By a combination of Sephadex gel filtration, CM-C and DEAE-C chromatography, 2 highly purified HCG fractions (dA: 18,000 IU/mg, dB: 20,000 IU/mg) were obtained from a crude preparation (2130 IU/mg) which was extracted from the urine of normal pregnant women in the first trimester. They were characterized by histological response of the ovary of hypophysectomized immature rats, bioassay (the Steelman-Pohley method and the OAAD method), amino acid analysis and determination of carbohydrate content, and their high homogeneity was proved by polyacrylamide gel electrophoresis, immunodiffusion and immunoelectrophoresis. The components confirmed were 17 amino acids, galactose, mannose, hexosamine and sialic acid. LH-like activity was dominant in fraction dA, and this fraction was richer in proline and valine than fraction dB and poorer in aspartic acid, glutamic acid, glycine, alanine, lysine and histidine. Fraction dB had dominant FSH-like activity in its biological character and was richer in acidic amino acids and sialic acid. Biological potency was particularly well correlated between LH-like activity of dA and hexose, and between FSH-like activity of dB and acidic amino acids and sialic acid. These results suggested that HCG is composed of 2 components. One has LH-like activity and the other has FSH-like activity. The ratio of these activities in HCG preparations differs according to the stage of pregnancy and biological and physicochemical properties of this hormone change throughout pregnancy. (Endocrinology 87: 233, 1970)

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