; . .... .... .... ... -....> . ../ \ > ; . . 7 v . • ' ■' i ■ 1 .. ' .. / ! . . .■ : . . . / ■ . ■ ■■ ■' ". ■ ■ ■ x .k . - . • ' J ' ... . y \ ' y ■y.' • ■■ y . THE URINE AND CLINICAL CHEMISTRY OF THE GASTRIC CON- TENTS, THE COMMON POISONS, AND THE MILK. BY J. W. HOLLAND, M.D., PROFESSOR OF MEDICAL CHEMISTRY AND TOXICOLOGY, JEFFERSON MEDICAL COLLEGE OF PHILADELPHIA. ILLUSTRATED. FIFTH EDITION, REVISED AND ENLARGED. PHILADELPHIA : P. BLAKISTON, SON & CO., 1012 Walnut Street. 1895. Copyrighted by P. Blakiston, Son & Co., 1895. PRESS OF WM. F. FELL &. CO., 1220-24 SANSOM ST., PHILADEL°HIA. PREFACE. This book is intended to be used as a syllabus for the laboratory. The text is made brief and to the point, so as to make a pocket volume, handy for reference. Pages are left blank for calculations, memoranda, or more extended notes to be made' by the student. For the convenience of those whose course of study is very short, the most important matter is printed in the larger type. Provision is made for more thorough study by the explanations and quantitative processes given in the small print. To this edition has been added a chapter on " the gastric contents," in which are given the detailed pro- cedures for chemical study of gastric indigestion. Much that is new has been incorporated with the text of the other parts, and the whole has undergone thorough revision. III TABLE OF CONTENTS. PAGE Composition of Healthy Urine, viii Reagents and Apparatus, viii Plan of Clinical Study, 9 EXAMINATION OF MORBID URINE. The Sample: Its Putrefaction, . . .. io Color, 11 Bile Pigment: Gmelin's Test, 12 Bile Acids: Oliver's Test, 12 Odor, 13 Specific Gravity, 13 Quantity, 13 Anuria, 13 Hydruria, .... 13 Polyuria, 14 Solid Urine 14 Reaction, 14 Chlorides: Tests for, 15 Sulphates: Tests for, 17 Phosphates: Tests for, 18 Deposits of, 20 Uric Acid, 22 Murexide test, 24 Deposits of, 24 Urates, 25 Test for, 25 Hippuric Acid, 26 Creatinine, 27 Calcium Oxalate, 27 Cystin, 28 Leucin and Tyrosin 29 Urea: Tests for, 30 Ureometry: Fowler's Method, 31 Russell and West's Method, 31 Lyon's " 32 Liebig's " 34 Sugar : Bottger's Test for, 35 Trommer's " " . . . . / 37 Glycerine Cupric " 37 Fehling's " " 38 Pavy's " " 39 IV TABLE OF CONTENTS. V PAGE Johnson's Picro-saccharometer, 40 Fermentation test for, 42 Indigo-carmine test for, 43 Phenyl-hydrazin test for, 43 Acetone 44 Albumin: Heller's Test for, 45 Robert's " " 45 Boiling " " 46 Picric Acid " " 46 Estimation of, 47 Esbach's albumimeter, . 47 Acetic Acid and Ferrocyanide, test for, 48 Trichloracetic Acid, « « 48 Acidulated Brine, " " 49 Potassio-mercuric Iodide, " " 49 Mucin, 49 Peptone, 49 Hemialbumose, 50 Hematuria, 5° HEMOGLOBINURIA: ALMEN'S TEST FOR, 51 Boiling, 51 Pyuria: Donne's Test for, 53 Epithelium, 53 Tube Casts 53 Spermatozoa, 55 Chyluria, 56 Micro-organisms, 56 Urinary Deposits, 57 Examination of Deposit, 58 Urinary Concretions, 59 Analysis of Concretion, 59 Composition of Gastric Juice, 60 Plan for Clinical Examination 61 Ewald's Test-breakfast, 62 Pepsin : Test for, 63 Estimation of, 64 Clinical import of, 64 Peptone, 64 Hydrochloric Acid, 64 Congo-red test for, 65 Tropjeolin test for, 65 Methyl violet test for, 65 Phloro-glucin Vanillin test for, 66 Boas' Resorcin test for, 66 Detection of Organic Acids, 67 Lactic Acid by Uffelmann's Test, . 67 Acetic Acid, 68 EXAMINATION OF GASTRIC CONTENTS, VI TABLE OF CONTENTS. PAGE Butyric Acid, 68 Total Acidity, 68 Estimation of free HC1 and Organic Acids, 70 Analysis by Evaporation, 71 Motor Power of the Stomach 72 Ewald's Salol test, . . . 72 EXAMINATION FOR COMMON POISONS. Classification of Poisons, 73 Preliminary Examination of Suspected Matters, 75 Acids and Alkalies, 77 Acidimetry, . , 77 Alkalimetry, 78 Nitric Acid, Hydrochloric Acid, 79 Uffelman's test for, 80 Nitro-muriatic Acid, 80 Sulphuric Acid, 80 Oxalic Acid, 81 Tests for Strong Mineral Acids, 82 Alkalies, 83 Potash 83 Soda, ' 83 Ammonia, 84 Volatile Poisons, 85 Carbolic Acid, • 85 Hydrocyanic Acid, ". 85 Phosphorus, 86 Chloroform, 87 Chloral Hydrate, 87 Alcohol, 88 Metallic Poisons, 89 Arsenic, 89 Fleitmann's test for, 90 Reinsch's test for, 91 Marsh's test for, 92 Antidote for, 92 Antimony, 93 Copper, 94 Mercury 94 Lead, 95 Silver, 96 Zinc, 97 Barium, 97 Analytical Table for a Common Metal, . 98 Alkaloids, 99 Strychnine, 99 Morphine, 99 Opium, 100 VII TABLE OF CONTENTS. PAGE Meconic Acid, 100 Atropine, 100 Analytical Table for Common Alkaloids, 101 STUDY OF NORMAL MILK. Composition of Normal Milk, 103 Reaction of " " 104 Spontaneous Change, 104 Preservation : Refrigeration, 105 Sterilization, 105 Pasteurization 106 Milk Sugar and Salts, 108 Butter, 108 Pepsin Curd, 108 Junket 108 Curds and Whey, 109 Rennet Curd, 109 Specific Gravity of Whole Milk, 109 " " Skim " no Milk Testing, no Lactometer, no Creamometer, no Feser's Lactoscope, in Milk Examination, 112 Determination of Water, 112 " " Solids, .... 112 " " Fat, 113 " " Solids not Fat, 114 Clinical Examination. Formulae of Hehner and Richmond, 114 COMPOSITION OF HEALTHY URINE. Healthy urine is a clear yellow fluid, slightly acid in reaction, and hold- ing in solution variable amounts of substances organic and inorganic. Average Composition of Normal Urine. Percentage Composition. Grains per Diem. Water, 96.0 50 fl. ozs. Solids, as tabulated below, 4-o 1000 grs. Urea, 2.000 500.0 Uric Acid, .040 10. Hippuric Acid, •075 15- Creatinine, •075 15. Pigment, Mucus, Xanthine, other extractives, etc., 1.000 170. Chlorides of Potassium and Sodium, 1.000 170. Sulphates of Potassium and Calcium, . 110 40. Phosphates of Potassium and Sodium, .... .120 45- Phosphates of Magnesium and Calcium, . . . .080 3°- Beside these there have been found traces of Indican, Diastase, Oxalic, Lactic and Carbolic acids, Unoxidized Sulphur and Phosphorus. REAGENTS AND APPARATUS FOR URINALYSIS. Acid, Nitric. - Burette with pinch-cock. " Sulphuric. Pipette, 5 c.c. " Hydrochloric. Capsule, 50 c.c. " Acetic. (Sp. gr. 1.048.) Graduated Cylinder, 50 c.c. " Picric, 5 gr. to Urinometer. Ammon. Hydrate, 20°. Ureometer. Potass. " (normal). Urinary Pipetie. Silver Nitrate (standard). Glass Rod. Copper Sulphate. (1 in 10.) Blowpipe. Potassium Chromate, neutral. (1 in 10.) Watch Glasses. Magnesia Mixture (MgSO4 10; NH4C1 10; Wire Gauze. Aq. Ammon, fort. 10; Water 80.) Platinum Foil. Sodium Hypochlorite (Labarraque). Funnel. Barium Chloride (standard). Filter Paper. Fehling's Solution. Tripod. Glycerin. Bunsen's Burner. Litmus Test Papers. VIII EXAMINATION OF MORBID URINE. PLAN FOR CLINICAL STUDY. 1st. Ascertain from patient the total daily discharge. 2d. Note if the color and odor are normal. If deep yellow, green or brown, test for biliary pigment, $ 2. If red or chocolate, test for haimoglobin, g 55-56. 3d. Determine the reaction with litmus paper, § 6. 4th. Take the specific gravity and calculate the solid urine, $ 4-5. 5th. Having allowed the urine to settle, carefully decant the clear part into a test tube until it is half full, and examine for albumin by boiling the upper part. One drop of acetic acid must be added if the reaction is not acid. If the flakes formed are white, pure albu- min is present. If greenish, probably bile pigment is the cause, g 2. If red-brown, there is probably blood, § 54. After 24 hours, note the height of albuminous layer. Confirm by test with cold nitric acid, § 44. 6th. Test for glucose by Fehling's and by Bottger's method, | 28, 32-34. Determine the amount. 7th. Estimate if the chlorides are absent or much reduced, $ 7. Determine the amount. 8th. Estimate the amount of urea. 9th. Note the naked-eye characters of the deposit, and also the microscopic appearances, $ 58. If amorphous, and the urine is acid, then the deposit is composed of mixed urates; if the reaction is alkaline, then the deposit is phosphatic. To make sure, test for urates and phosphates, § 59. 9 10 EXAMINATION OF MORBID URINE. In casual examination for the presence of abnormal constituents, the sample of urine may be voided at any hour of the day. If a single dis- charge is used, that made before breakfast is to be preferred. Cases which exhibit a trace only of albumin in the morning urine will sometimes show a larger quantity in a sample passed after active exercise. For accurate results, it is necessary to take a portion of the total quantity passed in the twenty-four hours. To obtain this, the patient is instructed to throw away the urine voided at an hour noted, say 9 A. M. Every evacu- ation of the bladder after this hour must be made into one vessel, such as a large " specie jar," which should be well cleaned daily and kept in a cool place. At the same hour of the following day the urine is passed into this col- lecting jar, and the amount measured and noted. (The mean daily dis- charge is 1250 c.c., or 40-50, or 2% pints.) About a half pint of this should be put into a clean glass bottle, with a fresh stopper, and examined within twelve hours at furthest. As a rule, it is best to examine within six hours after emission. The separation of sediment can be obtained in a few minutes by rotating the sample of urine in a centrifugal separator. All suspended matter is thrown to the bottom at once. Ordinarily it is allowed to subside spontaneously by standing several hours. Urine kept in a warm place soon passes into putrefaction, and then is not suited for analysis. Putrefactive changes can be retarded by adding to half a pint of urine as much salicylic acid as can be taken on the point of a penknife, which would be about three grains. Without putrefaction, some organized deposits, as blood, tube casts and renal epithelium, are altered, if not broken up, by urines of low density. If it is desired to pre- serve a specimen of these organized deposits the clear urine should be de- canted, leaving the sediment, which can be washed free of stale urine by pouring upon it a filtered strong solution of potassium acetate with a small quantity of salicylic acid dissolved in it. After standing long enough to allow the sediment to fall, the clear fluid should again be decanted and a fresh portion of the salicylated acetate of potassium solution be poured on. Instead of washing with salicylated acetate of potash, it is sometimes pref- erable to use in the same way a solution of chloral, 15 grains to the ounce of water. The specimen in chloral can be mounted on a microscope slide and will keep indefinitely. By repeating this operation several times all putrescible matter can be re- moved and the cells, etc., be left in a permanent medium. In examining urine THE SAMPLE. 11 EXAMINATION OF MORBID URINE. under the microscope for casts, epithelial cells and other organic bodies, difficulty is sometimes caused by suspended urates ; to obviate this diffi- culty and to preserve the specimen, Wendringer advises that the urine should be mixed with a nearly saturated solution of borax and boracic acid. This dissolves the urates and prevents fermentation. The solution is pre- pared by mixing twelve parts of powdered borax in one hundred parts of hot water, and then adding twelve parts of boracic acid, stirring the mix- ture well. It is filtered while ho£ The urine to be examined is put into a conical glass, and from a fifth to a third of its bulk of the boracic solution added to and agitated with it. The urine will be found to have become clear in a short time-i. e., if there is no cloudiness due to bacteria; and if a third of its bulk has been added, no fermentation takes place, even if the glass is left uncovered in warm places. Albumin, too, if it exists, is not coagulated. The organic elements-as epithelial cells, casts, blood corpuscles, etc.-subside quickly, without undergoing change. It some- times happens that this solution causes a fine amorphous and crystalline precipitate which obscures the view of casts and cells. 1. Experiment.-Having noted the characters of a sample of fresh urine, let it stand aside in a warm place for a few days, and then mark the change in odor, speci- fic gravity, reaction and sediment. A healthy urine may first become more acid, depositing urates and uric acid, but all samples ultimately become ammoniacal, putrid, and throw down a sediment of phosphates. COLOR. In recording color, it is well to take as a standard Vogel's scale, found in large treatises. The urine is pale yellow after free potations, after hysterical or epileptic attacks and in diabetes ; orange red by the presence of santonin when alkaline; reddish with scant potations, after heavy meals, much exercise, or copious sweating, after hemorrhage into any part of the urinary tract and during fever; after the administration of logwood; brownish in melanosis and after the administration of rhubarb, senna, gallic acid and tannic acid. When tar ointment or pitch plasters are applied, and when tar and carbolic acid are given internally, compounds of sulpho- carbolic acid appear in the urine. These oxidize in the air, and as the urine 12 EXAMINATION" OF MORBID URINE. turns alkaline change to greenish-black pigments. The presence of biliary coloring matter imparts an intense yellow or brown or olive-green hue ; it is characteristic of jaundice. Blood gives a scarlet or a smoky-brown tint. The presence of Indican or indigo- forming substance is shown by adding to normal urine its volume of hydrochloric or nitric acid. By boiling, or even by standing awhile, the color deepens or changes to pink, from the formation of in- digo-red. GMELIN'S TEST FOR BILE PIG- MENTS. 2. Put some yellow nitric acid (z. e., containing lower oxides of nitrogen) into a test tube and overlay it with biliary urine. At the line of junction there will rise, in the following order, bands of green, blue, violet, red and yellow. The green and red will be the most persistent. The same play of colors can be seen if the test is performed by bringing together on a white plate or piece of white filter paper the urine and the nitric acid. If the amount of bile color is very small, yielding a doubtful result, it can be con- centrated by shaking a quantity of urine with chloroform and then separating the chloroform by decanting the urine. If the chloroform is now overlaid with nitric acid the colors will appear at the line of contact. 3. Oliver's Test for the Biliary Acids.-The reagent is prepared by dissolving 30 grains of pulverized meat peptone, four grains of salicylic acid, and 30 minims of strong acetic acid, in sufficient water to make eight fluidounces. Preserved by the salicylic acid the peptone shows no signs of decomposition on keeping. To use the test, mix one fluidrachm of the reagent with 20 minims of urine, Fig. i. Graduated Cylinder for Measuring the Amount of Urine. Urinometer 13 previously diluted to a standard specific gravity of 1.008. A faint haze is produced after a minute by the trace of biliary salts in normal urine, but if these are in excess a milky cloud promptly appears, which will be more or less distinct, according to the proportion of bile salts present. Pettenkoffer's test is not practicable for urinary analysis. EXAMINATION OF MORBID URINE. ODOR. It is easy to detect a departure from the familiar odor of health. Some articles of food, as asparagus, and some aromatic medicines, impart a change. Saccharine urine has a fruity odor. Putrid urine has the odor of ammonia modified. SPECIFIC GRAVITY. 4. Test.-In taking the specific gravity, if the contain- ing vessel has a level lip, the most accurate reading is made by immersing the urinometer while the vessel is half-full of urine, and then carefully filling it to the brim. The exact line of the registry can be taken by sighting horizontally the surface of the urine, which rises slightly about the stem of the instrument. For scientific accuracy, the specific gravity bottle and balance are to be preferred. Import.-The healthy standard is between 1015 and 1025. If per- sistently below 1015, Bright's disease or diabetes insipidus may be suspected. If persistently above 1030, diabetes mellitus probably exists. Further examination is necessary to make its meaning clear. QUANTITY. The mean daily amount is about pints, or 40-50 fluidounces (1250 c.c.). It may vary in health between and 4 pints. Anuria.-Entire failure to pass urine may be due either to suppression of the secretion of the kidneys, or retention of the urine by obstruction to the bladder or other parts of the urinary passages. Hydruria.-In this condition the water is increased out of proportion to the solids. 14 EXAMINATION OF MORBID URINE. Polyuria.-This term is applied to cases of increase in the urine as a whole, both solids and water. Import.-Scanty urine is found in certain forms of Bright's disease, in cirrhosis of the liver, and in collapse. Hydruria is found in diabetes insipidus, and in a minor degree in hysteria and the middle period of atrophic nephritis. 5. Ro ughly estimate the solid urine in parts per fluidounce, by Bird's formula. The last two figures of the specific gravity nearly represent the grains of solids to the fluidounce. The same figures multiplied by 2 (Trapp's factor) will give, more accurately, parts per 1000. For example, if the specific gravity is 1025, then there are 25 grains of solids to the fluidounce of urine, or 50 parts in the thousand. Haeser's factor is 2.33, which in a large number of samples is no more accurate than Trapp's. The most exact method is to evaporate a given amount to dryness on a water bath and then weigh the residue. In practice this is found too tedious for clinical use. SOLID URINE. 6. Test.-First put a drop of the sample on a piece of blue litmus paper; if the blue turns to red, then the reaction is acid; if the paper is unchanged, then it will be alkaline or neutral. Now try it with red litmus paper; if it turns blue, the reaction is alkaline; if the paper is unchanged, the urine is neutral. Violet-colored " neutral litmus " paper alone can be used. It will turn reddish with acids and bluish with alkalies. The normal reaction is acid, due mainly to acid salts, such as acid sodium phosphate, NaH2PO4. The degree of acidity can be determined by the process described, p. 77, using decinormal solution of potassium hydrate (5.6 in 1000), and report - REACTION. 15 EXAMINATION OF MORBID URINE. ing in terms of oxalic acid. Each c.c. of the titrating solution represent- ing 0.006 gm. oxalic acid in the volume of urine used. If alkaline, it may be so from salts of potassium and sodium, such as Na2HPO4 (fixed alkali), or from ammonia (volatile alkali). If due to ammonia, then by exposure the ammonia vaporizes and the red color of the paper is restored; whereas, the blue from fixed alkali remains. While the normal reaction of the total daily discharge is acid, it can be made alkaline by the administration of alkaline medicines. Urine is made ammoniacal by the decomposition of urea. CON2H4 + 2H2O = (NH4)2.CO3. Urea. Water. Ammonium Carbonate. CHLORIDES. The chlorine in the urine is in combination principally with sodium as NaCl. The mean daily discharge is 200 grs., or 12 gras., amounting to more than all the other salts together. The chlorides diminish in acute fever when exudations are forming, and in croupous pneumonia may dis- appear entirely to reappear with convalescence 7. Test for Chlorides.-First, add a few drops of nitric acid to prevent the precipitation of phosphates, then on the addition gradually of solution of silver nitrate a heavy white precipitate of silver chloride forms if the chlorides are abundant, but if scanty then a milky appearance only. By comparison of this precipitate with that from a sample of normal urine a rough estimate can be made of the relative amount of chlorides. NaCl + AgNO3 = NaNO3 + AgCl. 8. Volumetric Determination.-See p. 77 for rules to be observed. Dilute ten c.c. of urine with 50 c.c. of water. Put this into a beaker or capsule, with a few drops of neutral potassium chromate. Add gradually from a burette (Fig. 2) the standard solution of silver nitrate until the urine is permanently orange red; then note the c.c. of reagent used, subtracting one c.c. for excess of silver solution. The standard solution contained AgNO3 29.06 gms. per 1000 equal to ten gms. NaCl, then one c.c. = 0.01 NaCl in the ten c.c. of urine used, or o. 1 per cent. 16 EXAMINATION OF MORBID URINE. Fig. 2. Graduated Burette. 17 By multiplying the number of c.c. noted by 10, the percentage can be obtained, or if rhultiplied by 45.57, the number of grains to the fluidounce of urine. Then, if 15 c.c. were used, 14 c.c. would be counted. 1 c.c. = 0.01 NaCl in 10 c.c. of urine. 14 c.c. = 0.14 " " " = 1.40 " 100 " " I.40 per cent, multiplied by 4.55 = 6.37 grains to the fluidounce. This method, which serves well enough for clinical study, cannot be con- sidered scientifically exact when the urine is high-colored, or putrid, or albuminous. To make it satisfactory, the organic compounds may be removed by boiling ten c.c. of the urine and adding to it, drop by drop, a solution of potassium permanganate, until a faint pink hue is left. The brown precipitate formed is separated by filtration. Before titration the pink tint should be removed by a few drops of oxalic acid, the urine made neutral with calcium carbonate, and filtered. EXAMINATION OF MORBID URINE. SULPHATES. The compounds of sulphur are derived from food and the oxidation of proteids of the body. They are discharged daily to the amount of about 30 grs. or two gms., mostly with alkaline bases (K2SO4 Na2SO4), but partly with organic compounds. 9. Test-Into the test tube containing the urine put a few drops of hydrochloric acid to prevent the formation of barium phosphate. On adding solution of barium chloride, a white precipitate falls, varying in density from a cloudiness to a creamy consistency. Compared with the results obtained from equal amounts of healthy urine acidified, one can judge as to whether the sulphates are greater or less than normal. 10. Volumetric Estimation.-Into a beaker on a sand bath put too c.c. of urine acidulated with hydrochloric acid, and boil it. From a burette drop in the standard solution of barium chloride as long as the precipitate forms. Set aside for subsidence of the precipitate. Now cautiously run one drop of the barium chloride solution down the side of the beaker to see if there will be further precipitation. Run in the barium chloride until EXAMINATION OF MORBID URINE. 18 the white cloud no longer increases. To determine if too much of the barium chloride has been used, filter a few drops out of the beaker and test it with a drop of sodium sulphate. A white precipitate indicates excess of the standard solution, and calls for a repetition of the titration. The standard solution contained 30.5 gms. BaCl2 in 1000 c.c. and one c.c. =0.01 gm. SO3 in the 100 c.c. of urine used. If 15 c.c. were needed, then there were present 0.15 gm. SO3 in the hundred. To get grains in the fluidounce multiply by 4.557. Import.