Part 70

=357. Separation with Lime.=—A quicker, though perhaps not as accurate a separation of the oleic from the palmitic and stearic acids, is accomplished by means of lime according to the method developed by Bondzyuski and Rufi. This process is used chiefly, however, to separate the free fat acids (palmitic, stearic) from the neutral fat and the free oleic acid. It probably has no point of superiority over the lead process.

=358. Separation of the Glycerids.=—The fact that olein is liquid at temperatures allowing palmitin and stearin to remain solid, permits of a rough separation of these two classes of bodies by mechanical means. The mixed fats are first melted and allowed to cool very slowly. In these conditions the stearin and palmitin separate from the olein in a crystalline form and the olein is removed by pressure through bags. In this way lard is separated into lard oil, consisting chiefly of olein, and lard stearin, consisting largely of stearin. Beef (caul) fat is in a similar manner separated into a liquid (oleo-oil) and a solid (oleo-stearin) portion. It is evident that these separations are only approximate, but by repeated fractionations a moderately pure olein or stearin may be obtained.

=359. Separation as Lead Salts.=—Muter’s process, with a special piece of apparatus, has already been described (=339=). For general analytical work the special tube may be omitted. In a mixture of insoluble free fat acids all are precipitated by lead acetate, and the resulting soap may be extracted with ether, either with successive shakings or in a continuous extraction apparatus. In this latter case a little of the lead stearate or palmitate may pass into solution in the hot ether and afterwards separate on cooling. When the operation is conducted on from two to three grams of the dry mixed acids, the percentage proportions of the soluble and insoluble acids (in ether) can be determined. The lead salt which passes into solution can be decomposed and the oleic acid removed, dried and weighed. Dilute hydrochloric acid is a suitable reagent for decomposing the lead soap. The difference between the weight of the oleic acid and that of the mixed acids before conversion into lead soap furnishes the basis for the calculation. For further details in respect of the fat acids the reader may consult special analytical works.

=360. Separation of Arachidic Acid.=—Peanut oil is easily distinguished from other vegetable glycerids by the presence of arachidic acid.

The method used in this laboratory for separating arachidic acid is a modification of the usual methods based on the process as carried out by Milliau. About twenty grams of the oil are saponified with alcoholic soda, using twenty cubic centimeters of 36° baumé soda solution diluted with 100 cubic centimeters of ninety per cent alcohol. When the saponification is complete, the soda is converted into the lead soap by treatment with a slight excess of a saturated alcoholic solution of lead acetate. Good results are also obtained by using dilute alcohol, viz., fifty per cent, instead of ninety per cent, in preparing the lead acetate solution.

While still warm the supernatant liquid is decanted, the precipitate washed by decantation with warm ninety per cent alcohol and triturated with ether in a mortar four times, decanting the ethereal solution in each instance. By this treatment all of the lead oleate and hypogaeate are removed and are found in the ethereal solution, from which they can be recovered and the acids set free by hydrochloric acid and determined in the usual way.

The residue is transferred to a large dish containing two or three liters of pure water and decomposed by the addition of about fifty cubic centimeters of strong hydrochloric acid. The lead chlorid formed is soluble in the large quantity of water present, which should be warm enough to keep the free acids in a liquid state in which form they float as a clear oily liquid on the surface. The free acids are decanted and washed with warm water to remove the last traces of lead chlorid and hydrochloric acid. The last traces of water are removed by drying in a thin layer in vacuo. Practically all of the acids, originally present in the sample except oleic and hypogaeic, are thus obtained in a free state and their weight is determined.

The arachidic acid may be separated almost quantitively by dissolving the mixed acids in forty cubic centimeters of ninety per cent alcohol, adding a drop of hydrochloric acid, cooling to 16° and allowing to stand until the arachidic acid has crystallized. The crystals are purified by washing twice with twenty cubic centimeters of ninety per cent and three times with the same quantity of seventy per cent alcohol. The residual impure arachidic acid is dissolved in boiling absolute alcohol, poured through a filter and washed with pure hot alcohol. The filtrate is evaporated to dryness and heated to 100° until a constant weight is obtained. From the above data, the percentages of oleic, hypogaeic, arachidic and other acids in the sample examined are calculated.

