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

98 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS TABLE 5--CHROMATOGRAPHY OF BEESWAX UNSAPONIFIABLE MATTER Sample "a," Sample "b," Sample "c," Sample "d," Gm. Gm. Gm. Gm• Unsaponifiable matter 1. 696 1.688 1. 687 1. 705 Unsaponifiable matter insoluble in heptane and petroleum benzin (insoluble beeswax alcohols) 0. 987 1.014 0. 983 0. 928 Unsaponifiable matter soluble in heptane and petroleum benzin (by difference) 0. 709 O. 674 O. 704 O. 777 First petroleum benzln fraction* 0.427(13.6%) 0.423 (13.5%) 0.422(13.4%) 0.423 (13.3%) Second petroleum benzin fraction 0.001 0.000 0.000 0.000 First hot alcohol fraction 0. 276 0. 246 0. 275 0. 325 Second hot alcohol frac- tion 0.003 0.003 0. 003 0.021 * Av. 13.5%. Sample "a"--rate 5 ml. p.b./minute. Sample ?' --rate 4 ml. p.b./minute. Sample c' --rate 4 ml. p.b./minute. Sample "d"-:-rate 31/a mi. p.b./minute. TABI, V. 6---SOLUBILITY or BEv. swAx ALCOHOLS IN COLD MV. THYL ALCOHOL Beeswax Alcohols, Gm. Soluble Material, Expt. 1 Expt. 2 Av. Insoluble Material (by Difference) Av., Gm. 0.100 0.200 O. 300 0.400 0.500 0.026 0.028 0.027 0.073 0.040 0.039 0.040 0.160 0.046 ,0.044 0.045 0.255 0.054 0.054 0.054 0. 346 0.061 0.062 0.062 0.438 (12-14%) (2). The unsaponifiable material insoluble in heptane and petroleum benzin is also beeswax alcohols which are removed to de- crease the amount of material neces- sary to chromatograph. ' - T•. ANAL¾S•S or B•.•.SWAX- SPERMACET! ALCOHOL MIXTURES An empirical methoc• based on the solubility differences in cold methyl alcohol can be used to analyze mix- tures of the two alcohols. The pro- cedure is to treat a weighed sample with methyl alcohol, chill the solu- tion, and filter off the insoluble ma- terial. Evaporate the methyl alco- hol from the tiltrate and weigh the residue. Calculate the insoluble material, mostly beeswax alcohols, by difference. By use of Fig. 1, compute the weight'of soluble bees- wax alcohols associated with the insoluble beeswax alcohols. The sum Of the insoluble material and the soluble beeswax alcohols is ap- proximately equivalent to the total beeswax alcohols in the sample.