A Study of Some of the More Important Biochemical Tests DISSERTATION SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN THE FACULTY OF PURE SCIENCE OF COLUMBIA UNIVERSITY BY CHESTER ARTHUR MATHEWSON, B.S., M.A. NEW YORK CITY 1912 Easton, Pa.: Eschenbach Printing Co. 1912 A Study of Some of the More Important Biochemical Tests DISSERTATION SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN THE FACULTY OF PURE SCIENCE OF COLUMBIA UNIVERSITY BY CHESTER ARTHUR MATHEWSON, B.S., M.A. NEW YORK CITY 1912 Easton, Pa.: Eschrnbach Printing Co. 1912 TABLE OF CONTENTS. I. Introduction 7 II. Historical 8 III. Method 15 IV. Procedure 21 V. The Compilation of the Data 22 VI. Tables of Interferences: 1. The Molisch Test 26 Glucose. Sucrose. Levulose. Lactose. Lintner's Starch. Inulin. 2. The Iodine Test 30 Potato Starch. Lintner's Starch. 3. The Fehling-Benedict Test 32 Glucose. 4. The Barfoed Test 34 Glucose. 5. The Seliwanoff Test 36 Levulose. 6. The Biuret Test 38 Egg Albumin. Witte's Peptone. Acid Albumin. Edestin. Ovomucoid. Yeast Nucleoprotein. 7. The Millon Test 41 The same proteins as for the biuret test. 8. The Hopkins-Cole Test 46 Egg Albumin, Witte's Peptone, and Acid Albumin. 9. The Xanthoproteic Test 48 The same proteins as for the biuret test. VII. Supplementary Studies on the Millon Test 50 VIII. Comment on the Tables of Interference 56 IX. A General Summary in Tabular Form 62 X. A Summary of General Conclusions 64 XI. Bibliography 66 XII. Biographical 67 TO MY WIFE ACKNOWLEDGMENT. This dissertation is the outcome of a suggestion on the part of Professor William J. Gies, made to the writer in the Fall of 1910. It is with an especial sense of gratitude that acknowledg- ment is made of Professor Gies's cooperation in overcoming difficulties in connection with the work. His numerous and timely suggestions, particularly in respect to materials and methods, were factors which contributed very largely to the accomplishment of this research. The writer wishes also to express his thanks for the cordial help of the members of the Biochemical Department. C. A. M. Laboratory of Biological Chemistry, College of Physicians and Surgeons, ^Columbia University, May, 1912. I. Introduction. It is merely trite to say that in all sorts of work, from the most elementary to that done in research laboratories, confus- ing or possibly anomalous results have been obtained when any of the common tests for proteins, carbohydrates or fats have been used. Nevertheless it is true that in spite of the fre- quent occurrence of such results they have received compara- tively slight attention from laboratory workers. The reason probably is that when interfering factors do occur they are eliminated at the earliest moment and in the easiest way, in order that there may be the least possible interruption to a piece of work. Interfering factors that are necessarily so common as to force themselves constantly upon one in the course of laboratory practice may or may not be well enough understood to be fore- seen and avoided. Isolated instances thus noted and verified are mentioned in such books as Hawk's "Practical Physio- logical Chemistry" and other laboratory guides. Thus there are memoranda of the inhibitory effect of zinc on the acetic acid and potassium ferrocyanide tests for proteins; of sodium chloride on the Barfoed test for monosaccharides, and many others. Now and then it happens that an investigator will pause and examine not merely the qualitative aspects of an interference but the quantitative as well, with a view to determining meth- ods of overcoming the difficulty. Such studies, however, have, as a rule, been merely incidental and rather isolated. There has hitherto been no especial attempt to attack systematically or in a comprehensive manner, the problem involved in establishing the existence or non-existence of interference in connection with common color reactions. Many interferences are well known either empirically or theoretically. These are necessarily reviewed in the present work. The chief aim of the work, however, has been, first, to demonstrate interferences which a priori could not be fore- told; and second, to demonstrate the absence of interferences in cases where one might on theoretical grounds be tempted to look for them. 8 In brief, then, the present investigation attempts to answer some of the questions constantly arising from interfering fac- tors; and attempts within certain narrow limits to open fur- ther fruitful fields for study, especially quantitative study. II. Historical. Many writers during the past fifty years have made the sub- ject of interference the basis of extended experiment, but in no case has the writer been able to find the record of a study even approximating the present one. No record was found of any such work having been done upon the xanthoproteic and Hopkins-Cole reactions for proteins, or upon the Sudan III test for fats. All of the other seven tests have been investi- gated to some extent from various points of view. Some of these studies touched upon facts noted in the present work, but most of them referred to matters not strictly within its province. Taking the various tests in order, the following review will serve to emphasize the pertinent facts already established. The review covers in all cases practically the en- tire time from the publication of each test up to the present. All of the references listed in the bibliography have been verified and reviewed with the exception of the few that are preceded by an asterisk. The Molisch Test.-Colasanti1 found that sodium and potas- sium thiocyanate and thiocyanic acid give the Molisch reac- tion, and pointed out that since urines usually contained the latter a positive Molisch reaction is not conclusive evidence of the presence of sugar in the urine. Reinbold,2 in a thorough- going paper dealing with this reaction, called attention to the fact that it was first used in 1886, and showed that experimental results with it vary according to the conditions. Nowhere in the literature has the writer found any reference to the inter- fering factors noted in the following tables, except such mat- ters as the darkening effect of sulphuric acid on certain organic compounds. The Iodine Test.-Of all the tests included in the present survey, that of iodine for starch has received by far the most attention. In the great mass of literature one significant fact 1 G. Colasanti: Gazetta Chimica, 1891, XX., pp. 299-305. 2 B. Reinbold: Pfliiger's Archiv, 1904, CHI., pp. 581-617. 