By Erich Heftmann (Eds.)
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Additional resources for Chromatography of Steroids
Sample text
An advantage of alumina over silica in TLC of A4-3-ketosteroidsis that they form fluorescent oxidation products when the chromatographic plate is heated to 150-180°C 16 PAPER A N D THIN-LAYER CHROMATOGRAPHY [484]. The reaction appears to be specific and permits the detection of nanogram quantities of these steroids. Another sorbent used in TLC of steroids is polyamide [ 1 1691. Polyamide plastic sheets, developed with hexane-acetone (4: 1) for the less polar steroids or with chloroform-acetone (4: 1) for the more polar ones, produce satisfactory separations.
Borohydride reduction or periodate oxidation [401]. 2 SULFURIC ACID REACTION OF STEROIDS [423] Abbreviations: BE = blue; BK = black; BN = brown; BT = bright; DK = dark; DL = dull; ED = emerald; GN = green; GY = grey; LC = lilac; LT = light; OE = orange; PE = pale; PK = pink; PU = purple; RD = red; VY = very; YW = yellow. 00 BN PKGY GY-PK GY GY-PK PK-PU PK BN OE YW YWGY GY GY LT-BN GY-PK GY -PK RD-BN RD GY BE-PU GY YW BN-RD OE BN DK-BE OE BN OE YW GY DKGN OE PU OE BN BN OE GY YW-BE VY-PE-BE BE LT-BE PE-BE BE YW YWCN BT-BEGN BT-BE BT-BE PE-BE BE LC VY -PE-BE VY -PE-BE DL-BE PE-BE PE-BE BT-BE OE BT-BE BE BT-BE BE DL-BE DL-OE YWGN BT-LC BT-BE BE BTGN BE RD BT-BE BTGN BE BT-BE BT-BE 5-Androstene-3p,l7a-diol 1lp-Hydroxy-4-androstene-3,17-dione 1la,l7a-Dihydroxy-4-androsten-3-one GY-BN GY-PK GY-PK PE-YW PE-PK PK PE-PK OE YW-PK PE-YW GY-YW PE-YWCY GY-PK YW GY-PK PK PK PK PE-PK GY-PK GN PE-YE YW YW GY GN PK YW LT-PK PE-YW GY GN PE-YW PK YW OE-PK YW GY-OE PEGN-YW (Continued on p .
The silylation reaction was studied in detail by Yasuda [ 11881,by Chambaz and Horning [ 1671, and, more recently, by Gleispach [359]. Neither pyridine nor tetrahydrofuran were found necessary. Incubation at room temperature for 24 h gave more reproducible results and less byproducts than heating at 60°C. 1 shows the results of incubating 100 pg of various steroids with 200 pl of various reagents (500 p1 in the case of HMDS) at room temperature for 24 h. Phenolic hydroxyl groups were readily silylated by all reagents.