-As yet no such correspondence has been detected between the amount of sulphates and any pathological process as to make an estimate of value in diagnosis. PHOSPHATES. The phosphoric acid of urine is united partly with the earthy bases, as acid calcium and magnesium phosphates (MgHPO4 Ca3(PO4)2), and partly with alkaline bases, as acid sodium phosphate and a trace of acid potassium phosphate (NaH2PO4KH2 POJ. The average daily output is 60 grs. or four gms., of which two-thirds are joined to alkaline bases and one-third to earthy. As the latter are insoluble in alkaline fluids, they are deposited whenever the urine is made alkaline by an internal course of such remedies, or by putrefaction, changing its urea into ammonium car-bonate. CON2H4 + 2H2O = (NH4)2COs. Urea. Ammonia Carb. 2(MgHP0J + (NH4)2CO3 = H2CO3 + 2(MgNH4PO4). Magnesium Phosphate. Triple Phosphate. 11. Test.-To the urine in a test tube add a few drops of potassium hydrate and boil. The earthy phosphates are thrown out and must be separated by filtration. 12. Test.-To the filtrate add one-third its volume of " magnesia mixture." The precipitate formed represents the phosphoric acid once held by alkaline bases, now in the form of ammonia-magnesium phosphate (triple phosphate). 13. Teissier's Method.-To get a fair idea of the quantity of phosphoric acid in urine, we can resort to the following easy method : Into a cylindri- cal vessel, graduated in c.c., measure 50 c.c. of urine. To this add "magnesia mixture " a sufficient quantity, at least 15 c.c. After shaking EXAMINATION OF MORBID URINE. 19 together set aside for 24 hours. As triple phosphate all the phosphoric acid will be thrown down in a compact white sediment, of which one c.c. in height represents 0.30 gm. per litre, or 0.03 per cent, of phosphoric acid. This is equivalent to 0.60 to 0.70 gm. of phosphates per litre, or 006 to 0.07 per cent.; multiplying this percentage by 4.55 will give the amount in grains to the fluidounce. 14. Volumetric Estimation.-Having measured 50 c.c of urine, pour it into a capsule resting on wire gauze or a sand bath. Add to this five c.c. of a saturated solution of sodium acetate, which has an excess of acetic acid. Heat to boiling, and slowly add from a burette the standard solution of uranium acetate. A precipitate forms, of uranium phosphate, insoluble in the acetic acid. With a glass rod stir the mixture, and at intervals let a few drops fall on a solution of potassium ferrocyanide in a white plate, to see if it will form a red-brown spot, indicating that excess of uranium acetate has been added. Note the number of c.c. of standard solution used up to that point. The standard solution of uranium acetate contained 31.1 gms. to 1000 c.c. equal to five gms. phos- phoric acid, P2O5. As 1000 c.c. = five gms. P2O5, then one c.c. of the test will represent 0.005 gm- P2O5 in 5° C.C. of urine taken. To get per cent., multiply the number of c.c. used by 0.01, which is the equiva- lent of 0.005 X 2. The per cent, multiplied by 4.55 will give grains to the fluidounce. Import.-The quantity of phosphates in the urine depends mainly upon the amount absorbed from the food, which varies greatly. A knowledge of the rela- tive amount of phosphates in a sample of urine is of no service in diagnosis or treatment unless an almost impossible determination be made of the amount absorbed from food. Fig. 3. Deposit in Ammoniacal Urine (alkaline fermentation), a, Acid ammonium urate; b, Ammonio-magnesium phosphate; c, Bacterium ureas. 20 EXAMINATION OF MORBID URINE. The clinical significance of the phosphates depends not on the amount, but on their state. When they are persistently thrown out from solution, there is reasonable ground to fear that in time a gravel or calculus may be formed. When retained in the bladder and when from cystitis the urine is puru- lent and rapidly putrescible, passing into ammoniacal fermentation, the alkaline reaction induced by the ammonia causes the precipitation of the earthy phosphates before micturition. Phosphatic Deposits.-The amorphous phosphate de- posit occurs whenever the urine becomes alkaline, is grayish-white and composed of the phosphates of calcium and magnesium. Under the microscope it appears as granules, without color or tendency to aggregate in masses of any particular shape. The granules of amorphous urates usually are pinkish, arranged in delicate moss-like forms, and disappear on the addition of potassium hydrate. Under this reagent the phosphates are unchanged, but they clear up when a drop of acetic acid is introduced under the cover glass. Triple or Ammonio-magnesic Phosphate.-This is the combination in which the phosphoric acid falls after putrefactive changes. To the naked eye it is a whitish sediment in a turbid fluid. Sometimes the crystals may be detected as brilliant white points on the sides of the glass, or as a shining film on the surface of the urine. Under the microscope most of these crystals are prismatic and have some resemblance to a " coffin lid." They are colorless, bright, relatively large, and easily distinguished from the spiny spherules of ammonium urate which often accompany them. Import.-Their presence denotes ammoniacal decomposition of the urine. If this condition exists at the time of micturition, it is evidence of cystitis or other disease of the genito-urinary tract. If persistent there is liability to the formation of phosphatic calculus. 21 EXAMINATION OF MORBID URINE. Fig. 4. Deposit from a Case of Inflamed Bladder (ammoniacal fermentation), a, Detached epithelium ; b, pus corpuscles ; c, triple phosphate ; d, microorganisms. The More Usual Forms of Triple Phosphate. X 3°°- 22 EXAMINATION OF MORBID URINE. URIC ACID, Uric acid, like urea, is nitrogenous. The quantity eliminated daily is 0.7 gm. or ten grs. In healthy urine it exists only in combination, as potassium and sodium urates. Acid and dense urine, on standing, will undergo change, becom- ing more acid and throwing down a deposit of uric acid, calcium oxalate, and amorphous sodium urate. Free uric acid may exist in the urine at the time it is voided, but in any but the smallest amount must be considered as pathological. It is prac- tically insoluble, requiring 18,000 parts of water to dissolve it. It is freely soluble in the alkalies and solutions of the alkaline carbonates. 15. Separation of Uric Acid.-To 100 c.c. of urine add ten c.c. of hydrochloric acid; let it stand for 48 hours, when the uric acid will be settled as fine crystals, looking like grains of Cayenne pepper. 16. Estimation.-Having obtained the crystals as above, decant the urine and stir the sediment with 30 c.c. of water, using for this purpose a glass rod with a piece of rubber tub- ing at the end. Throw the suspend ed crystals on a weighed filter, dry over a water bath and weigh. This method, though simple, is approximative only. 17. Hay era ft's method is to be pre- ferred where scien- tific precision is desired and a well furnished laboratory is at command; it is too compli- cated for the clinician and its accuracy is not absolute. C5H4N4O3. Fig. 5. Deposit in " Acid Fermentation" of Urine, a, Fungus; b, amorphous sodium urate; c, uric acid ; d, calcium oxalate. 23 The method in detail has been given as follows :- The following solutions must be prepared: I. Dissolve five gms. of nitrate of silver in 100 c.c. of distilled water, and add ammonia until the precipitate first formed redissolves. 2. Dilute strong nitric acid with about two volumes of distilled water; boil, to destroy the lower oxides of nitrogen, and preserve in the dark. 3. Dissolve about eight gms. of ammonic sulpho- cyanide crystals in a litre of water, and adjust to decinormal argentic nitrate solution, by diluting till one volume is exactly equal to a volume of the latter. Dilute the solution thus prepared with nine volumes of distilled water, and label " Centinormal ammonio-thiocyanate solution." 4. A satu- EXAMINATION OF MORBID URINE. Fig. 6. Uric Acid, a, Rhombic tables (whetstone form); b, barrel form ; c, sheaves ; d, rosettes of whetstone crystals. rated solution of ferric alum. 5. Strong solution of ammonia (sp. gr. 0.880). The uric acid estimation is conducted as follows: Place 25 per cent, of urine in a beaker with one gm. of sodic bicarbonate. Add two or three c.c. of strong ammonia, and then one or two c.c. of the ammoniated silver solution. If, on allowing the precipitate caused by the latter reagent to subside, a further precipitate is produced by the addition of more solution, the urine contains an iodide, and silver solution must be added till there is an excess. The gelatinous urate must now be collected, the following special procedure being necessary: Prepare an asbestos filter by filling a 24 four-oz. glass funnel to about one-third with broken glass, and covering this with a bed of asbestos to about a quarter of an inch deep. This is best managed by shaking the latter in a flask with water until the fibres are thoroughly separated, and then pouring the emulsion so made in separate portions on to the broken glass. On account of the nature of the precipi- tate and of the filter, it is necessary to use a Sprengel pump or aspirator in order to suck the liquid through. Having collected the precipitate of silver urate on the prepared filter, wash it repeatedly with distilled water, until the washings cease to become opalescent with a soluble chloride. Now dissolve the pure urate by washing it through the filter with a few cubic centimeters of the special nitric acid. Place in a beaker, and estimate the silver by Volhard's method, which can be employed in the presence of nitric acid. The process is carried out thus : Add to the liquid in the beaker a few drops of the ferric-alum solution to act as an indicator, and from a burette carefully drop in centinormal ammonic thiocyanate until a perma- nent red coloration of ferric thiocynate barely appears. The number of cubic centimeters used of the thiocynate solution multiplied by 0.00168 gives the amount of uric acid in the 25 c.c. One milligramme may be added to compensate for loss, and the whole multiplied by four gives the percentage of uric acid in the urine. The whole process depends on the fact that argentic urate fails to dissolve in ammonia, but is soluble in nitric acid, and is thus easily obtained in the pure state. By determining the amount of combined silver, the percentage of uric acid can readily be cal- culated. The addition of sodic bicarbonate prevents the otherwise inevitable reduction of the silver salt. 18. Murexide Test.-Put any substance supposed to be uric acid or urates into a watch crystal or a porcelain dish. Dissolve in strong, cold nitric acid, and at gentle heat evaporate to dryness. Touch the yellow or red residue with a drop of ammonia water, or simply expose it to the vapor of ammonia. A bright violet blue or purple red (murexide) indicates uric acid. Deposit of Uric Acid.-While a faint trace of free uric acid may be found in healthy urine, if present in noticeable amount, it is a pathological change. Once deposited, it remains undissolved in acid urine. It is colored by urinary pigment and easily made out by the microscope. To the naked eye it appears in minute reddish specks. Under the EXAMINATION OF MORBID URINE. 25 EXAMINATION OF MORBID URINE. microscope these specks are found to be modifications of rhombic crystals. The simpler lozenge forms have a resemblance to a whetstone. Some are like a drum, others are like rosettes, others, again, sheaf-like bundles. Import.-Beside its connection with the pathology of gout, uric acid is important as a deposit. If it is deposited soon after micturition, it is an indication of a tendency to form gravel. URATES. It has been said above that uric acid in health exists as neutral urates in combination with sodium principally, but with potassium partly. These urates, in ordinary amount, are soluble in normal urine at common tem- peratures. If healthy urine is kept in a cold place, it loses its solvent powers, and the mixed urates are precipitated as a loose pinkish powder at the bottom of the glass and as a film on the surface. The same thing happens at higher temperatures if the amount of urates is in excess of health and the reaction highly acid. This deposit, often re- ferred to as sodium urate, the brick-dust or lateritious sedi- ment, can be distinguished by its prompt solution on heating the urine. It dissolves readily in potassium hydrate. 19. Test.-Having ob- tained a specimen of urine that is turbid, fill with it a test tube until it is half full. Gently heat the upper half. It will clear up if the tur- bidity is due to urates. 20. Test.-To another portion of the same add potas- sium hydrate. The sediment of urates will dissolve. Fig. 7. Uric Acid, a, Rhomboids!, truncated, hexa.- hedral, and laminated crystals; b, rhombic prism, horizontally truncated angles of the rhombic prism, imperfect rhombic prisms ; c, prism with a hexahedral basic surface, barrel- shaped figure, prism with a hexahedral basal surface; d. cylindrical figure, prism with a superimposed group of crystals. 26 EXAMINATION OF MORBID URINE. Microscopically (Fig. the brick-dust deposit is composed of granules usually disposed in moss-like groups. Sometimes these granules have spiny radiations. The ammonium urate (Fig. 3-a) occurs in opaque, brownish spherules, with or without spines, and in company with crystals of triple phosphate. Import.-This deposit is not so important as it is commonly considered, occurring often from such physiological conditions as unusual exercise or sweating. Pathologically, it is associated with fever and chronic hectic diseases. HIPPURIC ACID, C9H9NO3, combined with alkaline bases, is a normal constituent. The average quan- tity eliminated daily is 15 grs. This amount is increased by excess of Fig. 8. Hippuric Acid. vegetables in the diet or by large doses of benzoic acid. If the urine be at the same time diminished in volume the hippuric acid may be deposited spontaneously. This is, however, an extremely rare occurrence, owing to the solubility of the acid and its salts. It occurs in rhombic prisms (Fig. 8), resembling somewhat the coffin-lid 27 EXAMINATION OF MORBID URINE. crystals of triple phosphate. It is to be recognized by the fact that it is precipitated from acid urine only, and does not dissolve when a drop of acetic acid is allowed to flow under the cover glass. The phosphates, on the other hand, are deposited in alkaline urine and clear up in the.presence of acetic acid. CREATININE, C4H7N3O, is a constituent of normal urine to the amount of about 15 grs. daily. It is derived from the creatin of muscle, is a strong base, alkaline in reaction, and owing to its free solubility in water is very rarely spontaneously depos- ited from the urine. It forms insoluble compounds with mercuric chloride, with silver nitrate, and with zinc chloride. The latter compound has a characteristic crystalline form, by means of which the presence of creati- nine may be determined. Under the microscope it is seen as needles Fig. 9. Oxalate of Lime, a, Octahedra J b, basal plane of an octahedron forming a rectangle ; c, com- pound forms; d, dumb-bells. Perfect Dumb-bell Crystals of Oxalate of Lime. arranged in rosettes or balls. It has the power ot reducing both cupric oxide and picric acid, and affects to a slight degree the accuracy of esti- mates of sugar made by reduction tests. CALCIUM OXALATE is present in extremely small amount in normal urine. As it takes 500,000 parts of water to dissolve it, the slightest increase over a minute quantity causes it to appear as a deposit. To the naked eye it appears as a scanty, grayish, mucus-like cloud in the middle or at the bottom of the glass. Microscopically, it is recognized by one of two lorms : the octahedra, usually appearing like bright, square " envelopes," and the dumb-bells or hour-glasses. 28 EXAMINATION OF MORBID URINE. Import.-The deposit sometimes follows the eating of "rhubarb plant" or sub-acid fruits containing oxalates. Usually it signifies imperfect oxida- tion of waste products or retarded metabolism. If persistent, it may, by accretion, form a mulberry calculus. CYSTIN, CgHjoNoSjO.i. This substance, in an excessively small amount, may be a normal con- stituent of the urine. It is nearly insoluble in water, and in the very rare cases where it is present in large amount it is in the form of a deposit. Fig. io. A, Crystals of cystin ; B, oxalate of lime ; c, hour-glass forms of B. To the naked eye the deposit is usually abundant and light, resembling urates. It is not dissolved by heat or vegetable acids, but readily by ammonia. The ammonia solution exposed on a glass slide deposits crystals which under the microscope are seen to be composed of hexagonal tablets. Iodoform escaping into the urine from surgical dressings has been mistaken for it. 21. Test.-When it decomposes, hydrogen sulphide is formed. If boiled with a solution of lead oxide in 29 EXAMINATION OF MORBID URINE. sodium hydrate, the black lead sulphide forms and is precipitated. Import.-The conditions of its formation are obscure. It occurs very rarely as a calculus. LEUCIN AND TYROSIN. These substances are formed in the intestines by the same processes of digestion, and in acute yellow atrophy of the liver, typhoid fever and phos- phorous poisoning, appear together in the urine. Fig. ii. a a, Leucin balls ; b b, tyrosin sheaves ; c, double balls of ammonium urate. When present, leucin will appear as a residuum left with tyrosin when the urine is allowed to evaporate on the glass slide. It occurs either as greenish-yellow globes with concentric markings or as radiating spines. If a small quantity is put in a watch crystal and saturated with nitric acid, by cautious evaporation to dryness a residue will be left which turns yellow or brown on the addition of solution of sodium hydrate. Tyrosin may be spontaneously deposited as greenish-yellow crystals com- posed of bundles of acicular needles arranged in various radiating forms. 30 EXAMINATION OF MORBID URINE. Tyrosin turns red when boiled with Millon's reagent (mercurous nitrate); but yields a violet color when first carefully warmed with sulphuric acid and treated with a drop of solution of ferric chloride. UREA, CO(NH2)2 Carbamide. Urea is the chief compound of nitrogen by which that element is elimi- nated after having served in the body as a constituent of the proteid principles. The average daily excretion of urea is 40 gms., or 500 grs. It is neutral, odorless, has a bitter taste, and plays the part of either acid or basic radical. Fig. 12. 1, Urea; 4, hexagonal plates ; and (3) smaller scales, or rhombic plates of urea nitrate. It is easily soluble in water and alcohol, but insoluble in ether. It can be prepared by synthesis, and is isomeric with ammonium cyanate. When crystallized slowly, it forms four-sided prisms with beveled ends. 22. Test.-Take a specimen of urine or any fluid suspected to contain urea, concentrate it to a syrupy consistence by evaporation on a water bath, add nitric acid. The nitrate of urea will form a crystalline precipi- tate of rhombic plates (Fig. 12). 31 EXAMINATION OF MORBID URINE. Estimation.-Several methods of quantitative analysis depend on the power of sodium hypobromite or hypochlorite to decompose urea. N2H4CO 3NaBrO == 3NaBr 4 CO2 + 2H2O 4 2N. The amount of. urea involved will be indicated either by the loss of sp. gr. of the urine or by the volume of nitrogen generated. 23. Fowler's Method.-First, with a urinometer having wide spaces and known to be accurate, note the sp. gr. of the urine, and of the liq. sodse chlorate about to be used. In practice the strength of liq. sodae chloratse is found to be uncertain. On this account it is best to fortify it by adding a few drops of free bromine. Multiply the sp. gr. of the mixture by 7 and add the product to the sp. gr. of urine. Divide the sum by 8 to get the mean sp. gr. Then to urine add liq. sodse chlorate After effervescence ceases, say in two or three hours, take the sp. gr. of the mixture. Deduct this last figure from the mean sp. gr. obtained in the first place, multiply the remainder by 0.77, and you get the per cent, of urea. To get grains per fluidounce multiply per cent, by 4.55. Example:- Sp. gr. of urine = 1020 X 1 volume == 1020 " " liq. sod. chlo. = 1025 X 7 " = 7175 10204-7175 =8195 8195 -4- 8 = 1024 the mean sp. gr. 1024-1022 (sp. gr. after mixing) = 2 2 X °-77 = !-54 % of urea. 24. Russell dr5 West's Method.-This method measures the nitrogen evolved by sodium hypobromite and uses apparatus which gives N in c.c., each of which represents 0.0027 gm. of urea, or apparatus graduated so as to give percentage (on the principle that five c.c. of a two per cent, urea solution will yield 37.1 c.c. of N). An apparatus often used is seen in Fig. 13. Solutions of sodium hypochlorite or hypobromite decompose so readily that it is best to use them freshly prepared. Knop's Solution.-Dissolve 100 gms. sodium hydrate in 250 c.c. of water and add 25 c.c. of bromine. So important is it that the solution should be fresh, that if any great interval is to occur between successive determinations, it is best to have the fluid in two parts, to be mixed as required. In a separate bottle keep 32 EXAMINATION OF MORBID URINE. the sodium hydrate solution (i to 25). To make Knop's fluid add 15 or 20 ex. of this to one c.c. of bromine. Method.-Fill the glass cylinder (Fig. 13) with water and immerse in it the graduated tube until the zero is at the surface. Into the flask measure 15 c.c. of hypobromite solution. A short test tube containing five c.c. of urine is put in the flask by forceps, so as not to spill. Having closed the flask tightly with a rubber stopper carrying a connection with the gradu- ated tube, tip it gradually, so as to pour out the urine into the hypobromite. Of the gas evolved CO2 is retained by the soda, while the N passes up the graduated tube. In ten minutes, having raised the col- lecting tube to the point where its contents are on a level with that of the cylinder, read off the c.c. of nitrogen. Each c.c. - 0.0027 gm. of urea in five c.c. of urine used. If it reads eight c.c. of N then were 8 X 0.0027 =0.0216 gm. urea in five c.c. of urine. This multiplied by 20 to get percentage = 0.432 per cent. Percentage multiplied by 4.55 gives grains to the fluidounce. Correction for Pressure to 760 mm. and Temperature to 0° C.-For perfect accuracy a correction must be made according to the following formula:- V' = V (b w) _ jn which 760 (1 4 0.00366 T) V' = volume required; v = volume observer, b = barometer in m.m. : w = tension of aqueous vapor. T = observed temperature Centigrade. 25. Lyon's Apparatus. * (Fig. 74.)-Into the bottle measure 40 c.c. of strong liq. sodae chloratae (or, better still, 20 c.c. of fresh Knop's hypobro- Fig. 13. Ureameter of Charteris, as made by W. Hume, of Edinburgh. * Made by Parke, Davis & Co., Detroit, Michigan. 