9 appears, -viz., that although the test has been in use for over half a century there has been, and still is, a controversy (to which many workers have been parties) as to the nature of starch iodide. Obviously this point is of some interest in de- termining factors of interference. It is not within the scope of this investigation to review this long controversy, but in the course of the studies made with a view to determining the na- ture of starch iodide many facts pertinent to the matter of in- terference have been brought to light. These will be treated chronologically. In 1863 Goppelsroder1 proved that the starch iodide reac- tion is "masked" by certain inorganic substances. By com- parative tests he proved that ammonium sulphate, magnesium sulphate, aluminium sulphate and potassium alum often in- hibit the reaction for a time. In 1872 DuClaux2 demonstrated that sodium carbonate and sodium hydroxide were interfering factors, but that sodium bicarbonate did not interfere. He also showed that sodium sulphide, as well as sodium sulphide plus sodium carbonate, have a similar effect, but the latter combination is more potent in causing interference with the iodine test than the single substance. He quotes Payen and Gobley to the effect that alcohol decolorizes starch iodide, and Goppels- roder and Schoenbein to the effect that salts, e. g., sulphates of potassium, ammonium, sodium, magnesium and aluminium have the property of masking to some extent the reaction of iodine with starch. Finally, he calls attention to the fact that an increase in the percentage of iodine solution present in a test raises the temperature at which decolorization occurs. Puchot,3 in 1876, showed that albumin in a starch iodide mixture, caused the blue color to vanish. The rate of such disappearance is conditioned by the relative percentages of iodide and albumin. He proved that the inhibitory action of the albumin continues until an excess of iodine solution pre- vents it and that this action was due to the union of albumin and iodine. Tschirikow,4 in 1891, in a paper on the effect of temperature 1 Fr. Goppelsroder: Poggend. Annalen, 1863, Bd. CIX., p. 57. 2 E. DuClaux: Ann. Chim. Phys., 1872, [4], XXV., pp. 264-284. 3 E. Puchot: Compt. rend., 1877, LXXXIII., pp. 225-226. 4 A. D. Tschirikow: PRu. 1891, Charkow, XXX., pp. 802-807. 10 upon the limits of sensitiveness of the starch iodide reaction, showed that the blue coloration is dependent to some extent on temperature and other external influences. His experi- ments showed that tests which were positive at 90 C. were nega- tive at 200 C. In 1894, Meinecke1 published a paper entitled "Studies on the Starch Iodide Reaction," in which he maintained that the sensitiveness of this test was increased by the use of many inorganic salts, e. g., iodides, chlorides, sulphates (cf. Goppels- rbder, and DuClaux, page 9) potassium alum and borates, their degree of effectiveness being in the order named. His conclusions were contrary to the views of the two investiga- tors mentioned, except as they referred to iodides and chlor- ides. In the same year Roberts2 called attention to the destruction of the blue color by iodic acid and by iodine chloride. She re- ported that one drop of weak silver nitrate solution destroyed the color of starch iodide. Her conclusion was that the loss of color seemed to be conditioned by a loss of hydriodic acid from the solution. The concluding remarks of her paper were: "If we assume that hydriodic acid is constantly being formed in traces in an iodine solution and remember that the reaction between hydriodic and iodic acids is not immediate in the presence of large amounts of water, we can readily understand why a considerable excess of iodic acid must be used in order to destroy completely the last traces of hydriodic acid and thus entirely prevent the formation of starch blue." In the same year (1894), Lonnes3 called attention to mer- curic chloride as an interfering factor in this test and also pointed out that hydriodic acid is an essential constituent of the blue compound. Marcel,4 in 1903, in a paper "On the Cause of Error in Test- ing for Iodine in Urines," showed that potassium cyanide interferes with or inhibits the iodine test. Grelot,5 in 1906, carried out a carefully planned series of ex- 1 C. Meinecke: Chem.-Ztg., 1894, XVIII., pp. 157-160. 2 Charlotte F. Roberts: Amer. Jour. Sei., 1894, [3], XL VII., pp. 422-429. 3 C. Lonnes: Zeit. anal. Chern., 1894, XXXV., pp. 409-436. 4 Guerbet Marcel: J. Pharm. Chim., 1903, [6], XVII., pp. 313-314. 5 P. Grelot: Jour. Pharm. Chim., 1906, [6], XXIV., pp. 154-161. 11 periments from which he concluded that certain organic sub- stances, notably reducing substances, diminished the sensi- bility of the starch iodide reaction "even in the cold." Sucrose, lactose, dextrose and gum arabic absorb iodine "in the cold" in proportion to the degree of concentration, duration of con- tact, and the amount of time of the experiment, according to his view. Riegler,1 in 1907, in a paper on a new method for the deter- mination of iodides in the presence of bromides and chlorides, proved that nitrites interfered, and suggested the possibility of the removal of these by urea and by boiling. In the same year Labbe and Chabriez2 demonstrated that the action of iodine on albumin3 may be divided into three phases: 1. A first phase of unstable compounds. 2. A phase of stable iodoamines which fails to give the biuret reaction. 3. A third phase of undialyzable, insoluble compounds depending upon the agencies and conditions of the reaction. Taken together then, these papers show that there is a wide variety of factors which may cause interference with the iodine test, and that there is not always unanimity of opinion regard- ing the effects of such factors. The Fehling-Benedict Test.-'This test worked out by Bene- dict4 has been used for only a couple of years and no published accounts of interferences in relation to it were found. Bene- dict mentions the fact that uric acid, chloroform and formalde- hyde do not reduce his reagent. Since it is based upon the old Fehling test, however, it was deemed worth while in this connection to review the literature of the latter test from the point of view of the present paper. The names of writers upon the Fehling test are many but the investigators who have dealt particularly with the phenomena of interference are few and far between. In 1875, Boivin and Loiseau5 noted the deposit of cuprous oxide on boiling Fehling's solution with distilled water. Comparison of this action with that of tap water (from the Seine) led them to note the absence of such reduction. 1 E. Riegler: Z. anal. Chem., 1907, XLVI., pp. 315-318. 2 H. Labbe and J. Chabricz: Compt. rend. soc. biol., 1907, LXIIL, pp. 32-34- 3 Cf. Puchot, Loc. cit. 4 S. Benedict: J. Biol. Chem., 1909, V., pp. 485-487. 5 E. Boivin and D. Loiseau: Compt. rend., 1875, LXIX., p. 1263. 