Halphen Reaction for Cottonseed Oil.
—Carbon disulfid, containing about one percent of sulfur in solution, is mixed with an equal volume of amyl alcohol. Mix equal volumes of this reagent and the oil under examination and heat in a bath of boiling brine for fifteen minutes. In the presence of as little as one percent of cottonseed oil an orange or red color is produced, which is characteristic.
Lard and lard oil from animals fed on cottonseed meal will give a faint reaction; also the fatty acids thereof.
This test is more sensitive than the Bechi test (nitrate of silver) and less liable to give unsatisfactory results in the hands of an inexperienced person. It is not affected by rancidity. The depth of color is proportional, to a certain extent, to the amount of oil present, and by making comparative tests with cottonseed oil some idea as to the amount present can be obtained, but it must be remembered that different oils react with different intensities, and oils which have been heated from 200° to 210° C. react with greatly diminished intensity. Heating ten minutes at 250° renders cottonseed oil incapable of giving the reaction.
Cottonseed oil also has the property of reducing silver in silver nitrate to a metallic state. When mixed with a solution of silver nitrate under proper conditions a blackening or precipitation of black metallic silver is observed. This is known as the Bechi test which is conducted as follows:
Bechi or Silver Nitrate Test for Cottonseed Oil.
—Reagent: Dissolve 2 grams of silver nitrate in 200 cubic centimeters of 95 percent alcohol and 40 cubic centimeters of ether, adding one drop of nitric acid.
Mix 10 c.c. of oil or melted fat, 5 c.c. of reagent, and 10 c.c. of amyl alcohol in a test tube. Divide, heat one-half in a boiling water bath for ten minutes, and then compare with portion not heated. Any blackening due to reduced silver shows presence of cottonseed oil.
Other oils which have become rancid, and lards which have been steamed or heated at high temperature, contain decomposition products which have a reducing action on silver nitrate. There were found in testing a large number of salad oils some which contained no cottonseed oil, according to the Halphen test, but gave a brown coloration with Bechi reagent, and in some cases reduced silver. These same oils on being purified gave no reaction. Hence the oils or fats should be purified before testing.
To purify the oils and fats, heat from 20 to 30 grams on water bath for a few minutes with the addition of 25 c.c. of 95 percent alcohol, shake thoroughly, decant as much of the alcohol as possible, and wash with two percent nitric acid, and finally with water. The oil or lard thus purified will give no reduction at all if it contains no cottonseed oil. Heating the oils or fats to 100° C. or simple washing with two percent nitric acid is not sufficient, except in a few cases.