(1)Saturated solution sulphanilic acid in 5 per cent. hydrochloric acid.
(2)0.5 per cent. aqueous solution sodium nitrite.
(3)Strong ammonia.

Mix forty parts of (1) and one part of (2). In a test-tube take equal parts of this mixture and the urine, and pour 1 or 2 c.c. of the ammonia upon its surface. If the reaction be positive, a garnet ring will form at the junction of the two fluids; and upon shaking, a distinct pink color will be imparted to the foam. The color of the foam is the essential feature. If desired, the mixture may be well shaken before the ammonia is added: the pink color will then instantly appear in that portion of the foam which the ammonia has reached, and can be readily seen. The color varies from eosin-pink to deep crimson, depending upon the intensity of the reaction. A doubtful reaction should be considered negative.

7. Pancreatic Reaction.—Cammidge has shown that in cases of pancreatitis a substance capable of forming crystals with phenylhydrazin can be developed by boiling the urine with a mineral acid, and has offered the following test as an aid in diagnosis of this obscure condition. The nature both of this substance and the antecedent substance from which it is derived is not known. As originally proposed, the test was complicated and probably not trustworthy, but with his improved and simplified technic, Cammidge has had very promising results. In 200 consecutive examinations in which the diagnosis was confirmed, postmortem or at operation, 67 cases of pancreatitis (65 chronic, 2 acute) gave positive reactions; 4 cases of cancer of the pancreas were positive, 12 negative; 4 cases in which no pancreatitis was found were positive, 113 were negative. Normal urines do not give the reaction. The difficulty and importance of diagnosis in pancreatitis warrant inclusion of the method here even though its true value cannot be definitely assigned. While the test is somewhat tedious, all the manipulations are simple and require no apparatus but flasks, test-tubes, and funnels.

Technic.—Careful attention to detail is imperative. An ordinary routine examination is first made. Albumin and sugar, if present, must be removed: the former, by acidifying with acetic acid, boiling, and filtering; the latter, by fermentation with yeast after the first step of the method proper. An alkaline urine should be made slightly acid with hydrochloric acid.

(1) Forty cubic centimeters of the urine, which has been rendered perfectly clear by repeated filtration through the same filter-paper, are placed in a small flask, treated with 1 c.c. concentrated hydrochloric acid and gently boiled on a sand-bath for ten minutes, a funnel with long stem being placed in the neck of the flask to act as a condenser (Fig. 28). After boiling, the urine is cooled in a stream of cold water and brought to its original bulk with distilled water; 8 gm. of lead carbonate are then added to neutralize the acid. The fluid is allowed to stand a few minutes and then filtered through well-moistened fine-grained filter-paper until perfectly clear.

FIG. 28.—"Pancreatic reaction" flasks fitted with funnel condensers on a sand-bath (Robson and Cammidge).

(2) The filtrate is shaken up with 8 gm. powdered tribasic lead acetate and filtered. The excess of lead is then removed by passing hydrogen sulphid gas through the fluid or by shaking well with 4 gm. finely powdered sodium sulphate, heating to boiling, cooling to as low a temperature as possible in a stream of water, and filtering as before until perfectly clear.

(3) Ten cubic centimeters of the filtrate are then made up to 17 c.c. with distilled water, and added to a mixture of 0.8 gm. phenylhydrazin hydrochlorate, 2 gm. powdered sodium acetate, and 1 c.c. 50 per cent. acetic acid in a small flask with funnel condenser. This is boiled on a sand-bath for ten minutes, and filtered while hot through filter-paper moistened with hot water into a test-tube with a 15 c.c. mark. Should the filtrate not reach this mark, make up to 15 c.c. with hot distilled water. Allow to cool slowly.

(4) In well-marked cases of pancreatitis a yellow precipitate appears within a few hours; in milder cases, it may not appear for twelve hours. The microscope shows this sediment to consist of "long, light yellow, flexible, hair-like crystals arranged in sheaves, which, when irrigated with 33 per cent. sulphuric acid, melt away and disappear in ten to fifteen seconds after the acid first touches them" (Fig. 29).

FIG. 29.—Improved "pancreatic reaction." Crystals obtained from a case of chronic pancreatitis with gall-stones in the common duct (X200) (from a photo by P. J. Cammidge).