12 They inferred that the latter phenomenon was due to the pres- ence of salts and showed that chlorides of calcium, barium, sodium and ammonium, ammonium nitrate, neutral potassium tartrate and caustic potash, even in small amounts, tended to hinder reduction. Rosenheim and Schidrowitz,1 in 1897, sug- gested means for varying the Fehling solution and reaction. In this connection they quote Boivin and Loiseau2 on the in- terference of chlorides and deny Jovitschitsch's claim that mineral acids and various alkaline salts of mineral acids will reduce Fehling's solution. The same statement is made about tartaric acid. Repiton,3 in 1908, maintained that very small amounts of free acids (both mineral and organic) in organic liquids caused a reduction in Fehling's solution. These facts show that there are quite a few phenomena of interference with the test, and the same may well be true of the Benedict test. The Barfoed Test.-In 1878 Muller4 published a paper deal- ing with the relation of normal urine to the Barfoed test. Later in the same year he, in collaboration with Hagen,5 published a similar paper. Both papers dealt with glucose in the urine and Hagen stated that lf to 1/64 per cent, solutions of glucose could be detected, and commended the reagent for its sensi- tiveness and specificity in determining glucose. At the close of his paper Muller says: "Before I entered upon the investi- gation of the relation of the other urinary constituents to cop- per salts I held it to be essential, first to make some tests upon the relation of uric acid and related alkalies to water solutions of copper sulphate and copper acetate and to the Barfoed re- agent in order to arrive at complete certainty regarding the re- lation of uric acid in the urine in this connection. These tests yielded uncertain results, so that a more exact study became necessary." The final remark is especially interesting in the light of the very high coefficient of interference demonstrated in the follow- 1 O. Rosenheim and P. Schidrowitz: Chem. News, 1897, LXXVI., pp 318-319; Ibid., LXXVIL, p. 97. 2 Boivin and Loiseau: Loc cit. 3 F. Repiton: Ann. de Chim. analyt. appl., 1908, XIII., pp. 269-270. 4 W. Muller: Pfliiger's Archiv, 1878, XVI., p. 562. 6 W. Muller and J. Hagen: Ibid., 1878, XVI., p. 567. 13 ing tables. Only one other investigator is on record as having studied the interference phenomena of Barfoed's reagent. Welker,1 in 1908, showed that sodium chloride produces a greenish white precipitate of unknown nature with the Barfoed reagent even when present in very small percentages. The Selrwanoff Test.-This test has received considerable attention in France and Germany. In 1903 Rosin2 showed that by adding sodium carbonate till ebullition ceased and then adding amyl alcohol (shaking vigorously) followed by a few drops of absolute alcohol the color of the Seliwanoff test is heightened to a pure rose-red, especially on cooling. R. and O. Adler,3 in 1905, in a paper on the reactions of some of the carbohydrates, mentioned the usefulness of the test in the recognition of ketoses and pointed out that aldoses do not give the test. In this paper is mentioned the inhibitory action of ferric chloride on the Tollens reaction. Malfatti4 mentioned the fact that in applying the Seliwanoff test to levulose in urines not only strong acids but also weak alkalies, carbonates, ace- tates, phosphates, etc., cause difficulty by changing dextrose to levulose. In 1909 Pieraerts5 in a paper entitled "Does the Seliwanoff Reaction Really Characterize the d-Fructose Group?" proved that this reagent is not a good one for detecting levu- lose in the presence of other sugars in mixtures. He found that when treated with resorcinol hydrochloride in the usual way pure dextrose gives the same tint as when mixtures of 98 per cent, dextrose and 2 per cent, levulose, 95 per cent, dextrose and 5 per cent, saccharose, and 90 per cent, dextrose with 10 per cent, raffinose were employed. He therefore concluded that the presence or absence of levulose and its anhydrides in a mixture of sugars cannot be definitely settled by the use of Seliwanoff's reagent. DeConinck,6 in a paper published soon after the above, expressed complete accord with the view of Pieraerts7 that hydrochloric acid with resorcin is not charac- 1 W. H. Welker: Proc. Amer. Soc. Biol. Chern., 1908, pp. 33-34. 2 H. Rosin: Ztschr. phys. Chern., 1903, XXXVIII., pp. 555-556. 3 R. and O. Adler: Pfliiger's Archiv, 1905, CVL, pp. 323-328. * H. Malfatti: Ztschr. phys. Chern., 1908, LVIII., pp. 544-546. J. Pieraerts: Bull. soc. chim., 1909, [4] V., pp. 248-250. 6 W. O. deConinck: Bull. soc. chim., 1910, V., p. 569; Compt. rend. soc. biol., LXVI, p. 509. 7 Pieraerts: Loc. cit. 14 teristic in its behavior toward levulose in sugar mix- tures. The Biuret Test.-In decided contrast to the great mass of references, both direct and indirect, dealing with the forma- tion of biuret, is the small number of references bearing on the problem of interference. When one considers the delicacy of the test this fact seems all the more striking. In 1898 Stokvis,1 in a paper on the meaning of the biuret reaction in the human urine noted that urobilin causes a coloration similar to that of the biuret reaction. He therefore concluded that "accord- ing to all the known methods of applying the test a positive reaction is possible in the absence of peptone and in the pres- ence of urobilin, hence the appearance of peptone in the urine has not been definitely established." In 1905 Feder,2 in a paper on the effect of alkaloids upon certain oxidation processes, noted that alkaloids have no effect upon the biuret reaction. He also called attention to the fact that by the use of hydrogen peroxide the interference of glucose in the biuret reac- tion might be overcome. Tschugaeff,3 in 1906, published two papers reporting the results of his experiments with the biuret test. He found it possible to obtain compounds related to those of the biuret reaction by using metals other than copper, but determined that copper is best. The influence of caustic alkalies was in the strengthening the basicity of the amino groups weakened by the proximity of CO and Cu. He noted that the substance formed during the biuret test is fairly stable in the solid form, but decomposes quickly in aqueous solu- tions, especially in the presence of acids, alkalies, alkaline carbonates, hydrogen sulphide and ammonium sulphide. Van Norman,4 in 1909, reported that an excess of uric acid, urates and creatinin makes it difficult to obtain the biuret reaction in concentrated urines. Gies5 has pointed out that the pres- ence of large amounts of compounds that react with alkali hydroxides causes interference. The Millon Test.-Although this reaction has been known 1 H. B. J. Stokvis: Z. Biolog., 1898, XXXIV., pp. 466-470. 2 E. Feder: Arch, der Pharm., 1905, CCXLII., pp. 680-704. 3 L. A. Tschugaeff: Jour. Russ Phys. Chem. Soc., 1906, VII., pp. 1083-1085. 4 K. H. Van Norman: Biochem. Jour., 1909, IV., pp. 127-135. 5 Wm. J. Gies: Multigraphed laboratory directions. 15 and used for nearly a hundred years, there are comparatively few references to it in the literature. No real study has been made of interferences with the test and only one author suggests improvements in the reagent. His ideas have had no apparent effect upon the manner of preparing the Millon reagent. Nasse,1 in 1901, suggested mixing mercuric acetate with a few drops of a 1 per cent, solution of an alkaline nitrite. This was to be used either with or without acetic acid, but Nasse maintained that nitric acid should be absent. In this connection he noted that hydrogen peroxide, alcohol, and large amounts of chlor- ides interfered and suggested that a small amount of the reagent and a moderate temperature were advisable. In criticizing the Millon reagent he said that many examples have been noted in which the coloration of the phenyl derivatives has varied with the use of the Millon reagent from a brownish-red through bluish-red to a pure blue. He maintained that this was fre- quently referable to the lack of uniformity in the reagent. In 1907 Repiton2 called attention to the fact that in reactions in urine analysis the use of Millon's reagent leads to grave errors as phosphates give a precipitate which carries down phenyl groups containing an OH group. He further quotes Arthus as mentioning that chlorides tend to give precipitates which are of the nature of tyrosin complexes, hence the conclusion that the red coloration is not positive proof of the presence of albu- minous material if Millon's test is used exclusively. Gies3 has called attention to the necessity for avoiding ex- cessive heating, and to interference due to urea, sulphates, alkalies, and excessive amounts of inorganic salts. III. Method. The method of the present research was the simplest possi- ble. Approximately seventy-five compounds and substances which are commonly met with in biological or chemical labora- tories were arbitrarily chosen. These seventy-five substances included representatives of every known group, both organic and inorganic, which might possibly cause interference in bio- chemical reactions. The following is the complete list. Not 1 O. Nasse: Pfliiger's Archiv, 1901, LXXXIII, pp. 361-368. 2 F. Repiton: Compt. rend. soc. biol., 1907, LXIL, pp. 339-340. 8 Wm. J. Gies: Multigraphed laboratory directions. 16 quite all of these were used in connection with every test but everything in the list was used in the course of the work: Agents of Interference I. Neutral Inorganic Salts. i. Ammonium sulphate. 2. Barium chloride. 3. Calcium chloride. 4. Calcium phosphate. 5. Magnesium chloride. 6. Magnesium sulphate. 7. Manganese sulphate. 8. Potassium chloride. 9. Potassium iodide. 10. Potassium nitrite. 11. Sodium chloride. 12. Sodium nitrate. 13. Sodium sulphate. 14. Sodium sulphite. 15. Zinc sulphate. II. Neutral Organic Compounds. 1. Halogen derivatives. Chloroform. 2. Alcohols: a. Ethyl alcohol. b. Glycerol. 3. Aldehydes and ketones: a. Acetone. b. Chloral hydrate. c. Formaldehyde. 4. Carbohydrates: a. Glucose. b. Sucrose. 5. Proteins: Egg albumin. 17 6. Hydrocarbons: Toluol. 7. Phenols: a. Phenol. b. Picric acid. c. Thymol. 8. Amides: Urea. III. Acids. 1. Inorganic: a. Boric. b. Hydrochloric. c. Nitric. d. Sulphuric. e. Phosphoric. 2. Organic: a. Acetic. b. Butyric. c. Benzoic. d. Citric. e. Formic. f. Lactic. g. Oxalic. h. Salicylic. i. Tannic. j. Tartaric. IV. Acid Salts. 1. Inorganic: a. Copper sulphate. b. Di-hydrogen sodium phosphate. c. Ferric chloride. d. Lead acetate. e. Mercuric chloride. 18 2. Organic: a. Acid potassium oxalate. b. Sodium acid tartrate. V. AlkaliES. i. Inorganic: a. Ammonium hydroxide. b. Calcium hydroxide. c. Sodium hydroxide. 2. Organic (very uncommon, therefore none used). VI. Alkaline Salts. i. Inorganic: a. Alum. b. Borax. c. Di-sodium hydrogen phosphate. d. Potassium thiocyanate. e. Potassium ferrocyanide. f. Sodium carbonate. g. Sodium arsenate. h. Sodium sulphide. 2. Organic: a. Basic lead acetate. b. Sodium acetate. c. Sodium benzoate. VII. Biological Mixtures, Etc. i. Beverages: a. Coffee. b. Milk. c. Tea. d. Cocoa. 2. Excretions: Urine. 19 3. Food Extracts: a. Beef tea. b. Meat extract. 4. Physiological saline. 5. Saliva. 6. Tissue extracts: a. Liver. b. Pancreas. c. Blood. • VIII. Miscellaneous. Soap solution. This investigation does not pretend to be fundamentally a quantitative one, although I well recognize that in individual cases of some problems in research a quantitative knowledge of interference is essential. Within the limits of time and expediency, due attention was paid, however, to approximating where possible quantitative notations in connection with the experiments. After deciding on the seventy-odd substances noted above, therefore, five per cent, (by weight) was fixed as the most favor- able solution for such a general survey as that contemplated in this work. All chemicals used were of the highest standard of purity and the solutions were all prepared with the utmost regard to maintaining this standard. Distilled water was used as the solvent wherever possible. The only exception was in the case of thymol, which was dissolved in alcohol. In all cases where a compound would not yield a five per cent, solution, e. g., HgCl2,PbAc, etc., a saturated solution was made. These solutions were kept in clean, well-stoppered bottles. In many cases where interference was found, e. g., in the Molisch test, Barfoed test, etc., confirmatory experiments were made, with solutions made from different "lots" of chemicals. This served as a double check on the results. Substances which could not be "made up" and kept indefinitely were used in the fresh condition-milk, urine, blood, etc. Most of these were used in the form in which they occur naturally. In the 20 case of coffee, tea and other solids, I used about one part to twenty (estimated). Molisch Test.-Five cc. of the carbohydrate solution were put into a test tube, then the five drops of the interfering substance, then the two drops of a-naphthol. Then 5 cc. of concentrated sulphuric acid were poured slowly down the side of the tube into the mixture. Iodine Test.-Five cc. of the starch paste put into a test tube, then the interfering substance, then the iodine solution was added, drop by drop from a burette. Fehling-Benedict Test.-Five cc. of the Benedict solution were put into a test tube, then the interfering substance, then 5 to 8 drops of the glucose solution. The mixture was then boiled about one minute. Barfoed Test.-Five cc. of the Barfoed reagent were put into a test tube, then the interfering substance, then the glu- cose solution-10 to 15 drops. In some cases of doubt the amount of glucose solution used reached 2-3 cc. The contents of the tubes were heated cautiously over a flame for about a minute and compared with the contents of other tubes heated on a boiling water bath for 3-8 minutes. Seliwanoff Test.-Five cc. of the reagent were put into a test tube, then the interfering substance, then 5-8 drops of the levulose solution. Boiling brought out the color. Biuret Test.