MIXTURES OF HYDROCARBONS, BEESWAX, AND SPERMACETI 99 Subtract the weight of the beeswax alcohols from the weight of sample taken the remainder is spermaceti alcohols. The results of a series of experiments employing this method are given in Table 7. All the results were in the range 100 •- 14%. It will be observed that the determinations of the major component of the mixture were ac- curate to •-5%. The minor com- ponent was determined with less accuracy. It will also be noted that with but one exception the re- coveries of spermaceti alcohol were somewhat low while those of the beeswax alcohols were high. In analyzing mixtdres of this type, it might be well to remember this and weigh the results accordingly. ANALYSI'S OF A MIXTURE OF HYDRO- CARBONS, BEESWAX, AND SPERM- ACETI Saponify and chromatograph the mixture of hydrocarbons, beeswax, and spermaceti according to the described experimental procedure. Combine the u.nsaponifiable alco- hols and treat a 0.5-gm. aliquot by the given solubility procedure. The data presented in Table 8 show the results obtained in a typical experi- ment by following procedure de- scribed. Estimation of Beeswax and Spermaceti The soluble alcohols (Table 8.) from a 0.5-gm. sample were foun'd to be 0.261 gm. The insoluble alcohols (by differ- ence) = 0.500 gm. - 0.261 gm. =. 0.239 gm.

100 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS From Fig. 1, the soluble beeswax alcohols which will be associated with the 0.239-gm. insoluble beeswax alcohols = 0.044 gin. ' Total beeswax alcohols in 0.5-gin. sample = 0.239 gm. + 0.044 gm. = 0.283 gm. But total alcohols = 0.906 gin. (Table 8). ' Beeswax alcohols in whole sample 0.906 - 0.5' X 0.283 = 0.513 gm. From Tables 4 and 5 beeswax alco- hols = 53.8% (total unsaponi- fiable material) - 13.5% (hy- drocarbons) = 40.3% of the bees- wax. 0.513 .'.Beeswax - 0.40• - 1.273 gm. (103.6% recovery). Total spermaceti alcohols = total alcohols - beeswax alcohols = 0.906 gm. - 0.513 gm. = 0.393 gm. and spermaceti alcohols = 49.7% of spermaceti (Table 2) 0.393 .: Spermaceti - 0.497 - 0.791 gin. (96.5 % recovery). Calculation of Non-Beeswax 'Hydro- carbons Total hydrocarbons = 5.693 gm. (Table 8). Beeswax hydrocarbons = 1.273 X 0.135 = 0.172 gm. ß Non-beeswax hydrocarbons = 5.693 gm. - 0.172 gm. = 5.521 gin. (99.5% recovery). EXPERIMENTAL PROCEDURE Saponification of Sample and the Chromatography of the Unsaponi- offable Fraction. S•/ponify sample by retluxing for two hours with 25 ml. of absolute alcohol, 1 gm. KOH, and 50 ml. of benzene. Transfer ß the saponified mixyure to a separa- tory funnel, add 75 ml. of hot water, shake well and draw off the aqueous layer.* Continue the ex- TABLE 8--ANALYTICAL DATA OF MIXTURE OF HYDROCARBONS• BEESWAX• AND SPERMA- CETI Gm. Sample: U.S.P. Mineral Oil ............... 5. 549 U.S.P. Beeswax ................. 1. 229 U.S.P. Spermaceti..: ............ 0. 820 Fatty acids ....................... 1. 034 Unsaponifiable matter ............. 6. 606 Unsaponifiable matter insoluble in heptane and petroleum benzin (insoluble beeswax alcohols) ...... .0.314 Unsaponifiable matter soluble in hep-. • tane and petroleum benzin (by difference) ...................... 6. 292 First petroleum benzin fraction* .... ' 5.690 Second petroleum benzin fraction... 0.003 First hot alcohol fraction ........... 0. 586 Second hot alcohol fraction ......... 0.006 Solubility of 0.5-gm. sample of com- bined alcohols in cold methyl alco- hol ............................ 0. 261 * Rate 4 ml. p.b./minute. traction with two additional 50-ml. portions of hot benzene. (Reserve the extracted aqueous solution.) Combine the benzene extracts wash with three 30-ml. portions. of 30% alcohol, and add the washings to the reserved extracted aqueous solu- tion. Filter the washed benzene •xtract through a' cotton plug into a 250-ml. tared beaker, evaporate the benzene on the steam bath, dry the residue in an oven at 100øC for 15 minutes, cool in a vacuum desiccator, and weigh as unsaponi- fiable m•.tter. Repeat the heating of the unsaponifiable matter in the oven until the weight is constant to 1-2 mg. • * Troublesome emulsions can be broken by fastening the stopcock of the separatory funnel with a rubber band and partially im- mersing the separatory funnel in a steam bath until the contents begin to boil.