Supremacy of Gradient Over Isocratic Elution in the Separation of Nine Pharmaceutical Active Ingredients Using Reversed-phase Separation and DryLab Modeling Software

Download or read book Supremacy of Gradient Over Isocratic Elution in the Separation of Nine Pharmaceutical Active Ingredients Using Reversed-phase Separation and DryLab Modeling Software written by Ahmad Alahmadi and published by . This book was released on 2018 with total page 314 pages. Available in PDF, EPUB and Kindle. Book excerpt: "In this project, nine pharmaceuticals drugs were randomly chosen, mixed, and dissolved in a 50:50 acetonitrile/water solvent and then separated using reverse phase HPLC. These chosen drugs are used commonly as relievers and antibiotics, as well as conditions such as migraines. The experiments used stock solutions for each drug with a concentration of 1000 ppm. Afterward, through trial and error, appropriate concentrations of aliquots for each drug were mixed to obtain good peak size for chromatographic purposes. Most of the research utilized various parameters in instrumentation and the mobile phase and were studied throughout the project to achieve optimum separation. These parameters include the selection of flow rates, UV spectra signals, injection volumes, isocratic and gradient percentages and time ramping rates, mobile phase solvents, solubility, buffers, and temperature changes. After each step of research, the experimental data was then tabulated and used with DryLab software to calculate and provide the optimum conditions for separation. Under these conditions, the final experiments were conducted and the data was then compared with the DryLab results. Comparison of using separation modes between gradient elution and isocratic elution was studied throughout this project in order to confirm that using gradient elution is the ideal mode to be considered chromatographic separation. At the end of the study, the optimum separation conditions were found with the use of an aqueous buffer of 25 millimoles of monobasic phosphate at a pH of 2.5. The temperature was set at 55 degree Celsius, and a linear gradient of blending MeoH and ACN (96:4) set at 5 to 55 percent with a run time of 30 minutes. The injection volume was set at 10 ul, flow rate at 1 ml per minute and the detection wavelength was set at 240 nm. Retention times were found to be from 4.056 to 27.747 minutes, and the average resolution was found to be 4.2 with a success rate 86.83%. Waters C18 (250 X 4.6mm, 5 μm) column used in this project provided an efficiency range from 7299 to 81,978 plates, and a tailing range from 0.995 to 1.2. The developed RP HPLC method was validated in terms of robustness and considered as robust."-- ‡c ProQuest database.