-Five cc. of the protein solution and 5 drops of the interfering factor were put into each test tube. Then by means of a burette the number of drops of biuret reagent indi- cated in the tables was allowed to flow into the tubes. The color was allowed to develop without heat. Millon Test.-This test was performed in precisely the same manner as the biuret test, with the exception that heat in varying degrees was applied to develop the color. Xanthoproteic Test.-Three cc. of the protein solution and 5 drops of the interfering substance were placed in each tube. To this was added about 1-2 cc. of nitric acid and after boil- ing about 2-3 cc. of ammonium hydroxide were added. Hopkins-Cole Test.-Two cc. of the protein solution and 5 drops of the interfering factor were put into a tube. To this were added 3 cc. of the reagent, glyoxylic acid solution. Five manner of performing the tests. 21 cc. of concentrated sulphuric acid were then used to stratify the solution and bring out the violet color. Sudan 111 Test.-The lard, butter, and olive oil tested were in each case triturated with the interfering substance. Then the special filter paper was smeared in the mixture. This paper was then immersed for ten minutes in the Sudan III solution and later washed in alcohol to determine decolorization, if any. No interfering substances were discovered, hence no separate tables for this test were prepared. IV. Procedure. Most of the tests were made in ordinary test tubes and in general the order of putting the substances into the tubes was as follows: i. The compound to be tested-the amount being 5 cc. or less, according to usual methods in the various tests. 2. The substances whose property of interference or non-inter- ference was to be determined. 3. The compound or compounds constituting the test, e. g., the Molisch reagents, the Millon re- agent, iodine, etc. The amounts of these used varied accord- ing to the experiment, and where the procedure is not fixed by common practice the amounts are specifically stated at the be- ginning of the tables to follow. The only constant amount was that of the interfering factors, this being uniformly five drops to every tube. The order noted above was of course not used in the experiments on reducing sugars. Here, as noted in the section on "Method," the reagent was put into the tube first, and this was followed by the five drops of interfering fac- tor. In this broad survey only approximations to definite amounts could be attained. Figuring approximately then, the percentage of interfering substance present was as follows: In the Molisch and Hopkins-Cole tests, about 0.15 per cent.; in the Xanthoproteic, about o. 2 per cent.; and in the others 0.3 per cent. For all factors of limited solubility in water the percentage was decidedly less, thus e. g., mercuric chloride, lead acetate, etc. The writer does not fail to recognize the im- portance of accurate knowledge of the percentages of these in- terferences. However, the underlying idea of this work was that of discovering the facts about interference due to sub- stances which might be present in biological materials in un- 22 known, small, amounts; therefore it was not practicable to do more than make this quantitative phase approximate. At a very early stage in the work discrepancies and difficul- ties were encountered which were traced to inefficient methods of cleansing the test tubes used. Accuracy of results was found to depend very largely on the success of the cleansing process. After trying various methods the following was finally adopted, and rigidly adhered to step by step: i. Rinsing several times in warm water. 2. Soaking overnight in strong alkali. 3. Rinsing several times in water. 4. Soaking from twenty min- utes to an hour in weak hydrochloric acid. 5. Thorough scrubbing with a test-tube cleaner. 6. Repeated rinsing in running water. The above proved efficacious as a rule, but occasionally it was found necessary to apply sapolio to get rid of gummy masses of protein or heavy precipitates. After cleansing, the tubes were inverted in test tube racks until used. Confusion of results was automatically prevented by having each hole in the test-tube racks permanently labelled with the name of one of the many interfering factors investigated. In the limited studies made upon fats there was necessarily a variation from the above procedure. The oils and fats were triturated in mortars with the interfering factors. The latter were used in the dry state in all cases where that is their natural condition. Then the filter paper used in the test was smeared in the mixture. This paper was then immersed for ten min- utes in a solution of Sudan III and later washed in alcohol to determine decolorization. The cleansing of the mortars and pestles used in triturating was done with an alcohol-ether-ace- tone solution followed by soap and hot water. Clean filter paper was used to complete the process. V. The Compilation of Data. In attempting to solve such a problem as the present one we are constantly being confronted with the difficulty of sharply delimiting the meaning of the expression "interfering fac- tor." In ordinary classroom procedure an interference is usually merely something that causes one to arrive at negative results in common qualitative tests. The mere knowledge of the ex- istence of possible factors of such interference is about all that one would care for in such work. To make easily accessible 23 the established facts in respect to these things, the experimental results of this research are put wherever possible into the form of tables. The interfering factors are arranged in groups be- ginning with the simplest salts and ending with "biological substances." Within the several groups either an alphabetical arrangement is used, or else the compounds are listed accord- ing to the order usual in texts. My chief aim in these tables has been to make reference to them a matter of ease and con- venience. The positive sign + , in the tables then means no in- terference', i. e., that the reaction is normal or approximately so. The zero, o, on the contrary is used wherever interference does occur, i. e., when the reaction is decidedly abnormal. The question mark (?) is used with the + and with the o in rare instances where definite judgment was impossible after repeated experimentation. Now interference may be slight, making necessary an in- crease in the amount of time for a reaction, or causing a slight variation from the normal color; or it may be absolute, causing complete inhibition of the reaction. Again, it may be due to the destruction of the compounds which are necessarily formed in a given test, e. g., K4Fe(CN)6, and the starch-iodide test. In all tests the control was performed with the minimum amount of reagent necessary to produce the proper result. This mini- mum amount is stated wherever necessary at the top of the tables, and it was adhered to throughout except where other- wise noted. Thus, e. g., in the iodine test on starch, one drop was used in the control and in each of the