OESTROGENIC SUBSTANCES IN COSMETICS 315 (1 ' 2 - 97). Spray until wet. Heat at 130øC for 3-5 min, until optimal colours have appeared. For approximate Rf values and characteristic colours see Table III. Table III. Approximate Rf values and characteristic colours Approximate Rf values Colours Solvents Compound 1 2 3 Initial Final (1) Trans-DES 0.38 0.50 0.52 Purple Purple (2) Cis-DES 0.35 0.45 0.35 Purple Purple (3) Oestradiol-17[• 0.18 0.28 0.20 Light green Ultra-marine-blue (4) Oestradiol-17[•-dibenzoate 0.29 0.32 0.42 Light green Ultra-marine-blue (5) Oestrone 0.30 0.35 0.40 Blue 'Berlinblue' (6) Oestriol 0 0.02 0.04 Grey Grey Solvent 1: n-hexane-ethylacetate (70: 30). Unlined tank. Solvent 2: n-hexane-benzene-ethylacetate (70:20: 10). Unlined tank. Solvent 3: n-hexane-ethylacetate (60: 40). Unlined tank. Thin layer chromatography (TLC)--two dimensional (G). Silica plates without fluorescent indicator (ready made), 10 x 20 cm. First direction' solvent 3. Second direction: solvent 1. Left section for reference spots. Reference spots Rrsl --- Figure 1. Thin layer chromatography of phenolic steroids. 3 = oestradiol- 17[• 4 == oestradiol-17[•-dibenzoate 5 = oestrone 6 = oestriol. Thin layer chromatography: visualization of DES (H). This method is very specific for DES. Expose to shortwave u.v. light (e.g. Philips TUV, 15 Watt) for 3 min. DES will be transformed into a yellow phenanthrene derivative, which gives a yellow-brown fluorescence under longwave u.v. light (360 nm). Gas chromatography (I). The hormonal compounds are first silylated before application to the column. A suitable extract containing ca 50-100 Ixg of hormonal compound is used. Evaporate solvent. Add 0.2 ml N,O-bis(trimethylsilyl)

316 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS Table IV. Claimed action No. Product class Biological assay Chemical assay Anti-wrinkle (for older 1 Face cream a a a* --•- -- women) 2 Face cream a a a -- -- 3 Throat cream a a a -- -- 4 Face cream a a a -- -- 5 Face cream a a a -- -- 6 Face cream a a a -- -- 7 Face cream a a a -- -- 8 Eye cream a a a -- -- 9 Face lotion a a a -- -- 10 Face cream a a a -- -- 11 Placenta cream a a a -- -- 12 Eye cream a a a -- -- 13 Night cream a a a -- -- 14 Night cream a a a -- -- 15 Face cream a a a -- -- 16 Face cream a a a -- -- 17 Placenta cream a a a -- -- 18 Face cream f b/d f + :1: 0.0025 •o DES 19 Placenta cream a a a -- -- 20 Throat cream a a a -- -- 21 D6collet6 cream a a a -- -- 22 Face cream a a a -- -- 23 Face cream a a a -- -- 24 Face cream a a a -- -- 25 Face cream a a a -- -- 26 Placenta cream a a a -- -- 27 Beautifying oil a a a -- -- 28 Face cream a a a -- -- 29 Face cream a a a -- -- 30 Royal yelly cream a a a -- -- 31 Placenta extract oil a a a -- -- 32 Idem aqueous a a a -- -- 33 Face cream a a a -- -- 34 Face mask a a a -- -- 35 Face cream a a a -- -- 36 Face cream a a a -- -- 37 Face cream a a a -- -- 38 Face cream a a a -- -- 39 Face cream a a a -- -- Development of the 40 Bust cream a a a -- -- female breast 41 Paper plaids a a a -- -- 42 Paper plaids a a a -- -- Depression of 43 Body cream f f f + 0.1 •o DES abnormal hairgrowth 44 Body cream-mild f f f + 0.01 • DES in the female 45 Body cream a a a -- -- * Cytological evaluation (see preparation and staining of cytological specimen). • -- ----Oestrogenic action absent + = oestrogenic activity present. • Results of quantitative biological assay see Table V.