In the above process, owing to the pasty state of the lead soaps, the trituration in a mortar with ether is found troublesome. The extraction of the lead oleate and hypogaeate is facilitated by throwing the pasty ethereal mass on a filter and washing it thoroughly with successive portions of about fifty cubic centimeters of ether. By this variation, it was found by Krug in this laboratory, that less ether was required and a more complete removal of the lead oleate effected. The solution of the lead oleate is completed by about half a dozen washings with ether as above described. The extraction may also be secured by placing the lead soaps in a large extracting apparatus and proceeding as directed in paragraph =40=. The residue is washed from the filter paper into a large porcelain dish and decomposed as already described with hydrochloric acid. After the separation is complete, the mixture is cooled until the acids are solid. The solid acids are then transferred to a smaller dish, freed of water and dissolved in ether. The ethereal solution is washed with water to remove any traces of lead salt or of hydrochloric acid. After the removal of the ether, the arachidic acid is separated as has already been described.

The melting point of pure arachidic acid varies from 73° to 75°.

=361. Detection of Arachis= (=Peanut=) =Oil.=—Kreis has modified the usual process of Renard for the detection of arachis oil, by precipitating the solution of the fat acid with an alcoholic instead of an aqueous solution of lead acetate, in a manner quite similar to that described above. The fat acids are obtained in the usual manner, washed with hot water and the acids from twenty grams of the oil dissolved in 100 cubic centimeters of ninety per cent alcohol. The solution is cooled in ice-water and the fat acids precipitated by the addition of fifteen grams of lead acetate dissolved in 150 cubic centimeters of ninety per cent alcohol. The precipitate, after standing for two hours, is separated by filtration through cotton wool and is extracted for six hours with ether. The residue is boiled with 250 cubic centimeters of five per cent hydrochloric acid until the fat acids appear as a clear oily layer upon the surface. The acids thus obtained are washed with hot water to remove lead chlorid, dried by pressing between blotting paper, dissolved in 100 cubic centimeters of ninety per cent alcohol, cooled to 15° and allowed to stand for several hours, after which time any arachidic acid present is separated by crystallization and identified in the usual manner.

When it is not important to obtain all of the acid present, the process may be simplified in the following manner:

The fat acids obtained from twenty grams of oil are dissolved in 300 cubic centimeters of ether and treated at the temperature of ice-water with a quantity of the alcoholic lead acetate solution mentioned above. Lead oleate remains in solution and the precipitate which forms after a few hours consists almost wholly of the lead salts of the solid fat acids. The precipitate is collected, washed with ether and identified in the usual manner.

=362. Cottonseed Oil, Bechi’s Test.=—Crude, fresh cottonseed oil, when not too highly colored, and generally the refined article, may be distinguished from other oils by the property of reducing silver salts in certain conditions. The reaction was first noticed by Bechi and has been the subject of extensive discussions.

The process as proposed by Bechi has been modified in many ways but apparently without improving it. It is conducted as follows: One gram of silver nitrate is dissolved in 200 cubic centimeters of ninety-eight per cent alcohol and forty cubic centimeters of ether and one drop of nitric acid added to the mixture. Ten cubic centimeters of the oil are shaken in a test tube with one cubic centimeter of this reagent, and then with ten cubic centimeters of a mixture containing 100 cubic centimeters of amyl alcohol and ten of colza oil. The mixture is divided into two portions, one of which is put aside for future comparison and the other plunged into boiling water for fifteen minutes. A deep brown or black color, due to the reduction of silver, reveals the presence of cottonseed oil.

In this laboratory the heating is accomplished in a small porcelain dish on which is often deposited a brilliant mirror of metallic silver. The white color of the porcelain also serves as a background for the observation of the coloration produced. In most instances a green color has been noticed after the reduction of the silver is practically complete. Unless cottonseed oil has been boiled or refined in some unusual way, the test, as applied above, is rarely negative. The reduction of the silver is doubtless due to some aldehydic principle, present in extremely minute quantities, and which may be removed by some methods of technical treatment. The silver nitrate test therefore is reliable when the reduction takes place, but the absence of a distinct reaction may not in all cases prove the absence of cottonseed oil.

=363. Milliau’s Process.=—Milliau has proposed the application of the silver salt directly to the free fat acids of the oil instead of to the oil itself. About fifteen cubic centimeters of the oil are saponified with alcoholic potash in the usual manner, 150 cubic centimeters of water added to the dish and the mixture boiled until the alcohol is evaporated. The fat acids are freed by the addition of decinormal sulfuric acid and as they rise to the surface in a pasty condition are removed with a spoon. The free acids are washed with distilled water. The water is drained off and the free acids dissolved in fifteen cubic centimeters of ninety-two per cent alcohol and two cubic centimeters of a three per cent solution of silver nitrate. The test tube containing the mixture is well shaken and placed in a water-bath, out of contact with light, and left until about one-third of the alcohol is evaporated. Ten cubic centimeters of water are added, the heating continued for a few minutes and the color of the supernatant fat acids observed. The presence of cottonseed oil is revealed by the production of a lustrous precipitate which colors the fat acids black. In some cases the process of Milliau gives better results than the original method of Bechi, but this is not always the case. It does away with the use of amyl alcohol and colza oil, but its manipulation is more difficult. In all doubtful cases the analyst should apply both methods.