seventy-odd tests of the series except where specifically stated in the table. Not in- frequently if a reaction is completely inhibited when only these minimum amounts are used, an increased amount of the reagent overcomes the interference. As a rule, an interference of that character is not to be considered a real one, and in the tables it is not indicated as such except for special reasons in indi- vidual instances. The notes accompanying the + and o signs often serve to explicate further questions as to varying degrees of interference. It is obvious that any extended study of interferences leads to the consideration of many complicated reactions-some readily understood, some partially understood, and others quite beyond explanation in the light of present knowledge. We 24 have to take into account the possibility of interrelations, re- actions and combinations between the compound studied, the interfering factor and the reagent involved in a given test. The degree of complexity of one or all of these is an index of the possibility of complications which may arise and need explanation. Usually such explanations will be found in con- nection with the tables, but it sometimes happens that no explanation is given because none is known. In work of this nature one is constantly under the necessity of arriving at one judgment after another in an endless array. Most of these judgments were of course quickly and easily made, but still there was a vast number of experimental results which were not so clear-cut as to point the way to what one could regard as safe conclusions. In order to overcome the personal and psy- chological element here involved, practically all negative re- sults, i. e., those showing interference, were studied a second time, or even a third or fourth time. Often complete new sets of materials were involved in such re- considerations. In order to further eliminate the personal element much of the repetition was performed only after the lapse of some months. In research work interference becomes a much more serious problem than in the usual classroom work referred to previously. In order to make these tables of greater service they are accom- panied sometimes by foot-notes. Such notes serve to indicate some of the possibilities and the need for further work, espe- cially studies of individual quantitative relationships not touched upon in this general survey. Many of these notes are particu- larly useful in showing experimentally the reactions among the compounds tested, the interfering factor and the com- ponent parts of reagents. Some supplementary tables are given which embody the re- sults of a few studies of a more detailed character-e. g., on the Millon reaction. At the outset of the work it was quite apparent that a great loss of time would ensue unless blank forms for recording re- sults were devised and used. Accordingly after several trials the form on the opposite page was adopted: 25 Table No. Test Substance added. Result. Remarks. This afforded a sure and convenient means for recording results rapidly. In case of doubt duplicate tables were compiled. Then by comparison of the two, errors or discrepancies were easily discovered. After completing such tables, those bearing on the same test were readily compared and thus the supplementary and comparative tables were made. Following this, further study often justified broader deductions which appear in connection with the various tables. The large majority of the tests naturally show negative re- sults so far as demonstrating interference is concerned. In judging the value of the tables which constitute the chief part of this dissertation, the author would ask that it be remembered that the experimental proof of the lack of interference is just as important as the finding of interferences. VI. Tables of Interferences. Test-Molisch: on o.i per cent, solutions of the carbohydrates indicated Table I. Results. <u si (U cn 0 bS-5 Substance added. o u 5 3 o b s tn 3 > U 0 Q a U S3 a .g 8 « U 3 a I. NEUTRAL INORGANIC SALTS. Ammonium sulphate + + + + + + Barium chloride + 1 + * + 1 + l + 1 + l Calcium chloride + + + + + + Calcium phosphate + + + + + + Magnesium chloride + + + + + + Magnesium sulphate + + + + + + Manganese sulphate + + + + + + Potassium chloride + + -f- + + + Potassium iodide +2 + 2 +2 +2 +2 + 2 Potassium nitrite O3 O3 O3 O3 o3 O3 Sodium chloride + + + + + + Sodium nitrate O4 O4 o4 o4 O4 o4 Sodium sulphate + + + + + + Sodium sulphite + + + + + + Zinc sulphate + + + + + + II. NEUTRAL ORGANIC COMPOUNDS. Chloroform + + + + + + Ethyl alcohol +5 + + + + + Glycerol +6 + + + + + Formaldehyde o7 O7 O7 o7 O7 O7 Chloral hydrate +8 + + + + + Acetone + + + + + + Glucose + + + + + Sucrose + + + + + Egg albumin + + + + + + Toluol + + + + + + Thymol + + + + 4* + Phenol . + + + + + Picric acid + 10 + 10 +10 +'° + 10 + ,# Urea + + + + + + III. ACIDS. Boric acid + + + + + + Hydrochloric acid + + + + + Nitric acid O11 OU Ou O11 O11 o" Sulphuric acid Phosphoric acid + + + + + + Acetic acid + + + + + + Butyric acid + + + + + + Benzoic acid + + + + + Citric acid + + + + + + Formic acid + + + + + + 1 Heavy white ppt. above ring. 2 Color of ring not sharp. Reddish ppt. 3 Heavy brown ppt. with one or two drops of KNO2 or NaNO2. 4 Heavy brown ppt. with one or two drops of KNO3 or NaNO3. 5 Color of ring darkened. 8 Brownish ppt. above the ring. 7 Rather heavy white ppt. Ring greenish rather than purple. 8 White ppt. above the ring. 9 Color of the Molisch ring darkened. 10 A heavy yellow ppt. tends to mask the reaction. 11 A heavy brown ppt. with one drop of HNOa. Test-Molisch: on o.i per cent, solutions of the carbohydrates indicated. Table I (Continued). Results. 8 V co o a 8 o 1 1 Q tjS-3 2-2* d Substance added. 0 a o U rt U § Lactic acid + 12 4-13 4- 4-14 4-14 4-14 Oxalic acid + 4- 4- 4- 4- 4- Salicylic acid + 4- 4- 4- 4" 4- Tannic acid + 1S 4-18 + 18 + 18 4-18 4-18 Tartaric acid + 4- 4- 4- 4- 4- IV. ACID SALTS. Copper sulphate 4- 4- 4- 4- 4- 4- Dihydrogen sodium phosphate. + 4- 4- 4- 4- 4- Ferric chloride o18 o" o18 o18 o18 o18 Lead acetate + 17 4-17 4-17 4-17 + 17 4-17 Mercuric chloride + 4- 4- 4- 4- 4- Acid potassium oxalate + 4- 4- 4- 4- 4- Sodium acid tartrate + 4- 4- 4- 4- 4- V. ALKALIES. Ammonium hydroxide 4- 4- 4- 4- 4- 4- Calcium hydroxide 4- 4- 4- + 4- 4- Sodium hydroxide 4- 4- 4- 4- + 4- VI. ALKALINE SALTS. Alum 4- 4- 4- 4- 4- 4- Borax 4- 4- 4- 4- 4- 4- Disodium hydrogen phosphate. 4- 4- 4- 4- 4- 4- Potassium thiocyanate 4- 4- 4- 4- 4- 4- Potassium ferrocyanide + 18 4-18 + 18 + 18 + 18 + 18 Sodium carbonate 4- + 19 4-19 4- 4' 4- Sodium arsenate 4- 4- 4- 4- 4- 4* Sodium sulphide | 20 | 20 [ 20 | 20 | 20 | 20 Lead acetate (basic) 4-21 4-21 4-21 +21 4-21 +21 Sodium acetate 4- 4- 4- 4- 4- 4- Sodium benzoate 4- 4- 4- + 4- 4- VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee 4- + 4- 4- 4- 4- Tea 4- + 4- 4- 4- 4- Cocoa 4- 4- 4- 4- 4- 4- Milk 4- 4- 4- 4- 4- 4- Urine 4- 4- 4- 4- 4- 4- Beef tea 4- 4- 4- 4- + 4- Meat extract [ 22 _|_ 22 4- 4- | 22 | 22 Physiological saline 4- 4- 4- 4- 4- 4- Saliva 4- 4- 4- 4- 4- 4- Liver extract 4- 4- 4- + 4- 4- Pancreas extract 4- 4- 4- 4- 4- + Blood _|_ 22 4- 4- 4- 4- _|_ 22 Soap solution + + | 22 | 20 4- 4- 12 Color appears slowly unless the tube is gently agitated. 