33 EXAMINATION OF MORBID URINE. MANUFACTURED BY PARKE, DAVIS & CO., DETROIT, MICH. Fig. 14. ''SU 34 EXAMINATION OF MORBID URINE. mite solution. As the liq. sodae chlorate is often not so strong as is needed, if used at all it is best to fortify it by adding two c.c. of bromine or ten c.c. of a strong solution of potass, bromide.) Fill the little test tube with urine to the four c.c. mark, and carefully lower it into the bottle so as not to spill. Hook the bent overflow tube on a bottle or tumbler, and having removed the little rubber cap, fill with water the graduated jar to slightly above zero. Put on the cap far enough to depress the surface of the water exactly to zero. Make all joints tight, and tip the bottle so that the urine slowly flows out, and shake it occasionally. In half an hour lower the end of the overflow tube to the level of the water in the cylinder, and read off the percentage of urea. This multiplied by 4.55 will give grains per fluidounce. 26. Correction for Pressure and Temperature.-The graduation of Lyon's instrument is based on the assumption that the temperature is 70° F. (210 C.) and the barometer 30 inches (760 mm.). Any variation from these standards will alter the volume of gas measured. Barometric changes have so little effect that ordinarily this correction need not be made. Temperature variations can be controlled by taking the observation when the thermometer is near 70° F. Within the usual range indoors the effect on the volume of gas may, for clinical purposes, be ignored. Greater accuracy can be obtained by the following rule : If the temperature is above 70° F., then subtract from the observed volume a cor- rection obtained by multiplying the number of degrees above 70° by 0.0028, and this product by the observed volume. To correct for barometric pres- sure, subtract from the above volume the amount obtained by multiplying the variation from 30 inches by 0.034, and this product by the observed volume. 27. Liebig's Volumetric method depends on the formation of an insoluble compound of urea and mercuric oxide. It is less easy than the hypobrom- ite method, but in the absence of the special nitrogen apparatus it may be resorted to with good results. It requires several solutions. 1st, the baryta solution, containing one volume of cold saturated solution of barium nitrate with two volumes of cold saturated solution of barium hydrate. 2d, a satu- rated solution of sodium carbonate. 3d, a standard solution of mercuric nitrate, which is best made by an expert chemist. It can be bought of leading druggists. Method.-Mix 20 c.c. of baryta solution with 40 c.c. of urine, to precipi- tate phosphates, sulphates and carbonates. Filter clear and put 15 c.c. of the filtrate into a beaker. Drop from a burette the standard mercuric nitrate solution, testing for excess of reagents by removing a drop at intervals and 35 EXAMINATION OF MORBID URINE. touching with it the sodium carbonate solution on a white plate. When a yellow spot appears the end point has been reached. Deduct two (for effect of chlorides) from the number of c.c. used and multiply by 0010. This gives the urea in grammes contained in the ten c.c. of urine operated upon. Again multiplied by ten gives percentage. Import.-As urea is highly soluble, it is never spontaneously deposited. It varies in amount with different diseased conditions, e. g., in febrile and inflammatory affections it is increased in the forming stage and diminished in that of defervescence. In diabetes mellitus and simplex it is excessive in the urine, while in acute yellow atrophy of the liver it may be absent entirely. In acute and chronic 'Bright's disease there may be a decided falling off from the healthy proportion, causing a lower specific gravity. In such cases there is more or less danger of uraemia. By its ready conversion into irri- tating ammonium carbonate it has pathological significance in all cases of retention of urine. While it is asserted by some that a minute trace of sugar can be detected in healthy urine by the use of tests of great delicacy, the weight of authority at present inclines to the opinion that normal urine usually contains no sugar. It is deemed probable that the reducing power possessed by normal urine (equal to that of a o.i per cent, solution of glucose) is due partly to uric acid and partly to creatinine. In diabetes mellitus this power of reducing metallic salts and other compounds is enormously increased, is easily appreciated by ordinary tests and is undoubtedly due to grape sugar, which may be present in the great amount of 50 grs. to the fluidounce, or ten per cent., though it passes four per cent, in severe cases only. In the application of any test but that of picric acid, it is necessary to make sure that the urine is free from albumin. If any is present, boil the sample with a drop of acetic acid, and filter. Reduction Tests.-The best tests for glucose are such as Bbttger's and Fehling's, which make use of its power to reduce metallic salts to lower oxides or the metallic state when boiled with them in alkaline solution. 28. Bottger's Test.-(First. Test for albumin; if present, separate it by acidulating the urine slightly, boiling and filtering). Put into a test tube equal volumes of sus- pected urine and potassium hydrate, with a pinch of SUGAR. 36 EXAMINATION OF MORBID URINE. bismuth subnitrate. On boiling for a few minutes, if sugar is present, the white powder turns gray, brown or black, from reduction to metallic bismuth. Fallacy: Albumin and sulphur compounds in the urine may cause a black precipitate. To make sure, repeat the test, using litharge instead of bismuth. If this blackens, then sulphur compounds have affected the bis- muth also, and some other test must be used. Nylander performs this test with a single alkaline bismuth oxide solution, made as follows :- R . Bismuth subnitr., 2 gms. Rochelle salts, 4 " Caustic soda (sol. of 8 per cent.), ioo " Mix. To ten c.c. of urine add one c.c. of test solution and boil. A brown or black color denotes sugar. TESTS BY COPPER SULPHATE. The reducing action of glucose on copper sulphate is subject to conditions which will be understood by performing the following experiments :- 29. In a test tube add to a solution of copper sulphate an equal amount of potassium hydrate ; a pale-blue pre- cipitate of cupric hydrate, Cu2HO, forms, insoluble in excess. On boiling, the blue hydrate changes to black oxide, Cu2HO = CuO -j- H2O. 30. Put into a test tube solution of copper sulphate and a few drops of glycerine (or instead of the glycerine a few grains of neutral potassium tartrate or of Rochelle salt). Now add potassium hydrate cautiously ; at first a blue precipitate falls, redissolved on the addition of more potass, hydrate. Boiling makes no change in the clear blue fluid. This property of holding the cupric hydrate in solution is shared by many organic compounds, as glycerine, the tartrates, and glucose; but, when boiled with it, glucose has the peculiar property of abstracting oxygen from the cupric hydrate, forming the red or cuprous oxide Cu2O. 37 EXAMINATION OF MORBID URINE. 31. Trommer's Test.-To a half-inch of urine in a test tube add an equal volume of potassium hydrate and a few drops of solution copper sulphate. Heat just to the first signs of boiling. If sugar is present, yellow or red cuprous oxide is precipitated. 32. Glycerine Cupric Test.-To an inch of potassium hydrate in a test tube add a few drops of copper sul- phate and a few drops of glycerine. Boil and add sus- pected urine by small amounts up to less than one inch in the tube. Sugar will throw out the cuprous oxide, red or yellow. For convenience and simplicity, the copper sulphate may be dissolved, 28 grs. in a fluidounce of pure glycerine, and a few drops used as above. This Glycerine Cupric solution keeps indefinitely and takes the place of two different fluids. 33. Approximative Method for Bedside Estimates.-In the absence of finely-graduated instruments, such as standard burettes, it is at times of value to use the ordinary apothecaries' vessels. The results obtained by this method are approximative only. It is found that, boiled in alkaline solution, one molecule of grape sugar (C6H12O6 = 180) will reduce to cuprous oxide five molecules of crystal- lized copper sulphate (CuSO4 5H2O = 249. 5X5- I247-5)- There- fore to make a copper solution in which we shall have sufficient copper sulphate in each fluidrachtn to be exactly equal to % (0.5) gr. of sugar- 180 : 1247.5 = : 0.5 : x = 3.47. Dissolve 3.47 grs. CuSO4 in a drachm (27.76 grs. in the fluidounce) of glycerine; then 15 minims (one c.c.) will be reduced by X gr- °f glucose. Method.-Put 15 minims (one c.c.) of glycerine cupric solution in a test- tube with 30 minims (two c.c.) of liquor potassse and 30 minims (two c.c.) of water. Boil, and add 30 minims or drops of urine, and boil again a few seconds. If the blue color is entirely lost, then there are at least two grs. of sugar in the fluidounce, or 0.4 per cent. If the blue color remains, there is less than two grs. to the fluidounce. Again, mix 15 minims of glycerine cupric solution with 30 minims of 38 EXAMINATION OF MORBID URINE. liquor potassee and 30 minims of water. Boil, and add one drop of the urine, and boil again for a moment. If the blue color is discharged, then there are at least 60 grs. to the fluidounce (12.0 per cent.). If the color is not discharged, add one drop more and boil. Entire loss of color indicates at least 30 grs. to the fluidounce (6.0 per cent.). If any color remains, add one drop more and boil. Entire loss of color denotes at least 20 grs. to the fluidounce (4 per cent.). If the amount of urine is insufficient to decolorize, increase it drop by drop, boiling after each addition until there is a sufficient amount to exactly discharge all the blue color. By dividing 60 by the number of drops used, we get the least number of grains of sugar to the fluidounce able to reduce the given quantity. If the amount is very large, the urine may be diluted with an equal volume of water, and then by dividing 120 by the number of drops used, we get the least number of grains to the fluidounce. 34. Fehling'sTest.-To obviate some of the defects of Trommer's test, Fehling's solution is employed. As it does not keep well,it is best to make it in two parts, not to be mixed until the time of use. A. Take copper sulph. 34.64 gms. and water enough to make 500 c.c. Mix. B. Pure Rochelle salt, 173 gms. Sol. sodium hydrate (sp. gr. 1.33) 100 c.c., and water enough to make 500 c.c. Mix. For use, mix equal volumes of A and B, and thus make Fehling's solution. Glycerine-Fehling's Solution.-Instead of Rochelle salt, glycerine can be used and the solution made as follows in one fluid which will keep some time though not indefinitely. Dissolve 34.64 gms. of copper sulphate in 500 c.c. of water. Add glycerine 130 gms., potassium hydrate 130 gms., and enough water to make one litre. Each c.c. will equal 0.05 gm. of glucose. 35. Method.-Boil in a test tube one inch of Fehling's sol. While hot, add urine and bring to a boil again. If no red or yellow color appears, add more urine until the total volume is inches. Again raise to boiling point and set aside. If yellow or red cuprous oxide appears the urine is saccharine. Precautions.-Let the volume of urine be less than that of the test solu- tion. Heat up to the boiling point without continuing the boiling. If these precautions are not observed, high-colored, acid, dense urines may reduce the copper sulphate without the presence of sugar. 39 EXAMINATION OF MORBID URINE. 36. Volumetric Estimation.-Measure into a capsule ten c.c. of Feh- ling's solution and 40 c.c. of water. Heat over wire gauze to boiling, and then from a burette containing a mixture of one volume of urine and nine of water, slowly let it run into the capsule until the color of the test solution disappears. Note the number of c.c. used, and then repeat the experiment with fresh materials to see if a less amount will not answer. Thus ten c.c. of standard solution is decolorized by 0.05 gm. glucose. If eight c.c. of dilute urine were used, then of the undilute 0.8 c.c. - 0.05 gm. glucose. If 1500 c.c. were voided in the day, then 0.8 : 0.05 :: 1500 : x = 93.75 gms. To calculate per cent. 0.8 : 0.05 : : 100 : x = 6.25. Parts per cent, multiplied by 4. 55 give grains to the fluidounce. 37. Pavy's Ammonia-cupric Process.-This is easy of application, and shows the end of the reaction more definitely than Fehling's method, which it closely resembles. Pavy's solution differs from that of Fehling in con- taining ammonia, which dissolves the cuprous oxideas soon as it is formed, yielding a colorless solution. There is no reddish precipitate to cloud the fluid and make the end point uncertain. The standard reagent is prepared as follows: Dissolve 356 grs. of Rochelle salt and the same weight of caustic potash in distilled water; dissolve separately 73 grs. of recrystal- lized cupric sulphate in more water with heat. Add the copper solution to that first prepared, and when cold add 12 fluidounces of strong ammonia (sp. gr. 0.880), apd distilled water to 40 fluidounces. The process is as follows: Dilute ten c.c. of this standard solution-equivalent to five milli- grammes of glucose-with 20 c.c. of distilled water, and place in a six or eight oz. flask. Attach this by means of a cork to the nozzle of an ordinary burette, filled previously with the urine diluted one part with nine of water. Boil the blue liquid in the flask resting on wire gauze. Allow the urine to flow into the boiling solution slowly until the blue color is exactly dis- charged. The number of cubic centimetres used will contain five milli- grammes of glucose. The urine should be diluted to such an extent that not less than four or more than seven c.c. are required to decolorize the solution, and the proportions necessary will be found to vary from one part of urine in two and a half to one in 30 or 40. If you wish the percentage of sugar, multiply 0.005 by 100, and divide the product by the number of cubic centimetres of dilute urine employed. The figure thus obtained, multiplied by the extent of dilution-i. e., if there is one of urine in ten, multiply by ten-gives the required percentage. The number of grains per fluidounce can be obtained by multiplying the percentage by 4.55. 38. Johnson's Picro-saccharometer.-This test is based on the observa- tion that glucose, in the presence of potassium hydrate, will reduce a bright- 40 EXAMINATION OF MORBID URINE. yellow solution of picric acid to a red solution of picramic acid. The value of this change for qualitative testing is much impaired by the fact that normal urine will cause some degree of redness, owing to the creatinine it contains. In quantitative work some allowance can be made for this in making a standard of color. The test is found practicable by students, and for comparative and clinical purposes may be depended on. The depth of the red color will correspond to the amount of glucose plus the effect of creatinine, which latter is usually equal to that of a solution of glucose containing o.l per cent. If albumin is present, the urine is simply made turbid and can be easily cleared by filtration. As a standard of red color, a solution of ferric acetate can be made by the following formula more easily than by Johnson's formula, which uses ingredients of a strength peculiar to the British Phar- macopceia:- K . Liq. ferri chloridi, U S. P., . . . fgj Ammon, carb.,gj Ac. acetic.,f£v Aq. destil.,ad . . M. This is put into a stoppered " weighing bottle," which is clamped to a graduated " mixing bottle " of the same diameter and of 100 c.c. capacity, or two cylindrical decimal graduates of equal size may be used. It is best to standardize the color solution by com- parison with that of a tested solution of glucose of known strength, made by rubbing up one grain of crystallized glucose in a fluidounce of urine. By this means we practically make allowance for the effect of the amount of creatinine in normal urine. In dia- betic urine the proportion of creatinine would be in most cases reduced. If the color of the ferric solution is not deep enough, add a few drops of liquor ferri chloridi, or if too deep dilute it with water. In the intervals of use the apparatus should be kept in the dark to prevent the bleaching action of sunlight. If constantly employed,* the color solution should be renewed or standardized every month. Johnson's method is to mix the suspected urine as follows:- Fig. 15. Picro-saccharometer of G. Johnson. 41 EXAMINATION OF MORBID URINE. Take of urine, fgj Liq. potassae, . f 3 ss Sol. picric acid (gr. 5.3 to f j), . . frpxl Water, ad . . Mix; put in a test tube, mark the level of its surface with a file scratch, boil sixty seconds, add a little water up to the mark, and cool. Measure into the graduated cylinder ten parts of this deeply-colored urine. If it is of the same depth of color as the standard, then the ten division denotes one grain of sugar to the fluidounce. If of greater depth, add water by small installments, shaking the bottle until it is like the standard. For every ten division of the diluted urine there is one grain of sugar to the ounce. If it measured 45, then there are 4.5 grs. to the fluidounce. If the sugar is very abundant, i. e., over five grs. to the ounce, dilute the urine one part in ten. Then each ten division stands for ten grs. and 45 would equal 45 grs. 39. To simplify the measurement of the ingredients of the urine mixture, it is customary in Jefferson Medical College laboratory to use but one delivery pipette of five c.c. This requires some change in the strength of solu- tions employed; the liq. potassae should be of lower specific gravity and the picric acid solution weaker, as stated in the following formula. Take of urine, 5 c.c. Liq, potassae (sp. gr. 1.036), . . 5 c.c. Sol. picric acid (gr. 3.5 to 5 c.c. Water 5 c.c. M. Boil 60 seconds, measure and dilute precisely as with the mixture given above. Having found the number of grains to the fluidounce the change to parts per cent, is made by multiplying with the factor 0.219. In all the reduction tests the reading is usually higher than is strictly accurate, owing to the presence of uric acid and creatinine. The aggregate proportion of reducing bodies in normal urine is relatively so small that for clinical purposes it may be disregarded. It is asserted that the reducing power of normal urine upon cupric oxide and picric acid is due to uric acid and creatinine. According to Johnson, both can be removed by adding to the amount of urine to be used its volume of cold saturated solution 42 EXAMINATION OF MORBID URINE. of sodium acetate, and then % its volume of a cold saturated solution of corrosive sublimate. After filtration, should the urine reduce picric acid, then sugar has done the work. 40. Fermentation Test for Quantity.-Roberts' Differ- ential Density Method. Take the sp. gr. of the urine and record it; mix well urine enough to float the urino- meter, say four fl. ozs. (120 c.c.) with half a cake of com- pressed yeast; set aside in a warm place (mantelpiece) for 24 hours. After the fermentation take specific gravity again and subtract from that taken before. Each degree of the remainder represents one grain of glucose to the fluidounce. Multiply by 0.219 to get percentage. Thus-:-Sp. gr. before fermentation, 1035 " after " 1015 1035 - 1015 = 20 degrees of density lost, or 20 grs. of sugar to the fluidounce. This test is very easily performed, and conclusive as to the presence of sugar, though it is not absolutely accurate as to quantity. 41. Fermentation Saccharometer.-None of the urinary substances which give reactions of reduction resembling those of sugar are susceptible of the vinous fermentation. When sugar is present and the urine is mixed thoroughly with brewers' yeast and kept in a warm place (about ioo°) for a day, it will ferment, giving off bubbles of carbon dioxide and forming alcohol by the following reaction :- ~ -ZCgHfiO 4 2CO2 Glucose. Alcohol. The fact of fermentation can be shown by collecting the carbon dioxide gas evolved. Fill a test tube with the urine to which yeast has been added, and invert it over a saucer containing some of the same mixture. Set aside in a warm place, and if after 24 hours gas has accumulated it proves the presence of sugar. A convenient vessel for this test is shown in Fig. 16. It is tipped on the side, filled with the urine and yeast mixture, ten c.c., and kept in a warm place. In 24 hours the gas collects in the upright tube, which is graduated to read from 0.25 to one per cent., or from one to five c.c. It will not show sugar when present to a less amount than o. 1 43 EXAMINATION OF MORBID URINE. per cent., and owing to the varying solubility of the gas in urines of variable reaction and density it cannot be considered accurate. Indigo Carmine Test.-A blue solution of sodium sulphindigotate (indigo carmine) made alkaline with sodium carbonate and boiled with saccharine urine, changes from blue to violet- red-straw-yellow. Cooling, or even agitation, will cause the restoration of the blue color; whereas, if the bleaching is due to strong alkalies, the color will not return. A very handy and reliable form for the test is that adopted by Parke, Davis & Co. in their pellets, each of which contains the indigo carmine and the sodium carbonate combined. The solution does not keep well. 42. Method.-Put in 30 minims of water one pellet of indigo car- mine and sodium carbonate ; heat gently to solution ; let fall from a pipette one drop of urine and boil quietly. A change to red or yellow indicates sugar. Precaution.-If more than one drop is added there is a chance that a reaction will occur from healthy urine. 43. Fischer's Phenyl- hy dr azin Test.-Into a test tube put nearly half an inch of dry phenyl-hydrazin hydrochlorate, add pulverized sodium acetate an equal volume, and then fill the tube one-half with the urine. Agitate until the sodium acetate is dissolved, gently heating up to the boiling point, and finally boil for 30 seconds. Set aside about 20 minutes for the sediment to fall, remove a portion of the sediment to the slide of a microscope and examine with a power of 200 diameters. If sugar be present, needle-shaped crystals of phenyl-glucosazon have formed, as yellow branching sprays, or sheaves, or radiating stars. When the glucose is present in very small amounts it may require 48 hours for the fine crystals to fall in sufficient quantities for recognition. The red globules and large yellow plates often seen are not significant. Geyer found that Fig. t6. Einhorn's Saccharometer. 44 EXAMINATION OF MORBID URINE. this test gave positive results with normal urine, and hence considers it unreliable. It has proved of so little practical value that physicians seldom resort to it. Import of Glycosuria.-If glucose is detected by any of the tests given above it is pathological. If abundant and persistent, and associated with excessive drinking and eating on the part of the patient, who is emaciating at the same time, it denotes typical diabetes mellitus. The urine in this disease is usually pale, of a fruity odor and very abundant. It may amount to over two gallons or 10,000 c.c. daily, and, notwithstanding the large volume, the presence of sugar makes it dense, the specific gravity ranging from 1025 to 1045. Temporary, slight glycosuria may follow an excessive saccharine diet, and reducing substances simulating glucose may appear in urine after the administration of chloroform, ether, chloral, amyl nitrite, turpentine, sali- cylic acid, benzoic acid, glycerine, camphor and carbolic acid; also after poisoning from strychnia, morphia, arsenic, phosphorus or carbon monoxide. These reducing derivatives do not, however, undergo yeasty fermentation, as glucose does. It may be incidental to various diseases of the brain and spinal cord, and accompany phthisis, pneumonia, asthma, cirrhosis, cholera, and even be intermittent, like malarial paroxysms. ACETONE. (CHS-CO-CH3). Sometimes in advanced diabetes, in fever, or even in health, when the diet is rich in meat, the urine has an ethereal odor due to acetone. Legal's Test.-To five c.c. of urine in a test tube add several drops of a freshly made, strong solution of sodium nitro-prusside and a few drops of sodium hydrate. A red color appears in the urine and soon changes to yellow. Holding the tube slanting a few drops of strong acetic acid are allowed to trickle down the side so as to form a separate layer. A change of color from the yellow to carmine red at the line of junction indicates acetone. On standing a blue-green color is developed. In addition to the blood albumin and globulin, which are congeners and have the same serious significance, other proteid principles of different import may occur in the urine, such as mucin (nucleo-albumin), peptone, hemialbumose. The albumin of the blood serum occurring in the urine ALBUMIN. 45 EXAMINATION OF MORBID URINE. is abnormal, and is nearly always evidence of grave disease. It is de- tected by the use of heat and of various reagents which throw it out of solution as a white precipitate. In case of turbid urine, it is necessary to filter the urine clear before testing. If the urine is turbid from urates fil- tration may be dispensed with if the boiling test is used; gentle heat first causes the disappearance of the urates, and as the temperature approaches the boiling point the albumin coagulates. The most highly valued tests are nitric acid and boiling heat, the most trustworthy single test being the boiling of an acid specimen. 44. Nitric Acid Test, Heller's Method.-Put one inch of urine in a test tube and gradually run y inch nitric acid down the side while the tube is inclined, so as to prevent mixing. The acid is heavier and forms a bottom layer. Wait some minutes for the appearance of a white belt or zone at the line of contact of urine and acid. This is a coagulum of albumin. Although this is not the most delicate test, it is highly convenient, and albuminuria is rarely serious unless decided enough to be shown by this " contact" method. In dense urines a haziness may appear in the upper part of the urine, due to acid urates. Mucin (nucleo-albumin), if very abundant, may make a haze about A of an inch above the line of contact. Heat will clear up the urate cloud, whereas albumin will remain undissolved. In case of doubt, try another sample diluted one-half. 45. Roberts' Nitric Magnesian Reagent.-This reagent has been devised in order to obviate objections to handling nitric acid, which, if undiluted, is a corrosive liquid not conveniently portable. It can be kept in a cork- stoppered bottle and does not stain the hands. To five volumes of a fil- tered saturated solution of magnesic sulphate, prepared by dissolving ten parts of the salt in 13 parts of distilled water, add one volume of strong nitric acid. A couple of drachms of bright filtered urine is allowed to float on an equal quantity of this solution in a test tube; care being taken that the contact-line is sharply defined. In a period of time varying from a few seconds to a quarter of an hour, according to the amount of albumin present, a delicate opalescent zone forms at the point of junction, and if mucin also is present a more diffused haze higher up in the urine. Special 46 attention should be given to the position of the opacity. In some concen- trated urines a belt of urates will appear a little above the line of demar- cation; but these dissolve on warming. 4<>. Boiling Test.-First test the reaction. If acid, proceed at once to apply heat. If the urine be alkaline, then to a tube containing three inches of urine add one drop of acetic acid. Heat the upper half to boiling. If contrasted with the lower half it has a turbid appearance, then albumin is present. Albumin coagulates in acid fluids at 70° C. (1600 F.), but remains dissolved if the urine is alkaline. Serum albumin and its associate globulin are the only proteids that coagulate at this tem- perature. This is the most trustworthy test. Precaution : Sometimes when acetic acid is added to a urine already acid, the albumin is converted into acid albuminate, a form non-coagulable even on boiling. The acetic acid should not be employed except in an alkaline urine and even then only a drop, just enough to neutralize the reaction or make it slightly acid. More than this might coagulate mucin. 47. Rough Estimation.-The urine acidulated by a drop of acetic acid is boiled for several minutes and set aside for 24 hours. White flakes fall in a well-defined layer, the depth of which may be expressed as a fraction of the whole fluid in the tube, thus: it is y(T or moist albu- minous layer. 48. Picric Acid Test.-A saturated solution is prepared by dissolving six grs. of recrystallized picric acid in a fluidounce of water with heat, and decanting the clear solution. Some of the urine is rendered perfectly clear by filtration ; the tube is held inclined, and an equal volume of the picric acid solutionis gently poured down the side so as to form a top layer. On gently shaking it, the yellow color will diffuse downward, and as far as it EXAMINATION OF MORBID URINE. 47 EXAMINATION OF MORBID URINE. extends, the picric acid will coagulate the albumin. In the presence of albumin a more or less distinct haze is produced, which on heating to the boiling point is rather intensified than otherwise. Fallacies : Peptones, if present, yield a similar haze, and quinin, morphin, or other alkaloids a more or less crystalline precipitate ; but in both these cases the opalescence is com- pletely dissipated by moderate heat. Mucin (nucleo-albu- min), another important constituent of most urines, is also coagulated, but much more slowly than the other proteids. Picric acid is a sensitive test for albumin, but its clinical value is impaired by the fact that it gives a faint but percep- tible reaction under conditions not serious. 49. Esbach's Albumimeter.-The only ac- curate way of estimating the quantity of albumin is to coagulate the albumin by boil- ing the acidulated fluid, then, after filtration, drying and weighing the precipitate. It is sometimes desirable to determine the proportion present at intervals, so as to study the progress of a case. For this comparative and clinical purpose, the easy method of Esbach is to be commended. A tube gradu- ated like that in Fig. 17 is to be employed.. If care is taken that the urine shall be pre- viously neutral or acid, Johnson's saturated solution of picric acid may be employed. Dr. Esbach's test solution is prepared by dis- solving ten gms. of picric acid and 20 gms. of citric acid in 900 c.c. of boiling distilled water, and then adding when cold sufficient water to yield one litre. The citric acid is only employed for the purpose of maintaining the acidity of the liquid, and Fig. 17. 48 EXAMINATION OF MORBID URINE. is not essential except for urines decidedly alkaline. Tt is open to objection from its power of precipitating mucin. The filtered acid urine is poured into the glass tube up to the mark U, and then the reagent is added till the level of the liquid stands at R. Avoid shaking lest air bubbles form and float the clots which it is desired to collect at the bottom. Mix the liquids thoroughly, without shaking, by slowly reversing the tube a dozen times, close with a cork, and allow it to stand upright for 24 hours. The height at which the coagulum then stands, read off on the scale, will indicate the number of parts per thousand, or grammes of dried albumin in one litre. This divided by ten gives the percentage. If the coagulum stands above the mark 4, try again after first diluting the urine with one or two volumes of water, and then multiply the resulting figure by two or three,as the case may be. If the amount of albumin be less than 0.5 gm. per litre it cannot be accurately estimated by this method. 50. Acetic Acid and Ferrocyanide Test.-Hydroferrocyanic acid yields a precipitate immediately in the presence of much albumin, and, if traces only are present, in the course of a few minutes. To apply the test, strongly acidulate with acetic acid, and then add a few drops of recently prepared potassic ferrocyanide solution. This is one of the most delicate tests known, but is open to the objection that it precipitates hemialbumose and mucin, though it does not affect peptone. Trichloracetic Acid.-This test is said to have more than six times the delicacy of metaphosphoric acid and more than three and a half times that of nitric acid. It may be used in the crystalline form or in saturated solu- tion. If used in the solid form, it is dropped into the specimen and a tur- bidity results if albumin is present. In the liquid form it is best used by superposition in the same way as Heller's nitric acid test. The white ring which forms at once is characteristic and unobscured by the colored zone of oxidized pigments as in the nitric-acid test. It is too sensitive for prac- tical purposes, showing mucin in healthy urine. 49 EXAMINATION OF MORBID URINE. Acidulatedt Brine Test.-A saturated solution of sodium chloride with five per cent, of ac. hydrochloric, dilut. may be used by the contact method. It sometimes gives a cloud with normal urine, probably due to mucin. 51. Potassio-Mercuric Iodide or Tanret's Test.-The reagent is com- posed of mercury bichloride, 1.35 gms.; potassium iodide, 3.32 gms.; acetic acid, 20 c.c.; distilled water enough to make 1000 c.c. Used by contact, it gives an opacity with all theproteids, including mucin, and some reaction with a large proportion of normal urines. On this account, like sodium tungstate and metaphosphoric acid, it is open to fatal objections as a clinical test. Import of Albuminuria.-If serum albumin is persistently present, found by nitric acid and boiling, and when deposited by the boiling test makes a layer as thick as half of the column of fluid, it indicates Bright's disease. It appears transiently in febrile diseases, blood poisonings, pregnancy, epilepsy. It is found in poisoning from lead and arsenic. Traces some- times appear in adolescents from trivial causes. To make the diagnosis of Bright's disease conclusive, when only small amounts are present, tube casts must also be found, or some of the general symptoms, such as anaemia, car- diac hypertrophy, dropsy and characteristic eye-lesions. MUCIN (Nucleo-albumin). It is not surprising that the urine flowing over a mucous membrane should contain even in health this constituent of mucus. Usually only a trace can be discovered by coagulating with acetic acid or trichloracetic acid. In catarrhal states of the urinary passages it is present in larger amount. Re- cently it has been stated that nucleo-albumin is mucoid but not mucin. Its importance depends upon the fact that it is often confounded with serum albumin when tests requiring the organic acids, citric, acetic, and picric, are employed. The differentiation is made certain by the boiling test, which does not coagulate any proteid but serum albumin. Peptone may be found in the urine with or without albumin. 52. Apply the nitric acid boiling test, § 46 ; if a negative result follows, then test a fresh portion with acetic acid cold. A cloudiness would justify the suspicion of peptone. 53. Biuret Test.-Having got rid of the albumin by boiling, after acidulating with nitric acid and then filter- PEPTONE. 50 EXAMINATION OF MORBID URINE. ing, pour slowly about a drachm of this clear urine down the side of a test tube which contains an equal quantity of Fehling's solution. At the line of contact of the two fluids a cloudy belt of phosphates will form, above which, if peptone be present, a rose-pink halo will appear. Hemialbumose gives the same result. The presence of albumin imparts a violet hue. Import.-Peptone is present mainly as a product of the breaking up of inflammatory effusions by suppurative process in various tissues and organs. HEMIALBUMOSE is a mixture of proteids intermediate between albumin and peptone. It appears in the urine under like conditions with peptone. It will form a cloud with cold nitric acid, but clears up on heating. It is thrown out by acetic acid and potassium ferrocyanide. HEMATURIA. If the blood of haematuria is derived from the kidney, it is well mixed with the urine, to which it imparts a smoky appearance, and microscopic tube casts can usually be found. If from the ureter, large vermicular clots may be found. If from the bladder, the blood is usually abundant and bright red, with stringy clots. 54. Microscopic Appearances.-The most satisfactory test for blood is the presence of the peculiar corpuscles. These corpuscles in ordinary acid urine retain their bi-concave shape for several days (Fig. 19). If the urine is dense, they become crenated (Fig. 18). When the urine is alkaline from ammonia, as in cystitis, the red corpuscles are shriveled and accompanied by numerous leucocytes. When the blood-coloring principle haemoglobin is excreted in the urine, diffused and not corpuscular, other tests than the microscopic must be resorted to. These tests indicate haemoglobin, either with or without the corpuscles. The microscope will serve to exclude the corpuscular element. HEMOGLOBINURIA. 51 EXAMINATION OF MORBID URINE. 55. Heat Test.-To a suspected sample add a drop of acetic acid, and boil. A red or brown coagulum indi- cates haemoglobin. 56. Almeris Test.-To suspected urine add a few drops of tr. guaiac, freshly made. A white or greenish turbid- ity ensues. Sliake well and add a few drops of solution of hydrogen peroxide or of an old ozonized sample of oil of turpentine. Haemoglobin will slowly change the color of the precipitate to blue. Turpentine slowly absorbs oxygen, and after standing on the chemist's shelves for some time will act for this test as an ozone solution. As saliva or potassium iodide will have the same effect as blood, these must be ex- cluded before a positive conclusion can be reached. Hellers Test.-Enough potassium hydrate is added to make the urine alkaline. On boiling the phosphates separate, and subsiding carry down the haematin as a flaky red or brown precipitate. In healthy urine the precipitate will be white.. After the administration of santonin, chrysarobin, rhubarb or senna, the urine contains a yellow principle which is reddened by the potassium hydrate and precipitated with the phosphates. To distinguish this from hsematin enough acetic acid must be added to neutralize the alkali. This will clear up all the red precipitate except hsematin. Import.-Hematuria indicates hemorrhage from some part of the genito- urinary tract. Hemoglobinuria occurs in various diseases, attended by " a dissolved state of the blood," as pyaemia, scurvy, typhus, purpura, poisoning from hydrogen arsenide, phosphorus, carbolic acid, chloral, potassium chlorate. There is a periodical form, the pathology of which is obscure. PYURIA. As pus is an albuminous fluid, its presence will cause the urine to respond to the tests for albumin. In addition, it contains white corpuscles, which are spherical, granular and opaque (Fig. 4). The mucus corpuscle is 52 EXAMINATION OF MORBID URINE. Fig. 18. Crenated Red Blood Corpuscles in the Urine. X 35°. Fig. 19. Colored and (a) Colorless Blood Corpuscles of Various Forms. 53 EXAMINATION OF MORBID URINE. indistinguishable from it, but mucin will not become hazy with the cold nitric acid test for albumin. 57. Donne's Test.-Let the suspected urine stand several hours for the pus to settle, and decant the clearer part. Into the deposit put a piece of potassium hydrate and stir with a glass rod; pus will grow thick, tough and gelatinous, while mucus will form flakes and become thin. Import.-Ar sudden appearance of pus in the urine would point to the opening of an abscess. Usually pyuria is an indication of catarrh or inflam- mation of some part of the genito-urinary tract. EPITHELIUM. The mucous membrane of any part of the genito-urinary passages may shed its cells into the urine. While there is a great variety in form, it is usually not difficult to distinguish renal epithelium from that of extra-renal origin. The accompanying Fig. 20 will give an idea of the microscopic appearance of the tessellated, the columnar and the transitional cells that appear at times from different parts external to the kidney. Free renal epithelium is spherical, granular, nucleated, with a very faint wall or no wall whatever. Its character is easily made out by the fact that careful search will always reveal renal tube casts in the same sample. TUBE CASTS. When either albuminuria or haematuria attends diseases of the kidney structure the deposit in the urine will be found to contain microscopic casts of the tubules. They may be found in cases in which neither albumin nor blood can be detected by ordinary testing. Sometimes they are made of coagulated fibrin, sometimes of mucoid exudation, sometimes of the plastic material of broken-down epithelial cells. I. Epithelial casts are composed of plastic matter with cells imbedded. They mark the process of desquamation (Fig. 21-0). II. Hyaline casts are either transparent, soft, and very delicate in outline, or transparent, well-defined and brittle. The former variety is called mucous Fig. 20. g a, Epithelium from the human urethra ; b, vagina ; c, prostate ; d, Cowper's glands ; e, Littre's glands ; f, female urethra ; g, bladder. Fig. 2i. a, Blood cast; i, granular cast with epithelial cells imbedded ; c, amyloid cast. 54 55 EXAMINATION OF MORBID URINE. (Fig. 22), and is not always an indication of nephritis; the latter, waxy (Fig. is evidence of serious nephritis. in. Granular casts are opaque and yellowish. They are mucoid or waxy, or cellular material which has undergone a granular change. They often contain cells and fat globules (Fig. 2i-<5). iv. Fatty casts are those which carry free fat globules along with the Fig. 22. Hyaline Casts. coagulated matter. They are usually considered proof of fatty degenera- tion of kidney. v. Blood casts are composed of coagulated blood with corpuscles im- bedded (Fig. 2l-«). They show that the haematuria originates in the secreting structure of the kidney. SPERMATOZOA. These may be present in the urine voided after coitus or an involuntary nocturnal emission of semen, or be an indication of spermatorrhoea. They form a viscid, mucus-like deposit, which, under the microscope, is seen to be composed of ciliated bodies, like a tadpole, inch long. In the urine they are motionless and resist decomposition for days. 56 EXAMINATION OF MORBID URINE. CHYLURIA. Under very rare conditions the urine contains chyle. It then presents the appearance of milk forming a cream at the surface, and owing to the fibrin of the chyle it may coagulate spontaneously like a white jelly. It responds to the various tests for albumin. If agitated with ether it clears up, the fatty matter is dissolved and separates as a layer at the top. Under Fig. 23. a, Spermatozoa; c, amorphous calcium carbonate; b, crystalline magnesium phosphate. the microscope the fat appears in a molecular state as small, bright granules, soluble in a drop of ether. The same substances appear in the urine when the filaria sanguinis hominis is in the blood. MICROORGANISMS. After emission, the urine may become the medium for the development of many low forms of life proceeding from germs that get into it from the air or from the containing vessels. Various moulds and the yeast fungzis (Saccharomyces cerevisiae) are commonly met with in putrid or saccharine urine after several days. Sarcince in quantities have been found in urine at the time of discharge, probably produced in the bladder. Local symp- 57 EXAMINATION OF MORBID URINE. toms are supposed to be due to their growth. These vegetations appear in microscopic cubes composed of distinct particles. Bacteria not only cause the ordinary decomposition of stale urine outside but sometimes set up like processes inside the bladder. It is likely that this form of bacteria gets access to the urine in the urinary passages by FtG. 24. a, Micrococci in short chains and groups ; b, sarcinse; c, fungi from acid fermentation; d, yeast cells from diabetic urine ; e, mycelium of a fungus. means of dirty instruments, such as catheters and sounds. They may steal down the urethra in the inflammatory products of old gonorrhoea. If, now, the urine is detained in the bladder, these organisms set up the ammoniacal fermentation which induces cystitis. URINARY DEPOSITS. Besides the different constituents of urine, healthy and morbid, which are seen in the deposits, extraneous substances are sometimes found. Some of these microscopic bodies, such as hair, or fibres of cotton and linen, may be mistaken for tube casts. A dirty vessel is often responsible for the presence of oil, starch, wood, etc. Healthy urine may form, at or near 58 EXAMINATION OF MORBID URINE. the bottom, a light cloud of epithelial debris suspended in mucus. But if on emission an acid urine is turbid throughout, excess of mucus or pus or blood is present. If an alkaline urine is turbid, then the precipitated phosphates may be the cause. 58. A specimen may be obtained for examination by allowing the urine to settle, and then by means of a pipette a drop or two of the sediment is conveyed to the glass slide and covered with a thin glass disk. Examine with a % -inch objective, or any combination giving a power of 200 to 300 diameters. If amorphous, it may be urates or phosphates. 1. Put a few drops of the deposit into a test tube with some urine and warm it; the urates will dissolve, the phosphates not. 2. To another portion add any acid, such as nitric or acetic ; the earthy phosphates will dissolve. If minute globular particles with bright centres, it may be milk, chyle or fat. Shake some of the urine with ether; if chylous, its milky appearance will disappear and the fat separate as a top layer. If crystalline, it may contain one or more of the substances already described and figured ; uric acid, Fig. 6 ; calcium oxalate, Fig. 9 ; triple phosphate, Fig. 4; calcium phosphate ; cystin, Fig. 10 ; leucin and tyrosin, Fig. 11. If organized bodies, they may be epithelial cells, Fig. 20; mucus or pus, Fig. 4 ; blood, Fig. 19 ; spermatozoa, Fig. 23 ; yeast fungus, mould, sarcinae, bacteria, Fig. 24. If a microscope is not at hand, the nature of an unorganized deposit may be determined by the following method :- 59. Rough Tests for Common Unorganized Deposits:- 1. Warm the deposit with some urine. It dissolves - urates. It does not dissolve = phosphates, or calcium oxalate, or uric acid. 2. Warm a fresh portion with acetic acid. It dissolves - phosphates. It does not dissolve = calcium oxalate, or uric acid. Warm the undissolved portion with hydrochloric acid It dissolves = calcium oxalate. It does not dissolve == uric acid. 59 EXAMINATION OF MORBID URINE. URINARY CONCRETIONS. About 80 in 100 of all urinary concretions are made of Uric Acid and Urates. Next in frequency are the Calcium Oxalate, or mulberry calculi. The rarer primary constituents are Blood Concretions, Cystin, Xanthin, Calcium Phosphate, Calcium Carbonate. Secondary to any of these there may appear at the last stage in the history of a calculus a deposit of Mixed Phosphates. These form a white layer, precipitated upon the calculus as a result of ammoniacal decomposition in the urine. As two or more deposits of different compositions may alternate in the formation, in examining a concretion it should be sawed through the middle, so as to expose its concentric layers. A small portion of each distinct layer should be examined by the following method:- 60. CASUAL ANALYSIS OF CONCRETION. First burn a portion of the powder on Platinum foil in a Bunsen burner or blowpipe fame. A. It chars greatly, leaving but little ash. = (Uric acid, urates, cystin, xanthin, blood.) It gives murexide reaction. See page 24. = (Uric acid or urates.) It dissolves in boiling water. - (Urates ) It does not dissolve in boiling water. = (Uric acid.) Cystin and xanthin are very rare; the first can be recognized by test on page 28. B. It chars very slightly, leaving very much ash. = (Phosphates, oxalate, carbonate of calcium.) 1. Treat a fresh portion with dilute hydrochloric acid. It is soluble with effervescence. = (Carbonates are present.) " " without " = (Phosphates or oxalate calcium.) 1. Treat with acetic acid. If it is soluble without effervescence, then it is phosphatic; it fuses into a bead on platinum foil (mixed phosphates) • it does not fuse (calcium phosphate). If it is insoluble, then it is calcium oxalate, and when cal- cined on platinum, leaves an ash that turns red litmus blue, or effervesces with HC1. PART II. EXAMINATION OF THE GASTRIC CONTENTS. By the term Gastric Contents, used in this manual, is meant the material found in the stomach one hour after a simple test-meal has been adminis- tered. At this time normal gastric digestion is about at its height. The meal to be referred to hereafter represents albuminoids and carbohydrates with the usual inorganic constituents of food. Before the expiration of one hour the food has been altered by digestion. The salivary ferment ptyalin has converted some of the starchy material into dextrine and glucose. Al- bumin has been converted to peptone. Part of the glucose has undergone a fermentation by the bacteria, which generate lactic acid. Immediately upon the reception of food the secretion of gastric juice begins. This is a clear, pale-yellow, thin fluid, acid in reaction, holding in solution about one per cent, of solids mostly organic in character. The quantity secreted daily is very large, averaging 20 pints. Its specific gravity varies between 1001 and 1010. Composition of Gastric Juice. Per Cent. Water, 99-44 Solids, as tabulated below, 0.56 Organic Substances (Pepsin and Peptones), 0.32 Free Hydrochloric Acid, 0.25 Sodium, Potassium and Calcium Chlorides, 0.21 Calcium, Magnesium and Ferric Phosphates, 0.02 60 61 EXAMINATION OF THE GASTRIC CONTENTS, a. Withdraw the gastric contents one hour after the administration of the "test breakfast" §62, noting if additional water was necessary for evacuation. b. Note the color, odor, and other physical characters. c. Filter one-half of the contents. f. Start the digestion test for pepsin, §64. 'g. Determine the total acidity with sodium hydrate, § 76. h. Test for free hydrochloric acid. i. Test for the organic acids. j. If the appearance warrants it, test for bile, § 2, and blood, § 56. 61. PLAN FOR CLINICAL EXAMINATION. REAGENTS REQUIRED, Congo-red paper. Tropseolin O O. Gunzburg's or Boas' solution. Egg albumin. Solution carbolic acid, four per cent. Phenolpbtalein, one per cent. ale. Sol. neutral ferric chloride. § 74 Hydrochloric acid. 62 EXAMINATION OF THE GASTRIC CONTENTS. The specimen to be examined should contain those most important con- stituents of the gastric juice, pepsin and hydrochloric acid. After taking food, for the first three quarters of an hour the hydrochloric acid present is not free but combined with proteid substances and does not interfere seri- ously with the amyloid digestion by the alkaline saliva. If the reaction at this first period should be acid the acidity is due to the organic acids, lactic, butyric, and acetic, produced by fermentative changes in the food. After an hour only a trace of lactic acid should be found, while the amount of hydrochloric acid increases for two or three hours. The sample for clin- ical examination is usually obtained after a test-breakfast. 62. Ewald's Test-Breakfast.-This consists of nine drachms (35 gms.) of dry bread (an ordinary roll of about this weight) and ten fluidounces (300 c.c.) of warm water or very weak tea without milk or sugar. In an hour this will be liquefied and can be expressed through the tube. The stomach-tube is made of soft, smooth, flexible rubber, long enough to reach the stom- ach and leave several feet free outside the mouth. It should have a large, free bore and several smoothly rounded windows near the end that enters the stomach. The patient sits erect in a chair with his head thrown back. After wetting the tube in warm water, it is passed back to the pharynx and held there in spite of gagging until the patient makes an effort at swallowing. By gentle pressure it can then be readily passed into the stomach. Sometimes the gastric contents are evacuated at once without effort. Pressure over the abdomen while the patient coughs will almost always force them out. At times it is necessary to wash out the stomach by in- serting a funnel into the outer end of the tube and fill- ing tube and funnel with warm water of known quan- tity. Then to make a siphon before all the water has THE SAMPLE. 63 EXAMINATION OF THE GASTRIC CONTENTS. run from the funnel the tube is lowered into a vessel on the floor. If ten fluidounces (300 c.c.) of water or tea have been given, about one and a half fluidounces (40 c.c.) of fluid should be obtained by the tube. By filtration a clear solution can be had for the application of the tests. 63. In some cases, such as suspected ulcer, and when the patient is highly nervous, a small amount will suffice for the necessary tests. To secure this Einhorn's little silver bucket is swallowed an hour after the test-breakfast. After five minutes the bucket is withdrawn by a cord. If resistance is felt when the bucket gets as high as the larynx a deep expira- tion will remove it. Pepsin is the ferment which changes proteid substances into peptones in an acid medium. Normally it is associated with hydrochloric acid; but the lactic acid found in the stomach, even when hydrochloric acid is absent will aid it to dissolve the proteids. To detect its presence the following procedure is used. 64. Test for Pepsin.-First boil an egg for four minutes. Having cut the white part into thin flakes, punch out small discs with a cork borer or a glass tube. (Such pieces as are not wanted can be kept indefinitely in glycerine and washed before using.) Into a test-tube or watch-glass containing two drachms or more of filtered gastric contents, put two drops of hydrochloric acid and a disc of albumin. Stand aside for a half-hour or more at a temperature of 104° F. (40° C.). If pepsin is present the disc will be made rough and smaller by partial solu- tion. (For practice the student can make an artificial gastric juice by dissolving 1 y? grains of pepsin in a fluid- ounce of water and adding five drops of hydrochloric acid.) PEPSIN. 64 EXAMINATION OF THE GASTRIC CONTENTS. Estimation of Pepsin Strength.-Cut the coagulated albumin into pieces inch (one m.m.) thick and punch out discs f inch (ten m.m.) in diameter. Into each of two test-tubes put two discs and drachms (ten c.c.) of filtered gastric contents. To one of these tubes add two drops of strong hydrochloric acid, and then stand both in an oven heated by a thermostat to ioo° F. (40° C.). If pepsin and acid are present in normal amount, the discs in both tubes will dissolve in one to two hours. If the acid is deficient, solution will occur in the tube only to which the hydro- chloric acid was added. Clinical Import.-Pepsin is very rarely absent even in serious disease of the stomach. Failure in gastric digestion is seldom due to the lack of it. If the flakes of albumin digest without adding hydrochloric acid, it does not prove a normal state of things, as digestion is possible by the presence of lactic acid alone, though the condition is not a healthy one. It is of the first importance to prove the presence of hydrochloric acid by tests to be given hereafter. 65. Peptone may be recognized by the biuret test (see § 53, page 49). It is seldom of any value to carry the investigation further, but if desired albumin may be detected in a fresh sample by a precipitate formed on boil- ing ; when this is separated by filtration, a precipitate obtained from the filtrate by adding nitric acid denotes propeptone; after a second filtration, the addition of solution of tannin will precipitate peptone. PEPTONE. HYDROCHLORIC ACID. In the gastric juice free hydrochloric acid is present 0.25 per cent. One hour after a test-breakfast, if hydrochloric acid is detected, we may be sure that the amount of lactic acid is so small as to be safely disregarded in casual analysis for clinical purposes. The hydrochloric acid exists partly in a loose combination with proteids and partly free. In various chronic diseases of the stomach a chemical examination of the gastric contents must be made to judge of the extent and nature of the morbid mischief. The main point is the. determination of the presence of hydrochloric acid and its amount, free and combined. 66. The Acid Reaction.-To detect the presence of free hydrochloric acid, litmus paper will not serve, as it is affected by acid salts and by the organic acids, lactic, butyric and acetic. It has been found convenient to 65 EXAMINATION OF THE GASTRIC CONTENTS. use paper colored with certain anilin dyes (Congo-red and tropaeolin are the best) which react to minute quantities of free hydrochloric acid, but which are unaffected by acid salts and by the organic acids, lactic, butyric and acetic only when present in 0.5 per cent., an amount never found in the gastric contents now being studiecj. The most delicate reaction for distinguishing hydrochloric acid from the organic acids is that given by phloroglucin vanillin. Detection of Free Hydrochloric Acid. 67. Congo-red Test.-Take a piece of Congo-red paper and put upon it a drop of the gastric contents. A dark blue spot will appear if free hydrochloric acid is present as much as 0.05 per cent. A pale blue or a violet spot or a blue ring only around the wet place signifies some free acid, either a trace of hydrochloric acid or some organic acid. 68. Tropceolin (dimethyl-orange) Test.-Have on hand saturated alco- holic solution of pure tropaeolin (00). Shortly before use, immerse in this solution some pieces of white filtering paper and dry them. (This paper loses its sensitiveness on keeping). Wet the prepared paper with a drop of the gastric contents. If hydrochloric acid is present to 0.05 per cent, a dark reddish-brown spot appears, which will change to lilac when gently heated on a watch glass held by the fingers over a small flame. The or- ganic acids as found in the gastric contents do not have any effect. In larger amounts they make a faint brown stain which does not show a lilac color when heated. 69. Methyl-violet Test.-Make a permanent concentrated solution of methyl-aniline-violet in water. Prepare the test fluid by adding to one or two drachms (five to ten c.c.) of water, enough of the strong solution (two or three drops) to give a distinct violet hue. Put half of this freshly diluted solution into a test tube with an equal quantity of filtered gastric contents. Compare the result with the unused remainder of test fluid. A change from violet to blue indicates hydrochloric acid when present in more than 0.03 per cent, and less than 0.5 per cent. Organic acids do not change the color unless present in amounts not found in the gastric contents. 66 EXAMINATION OF THE GASTRIC CONTENTS. 70. Phloro-glucin Vanillin {Gunzburg's) Test.-It is best to make the reagent freshly. It will serve for a while if kept in dark-colored bottles. Take of- Phloroglucin, 2 parts (30 grs.) Vanillin, I part (35 grs.) Absolute alcohol, 30 parts by weight (1 fl. oz.) or Alcohol, 80 per cent., 100 parts by measure. Mix and make a clear, pale yellow fluid. 71. Boas' Resorcin Solution is a reagent of almost equal delicacy, has greater stability, and is cheaper than Gunzburg's. To make it take of- Resorcin, pure, 5 gms. White sugar, 3 gms. Dilute alcohol, 100 c.c. These two tests are applicable to free mineral acids only, being unaf- fected by the amount of organic acids found in the gastric contents. The method is the same for both. 72. Method for Gunzburg*s or Boas' Tests.-Mix upon a watch crystal or porcelain dish about five drops of the test solution and, three of the gastric contents either filtered or unfiltered. Gently heat the dish held in the hand by cautiously moving it over a small flame, and if free hydrochloric acid is present to 0.005 Per cent, a bright red ring will form at the margin as the mixture dries. When several samples are to be tested, or when the reagent is used to indicate the end of a reaction, it is more convenient to put one c.c. of the reagent on a flat porcelain dish and dry it by constant motion, so as to spread it in a thin layer. The heat should not be above 1 io° C., or charring will ensue. A glass rod carrying a drop of the gastric fluid is touched to the prepared re- agent and the dish cautiously dried. A red ring will appear if so much as 0.003 °f ree hydrochloric acid is present. 67 EXAMINATION OF THE GASTRIC CONTENTS. Precaution.-The proteids in the gastric contents, if evaporated and dried with a strong heat, may show a deep red color at the centre. To avoid this fallacy, the evaporation must be done slowly or at a gentle heat. Hydrochloric acid will then cause a red color where drying first occurs, at the edges, or in streaks. 73. Approximate Estimate.-By diluting a portion of the gastric con- tents to or r or or until no reaction occurs on the dish prepared with dried Gunzburg's reagent, the amount of free hydrochloric acid can be estimated. As the limit of the reaction is about 0.005 Per cent, a red color with the dilution would indicate 0.05 per cent.; with the dilution it would mean as much as o. 1 per cent.; with the dilution as high as 0.15 per cent.; with the as high as 0.2 per cent. Clinical Import.-The absence of free hydrochloric acid in the contents taken one hour after the test-breakfast, de- notes either such a disorganization of the gastric mucous membrane as is found in atrophy and in amyloid degen- eration, or the dilatation which accompanies gastric can- cer and chronic catarrh. 74. Lac tic Acid, by the Carbolo-ferric or Uffelmann's Test.-Prepare Uffelmann's reagent freshly by mixing one drop of a dilute solution of fer- ric chloride (U. S. P.) with two and a half fluidrachms (io c.c.) of a four per cent, solution of carbolic acid and five drachms (20 c.c.) of water. Or if desired the following official preparations can be used :- K • Liq. ferri chloridi gtt. j Liq. acidi carbolici, gtt. iv Aqua, f 0 v- M. Sig.-Uffelmann's reagent. When first made the reagent has an amethystine blue color. Method.-Mix equal parts of Uffelmann's reagent and filtered gastric contents. If even so small an amount as 0.01 per cent, of lactic acid be present, the color changes to canary yellow or greenish yellow. By the other DETECTION OF ORGANIC ACIDS. 68 EXAMINATION OF THE GASTRIC CONTENTS. gastric acids, the color is not changed to the same depth of yellow, though the blue color may be dis- charged by them if the very unusual amount of 0.3 per cent, be present. In cases of doubt, to eliminate any chance of error, the lactic acid may be separated by shaking two and a half drachms (ten c.c.) of the gastric contents with ten drachms (40 c.c.) of ether. The test is then applied to the liquid residue left when the ether has evaporated. 75. Acetic and Butyric Acids.-The residue obtained by evaporation of the ethereal extract above mentioned contains all the organic acids. If they are present this residue will be an acid fluid showing the volatile acetic and butyric acids by their odor. Butyric Acid.-In the very rare cases when fermentation produces this acid in the large amount of over 0.5 per cent, it changes Uffelmann's blue reagent to a red brown. If a portion of the ethereal extract be diluted, and a piece of calcium chloride added, the light butyric acid will float like oil globules on the saline solution below. Acetic Acid.-Having neutralized the ethereal extract by carefully adding sodium or potassium hydrate, add one drop of solution of ferric chloride. Acetic acid will form deep red ferric acetate, which on boiling will precipi- tate as the brownish basic salt. TOTAL ACIDITY. 76. The hydrochloric acid is an anti-ferment. When it has been detected in the gastric contents one hour after a test breakfast, by some of the tests above named, it may be safely assumed that there is very little if any of the organic acids, lactic, acetic, and butyric, which are mainly pro- ducts of fermentation. Conversely, to discover the organic acids by Ufifel- mann's test is to establish the presumption against normal amounts of hydrochloric acid. The acidity of the small proportion of organic acids that may be present is so small as to be a negligible quantity in clinical studies. After ascertaining the presence of hydrochloric acid by Gunz- burg's or Boas' test, to determine the total acidity is practically to estimate the amount of hydrochloric acid present. The degree is expressed in terms of that acid. • Method.-Having diluted five or ten c.c. of the filtered gastric fluid with 20 or 40 c.c. of water, a few drops of 69 EXAMINATION OF THE GASTRIC CONTENTS. an alcoholic solution (one per cent.) of phenol-phthalein are added as indicator. The colorless liquid is put into a beaker or porcelain dish. A graduated burette is charged with the deci-normal solution of sodium hydrate and a few drops are run into the liquid in the dish until a reddish color appears. If ten c.c. of the gastric contents have been used and the amount of hydrochloric acid is normal, it will require from four to five c.c. of the sodium hydrate solution to change the color. As each c.c. of this alkali solution will neutralize 0.00364 gram of HC1; if 3 c.c. have been required, then the acidity is calculated thus : 3 X 0.00364 = 0.01092 of HC1. If ten c.c. of gastric liquid were used, then to get percentage 0.01092 X 10 = o. 1092 per cent. HCL If five c.c. were used, then 0.01092 X 20 = 0.2184 per cent. HC1. Sometimes the acidity is expressed by the number of c.c. of sodium hydrate which is required to neutralize 100 c.c. of the stomach liquid. An acidity of 45 per cent, would then mean that 100 c.c. of the gastric liquid required 45 c.c. of sodium hydrate to neutralize it. As stated above, the small fallacy due to the presence of organic acids may be safely ignored in most cases. If a marked reaction has been obtained by Ulfelmann's reagent, then before titration the organic acid may be separated by shaking the ten c.c. of gastric fluid with too c.c. of ether. 77. Clinical Import.-Acidity is diminished in some cases of anemia, fever, and atonic dyspepsia. Acidity is excessive in most cases of gastric ulcer, in some forms of nervous dyspepsia and gastric catarrh. Before drawing conclusions based on the degree of acidity, the total acidity must be determined. 70 EXAMINATION OF THE GASTRIC CONTENTS. ESTIMATION OF FREE HCl, THE ORGANIC ACIDS, AND ACID PROTEIDS. 78. Som% of the hydrochloric acid reported by the "total acidity" when phenolphthalein is used as indicator, is in reality not free but loosely com- bined with proteids. When so desired, the free hydrochloric acid may be estimated by the procedure recommended by Mintz, who uses Gunzburg's reagent as an indicator. He asserts that the alkali first combines with the free acid and later with the acid proteids. Method. First Stage, for Free HCl.-Put into a beaker or dish ten c.c. of filtered gastric fluid. From a burette add drop by drop the deci-normal solution of sodium hydrate. After each addition take a drop of the mixture on a glass rod and touch it to a prepared surface of dried Gunzburg's or Boas' reagent. The alkali is run in as. long as the crimson spot shows. When at last no reaction appears, the free acid is calculated by the amount of alkali solution used. Thus, if two c.c. of deci-normal solution of sodium hydrate have been re- quired to make the ten c.c. of gastric fluid cease to react with Gunzburg's reagent, then the free hydrochloric acid would be 2 X 0.00364 = 0.00728 in 10 cc., or 0.072 per cent. Second Stage, for Organic Acids.-Having reached the end of Gunzburg's reaction in the method just described, any acidity left must be from organic acids or acid pro- teids. Test with Congo-red paper a drop of the fluid already treated in the first stage. If no change occurs there is less than 0.02 per cent, of organic acid. If a blue spot appears, continue to run in the soda solution until the Congo-red is unaffected by the fluid. The total of soda used minus that employed in the first stage will give the free organic acid present, usually considered 71 EXAMINATION OF THE GASTRIC CONTENTS. as lactic acid, but expressed in terms of hydrochloric acid. Thus, if 0.5 c.c. of the soda solution were re- quired to neutralize this last portion, then the free organic acid would be 0.5 X 0.00364 = 0.0018 in ten c.c. or 0.018 per cent. Third Stage, for Acid Proteids.-At the end of Gunz- burg's reaction in the first stage, if no change is made on Congo paper, omit the second stage altogether and proceed at once to the third stage. Or if the end reac- tion of the second stage with Congo paper has been determined, add a few drops of phenolphthalein solution to the gastric fluid already treated in the first stage and run in the sodium hydrate solution until a faint pink hue appears. This last addition of sodium hydrate repre- sents the loosely combined acid. The whole amount of alkali required in all the stages (usually from seven to eight c.c.) gives the " total acidity." ANALYSIS BY EVAPORATION. 79. It is a familiar fact that free hydrochloric acid can be separated from the gastric contents by vaporizing with gentle heat. Recently it has been shown that when a portion is loosely united with the proteids of digestion the heat of a water bath will not volatilize this combined hydro- chloric acid. This is the basis of the following procedure for estimating the acid free and combined. Method. First Stage.-Apply Gunzburg's or Boas' test For free HC1. (§ 72.) Second Stage.-Estimate the "total acidity" in ten c.c. (§ 76.) Third Stage.-Evaporate ten c.c. to complete dryness in a water bath of any simple form (a short exposure in a warm oven will do, with care to prevent overheating). 72 EXAMINATION OF THE GASTRIC CONTENTS. Dissolve the residue in 40 c.c. of water and proceed to estimate the acidity as advised in § 76. If the first stage showed no free HCI, now apply Uffelmann's test (§ 74); if this showed no lactic acid, though there was an acid reaction, the total of which remained the same after as before evaporation, then the acid may be considered as all combined with proteids. If, however, the original gastric fluid gave the yellow reaction to Uffelmann's test, then this lactic acid can be removed from the gastric fluid by washing with ether. The remainder should be evaporated to dryness and titrated for acidity. This will represent the acid proteids, and if subtracted from the original total acidity will give the amount of lactic acid remaining. If evaporation of the remainder lessens its acidity, though no HCI is present, the loss of acidity will represent the volatile acetic and butyric acids. Ewald's Salol Test is the best for determining the sufficiency of the stom- ach in propelling its contents onward to the duodenun. Salol (phenol salicylate) in an hour passes unaffected by the acid contents of the stomach into the bowels, where it encounters the alkaline pancreatic fluid and is split into salicylic acid and phenol. These are absorbed and appear directly in the urine. Method.-Give io or 15 grs. of salol in a capsule or coated pill. In an hour test the urine for salicylic acid by wetting a piece of filter paper with urine and touching the moistened paper with a drop of a ten per cent, solu- tion of ferric chloride. A trace of salicylic acid is sufficient to develop a violet ring around the drop. A more conclusive result is obtained if the urine be examined after 30 hours. The salicylic acid should all have been absorbed and eliminated and therefore absent from the urine before that time. If the violet reaction should appear at this time there is proof of sluggish action of the stomach. 80. MOTOR POWER OF THE STOMACH. PART III. EXAMINATION FOR COMMON POISONS. CLASSIFICATION OF POISONS. The following simple arrangement of commonly occurring poisons is adopted as suited for the preliminary tests of the practitioner of medi- cine:- A. Acids. Nitric, HNO3; Hydrochloric, HCl; Sulphuric, H2SO4; Oxalic, H2C2O4. B. Alkalies. Potassa, KHO ; Soda, NaHO ; Ammonia, NH4HO. C. Volatile Matters. Carbolic Acid, C6H5OH ; Hydro- cyanic Acid, HCN; Phosphorus, P; Chloroform, CHC13; Chloral, C2HC13O ; Alcohol, C2H5HO. D. Metallic Poisons, subdivided into- 1. Those precipitated yellow or orange from an acid solution by hydrogen sulphide, H2S; Arsenic, As.; Antimony, Sb.; Cadmium, Cd ; Tin, Sn. 2. Those precipitated black from an acid solution by hydrogen sulphide, H2S ; Copper, Cu; Mercztry, Hg ; Lead, Pb; Silver, Ag; Bismuth, Bi ; Plati- num, Pt; Gold, Au. 73 74 EXAMINATION FOR COMMON POISONS. 3. Those precipitated by adding ammonia, NH4HO with ammonium sulphide, (NH4)2S; Zinc,Zn; Nickel, Ni ; Cobalt, Co; Iron, Fe; Chromium, Cr; Manganese, Mn ; Aluminium, Al. 4. Those precipitated white by adding ammonia, NH4HO and sodium phosphate, Na2HPO4; the metals of the alkaline earths ; Barium, Ba ; Cal- cium, Ca ; Strontium, Sr ; Magnesium, Mg. 5. Those not precipitated by preceding reagents: the alkaline metals, Potassium, K, and Sodium, Na. E. Vegetable Alkaloids. Morphine, Strychnine, Atro- pine. REAGENTS FOR TOXICOLOGY. In addition to the reagents and apparatus mentioned on p. viii, the fol- lowing solutions are required for the experiments described in this division. Acid Oxalic (normal solution). Potassium Ferrocyanide, (i in io.) Calcium Hydrate (Lime Water). Ferrous Sulphate, (i in 20.) Potassium Iodide. (1 in 20.) Ferric Chloride. (1 in 10.) Potassium Bichromate. (1 in 10.) Ammon. Sulphhydrate. Ammonium Chloride. (1 in 10.) Ferrous Sulphide in lumps for mak- Ammonium Carb. (1 in 20.) ing H2S. 75 EXAMINATION FOR COMMON POISONS. PRELIMINARY EXAMINATION FOR POISONS IN VOMITED OR OTHER SUSPECTED MATTERS. a. Note stains on the clothing or about the lips and mouth. b. Put vomited matters in a white dish and examine for odor, seeds, wing cases of cantharides, crystals, or powders. c. Take a small measured portion, say one-sixth or one-third ; put in a clean jar and stand in a bowl of warm water. d. TkvZ for volatile poisons. HCy by odor and by action of vapor on reagents ap- plied to the under side of a glass cover to the jar. § 118, 119, 120 . Phosphorus by odor and luminous vapor evolved in a dark room, by boiling a portion in a test tube. § 121, 122, 123. Carbolic acid by odor. §113, I14, 115, 120. e. Test for corrosive acids by litmus paper, by exam- ining stains on clothing, and by analyzing a sample of suspected substance. f. Test for corrosive alkalies by litmus paper or direct testing of suspected sample. g. Test for metallic irritants. Apply Reinsch's test, §136, for As, Sb, Hg ; if there is a deposit on the copper, then make a subli- Fig. 25. 76 EXAMINATION FOR COMMON POISONS. mate in an open glass tube. Examine sublimate with lens, wash out with HC1, evaporate, and test residue with H2S. If there is no deposit on the copper acidulate a portion with HC1 and add H2S. If it forms a black pre- cipitate test for Cu and Pb. § 146, 154, 155. h. If the patient has narcotism or convulsions, apply Stas' method for morphine or strychnine by mak- ing a portion of the fluid vomit alkaline with sodium carbonate, which sets the alkaloid free. The alkaloid is then separated from the liquid which holds it suspended by agitating it with four volumes of ether or amylic alcohol. By letting the mixture stand, the ether floating will hold the alkaloid in separate solution, and may then be removed by a pipette. (Fig. 26.) If the ether is allowed to evaporate on a saucer or watch-glass the residue may be ex- amined for strychnine, §166-169, and morphine, § 170-172. If the indications point to morphine, then agi- tate a fresh portion of the original vomited fluid with amylic alcohol, as it is a better solvent for that alkaloid, separate, evaporate, and test, § 170-172. Note.-The methods indicated above are " rough and ready," and are such as might be applied quickly to vomited matter by a practitioner be- tween his visits. They may serve to clear up a diagnosis before it is too late to help the patient. They would not answer for a juridical trial. The methods applied by expert analysts are far more searching, and require much more apparatus, time, and special knowledge. Before the analyst begins testing for the metallic irritants he first gets rid of all organic matter by some such process as that of Fresenius. The finely divided substance is boiled with pure hydrochloric acid, while potas- Fig. 26. 77 EXAMINATION FOR COMMON POISONS. sium chlorate is added from time to time until the solids are reduced to a straw-yellow fluid. This fluid is then treated with excess of sodium bisul- phite and saturated with hydrogen sulphide until the metals are thrown down as sulphides. These sulphides are then#treated with various tests for the metals. CORROSIVE ACIDS AND ALKALIES. The mineral acids and the alkalies corrode the tissues with which they come in contact and impart a strong corresponding reaction to the vomited matters. If alkaline antidotes have been administered the reaction may be neutral, or even alkaline, and if alkaline poison has been treated by vinegar or lemon juice the reaction may be changed to neutral or acid. The acids may have been converted into nitrates, chlorides, or sulphates. A complete examination of suspected material in the case of chlorides or sul- phates'requires an estimation of the quantity, to determine if these salts are in excess. As nitrates are not found in the normal viscera or the ordinary gastric contents, their bare presence in notable amounts is very suspicious. Besides determining the fact of acidity or alkalinity of the vomited matters, it may be necessary to determine the degree. This is done by measuring the amount of a standard test solution necessary to neutralize the sample, using litmus or phenolphthalein as an indicator of the neutral point. Rtiles for Volumetric Analysis.- I. Let some of the titrating solution run through the burette to wash it out. 2. Fill the burette to the brim at first, then let it run out into the supply bottle until the upper surface stands at the o mark. 3. At the first titration let the flow be drop by drop, mak- ing frequent trials with the indicator. 81. Acidimetry Experiment.-Measure one c.c. of nitric acid, HNO3, and fill to 50 c.c. with water. Put this dilute acid into a capsule or beaker and add a few drops of red litmus solution, or two drops of solution of phenolphtha- lein (1-100 alcohol). Fill a graduated burette (Fig. 2) to o with normal solution of potassium hydrate, KHO (56 gms. to 1000). Now, let this alkaline solution run, by drops, into the capsule until a violet color appears. 78 EXAMINATION FOR COMMON POISONS. Note the c.c. of KHO solution used, and calculate the amount of HNO3, official sp. gr. 1.42, neutralized by multiplying the c.c. of KHO used with 0.0908. If phe- nolphthalein be used as indicator, neutralization is denoted by a loss of the red alkaline color. HNO3 + KHO = KNO3 + H2O. Potassium nitrate and water are formed. Repeat this experiment with acids of different strength. 82. Alkalimetry.-This procedure is like acidimetry, using blue litmus as an indicator and a normal solution of oxalic acid (63 gms. to 1000) for testing. Each c.c. = 0.056 gm. of KHO. Nitric Acid, HNO3. Aqua Fortis. Acidum nitricum, sp. gr. 1.42 - 69.4 per cent. HNO3. Acidum nitricum dilutum (6 to 1) 10 per cent. HNO3. It causes death from acute laryngitis with oedema, or by perforation ot the stomach, or by secondary starvation. The tongue is stained yellow. 83. Test reaction with blue litmus paper. 84. Pour HNO3 on copper slips in a test tube and look for red-brown nitrous fumes, the liquid turning blue. 3Cu + 8HNO3 = 3(Cu2NO3) + 4H2O + N2O2. Copper nitrate, water, and nitric oxide are formed, and nitric oxide in contact with air changes to red fumes of higher oxides. 85. Touch the skin and the clothing with a drop of HNO3, and note in each case a yellow stain of picric acid, not removed by aqua ammonia. 86. HNO3 turns a crystal of morphia yellow, but one of brucia red. 87. To dilute HNO3 or a solution of a nitrate add a green crystal of ferrous sulphate and a drop or two of 79 EXAMINATION FOR COMMON POISONS. sulphuric acid. The crystal turns red-brown, ferric sul- phate being formed. Antidotes.-Soapsuds, magnesia, chalk suspended in milk, raw eggs. The stomach pump must not be used, as the tube may perforate the softened tissues of the stomach. Hydrochloric Acid. HC1. Spirit of Salt. Muriatic acid. Acidum hydrochloricum, 31.9 per cent. Acidum hydrochloricum dil. (3 to 13). It is the natural acid of the gastric juice in the proportion of 0.25 per cent. It causes death by corrosion, inflammation of the larynx, perforation of the stomach, or secondary starvation. 88. Test with blue litmus paper. 89. Add HC1 to a few grains of manganese dioxide, MnO2 in a test tube. Warm and note that chlorine gas forms and will bleach moist litmus paper held at the mouth of the tube. MnO2 + 4HCI = MnCl2 -j- 2lI2O + Cl2. Manganese chloride, water, and chlorine are formed. 90. Hold side by side the open mouths of bottles of HC1 and of ammonia, NH4HO, observing the white fumes of NH4CI formed. 91. Add to dilute HC1 or a solution of sodium chloride, NaCl, a drop of silver nitrate, AgNO3. A white precipi- tate falls, soluble in ammonia but insoluble in HNO3. NaCl + AgNO3 = NaN03 + AgCl. Sodium nitrate and silver chloride are formed. Mercurous salts will yield with HCI a white precipitate of Calomel, Hg2Cl2, insoluble in HN03 and blackened by NH40H. Lead salts give a white precipitate of lead chloride, PbCl2, insoluble in nh4oh. 80 EXAMINATION FOR COMMON POISONS. 92. Put into a test tube a solution of methyl violet, distinctly violet in hue ; add a few drops of filtered vomit or other matter; if free mineral acid is present the violet changes to blue, the vegetable acids being without effect or perhaps changing the violet to dirty yellow. HC1 will also change Congo-red paper a decided blue, while lactic acid of one per cent, or less, as it occurs in the gastric contents, causes no material change. 93. Uffelmanri s solution to distinguish HO from lac- tic acid in gastric contents is made by adding a drop of neutral ferric chloride solution to a mixture of ten c.c. of a four per cent, solution of carbolic acid and twenty c.c. of water. This steel blue liquid is turned yellow by lactic and butyric acids, but not affected by HC1 when diluted to the strength of gastric juice (0.25 per cent.). Reactions 92 and 93 are used clinically to determine the presence or absence of HC1 in vomit. HC1 is nearly always absent from gastric juice in cancer of the stomach. The value of these tests is impaired by interference from peptones, chlorides, and phosphates. (See Part II, Gastric Contents.) Antidotes.-Soapsuds, magnesia in milk, raw eggs. Nitro-muriatic Acid. Aqzia Regia. Acid, nitro-hydrochloricum, 4 of HNO3 to 15 of HC1. Acid, nitro-hydrochloricum dil. (100 to 76 of H2O). An unstable mixture, owing to the HNO3 yielding O to the HC1. 2HCI -J- 2HNO3 = Cl2 + N2O4 + 2H2O. It is detected by tests for HC1 and for HNO3. It dissolves gold foil. An exceedingly corrosive poison, with the same symptoms and antidotes as HNO3. Sulphuric Acid, H2SO4. Oil of Vitriol. Nordhausen acid, H2S2O7. Acidum sulphuricum, sp. gr. 1.84 = 96 per cent., H2SO4. Acid, sulph. dil., sp. gr. - 10 per cent., H2SO4. Acid, sulph. aromatic, 20 per cent., H2SO4, elixir vitriol. 81 EXAMINATION FOR COMMON POISONS. A powerful corrosive poison, causing oedema of larynx, perforation, and secondary starvation. Antidotes.-Soapsuds, magnesia in milk, raw eggs. Burns on the lips and body treated with sodium carbonate and protectives. 94. Test with blue litmus paper. 95. Put a few drops of H2SO4 with a drop of water on a sheet of paper and warm gently until dry; a charred spot appears. 96. To dilute H2SO4, ora soluble sulphate, add barium chloride, and a heavy white precipitate falls. H2SO4 + BaCl2 = 2HC1 + BaSO4. Hydrochloric acid and barium are formed. 97. A drop of H2SO4 applied to the black lining of the coat causes in time a moist red stain, which is re- moved by ammonia. Strong H2SO4 when heated with Cu generates color- less suffocating fumes of SO2:- Cu 4- 2H2SO4 = CuSO4 + SO2 + h2o. Oxalic Acid, H2C2O4. Acid of Sugar. Potass, binoxalate, C2HKO4 (salt of sorrel or salt of lemon; ink-stain remover). Oxalic acid and soluble oxalates when given in strong solution irritate the stomach, but rarely cause laryngitis or perforation. When absorbed they cause death by nervous symptoms, collapse, heart failure, and stupor. 98. Oxalic acid added to lime water gives a white pre- cipitate of calcium oxalate. 99. Heat a crystal of oxalic acid in a test-tube. It is volatile and condenses in the cool part of the tube. 100. Precipitate a solution of oxalic acid with silver nitrate. It is white, and when separated by filtration and dried, it will disperse in vapor if heated on platinum foil. 82 EXAMINATION FOR COMMON POISONS. 101. Repeat the experiment with silver nitrate and divide the white mixture between two test-tubes. The precipitate of one will dissolve in nitric acid, that of the other in ammonia, whereas silver chloride is insoluble in nitric acid. Antidotes.-Calcined Magnesia suspended in milk is the best. If this cannot be had, then prepared chalk, whiting or plaster from the wall, or eggs. For the mineral acids, soap or soapsuds may be used, but they are not antidotes to oxalic acid. The stomach tube would probably injure the corroded tissues. SHORT SYSTEM OF TESTS FOR THE STRONG MINERAL ACIDS. Test for each acid with a fresh portion of the sample:- Add BaCl2; white prpt., insoluble in HNO3, shows ........ H2SO4. If Hg2, Ag, or Pb are present, use BaNO3 instead of BaCl2. Add AgNO3. 1. White prpt., soluble in NH4HO and repre- cipitated by HNO3, shows .... HC1. 2. Yellowish prpt., insoluble in NH(HO, shows HI. HC1 may also be present. Filter; to filtrate add HNO3; prpt. shows .... HC1. Add strong H2SO4, then strong, fresh solution of FeSO4, in such a way that the two fluids do not mix ; shake them very gently ; a brown color at junction shows ..... HNO3. Add HC1. 1. Effervescence and smell of H2S shows . H2S. 2. Effervescence and no smell shows . H2CO3. 83 EXAMINATION FOR COMMON POISONS. CAUSTIC AND CARBONATED ALKALIES. Potassium Hydrate, KHO. Caustic Potash. Liquor potassse, KHO. Potassii carbonas, K2CO3. Caustic potash is a deep corrosive of great energy; causes a soapy taste, pain in mouth, throat, and stomach, vomiting and purging. Death may en- sue from perforation of the stomach or from starvation due to closure of the oesophagus. 102. Test with red litmus or turmeric paper. 103. Dilute the KHO with an equal part of water, and add HC1 until the reaction is neutral to litmus paper. KHO + HC1 = H2O + KC1. Water and potassium chloride are formed. 104. To this potassium chloride add platinum chlo- ride ; a yellow granular precipitate falls slowly. Ammo- nium salts give a similar result. 2KCI + PtCl4 = PtCl4.2KCl. A double salt, potassio-platinic chloride, is formed. 105. Heat platinum foil or wire in a Bunsen's burner until the flame is colorless; then touch it with moist KHO and heat again ; a violet tint appears. If mixed with yellow, observe through deep blue glass, which shuts out the yellow rays and leaves the violet. Antidotes.-Weak vinegar, lemon juice, milk, and oils (castor or olive). Sodium Hydrate, NaHO. Caustic Soda. Liquor sodse, NaHO. Concentrated lye. Sodii carbonas. Sal soda, washing soda, corrosive in all respects like potash. 84 EXAMINATION FOR COMMON POISONS. 106. Test for alkaline reaction. 107. Add HC1 to neutralize. 108. Heat NaHO on platinum foil; the flame is colored yellow. This test is so delicate as to reveal a trace of sodium present almost every- where. Antidotes.-Weak vinegar, lemon juice, milk, and oils. Ammonium Hydrate, NH4H0. Ammonia Water. Aqua ammonia fortior, 28 per cent, by weight. Aqua ammonia, ten per cent. Hartshorn. Spiritus ammoniae. It causes suffocation, with inflammation of mouth, larynx, oesophagus, and stomach. 100. Note the odor of aqua ammonia or ammonium carbonate. 110. Test the fumes with moist red litmus paper ; they turn it blue. 111. Neutralize with HC1; observe the white fumes of ammonium chloride, NH4Cl, and note that the ammo- niacal odor ceases. 112. Boil the last product, NH4C1, with KHO ; the odor of free ammonia may be perceived. NH4C1 + KHO = KC1 + NH4H0. Potassium chloride and ammonia are formed. Antidotes to the Alkalies.- Vinegar and Water (dilute acetic acid), lemon juice, oils, butter, milk. Do not use the stomach pump. 85 EXAMINATION FOR COMMON POISONS. VOLATILE POISONS. Under this head are included carbolic acid, hydrocyanic acid, and phos- phorus as the most important, and chloroform, chloral, ether, alcohol, nitro-benzene, besides some others of less importance in toxicology. Peculiar odors are distinctive of nearly all the volatile poisons. Acid Carbolic, C6H5HO. Phenol. Phenyl Hydrate. It is an escharotic and cardiac depressant, causing pain in the stomach, coma, collapse. 113. Note the odor. 114. A drop applied to blue litmus paper leaves a greasy stain, but does not redden the paper. 115. To a dilute solution of acid carbolic add a few drops of nitric acid and warm gently; it turns yellow from the formation of picric acid. 116. Strong bromine water causes a whitish-yellow precipitate. Antidotes.-Saccharate of lime, lime water, magnesia, alkaline sul- phates, eggs. The stomach pump may be used freely. Treat symptoms by stimulants. Acid Hydrocyanic, HCN or HCy. Prussic Acid. Acidum hydrocyanicum dil., 2 per cent. HCy. . Potassium cyanidum, KCN. Oil of bitter almonds has 5-14 per cent. HCy. Laurel water, peach, cherry, and plum kernels, etc. The symptoms produced by it are convulsions, coma, and collapse. It is not a corrosive poison, but a syncopant or hyposthenisant. 117. Note the odor of bruised peach kernels. 118. Put HCy or potassium cyanide solution, KCN, into a test-tube, and add a few drops of acid sulphuric. Cover with a watch crystal or saucer wet with one drop of 86 EXAMINATION FOR COMMON POISONS, silver nitrate and warm gently. A white spot appears, which is soluble in warm nitric acid. KCN +>AgNO3 = KNO3 + AgCN. Potassium nitrate and silver cyanide are formed. 119. Cover with another watch crystal wet with potas- sium hydrate. After a few minutes touch with ferrous sulphate and with ferric chloride, and finally with one drop of HCI. A precipitate of Prussian blue (ferro- cyanide of iron) forms. 120. Cover with another watch crystal or saucer wet with ammonium sulphide. HCy + NH4HS = 2H + NH4SCy. White ammonium sulpho-cyanide is formed. Now touch this with ferric chloride, Fe2Clti, and it turns to blood-red sulpho-cyanide of iron. 6NH4SCy + Fe2Cl6 = 6NH4C1 + Fe2 (SCy)6. Antidotes.-Affusions of cold water, ammonia, chlorine and a mixture of ferrous sulphate, ferric chloride and magnesia. Phosphorus, P. Oleum phosphoratum, i per cent, of P. Heads of matches. Rat paste contains 5 per cent. P. Its poisonous effects upon the stomach are those of a local irritant. When absorbed it causes general fatty degeneration. The liver and heart are markedly affected, causing jaundice and syncope. 121. Boil a match head in water with a few drops of some acid, and in a dark closet note the luminous appearance. 122. Observe that it has an odor like garlic. 123. Clasp over the mouth of the tube filter paper 87 EXAMINATION FOR COMMON POISONS. moist with silver nitrate. The phosphorus vapor will cause a dark stain of phosphide of silver. Antidotes.-Crude turpentine, magnesia in milk. Antal's antidote is potassium permanganate in weak solution given freely. Copper sulphate is of doubtful value. Chloroform, CHC13. Trichlor methane. It is an anaesthetic and causes death by failure of respiration and heart's action. 124. A strip of paper wet with chloroform and burned in the Bunsen flame imparts a green tint and yields acid fumes which redden moist blue litmus paper. If the glass rod wet with ammonia is held above the flame, white clouds of ammonium chloride show that HC1 is a product of combustion. 125. To an alcoholic solution of potassium hydrate add a few drops of aniline, C6H5.NH2, and one of chlo- roform. Immediately, or perhaps after gentle heat, a penetrat- ing, disagreeable, and peculiar odor is emitted. CHC13 + 3KHO + C6H5.NH2 = C6H5.NC + 3KCI + 3H2O. The offensive benzo-isonitril and potassium chloride and water are formed. Antidotes.-Treat symptoms by lowering the head, drawing out the tongue, cold affusion, artificial respiration, electricity. Chloral Hydrate, C2HC13O. Trichloraldehyde. A narcotic poison causing coma and syncope. 126. Dissolve a small piece of chloral hydrate in water, add potassium hydrate, and boil. C2HC13O 4- KHO = CHC13 4- KCHO2. Chloroform and potassium formate are formed. 88 EXAMINATION FOR COMMON POISONS. The chloroform vapor may be recognized by its odor or recovered by distillation, and the tests for chloroform applied to the distillate. 127. Apply the aniline test, §125, to a portion of the mixture in the test-tube, and benzo-isonitril may be detected by its offensive odor. 128. Remove the chloroform from the above mixture, §126, by boiling, then add ammonia and a crystal of silver nitrate. The silver is reduced with formation of a metallic mirror on the tube. Emetics, such as apomorphina, grain hypoder mically. Antidotes.-Wash out the stomach with tea and coffee and treat symp- toms with cold affusions, sinapisms to epigastrium, artificial respiration, and electricity. Alcohol, C2H5.HO. Present in malt, vinous, and spirituous liquors. It is an inebriant. 129. To solution of potassium bichromate, K2Cr2O7, add a few drops of acid sulphuric, and dilute with water. If alcohol be now added and the admixture gently heated, it will turn green and aldehyde vapor will be recognized by its odor. 3C2H5OH -f- K2Cr2O7 4- 4H2SO4 = 3C2H4O6 + K2SO4.Cr2(SO4)3 + 7H2O. Aldehyde and green potassio-chromic alum are formed. 130. Dissolve a crystal of iodine in some alcohol. Add potassium hydrate until the brown color disap- pears, and after gentle heat a yellow precipitate of iodo- form falls. C2H5OH 4- 6KH0 4- I8 = CHI3 4- CHO2K 4- 5KI 4- 5H2O. Iodoform, potassium formate, potassium iodide, and water are formed. Antidotes.- Emetics or stomach pump. Cold affusions, ammonium carbonate. 89 EXAMINATION FOR COMMON POISONS. Arsenum, As. Elementary Arsenic. Fly paper contains As in a paste. Hydrogen arsenide, arsenia AsH3, is the most poisonous form and must not be inhaled. Arsenic trioxide, As2O3. Aciduni arsenosum, white arsenic, ratsbane. It is usually called arsenic, and is the most commonly used poison for homicidal purposes. " Aqua Toffana," a boiled solution of As2O3. " Rough on Rats " contains As and As2O3. " Buffalo Carpet Moth Annihilator" contains As2O3. Liquor acidi arsenosi, one per cent. As2O3. " potass, arsenitis (Fowler's), one per cent. As2O3. " sodii " (Harle's), one per cent. As2O3. " " arseniatis (Pearson's), one per cent. Na2HAsO4. " arseni et hydrarg. iodid (Donovan's), one per cent. Asl3. Acidum arsenicum, As2O5, arsenic pentoxide. Realgar, As2S2. Orpiment, As2S3, King's yellow. London purple, arsenical dye refuse. Paris green, Schweinfurth's green, Cu(C.2H3O2)2 3(CuO4As2), a very common and dangerous poison. Scheele's green, copper arsenite and hydroxide. The arsenical compounds are irritant poisons, inflaming living tissue by contact, Usually in about half an hour causing symptoms of inflammation of stomach and bowels, such as epigastric pain, vomiting, thirst, and purg- ing. Sometimes they cause profound shock and death by coma. Slow or chronic poisoning induces cachexia, diarrhoea, paralysis, and death by wasting. 131. Put into a reduction tube a small quantity of As2O3 mixed with six times its bulk of dried flux (made of dry sodium carb. 3 parts to potass, cyanide I part). Heat the mixture and observe the formation of a dark mirror ring of As on the tube (I7ig. 27). Break off the closed end of the tube and heat the ring. A current of air plays through the tube and oxidizes the As to As2O3, METALLIC IRRITANTS. 90 EXAMINATION FOR COMMON POISONS. which is deposited in the cool part of the tube as a white ring of minute octahedral crystals (Figs. 28 and 29). 132. To a solution of As2O3 add a few drops of HC1 and pass through it hydrogen sulphide, H2S; a lemon- Fig. 27. Fig. 28. Fig. 29. yellow precipitate falls, soluble in ammonia, and in NH4HS, but not in HC1. 2H3AsO3 3H2S = As2S3 -f- 6H2O. Arsenic trisulphide and water are formed. 133. To a neutral, alkaline, or feebly acid solution of As2O3 add silver ammonio-nitrate; it throws down a canary-colored precipitate of silver arsenite, Ag3AsO3. 134. To a solution of As2O3 add copper-ammonio- sulphate; it throws down a green precipitate of copper arsenite, CuHAsO3. 135. Fleitmanris Test.-Into a test-tube put some strong potassium hydrate, and a few pieces of pure zinc; clasp 91 EXAMINATION FOR COMMON POISONS. over the mouth of the tube paper wet with silver nitrate, and boil. If the paper is not stained the materials used for this test are free from arsenic. Zn + 2KHO = H2 + K2ZnO2. Hydrogen and potassium zincate are formed. Now add a few drops of arsenical solution to the same mixture- and boil as before; a black stain appears on the paper. AsH3.+ 6AgNO3 4- 3H2O = 6Ag H3AsO3 -f- 6HNO3. Black metallic silver, arsenious acid, and nitric acid are formed. 13G. Reinsch's Test.-Into a solution of As2O3 in a test- tube put a few drops of HC1 and a clean slip of copper; then boil for a few minutes. When the slip has a steel- gray coat of As, remove it; dry with blotting paper, and heat in an open glass tube. Note the white ring of As2O3 crystals. 137. Marsh's Test.-Into a hydrogen-generating appa- ratus put some pure zinc and add dilute sulphuric acid (H2SO4 part one to H2O parts four). Wait fifteen minutes, until the explosive mixture of air and hydrogen has escaped, then light the H gas at the jet and hold in the flame a piece of cold porcelain. If no stain is made then arsenic and antimony are absent from the materials. Now add solu- tion of arsenious acid and try the flame again with the Fig. 30. 92 EXAMINATION FOR COMMON POISONS. porcelain. A brownish-black metallic mirror of As appears. 3Z11 3H2SO4 T H3AsO3 = 3ZnSO4 3H2O -f- AsH3. Zinc sulphate, water, and hydrogen arsenide are formed in the generator. When the AsH3 is burned it is decomposed, forming H20 + As. The arsenic, when deposited on the cold porcelain, is recognized by being quickly dissolved when treated with liq. sodae chloratae or a solution of calx chlorata. If treated with nitric acid and gently evapor- ated, it is oxidized to arsenic acid as a whitish deposit. The arsenic acid touched with a drop of strong solution of AgNO3 gives red silver arseniate. The original As deposit on porcelain will dissolve slowly in ammonium sulphide, leaving a yellow residue on evaporation. This residue is soluble in NH4H0. 138. Use a fresh hydrogen apparatus with solution of an antimony salt, say tartar emetic, repeating the same procedure. On the porcelain a sooty stain appears, which is not removed on adding a drop of liq. sodae chloratae. It is soluble in nitric acid, but does not after- ward turn red with silver nitrate. It is freely soluble in ammonium sulphide with an orange-red residue insolu- ble in NHJIO. 139. Make the antidote, ferric hydrate, by adding to tinctura ferri chloridi some calcined magnesia or aqua ammonia in excess and shake well together. If three fluidounces of tinct. ferri chlor, be mixed with three drachms of magnesia suspended in water, it yields anti- dote for ten grs. of arsenious acid. 2(Fe26HO) As2O3 = Fe32AsO4 -f- 5H2O -f- Fe2H0. Antidotes to Arsenic.-Milk, eggs, hydrate magnesia, dialysed iron, moist ferric hydrate, ferri oxidum hydratum cum magnesia. 93 EXAMINATION FOR COMMON POISONS. Antimony, Sb. Tartar Emetic, K(SbO) C4H4O6. Antimony oxide, Sb2O3. Antim. et potass, tartras, tartar emetic. Vinum antimonii, 0.4 per cent, of tartar emetic. Syr. scillse comp, contains tartar emetic. Tartar emetic has a faint metallic taste, and acts as an irritant poison with symptoms like those produced by arsenic. 140. Pass hydrogen sulphide into a solution of tartar emetic, acidified with dilute HC1 or tartaric acid. An orange-red precipitate of antimony sulphide forms. Divide this precipitate in three portions; one is insolu- ble in aq. ammonia, one is soluble in ammonium sul- phide, and the third soluble in hydrochloric acid. Arsenic sulphide is insoluble in HC1. 141. Repeat Reinsch's test, § 136, using solution of tartar emetic instead of arsenic. The deposit on copper is bluish, and when heated in an open glass tube yields a white amorphous sublimate. If this sublimate is dis- solved in HC1 and evaporated, the residue turns orange with H2S. 142. Repeat Marsh's test, §137, using solution of tartar emetic instead of arsenic. The stain on porcelain is sooty, barely soluble in NaClO but soluble in NH4HS, leaving an orange-red residue. 143. Put a drop of solution of tartar emetic on plati- num foil, add a drop of HC1, and touch the platinum through the drop with a slip of zinc. A black spot of antimony appears which turns orange-yellow with ammo- nium sulphide. Antidotes to Antimony.-Acid tannic, tea, vegetable astringents, dia- lysed iron, moist ferric hydrate, stomach pump. Copper, Cu. Copper sulphate, CuSO4, Blue vitriol. Copper subacetate, Verdigris. The salts of copper are irritants in full doses. If impure, they are apt to contain traces of arsenic. 144. Treat acidulated solution of copper sulphate, Cu- SO4, with hydrogen sulphide, and a brown-black pre- cipitate falls. 145. Dip into solution of CuSO4 a bright steel needle ; it soon has a red coat of metallic copper. 146. Treat a weak solution of CuSO4 with potassium ferrocyanide ; a brown precipitate falls. 147. To solution of CuSO4 add a drop of ammonia ; a bluish precipitate falls, but on addition of more ammonia this clears into a deep blue solution. CuSO4 + 2NH4H0 = CuSO4.2NH3 + 2H2O. Blue ammonio-sulphate of copper and water are formed. Antidotes.-Warm milk, raw eggs, sodium bicarb, followed by emetic. Mercury, Hg. Corrosive Sublimate, HgCl2. Mercuric chloride, HgCl2, Bichloride. Mercurous chloride, Hg2Cl2, Calomel. Mercuric oxide, HgO, red and yelloiv. Mercurous oxide, Hg2O, black. Mercuric iodide, Hgl2, red. Mercurous iodide, Hg2T2, green. The symptoms caused by poisonous doses of the mercuric salts are those of active irritants, i.e., pain, vomiting, thirst, purging, depression. Corrosive sublimate is more prompt in its effects than arsenic, having a strong taste and corroding the tissues at once. Chronic poisoning is characterized by salivation, tremors, wasting, debility. 94 EXAMINATION FQR COMMON POISONS. 95 EXAMINATION FOR COMMON POISONS. 148. Treat calomel, Hg2Cl2, with lime water; it turns to black oxide, Hg,O. Hg2Cl2 + Ca2HO = Hg2O + CaCl2 -p H2O. Black mercurous oxide, calcium chloride, and water are formed. 149. Treat corrosive sublimate HgCl2, with lime water; it turns to yellow oxide, HgO. 150. Perform Reinsch's test, § 136, with corrosive sub- limate solution without boiling ; the copper takes a gray coat of metal, Hg. • Dry the copper and heat it in an open tube; a sub- limate of minute gray globules forms. Heat this sub- limate with iodine; it forms red mercuric iodide, Hgl2. Galvanic Test.-On a gold ring or coin put a drop of solution of corrosive sublimate; with a steel needle or knife touch the gold through the drop, and a silver-like spot of metal, Hg, appears. This stain is removable by gentle heat over a flame. Antidotes.-Raw eggs, flour in milk, emetics. Lead, Pb. Plumbi oxidum, PbO, Litharge. Plumbi oxid. rubrum, Pb3O4, Red lead. Plumbic oxychloride, Pb4O3Cl2, Turner's yellow. Plumbi acetas, Sugar of lead. Liq. plumbi subacetatis, Gotdard's Extract. Plumbi chromas, PbCrO4, Chrome yellow. Plumbi carbonas, (PbCO3)2Pb(HO)2, White lead. All the salts of lead are poisonous except, perhaps, the sulphate and the sulphide. The carbonate is the most active. Lead poisoning is characterized by obstinate colic and constipation, with a blue line on the margin of the gums. Later on appear palsy and wasting of the extensor muscles of the forearm, cachexia, joint-pains, mania, or convulsions. 96 EXAMINATION FOR COMMON POISONS. 151. Put a crystal of lead acetate on a piece of char- coal and heat with the tip of a blowpipe flame. A globule of metallic lead soon forms with a yellow margin. 152. To a solution of a lead salt add hydrogen sul- phide ; a black precipitate of lead sulphide, PbS, falls; insoluble in KHO and NH4HS, but soluble in boiling dilute HNO3. 153. To another portion of lead solution add acid sul- phuric; a white precipitate of lead sulphate, PbSO4, falls. PbCO3 + MgSO4 = PbSO4 + MgCO3. 154. To another portion add potassium iodide; a yellow precipitate of lead iodide, Pbl2, falls. 155. To another portion add K2CrO4; a yellow pre- cipitate of lead chromate appears. Antidotes.-Magnesium sulphate and other soluble sulphates with milk and opium for colic, potassium iodide and iron for chronic poisoning. Silver, Ag. Argenti nitras, AgNO5. Lunar causticxs an irritant poison. Prolonged administration causes a slaty-blue color of skin. 156. To a solution of silver nitrate add H2S; it pre- cipitates black silver sulphide, Ag2S, insoluble in dilute acids, alkalies, alkaline sulphides, but soluble in boiling hno3. 157. To another portion cautiously add NH4HO ; it gives a brown precipitate of Ag2O, which readily dis- solves in excess of NH4HO. 158. HC1 gives white curds of AgCl, soluble in NH4HO. KI gives yellow ppt. of Agl, insoluble in NH4HO. II2SO4, yields no precipitate whatever. Antidotes.-Common salt, raw eggs and milk. 97 EXAMINATION FOR COMMON POISONS. Zinc, Zn. Zinci oxidum, ZnO, zinc white. Liq. zinci chloridum, ZnCl2, Burnett''s disinfectant, also used in a solder- ing fluid and as preservation injection for bodies in the dissecting room. Zinci sulphas, ZnSO4-|- 7H2O, white vitriol. The salts of zinc are irri- tant poisons, inducing secondary depression of the nervous system. 159. To solution of a zinc salt add ammonium hydrate. A white precipitate of zinc hydrate, Zn(OH)2 falls, solu- ble in excess of the ammonium hydrate. 160. To another portion add first ammonium chloride and then ammonium hydrate. There is no precipitate. Now add hydrogen sulphide. White zinc sulphide is thrown down. 161. To a neutral or alkaline solution of zinc salt add ammonium sulphide. A white precipitate, ZnS, is thrown down. Antidotes.-Raw eggs, milk, tea, vegetable astringents. Barium, Ba. The chloride and nitrate are irritant poisons. 162. From solution of a barium salt, potassium hydrate will throw out white barium hydrate, Ba(OH)2. Ammonium hydrate has no such effect when added to another portion. 163. To the clear solution with ammonia add sodium phosphate, Na2HPO4; it forms a white precipitate of barium phosphate, BaHPO4. 164. To a solution of barium salt add acid sulphuric or any soluble sulphate. A white precipitate, BaSO4, falls. Antidotes.-Magnesium sulphate or any soluble sulphate, eggs, and milk. 98 EXAMINATION FOR COMMON POISONS. 105. ANALYSIS OF A NEUTRAL OR SLIGHTLY ACID SOLUTION OF ORDINARY SALTS OF ONE OF THE COMMON METALS. (AUfield.} Add hydrochloric acid. Precipitate Hg(ous) Pb Ag. Collect, wash, and add NH4HO. Hg ppt., blackened. Pb ppt., still white. Ag ppt., dissolved. Sb and Bi may also be precipitated by HC1, but are dissolv- ed on adding more HC1. If HC1 gave no precipitate the metal is still in the liquid; pass H2S through the solution. Precipitate Cd Cu Hg(ic) Pb Bi As Sb Sn Au Pt. Collect, wash, add NH4HS. Insoluble. Soluble. Cd, yellow. As(ous&ic) ) Cu 1 c \ r - Hg(ic) 1 Sn(1C) Pb Sb, orange. Bi J Sn(ous) ) Au I Pt J S Apply special tests for each to the original solution. For these, see the previous pages. If H2S gave no precipitate add NH4C1,NH4HO and NH4HS. Precipitate Zn Mn Co Ni Al Fe Cr. Al }white' Cr, green. Mn, skin-tint. Ni j Co > black. FeJ Test specially for each in original solu- tion. See previous pages. If NH4HS gave no precipitate, add (NH4)2CO3. Precipitate Ba Sr Ca. Collect, wash, dis- solvein HC2H3O2, add K2CrO4. If (NH4)2CO3 gave no precipitate, add (NH4)2HAsO4. Ppt. Mg. If no precip- itate, test orig- inal solution in flame on loop* of Pt wire. L, crimson. Na, yellow. K, violet. If neither, test orig. sol. for NH4. Ppt. Ba. Sol. Sr Ca Add dil. H2SO4. Ppt. Sol. Sr.-Ca. 99 EXAMINATION FOR COMMON POISONS. NEUROTIC ALKALOIDS. Strychnina, C21H22N2O2. Alkaloid from Strychnos Nux Vomica. Strychnina sulphate and nitrate. Tincture and extract of nux vomica. " Battle's Vermin Killer " has 23 per cent. " Butler's " " has 5 per cent. " Gibson's " " has 0.5 gr. in each. All the above preparations have a very bitter taste, and in poisonous dose cause spinal symptoms, such as convulsions, epigastric pain, and death by exhaustion or prolonged spasm of respiratory muscles, the brain remaining clear. 166. Note the intensely bitter taste of one drop of the solution. 167. On a white dish put a crystal of strychnine and add to it a drop of acid sulphuric. It dissolves without change of color. Near by upon the same dish put a crystal of potassium bichromate and let the acid solution flow over it. At the line of contact a transient play of colors will be seen in the following order : blue, violet, red, and yellow. 168. Nitric acid makes with it a colorless solution. If brucine be present as an adulteration a red color is pro- duced. 169. Insert a few drops of strychnine solution under the skin of a frog's back. Within a few minutes it will be convulsed. Antidotes.-Emetics (such as apomorphina, grain hypodermically), draughts of tea or other vegetable astringents, chloroform by inhalation, chloral, and potassium bromide by enema. Morphina, C17H19NO3. Alkaloid from Opium Poppy. Morphina acetate, hydrochlorate, and sulphate. Opium and all its preparations contain it. It is present in the proprietary 100 •EXAMINATION FOR COMMON POISONS. opiates sold to relieve pain and induce sleep, as Godfrey's, Dalby's, Mrs. Winslow's, Battle's, Nepenthe, Chlorodyne, and various opium cures. It is a narcotic poison, causing stupor with contracted pupils and slow respiration, deepening into coma and death by arrest of respiration. 170. On a white dish put a crystal of morphine sul- phate and touch it with a drop of nitric acid; it turns at first orange-red and fades to yellow. 171. To a neutral solution or a crystal of morphine sulphate add pure neutral ferric chloride, Fe2Clc; a deep blue color appears. 172. To morphine solution add iodic acid in solution; a yellow color of reduced iodine appears. If the yellow liquid is shaken with chloroform, the iodine separates in a violet-colored layer. Antidotes.-Emetics (such as apomorphina, TXg grain hypodermically), washing out the stomach with strong tea, electricity, flogging, artificial res- piration, brandy, potassium permanganate freely in weak solution (four grains to four ounces of water). Opium and its Characteristic Meconic Acid. 173. To a portion of solution of meconic acid or a decolorized infusion of tincture of opium add ferric chloride, Fe2Cl6 (pure and neutral); a brown-red color appears. This color persists even if treated with mer- cury bichloride, HgCl2, or after boiling with dilute HC1. Atropina, C17H23NO3. Alkaloid of Atropa Belladonna. Daturina an alkaloid of Datura stramonium. Hyoscyamina an alkaloid of Hyoscyamus niger. Duboisina an alkaloid of Duboisia myoporoides. All of the above are practically identical. They are active deliriant poisons, causing dilatation of the pupils, flushing of the face, dryness of the throat, muscular weakness, dizziness, delirium, and coma. 101 EXAMINATION FOR COMMON POISONS. 174. A small portion of atropine, if heated with strong acid sulphuric, does not change color, but gives off an odor of roses. 175. Into a test-tube put a few drops of acid sulphuric and a crystal of potassium bichromate, and heat them. If a fraction of a grain of atropine is now dropped in, a green color and an aromatic odor may be perceived. Antidotes.-Emetics, washing out the stomach with strong tea or other preparation containing tannin. 176. ANALYTICAL TABLE FOR COMMON ALKALOIDS.-(Senter's ) PART I. Morphine. Brucine. Salicin. Strychnine. To a small quantity on a white plate add strong nitric acid. If no Morphine or Brucine,moisten a small quantity on a white plate with strong sulphuric acid. An Orange Color, decolorized by Sn Cl,, Na,S,O3, or NallS = Mor- Confirm by-Fe2CI6 (neut which gives with morphine or its salts a blue color. HIO3 is decom- posed by morphine or its compounds, with liberation of iodine, which may be recognized by starch. A Red Color. To a few grains in a test-tube add a drop or two of HNO3; warm to boiling and di- lute with water; then add a few drops of stannous chloride. A violet color = Brucine. A deep Red Col- or - Salicin. Confirm by boil- ing the substance with water to which has been added a few drops of dilute H2SO4. Then make solution alkaline and ex- amine for glu cose. No Color, or only a slight Col- or. Draw a mois- tened crystal of K2Cr2O7 across the acid film when a transient play of colors, violet to red = Strychnine. * Strychnine of commerce often gives an orange or red color, due to contamination with brucine. 102 EXAMINATION FOR COMMON POISONS. 177. ANALYTICAL TABLE FOR COMMON ALKALOIDS. gART II. Quinine. Quinidine. Cinchonine. ClNCHONIDINE. If Morphine, Brucine, Salicin, and Strychnine are absent, make a saturated solution in hot water and cool down to about 6o° F., removing the crystals which have deposited by filtration. If the substance is an alkaloid or an acid salt, it must be neutralized. To one volume of this solution in a test-tube add exactly half a volume of ether and then ammonia solution in slight excess. Agitate and set aside for a quarter of an hour. Precipitate entirely solu- ble in the ether. To a portion of the satur- ated aqueous solution add rather more than an equal bulk of ammonia solution (sp. gr. 0.96) and agitate. The precipitate dissolves completely = Ordinary Sulphate of Quinine. If precipitate does not dis- solve completely, add to another portion of the aque ous solution (NH4)2C2O4 and agitate (no precipitate at once with ordinary sul- phate of quinine). A bulky crystalline pre- cipitate = Quinine. A crystalline layer be- tween the ethereal and aqueous solutions. To a portion of the aque- ous solution add Na2C4- H4O6 and agitate. A bulky crystalline precipitate = Cinchonidine. If no precipitate or only a trace, add to another por- tion of the aqueous solution KI. A heavy white pre- cipitate = Quinidine. If the precipitate be semi- liquid, it contains cinchoni- dine or other alkaloids, the hydriodates of which are soluble in weak spirit. Precipitate in- soluble in ether = Cinchonine. Confirm b y other reactions. Chlorine and ammonia give a yellow precipi- tate. PART IV. STUDY OF NORMAL MILK AND MILK EXAMI NATION. Milk is a sweet, opaque, bluish-white fluid with a peculiar odor holding in solution casein, albumin, sugar, and mineral salts. Its turbidity is due to the minute fat or butter globules which are suspended in it. COMPOSITION OF NORMAL MILK Percentage Composition of Normal Milk. Cow. Human. Water, 85.70 88.90 Solids, as tabulated below, I4-3O 11. IO Casein, 4.82 3-90 Albumin and Globulin, 0.58 Butter or Fat, 4-3° 2.65 Milk Sugar, 4.00 4-35 Salts, 0.50 c.20 The composition is somewhat variable, the proportion of casein rising when the breasts are emptied frequently, and the last portions obtained containing more butter. Microscopically, the milk is found to be composed of minute brilliant oil globules in thin envelopes of casein suspended in the clear milk plasma. Immediately after delivery of the young, the milk is relatively poor in casein but rich in fatty matter, which exists to a considerable extent in the form of colostrum masses. (Fig. 31.) 103 104 MILK EXAMINATION. MORBID MILK. The constitution and the food value of human milk are impaired when the mother is not healthy. It is affected injuriously by undue emotional excitement. It will contain an appreciable amount of certain medicines given to the mother, and thus the infant be affected secondarily. Cow's milk is subject to the same laws. A sickly cow gives unwholesome milk. Serious injury may be caused by the use of toilk taken from a tuberculous cow, or one affected by " foot and mouth disease." 178. Reaction.-With red litmus paper test human milk; it turns blue, showing alkalinity. Cow's milk is Fig. 31. usually alkaline or neutral when fresh, though some- times acid, and occasionally a sample will be found that is amphoteric, i. e., reddens blue litmus paper, and turns blue litmus paper red. 179. Quantity.-The average daily amount secreted by a woman is I liter, or 2 pints. 180. Spontaneous Change.-Let milk stand for several days in a warm place; it coagulates and " sours]' i. e., 105 MILK EXAMINATION. turns acid. At the same time it would be advisable to conduct experiments §§ 181, 182, 183. This change is due to the conversion of the milk sugar (lactose) into lactic acid by a minute organism, the bacterium lactis. The casein is normally held in solution by alkaline phosphates. Acids change the reaction and thus cause the casein to be precipitated. 181. To half a test-tube of diluted milk add a drop or two of hydrochloric or acetic acid and gently warm it; an abundant precipitate falls. This precipitate is the curdled casein, carrying with it most of the fat. According to recent studies casein exists in solution as caseinogen, a form of union with calcium phosphate. The phosphate being removed by acids the insoluble casein is precipitated. PRESERVATION OF MILK. 182. Refrigeration.-Having tested with litmus paper, put a sample of fresh milk in a refrigerator kept at or below 50° F. (io° C.). After several days examine it again with litmus paper. The reaction will be but little changed, the milk having been kept sweet and un- curdled. In spite of the temperature the lactic ferment will in time develop to such a degree as to impair the digestibility of cow's milk used for infants' food. While the growth of bacteria is checked by cold the specific forms which cause disease are not destroyed by it. 183. Sterilization.-Put some fresh milk in a test-tube and boil it for 15 or 20 minutes. A scum will form, due to coagulation of the lact-albumin and globulin. Close it with a tight cork or a plug of cotton and stand it aside for several days. It will keep sweet and uncurdled 106 MILK EXAMINATION. owing to the death of the bacteria which cause lactic fermentation. At the same time, the fats, sugars, casein, and albumin are altered in such a way as to make boiled milk less digestible and nourishing than raw milk. Cow's milk delivered in cities contains numbers of bacteria. It is usually 24 or 48 hours old before it is delivered, and in that time the bacteria have a chance to multiply rapidly. Most varieties of these microbes are entirely harmless, as is proved by the fact that mankind has been consuming milk for ages without injury from them. Without doubt cholera, diphtheria, scarlatina and typhoid fever have been communicated by milk containing the specific germs or bacteria of these diseases. It is probable that tuber- culosis can be conveyed in the same medium, the bacillus of tubercle having been found in the milk of tuberculous cows. The danger of its distribution is materially lessened by the circumstance that this germ cannot multiply in milk. A small portion of tuberculous milk is simply diluted by being mixed with a large quantity that is not tuberculous, and thus the number of bacilli taken by any individual is relatively small. Experiment shows that this particular germ must be introduced in such numbers at a time, in order to cause disease, that the chances are against infection by it through ordinary mixed city milk. Except in persons of low vitality or inherited susceptibility the danger from this source is not worth noticing. Other bacteria produce certain intestinal troubles, such as summer diar- rhoea and cholera infantum. They have the power of multiplying in milk and developing ptomaines or toxalbumins which poison the stomach and intestines directly. A highly poisonous substance called diazo-benzene or " tyrotoxicon " is sometimes developed in milk used in various foods, such as ice cream. These poisons are not present in fresh milk, which is there- fore harmless so far as summer diarrhoea is concerned. Their production is prevented by boiling the milk when fresh. At the same time the high heat of 2120 F. (ioo° C.) injures the milk as an assimilable food, and where it is convenient a method of lower heating known as Pasteurization is preferable. 184. Pasteurization.-Put some fresh milk in a test- tube or clean glass bottle, closing the neck with a plug of cotton. Stand the tube or bottle in a vessel of water, 107 MILK EXAMINATION. and heat the water to i6o° F. for a few minutes, carefully observing the temperature by an immersed thermometer. Set it aside stopped with the plug of cotton, and it will be found to keep for twenty-four hours at least. Milk heated above 167° F. (750 C.) is so changed that children fed on it exclusively do not thrive. Adults find it less palatable and less digestible, the curds forming in whole masses instead of in flakes. At and below 167° F. (750 C.) it does not lose the taste of fresh milk nor become less digestible. At this temperature and as low as 150° F. the matured disease germs are killed and the spores so much weakened in vitality that all lia- bility to cause intestinal disorder is removed if the milk be used within twenty-four hours of the treatment. It must be noted that if the milk is not tolerably fresh poisons may have become developed already. Pasteurizing will not destroy the toxalbumins or dissolved poisons when once produced, nor render stale milk harmless. 185. Pasteurizing Apparatus. - More or less elaborate apparatus, such as the Arnold Sterilizer, can be purchased, but the following form is easily con- structed and the method simple in operation and sure. Take a pail made of tin or iron or copper, and have a tinner cut a disc of tin a little smaller than the bottom of the pail with some strips of metal left radiating from the disc, which, when turned back, will serve as legs half an inch high. The disc must be perforated like a colander, so as to allow of free cir- culation of water when this false bottom is put into the pail. The bottle of milk is closed with a stopper, which has a glass thermometer fitted through a hole, or by a plug of cotton holding the thermometer with firm pressure. The bulb of the thermometer dips into the milk, while the scale can be read outside. Set the bottle full of milk on the false bottom and fill the pail with water as high as the surface of the milk in the bottle. Heat the pail of water until the temperature of the milk reaches 1550 F. Remove the bottle of milk and allow it to cool gradually, or the milk can be kept at 1600 for ten minutes and then cooled rapidly. When the thermometer is not at hand, at a pinch resort may be had to the following rough method. Put the fresh milk in clean glass bottles stopped with cotton. A basin containing several inches of water is placed on a slow fire and the bottles of milk set in it. After boiling the water for ten minutes the milk, which has not boiled but only simmered, is removed and cooled before using. 108 MILK EXAMINATION. 186. Milk Sugar and Salts.-Having filtered the whey from the acid curd, § 181, test the whey with Fehling's solution. The milk sugar reduces the cupric oxide. To another portion add magnesia mixture ; the phosphates are precipitated. To another portion add silver nitrate; the chlorides are precipitated, insoluble in nitric acid. 187. Butter.-To a test-tube one-third full of milk add half its volume of potassium hydrate and half of ether ; shake the mixture and stand in a warm place. The milk clears up, and the butter dissolved in the ether floats at the top. On separating the ethereal layer and evaporating it a residue of butter is left. In cow's milk ether will not dissolve the fat globules unless they are lib- erated by dissolving their envelopes with acetic acid or caustic potash or soda. With human milk it suffices to agitate with ether alone. In ordi- nary churning the envelopes of casein are ruptured mechanically, and the fat globules cohere in large masses of butter. This process does not sepa- rate all the fat as butter. The residue called " buttermilk " still contains about one per cent, of fat. 188. Pepsin Curd.-Into a test-tube about one-third full of milk, put a few drops of neutral essence of pepsin (Fairchild's). Mix gently, warm to the temperature of the body, and keep at 104° F. (40° C.). A solid curd forms in 10 or 12 minutes, so that the tube can be in- verted without losing the milk. In a short time a ivhey separates from the clot. 189. Junket, or Curds and Whey.-To make this delicacy for invalids, take one-half pint of fresh milk heated as hot as can be agreeably borne by the mouth, add one teaspoonful of rennet, or essence of pepsin (Fairchild), and stir just enough to mix. Let it stand till firmly curdled; may be served plain or with sugar and grated nutmeg. An egg beaten to a froth and sweetened with two teaspoonfuls of sugar may previously be added to the half pint of milk, forming a highly nutri- 109 MILK EXAMINATION. tious and smooth jelly. The essence will curdle milk with egg as readily as plain milk. 190. Whey.-Curdle warm milk with the pepsin essence as above directed; then beat up with a fork until the curd is finely divided, now strain and the whey is ready for use. Whey is useful in diet for the sick. It is frequently resorted to as a food for infants to tide over periods of indigestion. 191. To the curd obtained, § 184, add a few drops of dilute hydrochloric acid, so as to make with the pepsin an artificial gastric juice, and set aside at 104° F. for two or three hours. The curd is digested and gradually dis- solves to make a yellowish fluid with the peculiar odor and bitter taste of "peptonized milk.'' 192. Rennet Curd.-To a small quantity of fresh milk in a test-tube add a few drops of com- mercial extract of rennet, warm to 104° F., and set aside. In a few minutes a solid coagulum forms, and later on a sweet whey separates. If the milk be previously boiled, or if the rennet be boiled, the ferment will not work. It acts best at a temperature that can be borne in the mouth, i. e., about 104° F. 193. Specific Gravity.-Shake well a specimen of fresh or " whole milk," and pour into a cylindrical glass vessel until two-thirds full. Slowly immerse a hy- drometer; it will record at the surface somewhere between 1029 and 1034 (Fig. 34 194. The specific gravity of dairy milk, the product of a number of cows, should never fall below 1029. When lower than this it is usually due to adulteration with water, but very rarely the low density is due to excess of cream in very rich milk. Fig. 32. Hydro- meter (Starr). Crea mo- meter (Starr). 110 MILK EXAMINATION. 195. Let the milk stand for 24 hours, "skim" and remove the " cream " which has risen to the surface. Now take the sp. gr. : it will range between 1032 and 1040. The fat globules of the cream in "whole milk" lower the gravity, because fat is lighter than water. " Skim milk " is heavier than fresh or "whole milk," but this difference may be removed by adding water, which is also lighter than milk. 196. Correction for Temperature.-As cold milk is heavier than warm milk-i. e., has a higher sp. gr.-a correction for temperature is necessary. For ordinary hydrometers, adjusted at 6o° F., an approximately correct sp. gr. is obtained by subtracting one from the hydrometer reading for each io° F. below 6o° F., and by adding one to the reading for each io° F. above 6o° F. For variations of less than io° F., the proportionate frac- tion of one must be added or subtracted, as the case may be. MILK TESTING. 197. (fi) Take the specific gravity with a hydrometer and correct for temperature. (Fig. 32.) Any milk that stands above 1033 (33 of the scale) is almost sure to be skimmed, while that falling below 29 of the scale is nearly always watered. The only exception to this rule is in the case of a very rich milk contain- ing an excess of cream. This excess would be apparent to the unaided eye by the consistency and the "creamy" appearance. There is a trick known to the milk trade by which the indications yielded by the sp. gr. may prove fallacious. Some of the cream is skimmed off of fresh milk with a sp. gr. of about 1030, leaving a milk of higher sp. gr. about 1035. By adding water in proper amounts the sp. gr. may be lowered again to that of the original whole milk. The trained eye may detect a change in color and consistency, but we cannot depend on the creamometer (a grad- uated cylinder by which we measure the layer of cream that rises in a cer- tain time). A milk that has been watered will in a few hours separate a thicker layer of cream than would the same amount of the unwatered. The separation takes place with greater rapidity in the diluted than in the whole milk. The lactometer of the N. Y. Board of Health is a hydro- meter which has a scale on which ioo° stands for a sp. gr. of 1029 (the minimum density of pure milk), while o° stands for the sp. gr. of water, 111 MILK EXAMINATION. and 120° for 1034, the maximum range of pure milk. On this instrument t° is read as one per cent, of milk in the sample. 198. (f) Note the color and consistency. For clinical study of mother's milk or ordinary sanitary testing of cow's milk Feser's lactoscope gives valuable results. A rich milk looks like cream, and is thicker than a skimmed milk. If the color is bluish or less creamy, and the fluid less opaque, while the sp. gr. is less than 1029 or under ioo° of the New York lactometer, then it is safe to assume that the milk has been watered. A pure, rich milk is perceptibly more opaque than a skimmed or watered sample. The opacity or white color of the milk is due to the suspended fat globules, and is proportionate to the number of them. By measuring this opacity an approximate estimate can be made of the percentage of fat. For making this estimate roughly, Heeren's pioscope may be used, but Feser's lactoscope (Fig. 33), is very con- venient and more accurate. In the axis of a cylindrical, clear glass vessel, and at its lower part, A, is a smaller cylinder of white glass, marked with a few black lines. In testing with this instrument four c.c. of milk are intro- duced with the graduated pipette ; the black lines are entirely concealed. Pure water is gradually added, while shaking, until the milk clears up sufficiently to make the black lines visible. By the graduation on the vessel the surface level of diluted milk can be read as percentage of fat in the original sample. The microscope having determined the absence of chalk, starch, or other suspended adulterants, a sample showing three per cent, and over is judged pure. Some rich Jersey milk shows six per cent. Any one experienced in its use will be accurate to within one-fourth of one per cent. Having obtained the specific gravity by the lactometer, and the percentage of fat by the lactoscope, experiment shows that the proportion of total solids can be calculated by the formula of Hehner and Richmond given on page 114. 199. (r) Measure the amount of cream. (Fig. 32.) For this purpose a creamometer or a 100 c.c. glass cylinder graduate may be used. Having mixed the milk thoroughly, a sample is poured into the Fig. 33. Feser's Lactoscope {Queen). 112 MILK EXAMINATION. vessel up to the highest mark. After standing in a cool place for 24 hours the depth of cream layer thrown up is read off. It ought to show from 10 to 20 volumes in the hundred. The average sample would be 12 per cent. If the cream forms 20 per cent, of the column, the sample would probably also show a low specific gravity. The accuracy of this test is affected by the length of time since milking, by the amount of previous agitation of the milk, by the fact that dilution causes a more rapid separation of the cream, by the temperature and other variable conditions. It may serve a useful purpose when taken in consideration with other observations. Less than ten per cent, of cream in a milk of specific gravity above 1033 denotes skimming. Less than tiventy per cent, of cream, if joined to a specific gravity less than io2q, indicates ivatering. DETERMINATION OF WATER, SOLIDS, ASH, AND FAT. Milk Standards.-In a number of States a chemical standard of purity has been fixed by law. All samples of cow's milk falling short of this are assumed to be adulterated. By the U. S. Treasury Department, it should contain by weight not less than 13 per cent, of solids and not less than 3.50 per cent, of fat. In Philadelphia the ordinance of 1890 requires not less than 12 per cent, of solids nor less than 3.50 per cent, of fat. In the State of Pennsylvania it must contain not less than 12.50 per cent, of solids and not less than 3.00 per cent, of fat. In the States of New York and New Jersey it should contain not less than 12 per cent, of solids, nor less than three per cent, of fat. 200. Total Solids.-Into a tared dish, preferably of platinum, though a watch glass will answer, five gms. of milk are weighed or five c.c. measured. The dish is then exposed to the heat of a water bath for three hours. As evaporation is nearly done, it is now put into a water oven, and .at intervals weighed until it ceases to lose weight. This constant weight, less the weight of the capsule, gives the total solids. The difference between the five gms. and the constant weight of the dry solids represents the water. By carefully incinerating the 113 MILK EXAMINATION. solids to a grayish-white color the ash or inorganic salts are determined. In pure milk the amount ranges from 0.70 to 0.80 per cent. A watered milk will show a re- duced amount both of solids and of ash. 201. Ah/ by the Werner Schmid Process.-This is an easy, rapid, and quite accurate method. Into a long test- tube with a capacity of 50 c.c., and graduated to show c.c. in tens, measure 10 c.c. of milk and 10 c.c. of strong hydrochloric acid. (A large common test-tube can be used, and the measurement made by pipettes or other graduated glasses.) The mixture of acid and milk is boiled I j/a minutes, or the tube may be corked and heated in a water-bath for five or ten minutes, until the liquid turns a deep brown, but not black. Having cooled the tube and its con- tents in running water, 30 c.c. of well washed ether must be added, the tube corked, the mixture well shaken, and finally stood aside. When the line of separation between ether and acid is distinct, a wash-bottle cork stopper with its tubes is substituted for the plain stopper (see Fig. 34). The lower end of the exit tube has a short curve, which is adjusted so that its opening is just above the line of separation. A weighed flask or beaker is held so as to receive the ethereal layer when it is blown out by the lips at the upper tube. In suc- cession two additional portions of washed ether, 10 c.c. each, are shaken with the acid and blown out into the Fig. 34. 114 MILK EXAMINATION. weighed flask. The ether is then distilled off or evap- orated, and the fat residue dried in a water oven and .weighed. It is the amount contained in io c.c. of milk. 202. Fat.-To determine the butter by the gravimetric method, ten gms. of milk are weighed into a tared dish containing a weighed amount of dry sand. The milk is evaporated on a water bath and last on a water oven, with constant stirring. The residue is washed a number of times with warm ether or petroleum naphtha of 70° Baumd and the washings passed through a small filter. The filtrates are all received in a tared beaker and carefully evaporated to a constant weight. The residue is fat. This subtracted from the amount of total solids gives the solids not fat. 203. Fat by Adams' Method.-This is the standard pro- cess in use by official chemists who have well equipped laboratories. The milk is absorbed by strips of pure, fat-free paper which distributes the milk fat in a thin layer. The coiled strip is dried in a water oven and then placed in the middle chamber of a Soxhlet extractor (Fig. 35). The tared flask containing 75 c.c. of ether is heated on a water bath. Ether vapor condenses in the upper apparatus, flows back upon the coil of paper and returns to the flask. After ten such washings, the flask containing the ether is detached and connected with a condenser. After distillation the fat residue is dried in an air oven, cooled, and weighed. CLINICAL EXAMINATION. Formula of Helmer and Richmond.- These are used to facilitate analysis of mother's milk for clinical pur- poses, and sometimes in sanitary sorting of cow's milk, when rigid accuracy is not required, but time must be saved. By these formulae any one of the three data, specific gravity, fat,mand total solids, can be calculated Fig. 35. 115 MILK EXAMINATION. when the other two are known. Having ascertained the specific gravity with the lactometer and the fat with Feser's lactoscope (see p. in), or by the Werner-Schmid process, the analyst can be spared the tedious process of determinirig the total solids by evaporation. When the fat and the specific gravity are known, the formula for estimating total solids is- F 4- 0.2186 G 0.859 in which F stands for percentage of fat, T the percent- age of total solids, and G the specific gravity expressed in the last two units and any decimal; thus, if the specific gravity is 1028.5 then G stands for 28.5. For example, if a specimen of milk had a specific gravity of 1030, and the percentage of fat was four, then- Total solids = 4 + (0.2186 X 30) I2 cent. 0-859 When the specific gravity and the total solids are known, then for ordinary whole milk- F - 0.859 T - 0.2186 G. For poor skim milk there is a possible error of small amount, and when exceeds 2.5 some change is neces- sary :- F = 0.859T - 0.2186G - 0.05 -2.5). A sliding rule has been devised by Richmond for ready calculations. It has a scale for fat and one for total solids fixed, while that for specific gravity slides. Having adjusted the arrow point to the fat ascertained by the lactoscope or other process, the total solids will be found opposite the known specific gravity.