Abstract
Two couples of biological guaiacol estrogen isomers and a couple of synthetic ones were separated satisfactory by thin-layer chromatography by using continuous developing technique. The chromatographic plates were Merck silica gel 60 F254 (No. 5715) and spotted plates were developed continuously. Following solvents were used: benzene for the separation of 2-methoxy-estrone (II) and 2-hydroxyestrone-3-methyl ether (III), 2% acetone in benzene for the mixture of 2-methoxyestradiol(V) and 2-hydroxyestradiol-3-methyl ether (VI), and 5% acetone in benzene for 2-methoxy-3, 17 β-dihydroxyestra-1, 3, 5(10)-trien-6-one (VIII) and 3-methoxy-2, 17β-dihydroxyestra-1, 3, 5 (10)-trien-6-one (IX). Combination of the present separation technique with thin-layer chromato-scanner made possible for the estimation of guaiacol estrogens. Calibration curves between the spotted steroid amounts and peak area on chromatograms were shown to be linear in the range of (0.510.0) μg for II, III, V, and VI, and (0.210) μg for VIII and IX. Estimation of radioactive guaiacols obtained by the incubation of catechol estrogens with rat liver catechol O-methyl transferase in the presence of S-adenosyl-L-methionine (3H3C) was also possible by using radio thin-layer chromato-scanner.