=364. Detection of Sesame Oil.=—Milliau has pointed out a characteristic reaction of this oil which may be used with advantage in cases of doubtful identity. The identification is based on the fact that the free acids of sesame oil, or some concomitant thereof, give a rose-red color when brought in contact with a solution of sugar in hydrochloric acid.

The analytical process is conducted as follows: About fifteen grams of the oil are saponified with alcoholic soda and when the reaction is complete treated with 200 cubic centimeters of hot water and boiled until the alcohol is removed. The fat acids are set free with decinormal sulfuric acid and removed with a spoon as they rise to the surface in a pasty state, in which condition they are washed by shaking with water in a large test tube. When washed, the acids are placed in an oven at 105° until the greater part of the water is evaporated and the acids begin to become fluid. At this point they are treated with half their volume of hydrochloric acid saturated with finely ground sugar. On shaking the mixture, a rose color is developed which is characteristic of the sesame oil. Other oils give either no coloration or at most a yellow tint.

=365. The Sulfur Chlorid Reaction.=—Some vegetable oils, when treated with sulfur chlorid, give a hard product similar to elaidin, while lard does not. This reaction is therefore helpful in discriminating between some vegetable and animal glycerids. The process which is described by Warren has been used with some satisfaction in this laboratory.

Five grams of the oil or fat are placed in a tared porcelain dish and treated with two cubic centimeters of carbon bisulfid and the same quantity of sulfur chlorid. The dish is placed on a steam-bath and its contents stirred until the reaction is well under way. The heating is continued until all volatile products are evaporated, the hard mass being well rubbed up to facilitate the escape of imprisoned vapors. The powdered or pasty mass is transferred to a filter and washed with carbon bisulfid to remove all unaltered oil. The washing with carbon bisulfid is hastened by pressure and about 200 cubic centimeters of the solvent should be used. After drying, the weight of insoluble matter is obtained and deducted from the total weight of the sample used.

The color and tenacity of the hard, insoluble portion are characteristic. The quantitive part of the operation appears to have but little value, but applied qualitively in this laboratory it produces hard, leathery masses with cotton, olive and peanut oils, and but little change in lard and beef fats. Qualitively applied, the process is conducted as described above but without making the weighings. In this instance it is as easy of application as the process of Bechi and is deserving of greater attention than has been given it by analysts.

In the combination which takes place between the sulfur and the fat it is probable that only addition products are formed, since the quantity of alkali required for saponification is not diminished by previously treating the fat with sulfur chlorid. The reactions which take place are probably well represented by the following equations, in which oleic acid is treated with sulfur chlorid:

C₁₈H₃₄O₂ + S = C₁₈H₃₄S.O₂. C₁₈H₃₄S.O₂ + NaOH = C₁₈H₃₄SO₂Na + H₂O.

=366. Detection of Cholesterin and Phytosterin in Glycerids.=—Cholesterin is often found in animal glycerids and a corresponding body, phytosterin, is sometimes found in oils of a vegetable origin. When one of these two bodies is present it may be useful in distinguishing between animal and vegetable glycerids. They are detected as follows: Fifty grams of the glycerids in each case are saponified with alcoholic alkali, preferably potash, in order to have a soft soap. After saponification is complete, the alcohol is evaporated and the residual soap dissolved in two liters of water. The mixture is shaken with ether and the ethereal solution evaporated to a small bulk. The residue, which may contain a small quantity of unsaponified fat, is again treated with alcoholic potash and subjected a second time to the action of ether, as indicated above, with the addition of a few drops of water and of alcohol if the emulsion separate slowly. The ethereal extract finally secured is allowed to evaporate slowly and the cholesterin (phytosterin) is obtained in a crystalline form. The melting point of the cholesterin crystals is 146° and that of the phytosterin 132°.

Cholesterin crystallizes in thin rhombic tables while phytosterin separates in stellar aggregates or in bundles of long needles.

When dissolved in chloroform the two products show different color reactions with sulfuric acid, cholesterin giving a cherry and phytosterin a blue-red tint. In a mixture of animal and vegetable glycerids the two products are obtained together and the melting point of the mixture may afford some idea of the relative quantities of each present. It is evident, however, that no reliable judgment can be formed from these data of the relative proportions of the two kinds of glycerids in the original sample.

=367. Cholesterin and Paraffin in Ether Extracts.=—In ethereal extracts of some bodies, especially of flowers of the chrysanthemum, paraffin is found combined with cholesterin. The two bodies may be separated as follows: