Sample purification using a C18-bonded reversed-phase cartridge for the quantitative analysis of corticosteroids in adrenal cell cultures by high-performance liquid chromatography or gas chromatography--mass spectrometry.
Quantitative extraction and subsequent purification of small biological samples often involve cumbersome procedures. We have devised a short and efficient method for the quantitative extraction of the corticosteroid and the 20 alpha reduced steroid series from culture medium containing 20% sera in a single, pure fraction with separation from cholesterol. Passage through a C18-bonded reversed-phase Sep-Pak cartridge of the acidified culture medium and subsequent extraction of the steroid fraction with methanol yields a single fraction containing all steroids in 90% recovery and reduced quantities of cholesterol down to 30%. The extract can then be used without further purification for quantitative analysis by high-performance liquid chromatography or derivatized and analyzed by gas chromatography and gas chromatography--mass spectrometry.
D000305:Adrenal Cortex Hormones / Q000302:isolation & purification*; D000311:Adrenal Glands; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation; D008401:Gas Chromatography-Mass Spectrometry; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D000305:Adrenal Cortex Hormones