Rp Uppc Case Study

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To optimize the RP-UPLC parameters and reach a good resolution and peak Tailing for Lamivudine,Abacavir and Dolutegravir, many chromatographic parameters were tested. Several mobile phases of different ratios were analysed to get that good resolution ,peak shape ,provided sufficient selectivity for the drugs. The phosphate buffer provided a higher sensitivity and selectivity than other buffers did. Using methanol and acetonitrile as organic components shown results in higher sensitivity, but varying the amounts of methanol and acetonitrile in the mobile phase affected the resolution , tailing factor theoretical plates and run time. Varying the pH of the mobile phase resulted in poor peak shapes, not showing good resolution. So we introduced potassium dihydrogen phosphate and di potassium hydrogen phosphate into the mobile phase to…show more content…
The limit of quantification (LOQ) and limit of detection (LOD) were determined based on the slope and the standard deviation of the response using the signal-to-noise ratio (S/N) as per ICH Guidelines Q2(R1) 2005. The LODs for Lamivudine, Abacavir and Dolutegravir were found to be 0.021, 0.330 and 0.038µg/mL respectively, and the LOQs for Lamivudine, Abacavir and Dolutegravir were 0.056, 1.320 and 0.095 µg/mL respectively. (table 6 ) Robustness Robustness of the method was performed by making slight deliberate changes in the analytical methodology like flow rate and solvent ratio. It was observed that this method did not significantly affect in system suitability parameters like USP tailing factor, theoretical plates and resolution, which confirmed that the developed HPLC method is

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