• B. Shirisha
• D. Ashlesha
• N Mamatha

The UV-spectroscopic method and RP-HPLC method were developed and validated for the estimation of Lamivudine/Raltegravir per ICH guidelines. Buffer(opa): Acetonitrile (50:50) was used as the solvent. The λmax of Lamivudine/Raltegravir was found to be 302 nm and it was proved linear in the concentration range of Lamivudine 1-8μg/ml and for Raltegravir 2-16μg/ml with a correlation coefficient value of 0.999. Accuracy studies of UV-spectroscopy method was performed at three different levels, i.e., 50%, 100%, and 150% and recovery was found to be in the range of 99.6 to 100.8% for Lamivudine and the range of 98.3 to 101.2% for Raltegravir respectively. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.117 and 0.357 µg/ml for Lamivudine and 0.19 and 0.583 for Raltegravir. A simple, fast, accurate and precise RP-HPLC method was developed by using Acetonitrile: water, 0.1% ortho phosphoric acid(60:40) .The method was developed by using Kromosil 250 column (250 mm × 4.6 mm, 5μm particle size).and the mobile phase was pumped with Acetonitrile and water (hplc grade water)and PH was adjusted to 3.2 by using ortho phosphoric acid and the mobile phase was pumped at 1ml/min flowrate and the temperature was maintained at 30˚c.the retention time for Lamivudine/raltegravir were found to be 2.110 and 4.617 and the total run time was found to be 8min,the no of Theretical plates and Tailing factor of Lamivudine and Raltegravir were found to be 2584,1.36 and 3448,1.09, RP-HPLC method was found to be Linear in the range of Lamivudine/Raltegravir is 37.5-225µg/ml and 75-450µg/ml with a correlation coeffiecient of 0.999 the accuracy studies of RP-HPLC method was performed at three different levels i.e.,50%,100%,150% and recovery was found to be 99.24 to 100.13% for Lamivudine and 98.18 to 99.27% for Raltegravir respectively.Repeatability and intermediate results of Lamivudine/Raltegravir were found to be 0.55,1.33 and 0.8,0.4 Robustness of this method was performed by changing the flowrate at 0.9ml/min,1ml/min,1.1ml/min.the organic strength in the mobile phase was changed to 55:45v/v and 65:35 and results were found to be with in the limts.the % RSD was less than 2 the solution stability was determined at 0hr&24hr the %RSD of solutions were less than 2.the Ruggedness values were found to be 0.092 and 0.184 respectively. the %purity of the Dutrebis(label claim 150mg,300mg) was found to be 99.64 for Lamivudine and 100.5 for Raltegravir. The limit of Detection(LOD) and limit of Quantification(LOQ) were found to be 0.441 and 1.336µg/ml for Lamivudine and 0.031 and 0.093µg/ml for Raltegravir for RP-HPLC method. the sample was degraded in acidic,basic,peroxide,heat,photolytic,neutral and the results of Lamivudine/Raltegravir were found to be 5.85,2.6,3.66,1.88,0.83,0.92 and 5.66,2.76,4.89,1.81,0.78,0.69 respectively the above method was a rapid tool for routine analysis of Lamivudine/Raltegravir in the bulk and Pharmaceutical Dosage form.

METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF LAMIVUDINE AND RALTEGRAVIR BY UV SPECTROPHOTOMETRY AND RP-HPLC

"METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF LAMIVUDINE AND RALTEGRAVIR BY UV SPECTROPHOTOMETRY AND RP-HPLC", International Journal of Emerging Technologies and Innovative Research (www.jetir.org), ISSN:2349-5162, Vol.9, Issue 11, page no.g89-g99, November-2022, Available :http://www.jetir.org/papers/JETIR2211647.pdf

"METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF LAMIVUDINE AND RALTEGRAVIR BY UV SPECTROPHOTOMETRY AND RP-HPLC", International Journal of Emerging Technologies and Innovative Research (www.jetir.org | UGC and issn Approved), ISSN:2349-5162, Vol.9, Issue 11, page no. ppg89-g99, November-2022, Available at : http://www.jetir.org/papers/JETIR2211647.pdf