine using sodium nitrite and sulfamic acid under acidic condition (2 % H2SO4) for 5 minutes followed by coupling with paratoluidine reagent leading to the formation of yellow chromogen which was allowed to stand for 1 hour for complete colour development, then scanned using spectrophotometer and was observed to have a λmax of 282 nm. The proposed methods were used to prepare a calibration curves for lamivudine and also to assay sample of standard lamivudine powder and three different brands of lamivudine tablets and compared with International Pharmacopoeial method for assay of lamivudine. The three proposed methods obeyed Beer’s law within a concentration range of 2.5 to 15.0, 5.0 to 35.0, and 2.5 vi to 12.5 µg/ml respectively. Correlation coefficients for the respective methods are 0.9997, 0.9988, and 0.9975. The precision (% coefficient of variation) and accuracy (% relative error) for the respective methods are 1.7, 1.8, 0.5 and 2.1, 4.0, 2.0 %. Percentage recoveries for the three methods obtained were 99.4, 98.9 and 99.6 % respectively. Limits of detection and limits of quantitation for the respective methods were 0.25, 1.3, 1.5 and 0.77, 3.8, 4.5 µg/ml. The percentage content of lamivudine in the standard powder and the three different tablet brands assayed in all the proposed methods were within the BP range of 97.5% to 102.0%. No statistically significant difference was observed between the percentage drug content of the proposed methods and International Pharmacopoeia method at P < 0.05. The proposed methods can be interchangeably used with the International Pharmacopoeia method for quantitative estimation of lamivudine in pure and tablet dosage forms.
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