Steroids Retained and Separated with a Unique Separation Strategy
Polarity of Anti Inflammatory and Birth Control Steroids and their chemical shapes were used to analyze them with HPLC. Retention Time profiles with 3 different Mobile Phases were compared to assess the likely impact of varying Acetonitrile and Methanol ratios in the Mobile Phase. The Organic Solvent percentage and Temperature dependence were studied for optimal Separation.
Temperature changes above 15°C had little effect on the Polarity based, Reversed Phase Selectivity and negative effects on Shape Selectivity. As a result 15°C was used.
The Chromatograms below show that Relative Retention Times (RRT) and Elution Order can vary when the ratios of the Selectivity Mechanisms are modified with Acetonitrile and Methanol changes.
4. Ethinyl Estradiol
Column: Cogent UDC-Cholesterol™, 4μm, 100Å
Catalog No.: 69069-75P
Dimensions: 4.6 x 75mm
—Chromatogram A. 56:20:24 DI Water with 0.1% Trifluoroacetic Acid (TFA) / Methanol / Acetonitrile
—Chromatogram B. 53:27:20 DI Water with 0.1% Trifluoroacetic Acid (TFA) / Methanol / Acetonitrile
—Chromatogram C. 56:24:20 DI Water with 0.1% Trifluoroacetic Acid (TFA) / Methanol / Acetonitrile
Flow rate: 1mL / minute
Detection: UV @ 240nm
No 27 Combining Polarity and Shape Recognition in HPLC pdf 0.2 Mb Download File