13 A yellowish ring forms unless the test js done with extreme care. 14 A yellow substrate formed. 15 A white persistent ppt. above the ring. 16 A lavender ppt., due to the reaction between Fe2Cl6 and a-naphthol. 17 A heavy white ppt. tends to mask the reaction, especially in the begin- ning. 18 A slight white ppt. above the ring. 18 A slight white ppt. above the ring. 20 Usually a heavy white ppt. above the ring. 21 Heavy white ppt. is formed. 22 A slight white ppt. above the ring. 28 Table II.-Supplementary Table of Interferences with the Molisch Reaction. Note.-As usual, the zero, (o), is used to indicate a negative result. Substance added. Glucose. Sucrose. Levulose. L actose. Starch. Inulin. NaNO2l O O O Brown ppt. with all carbohydrates o O O KNO2' o o o Brown ppt. with all carbohydrates O o O KNOj2 o o o Brown ppt. with all carbohydrates o O O HNO,2 o o o Brown ppt. with all carbohydrates O o O Fe2Cl63 o o o Lavender ppt. with all carbohydrates O o O Formaldehyde4.. . . o o o o o Green ring Green ring Green ring Green ring Green ring white PPt. O Green ring. Both rings turn brown 1 It seems probable that the interference may be caused as follows: The nitrites and sulphuric acid yield nitrous acid, and sodium sulphate; the nitrous acid in turn unites with the a-napthol to form a nitrosonaphthol, C10H6(OH)NO. There is thus no chance for compounds of a-naphthol and furol (upon which the Molisch test depends) to form. 2 Nitrates + H2SO4 yield colorless solutions. HNO3 + H2SO4 yield colorless solutions. Either of these combinations yields, with a-naphthol a heavy brown ppt. in large amounts. The inhibition is therefore somewhat analogous to that noted under footnote h 3 Fe2Cl6 + a-naphthol = lavender ppt. in large amounts. Fe2Cl6 + H2SO4 = yellow to green liquid and no ppt. Fe2Cl0 + sucrose = yellow liquid and no ppt. a-naphthol + H2SO4 = reddish liquid. a-naphthol + sucrose = pink liquid and reddish ppt. (a-naphthol). H2SO4 + sucrose = amber liquid and a reddish zone near bottom. The substitution of various other carbohydrates for sucrose in the above tests made no marked differences. There were merely minor variations in color. The inhibition is therefore due to an iron derivative of the naphthol or dinaphthol. * H-CHO + H2SO4 = clear solution. H-CHO + a-naphthol = clear solution. H2SO4 + a-naphthol = reddish liquid. Starch + a-naphthol = clear solution. Test-Iodine: (1 drop) on starch (Lintner's soluble)-0.1 per cent, solution, starch (potato)-0.1 per cent, paste. Table III. Substance added. Result. Remarks. I. NEUTRAL INORGANIC SALTS. Ammonium sulphate Barium chloride Calcium chloride Calcium phosphate Magnesium chloride Magnesium sulphate Manganese sulphate Potassium chloride Potassium iodide . + . + . + . + . + . + . + . + . + Potassium nitrite Sodium chloride Sodium nitrate Sodium sulphate + Nitrites in excess cause total in- hibition of the reaction. . + . + . + n. Sodium sulphite Zinc sulphate NEUTRAL ORGANIC COMPOUNDS. Chloroform Ethyl alcohol Glycerol Formaldehyde Chloral hydrate Acetone Glucose Sucrose . + Two drops usually necessary to get a good reaction. . + . + . + . + . + . + . + . + Egg albumin . Toluol + An excess of albumin, peptone, acid albumin, edestin, and ovomucoid causes the blue color to fade rapidly and vanish. + Thymol.. ? + From two to three times the usual amount of iodine needed to get proper reaction. Phenol + Reaction inhibited unless 2-3 drops of iodine used-phenol and starch react. hi. Picric acid Urea ACIDS. Boric acid Hydrochloric acid .. Nitric acid Sulphuric acid Phosphoric acid Acetic acid + Iodide ppt. is very slight. + + + + + + + Butyric acid Benzoic acid Citric acid + + + Formic acid Lactic acid Oxalic acid Salicylic acid + + + + IV. Tannic acid Tartaric acid ACID SALTS. Copper sulphate Dihydrogen sodium phosphate. Ferric chloride + 1 Reaction inhibited unless 4-10 drops of iodine are used. Coloration yellowish. + + + + Lead acetate 4- Pbl2, a bright yellow ppt. is formed in large amounts. Greenish gray ppt. Mercuric chloride 0 With excess of iodine a drab ppt. Total inhibition due to mer- curic iodide. Table III {Continued}. Test-Iodine: (i drop) on starch (Lintner's soluble)-o.i per cent, solution, starch (potato)-o.i per cent, paste. Substance added. Result Remarks. Acid potassium oxalate + Sodium acid tartrate + V. ALKA MES. Ammonium hydroxide o?2 Absolute inhibition, unless excess Calcium hydroxide O?2 of iodine present. Good reaction with 3 drops iodine. Sodium hydroxide o?2 Absolute inhibition, unless 40-50 times the usual amount of iodine present. VI. ALKALINE SALTS. Alum + Borax + Disodium hydrogen phosphate. + Potassium thiocyanate o? Will interfere unless there is an Potassium ferrocyanide excess of iodine unreduced. + ?3 The yellow color of reagent masks Sodium carbonate + ? blue. If formed, blue disap- pears at once. No blue color unless 2 drops of Sodium arsenate + iodine used. Sodium sulphide . o?4 Absolute inhibition, unless 40 Lead acetate (basic) . + times the usual amount of iodine is present. Pbl2 which is a bright yellow ppt. Sodium acetate + is formed in large amounts. Greenish gray ppt. Sodium benzoate + VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee + Double quantity of iodine neces- Tea . + sary. Double quantity of iodine neces- Cocoa + sary. Double quantity of iodine neces- Milk + sary. Urine + 5 drops necessary to get a strong Beef tea . + blue. Meat extract + Physiological saline + Saliva. o Starch digested. Liver extract ? Pancreas extract ? Blood + Soap solution . o?2 Inhibition caused by the alkali in soap. 1 If tannic acid and iodine are allowed to react before being put into the test tube with starch, inhibition is instantaneous (unless there is an excess of iodine). But if this interaction does not take place in the absence of the starch the blue color appears and then disappears. This is contrary to what happens in the case of phenol. The inhibitory action of the phenol is due to a reaction between it and starch, rather than between it and the iodine. Tannates, gallates, and salts of heavy metals all interfere if present in sufficient amounts. 2 Vide comments on the iodine reaction in general. 