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High‐pressure liquid chromatography and ultraviolet spectrometry of ketonic C 27 sterols
Author(s) -
Hunter Irving R.,
Walden Mayo K.,
Bailey Glen F.,
Heftmann Erich
Publication year - 1979
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02533456
Subject(s) - chemistry , dichloromethane , chromatography , elution , ethyl acetate , hexane , silica gel , high performance liquid chromatography , ultraviolet , absorbance , mass spectrometry , organic chemistry , solvent , physics , quantum mechanics
A method for the separation of ketonic C 27 sterols was devised, based on high‐pressure liquid chromatography (HPLC) and ultraviolet absorption (UV). The adsorption column contained silica gel, particle size 10 μm, and the eluents were dichloromethane/ n ‐hexane/ethyl acetate (94∶5∶1) and dichloromethane/ethyl actate (99∶1) followed by dichloromethane/ethyl acetate (3∶1). The 5β‐sterols were eluted before the 5α‐analogs, sterols with isolated double bonds before conjugated carbonyl compounds, and ketones before hydroxy ketones. The effect of carbonyl groups on polarity depends on the position in the molecule and decreases in the order C‐3>C‐6>C‐7. The ultraviolet absorption spectra of eleven sterols were determined, and their absorbance at 254 nm and at 280 nm was used for analyzing the column effluent with a dual detector system.