Reversed-phase Separation : Separation of Ten Pharmaceutical Active Ingredients Using Gradient Elution and DryLab® Modeling Simulation
Author: Phillip R. Onagan
Publisher:
Published: 2017
Total Pages: 254
ISBN-13:
DOWNLOAD EBOOK"In this study ten pharmaceuticals were randomly selected, mixed, and dissolved in a 50:50 acetonitrile–water solvent and then separated through reverse phase HPLC. Stock solutions for each drug with a concentration of 1000 ppm were prepared for the experiments. 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 involved various parameters in instrumentation and the mobile phase and were studied throughout the project to obtain optimum separation. These include the selection of UV spectra signals, flow rates, injection volumes, isocratic and gradient percentages and time ramping rates, solubility, mobile phase solvents, buffers, and temperature changes. After each phase of research the experimental data was then tabulated and used with DryLab software to calculate and provide the optimum conditions for separation. Under those conditions the final experiments were conducted and the data was then compared with the DryLab results. At the end of the study the optimum separation conditions were found with the use of an aqueous buffer of 25 millimoles of dibasic phosphate at a pH of 6.05. The temperature was set at 55 degree Celsius, and a linear gradient set at 12 to 90 percent acetonitrile with a run time of 26.2 minutes. The injection volume was set at 10 ul and flow rate at 1.5 ml per minute. Retention times were found to be from 4.42 to 20.56 minutes, and the resolution ranges were from 9.24 to 32.24. The Supelco C18 column used in this project provided an efficiency range from 13,791 to 273,578 plates, and a tailing range from 0.703 to 1.931."--