(1) Gerhardt's Test.—To a few cubic centimeters of the urine add solution of ferric chlorid (about 10 per cent.) drop by drop until the phosphates are precipitated; filter and add more of the ferric chlorid. If diacetic acid be present, the urine will assume a Bordeaux-red color which disappears upon boiling. A red or violet color which does not disappear upon boiling may be produced by other substances, as phenol, salicylates, and antipyrin.
(2) Lindemann's Test.—To about 10 c.c. of urine add 5 drops 30 per cent. acetic acid, 5 drops Lugol's solution, and 2 or 3 c.c. chloroform, and shake. The chloroform does not change color if diacetic acid be present, but becomes reddish-violet in its absence. This test is claimed by its advocates to be more sensitive and more reliable than Gerhardt's.
(3) Oxybutyric acid has much the same significance as diacetic acid, but is of more serious import. There is no satisfactory clinical test for it.
4. Bile.—Bile appears in the urine in all diseases which produce jaundice, often some days before the skin becomes yellow; and in many disorders of the liver not severe enough to cause jaundice. It also occurs in diseases with extensive and rapid destruction of red blood-corpuscles. Both bile-pigment and bile acids may be found. They generally occur together, but the pigment is not infrequently present alone. Bilirubin, only, occurs in freshly voided urine, the other pigments (biliverdin, bilifuscin, etc.) being produced from this by oxidation as the urine stands. The acids are almost never present without the pigments, and are, therefore, seldom tested for clinically.
Detection of Bile-pigment.—Bile-pigment gives the urine a greenish-yellow, yellow, or brown color, which upon shaking is imparted to the foam. Cells, casts, and other structures in the sediment may be stained brown or yellow. This, however, should not be accepted as proving the presence of bile without further tests.
(1) Smith's Test.—Overlay the urine with tincture of iodin diluted with nine times its volume of alcohol. An emerald-green ring at the zone of contact shows the presence of bile-pigments. It is convenient to use a conical test-glass, one side of which is painted white.
(2) Gmelin's Test.—This consists in bringing slightly yellow nitric acid into contact with the urine. A play of colors, of which green and violet are most distinctive, denotes the presence of bile-pigment. Colorless nitric add will become yellow upon standing in the sunlight. The test may be applied in various ways: by overlaying the acid with the urine; by bringing a drop of each together upon a porcelain plate; by filtering the urine through thick filter-paper, and touching the paper with a drop of the acid; and, probably best of all, by precipitating with lime-water, filtering, and touching the precipitate with a drop of the acid.
Detection of Bile Acids.—Hay's test is simple, sensitive, and fairly reliable, and will, therefore, appeal to the practitioner. It depends upon the fact that bile acids lower surface tension. Other tests require isolation of the acids for any degree of accuracy.
Hay's Test.—Upon the surface of the urine, which must not be warm, sprinkle a little finely powdered sulphur. If it sinks at once, bile acids are present to the amount of 0.01 per cent. or more; if only after gentle shaking, 0.0025 per cent. or more. If it remains floating, even after gentle shaking, bile acids are absent.
5. Hemoglobin.—The presence in the urine of hemoglobin or pigments directly derived from it, accompanied by few, if any, red corpuscles, constitutes hemoglobinuria. It is a rare condition, and must be distinguished from hematuria, or blood in the urine, which is common. In both conditions chemic tests will show hemoglobin, but in the latter the microscope will reveal the presence of red corpuscles. Urines which contain notable amounts of hemoglobin have a reddish or brown color, and may deposit a sediment of brown, granular pigment.