Chromatography of Steroids by Erich Heftmann (Eds.)

By Erich Heftmann (Eds.)

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The effects of various treatments on the surface properties of glass capillaries have been studied in detail [15]. Instead of etching the glass capillaries, German and Horning [353] and German et al. [354] have conditioned the walls by adding very fine silanized silica particles to the stationary liquid. This produces a thermostable film that does not break up into micro droplets when the column is repeatedly heated and cooled.

They have desirable properties, not only in GC, but also in MS. 5 ml of pyridine at 60" for 3 h [469]. Steroids containing several hydroxyl groups in the molecule are not heated, but kept at room temperature overnight [294]. 2 ml BSA in the presence of 50-100 p1 of TMCS at room temperature for 3-5 h , if silylation of hindered groups is desired (see p. 31) 12811. In many cases, the syn and anti isomers of the methoximes are separated by GC as well as TLC [247]. The benzyl oximes (BO) are prepared by using 0-benzylhydroxylamine hydrochloride instead of 0-methylhydroxylamine, in analogy to the MO derivatives [231].

3). Ordinary thin-layer plates can be made fluorescent by spraying or predevelopment with fluorescent organic compounds. A spray reagent suitable for photography at 366 nm is prepared from 100 mg Rhodamine B, 35 mg 2’,7’-dichlorofluorescein, 150 ml diethyl ether, 70 ml95% ethanol, and 16 ml water [524]. 2% 2,2’-p-phenylene-bis(4-methyl-5phenyloxazole) in methanol, kept in the dark] can be used either as a spray (stock solution diluted ten times with a volatile solvent) or by predeveloping the plate with a dilute solution ( 5 ml of stock solution made to 100 ml with chloroform-methanol (1 : l ) ] to detect microgram quantities of most steroids under a short-wave W lamp [55].

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