By Erich Heftmann (Eds.)
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Additional resources for Chromatography of Steroids
Solution of NH4HS04 in abs. ethanol for 30 sec the adsorbent is modified in such a way that steroids can be detected in daylight or long-wave UV light by heating the plate at 140°C for 15 min [ 10671. Moreover, the modified adsorbent permits the resolution of the two 1 6 , 1 7 4 s epimers of estriol, which are unresolved on ordinary silica gel. Thin-layer plates prepared by mixing 10 g Silica Gel G with 22 ml of 5% aqueous cupric sulfate solution are suitable for the detection of steroids by charring at 180°C for 1 h .
Rhodamine 6G is generally less sensitive [ 1 1361. 6. Other additives have been suggested. For instance, silica gel plates prepared with 1% (NH4)2S04 solution instead of water have unaltered chromatographic properties, but when they are heated at 200°C for 20 min, sterols and other lipids are revealed as charred zones . By laying a silica gel plate into a satd. solution of NH4HS04 in abs. ethanol for 30 sec the adsorbent is modified in such a way that steroids can be detected in daylight or long-wave UV light by heating the plate at 140°C for 15 min [ 10671.
4 mg of steroids, dissolved in 200 pl dry pyridine, may be left overnight in a mixture of 200 p1 HMDS and 40 p1 TMCS [ 1 131. 5 ml tetrahydrofuran with twenty drops of HMDS and one drop of TMCS at 56°C for 3 h . O ml of N,O-bis(trimethy1silyl)acetamide [BSA, MeC(OSiMe,):NSiMe,] in a tightly stoppered vessel at room temperature overnight . 2 ml) may be added to promote solution. In any case, the excess reagents are later evaporated below 50°C. The silylation reaction was studied in detail by Yasuda [ 11881,by Chambaz and Horning [ 1671, and, more recently, by Gleispach .