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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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Purpose: The purpose of this lab is to titrate an unknown solid acid (KH2PO4) with a standardized sodium hydroxide solution. After recording and plotting the data, the acid’s equivalence point will be recorded once the color changes. Using the equivalence point, the halfway point will be calculated, which is used to determine the acid’s equilibrium constant. The acid’s calculated equilibrium constant will be compared with the acid’s established pKa value.

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This addition aids in controlling the reproducibility and retention. Separation of the mixture via RP-HPLC can be done using continuous gradient or stepwise to move out the sample components. For every separation, the ideal gradient and volume must be

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TLC was used to identify the actual unknown product as well as other products/reactants present in the filtered solution. The procedure was conducted by placing a TLC plate in a developing chamber that is filled with a small amount of solvent. The solvent cannot be too polar because it will cause spotted compounds on the TLC plate to rise up too fast, while a very non-polar solvent will not allow the spots to move. The polarity of the spots also determines how far it moves on the plate; non-polar spots are higher than polar ones. After spots on the TLC form, the Rf values are calculated and used to analyze the similarity of the compounds.

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It forms a complex with HBr and extracts it from the aqueous phase into the organic phase where the alkene is. This dehydrates the acid, making it more reactive so that the addition reaction is possible. Rapid stirring is required in order to maximize the surface area

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The mobile phase used was a mixture of ammonium acetate buffer and acetonitrile at a ratio of 400:600. A flow rate of 1 mL/min was maintained, and the detection wavelength was 292 nm (22). The required studies were carried out to estimate the precision and accuracy of the HPLC method and were found to be within limits [percent coefficient of variation was less than 15%]. Sample preparation briefly involved 0.4 μ membrane filter through which the sample was filtered, diluted with mobile phase, and 10 μL was spiked into

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The substituents should therefore be hydrophobic and electron donating for maximum activity. • log 1/C is the concentration of drug that has a defined biological activity. The bigger the value of log 1/C , the smaller the value of c, the better the drug. • P is a measure of overall hydrophobicity. • π is the substituent hydrophobicity constant and measures the hydrophobicity of a specific region on the drug’s skeleton.

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The NaH2PO4 and Na2HPO4 powdered were weighted by using weighing machine, followed the mass that has been calculated in step (3). The NaH2PO4 and Na2HPO4 powdered were mix in a 500 mL beaker. 500 mL of distilled water were measured by using a 500 mL measuring cylinder, then is poured inside the 500 mL beaker containing both the powdered.

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