Doctor of Philosophy in Clinical-Bioanalytical Chemistry, Cleveland State University, 2024, College of Arts and Sciences
This dissertation explores the influence of mobile phase (MP) polarity on the separation of steroid enantiomer pairs in reversed-phase high-performance liquid chromatography. Previous studies varied the concentration of a single organic modifier in binary MPs, affecting both the MP strength and polarity. This work isolates the effect of MP polarity on separation alone by using a ternary MP. In this work, MP strength is kept constant by varying the percent water, while the MP polarity is changed by varying the ratio of the two organic modifiers.
The combined ET(30) and Kamlet-Taft polarity model for MP polarity components of acidity, basicity, and dipolarity/polarizability was studied. The bile acid enantiomer pairs analyzed were alpha-muricholic acid and beta-muricholic acid, and omega-muricholic acid and gamma-muricholic acid. The separation factor (α) did not correlate with the total MP polarity, but did with a specific combined component polarity. The α versus the summed MP dipolarity/polarizability and basicity plot for all non-acetonitrile (ACN) ternary MPs experiments considered as a whole (six different organic pairs of methanol, isopropanol, dioxane, tetrahydrofuran in water), showed good correlation, except for the dioxane-containing MPs. Examination of previously obtained data of five other steroid enantiomer pairs (3β5β+3α5β-, 3β5α+3α5α-, 5α+5β-abiraterones, 17α- + 17β-estradiol, 11α- +11β-hydroxyprogesterone) showed good correlation with component polarity, excluding different aberrant organic modifier (isopropanol or methanol) MPs for several of these pairs. ACN-containing MPs demonstrated different separation characteristics, showing no correlation with non-ACN phases.
The dissertation also reports the separation versus component polarity of non-enantiomer compounds (theophylline, caffeine), revealing that changing the polarity of the non-ACN-containing MPs did not result in a significant change in separation. Plots for the ACN-containing MPs wer (open full item for complete abstract)
Committee: David Anderson (Advisor); Xue-Long Sun (Committee Member); Adam McShane (Committee Member); Yana Sandlers (Committee Member); Aimin Zhou (Committee Member)
Subjects: Analytical Chemistry