96 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS Filtrate I . Hydrocarbons Spermaceti alcohols Soluble beeswax alcohols (Evaporate solvent and dissolve residue in petroleum benzin) CHART 1-•PREPARATION OF MD•:TURE FOR CHROMATOORAPHY Mixture Saponify and extract with CeHe C•IJ H•O Hydrocarbons Fatty acids Beeswax alcohols Spermaceti alcohols Treat with heptane and petroleum benzin R!sidue Insoluble b•eswax alcohols CHART 2--CHROMATOGRAPHY OF HYDROCARBONS, SPERMACETI ALCOHOLS, AND SOLUBLE BEESWAX ALCOHOLS Petroleum benzin solution of hydrocarbons, spermaceti alcohols, and soluble beeswax alcohols Pass through AltOs column using petroleum benzin as solvent Filtrate I Hydrocarbons I On AhOa column I Soluble beeswax alcohols Spermaceti alcohols I Pass hot ethanol through column I Flitrate: Spermaceti alcohols and soluble beeswax alcohols (Combine wi•h insoluble beeswax alcohols (Chart 1) and evaporate solvent) , CHART 3--SOLUBILITY OF COMBINED TOTAL BEESWAX AND SPERMACETI ALCOHOLS IN METHYL ALCOHOL Total beeswax and spermacetl alcohols Treat weighed sample with methyl alcohol ß I Filtrate I Soluble beeswax alcohols Spermaceti alcohols Evaporate methyl alcohol and weigh residue I Residue I Insoluble beeswax alcohols

MIXTURES OF HYDROCARBONS, BEESWAX, AND SPERMACETI 97 are 49.7% unsaponifiable matter and' 53.4% fatty acids. On chro- matographing the unsaponitiable matter, it will be adsorbed by the AlcOa from the' petroleum benzin solution and stripped from the AlcOa by hot ethanol (Table 3). The spermaceti 'alcohols obtained from the hot ethanol tiltrate will be al- most completely soluble in cold methyl alcohol. Although Table 7 indicates that 0.008 gm. out of 0.500 gm. of the spermaceti alcohols is insoluble, the 'value of 0.008 gm. TABLE 3--CHROMATOGRAPHY OF SPERMA- CETI UNSAPONIFIABLE MATTER Sample Sample "a," Gm. ø "b," Gm. Unsaponifiable matter 1. 894 1. 903 Unsaponifiable matter insoluble in heptane and petroleum benzin Nil Nil Unsaponifiable matter soluble in heptane and petroleum ben- zin (by difference) 1.894 1.903 First petroleum ben- zin fraction* 0.004 0.003 Second petroleum benzin fraction 0. 001 0.002 First hot alcohol frac- tion 1. 871 1. 879 Second hot alcohol fraction 0.006 0. 004 * Sample "a"--rate 4 mi. p.b./minute. Sample "b"--rate 3.6 ml p.b./minute. TABLE 4--THE SAPONIFICATION OF BEES- WAX Sample U.S,P. White Unsaponifi•ble Fatty Beeswax, Matter, Acids, Gm. Gm. Gm. 3.147 (a) 1.696(53.9%) 1.516(48.2%) 3.134(b) 1.688 (53.9%) 1.511 3.139 (c) 1.687(53.7%) 1.515(48.3%) 3.179 (d) 1.705 (53.6%) 1.534(48. Av. (53.8%) Av.,(48.3% i includes the errors of the manipula- tive procedure all of which tend to make the result high. Beeswax. The saponification con- stants of beeswax are 53.8% unsa- ponifiable matter and 48.3% fatty acids (Table 4). On chromato- graphing the beeswax unsaponifiable matter soluble in heptane and pe- troleum benzin, 13.5% (based on original weight of beeswax sample) will not be adsorbed by the AI=Oa from the petroleum benzin solution. The adsorbed material will be sub- sequently stripped by hot ethanol (Table 5). With the solubility pro- cedure used the combined beeswax alcohols (the ones insoluble in hep- tane and petroleum benzin added to the ones stripped from the column with hot ethanol) will vary in solu- bility in cold methyl alcohol from 12% for a 0.5-gm. sample to 27% for a 0.1-gm. sample (Table 6 and Fig. 1). • 0.07 0.06 0.05 • 0.04 x • 0.03 o• • o.oa m Lq, 0.01 133 o I I . INSOLUBLE BEESWAX ALCOHOLS, GRAMS Figure 1 It is known that hydrocarbons (1) are readily washed through an A1,.Oa column with' petroleum benzin and the value of 13.5% unadsorbed material falls within the range of hydrocarbons reported for. beeswax