3 Potassium ferrocyanide exhibits a relation to iodine similar to that noted in connection with tannic acid (cf. note i). If potassium ferrocyanide and iodine are allowed to react before being mixed with the starch no blue color appears (unless there is a great excess of iodine). If the three compounds are all put into a test tube together the blue iodide is formed and then rapidly disappears. Whether this is due to the substitution of iodine for the CN radical or to the formation of an addition compound is not clear. * When Na2S is present, both the bisulphide and the iodide of sodium are formed in large amounts. This makes a great excess of iodine necessary if the iodide of starch is to be produced. Test-Fehling-Benedict: on glucose-0.1 per cent, solution. Table IV. Substance added. Result. Remarks. I. NEUTRAL INORGANIC SALTS. Ammonium sulphate A clear blue solution. No ppt. Barium chloride . + Calcium chloride . + Calcium phosphate . + A white phosphate ppt. tends to Magnesium chloride . + mask the reaction. Magnesium sulphate . + Manganese sulphate . + Usually a dark solution and only Potassium chloride . + a slight reduction. Potassium iodide . + Potassium nitrite . + Sodium chloride . + Sodium nitrate . + Sodium sulphate . + Sodium sulphite . + Abundant white ppt. Reduction Zinc sulphate slow and small in amount. II. NEUTRAL ORGANIC COMPOUNDS. Chloroform Ethyl alcohol . + Glycerol . + Formaldehyde . + Chloral hydrate . + Acetone . + Glucose Sucrose . + Egg albumin . + Toluol . + Thymol . + Phenol . + Picric acid - . + Urea . + III. ACIDS. Boric acid . + Hydrochloric acid . +? Reduction is very slow and ap- Nitric acid . + pears only after cooling (30 minutes). Reduction slow Sulphuric acid . + Reduction slow Phosphoric acid . + Acetic acid . + Slight reduction. Butyric acid . + Benzoic acid . + Citric acid . + Formic acid . + Slight reduction. Lactic acid . + Oxalic acid . + Salicylic acid . + 1 Na2CO3 decomposes ammonium salts, resulting in the formation of am- monium copper compounds which are soluble. CuSO4 + (NH4)2SO4 = a clear solution, Benedict reagent + (NH4)2SO4 = a clear solution, Bene- dict reagent + sugar = a. clear solution, Na2CO3 4- (NH4)2SO4 = a clear solution. Ammonium sulphide inhibits more strongly because of precipitations as follows: CuSO4 + (NH4)S2 = brown to black ppt. and a bluish white ppt. Na2CO3 + (NH4)S2 = yellowish solution. Sodium citrate + (NH4)S2 = yellowish solution. Test-Fehling-Benedict: on glucose-o.i per cent, solution. Table IV (Continued). Substance added. Tannic acid Result. Remarks. o2 Absolution inhibition owing to pptn. of sodium tannate. Dark coloration. Tartaric acid IV. ACID SALTS. Copper sulphate Dihydrogen sodium phosphate. Ferric chloride + o Total inhibition due to the forma- Lead acetate + tion of Fe2(HO)6 which later forms a carbonate. Heavy white ppt. tends to mask Mercuric chloride o3 reaction. Total inhibition due to formation V. Acid potassium oxalate Sodium acid tartrate ALKALIES. Ammonium hydroxide + + + ? of a hydrated mercuric oxide (reddish). No inhibition because of re- VI. Calcium hydroxide Sodium hydroxide .... ALKALINE SALTS. Alum + + + latively small amount of ammonia in NH4OH. A slight increase in the amount causes inhibition. Reduction slight. Borax + Reduction slight. Disodium hydrogen phosphate. Potassium thiocyanate Potassium ferrocyanide + + o Clear solution. Total inhibition Sodium carbonate Sodium arsenate Sodium sulphide + + + due to formation of Na4Fe(CN)6 light yellow. In larger percentages this causes Lead acetate (basic) + inhibition, due to pptn. of the copper. Heavy white ppt. tends to mask VII. reaction. Sodium acetate + Sodium benzoate + . BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee o Slight interference due to presence Tea Cocoa Milk Urine Beef tea Meat extract Physiological saline Saliva Liver extract Pancreas extract Blood Soap solution o + + + + + + + + + + + of tannic acid in both of these. 2 This inhibition is due to a reaction between the Na2CO3 and tannic acid. There is a brown solution with a blackish ppt. 3 The reaction here is between the Na2CO3 of the Benedict solution and HgCl2 thus-2HgCl2 + 2Na2CO8 + H2O = zHgO.OH + 4NaCl + 2CO2 + H. Table V. Substance added. Result. Remarks. I. NEUTRAL INORGANIC SALTS. Ammonium sulphate . O Slight bluish white ppt. when Barium chloride . o heated. White ppt. after heating. Calcium chloride . O White ppt. after heating. Calcium phosphate . O Total inhibition due to the forma- Magnesium chloride . O tion of insoluble cupric phos- phate. White ppt. after heating. Magnesium sulphate . O White ppt. after heating. Manganese sulphate . o? Slight grayish ppt. and usually a Potassium chloride . O very slight reduction after heat- ing. White ppt. after heating. Potassium iodide . o Total inhibition due to the forma- Potassium nitrite . + tion of cupric and cuprous iodides. Good reduction. Sodium chloride . O White ppt. after heating. Sodium nitrate . + Perfect reduction. Sodium sulphate . o Bluish white ppt. after heating. Sodium sulphite . o Slight bluish white ppt. on heat- Zinc sulphate . O mg. Slight bluish white ppt. on heat- II. NEUTRAL ORGANIC COMPOUNDS. Chloroform ...+ mg. Ethyl alcohol . + Glycerol . + Formaldehyde . + Chloral hydrate . O A clear sol. results. The in- Acetone . + hibition seems due to the dis- solving of Cu2O. Glucose Sucrose . + Egg albumin . + Toluol . + Thymol . + Phenol . + Picric acid . + Urea . + III. ACIDS. Boric acid . o1 Hydrochloric acid . O1 Nitric acid O1 Sulphuric acid . o1 Phosphoric acid . o Total inhibition due to abundant Acetic acid . O1 ppt. of insoluble cupric phos- phate. Butyric acid . ol Benzoic acid . + Citric acid . o' Formic acid . o' Lactic acid . O1 Test-Barfoed: on 0.4 per cent, glucose solution. 1 The mineral acids and some organic acids form copper compounds which dissolve freely. A clear solution is the usual result of adding even small quantities of acid in making the Barfoed test. Table V {Continued). Substance added. Result. Remarks. Oxalic acid O Total inhibition due to abundant Salicylic acid . + ppt. of cupric oxalate. Tannic acid . O Heavy brown ppt. Tartaric acid . O1 IV. ACID SALTS. Copper sulphate o Slight white ppt. Dihydrogen sodium phosphate . o . Inhibition due to formation of Ferric chloride o large amount of cupric phos- phate. Brown sol. Inhibition probably Lead acetate . + due to formation of soluble ferric acetate. Mercuric chloride o Slight white ppt. Some mercuric Acid potassium oxalate o acetate is formed. Inhibition due to formation of Sodium acid tartrate . + large amount of cupric oxalate. V. ALKALIES. Ammonium hydroxide . o Acidity of reagent neutralized. Calcium hydroxide . + Good reduction. Sodium hydroxide o Acidity of reagent neutralized. VI. ALKALINE SALTS. Alum . . .. o Large amt. of white flocculent Borax . +? ppt. before heating. Cupric borate tends to interfere. Disodium hydrogen phosphate o Inhibition due to cupric phos- Potassium thiocyanate o phate. Inhibition due to formation of Potassium ferrocyanide o cuprous and cupric sulfo- cyanates. Grayish ppt. Inhibition due to heavy reddish Sodium carbonate + brown ppt. of cupric ferro- cyanide. White ppt. Fair reduction. Sodium arsenate o Inhibition due to white ppt. of Sodium sulphide o cupric arsenate. Black ppt. of copper sulphide on Lead acetate (basic) + heating. Sodium acetate + Sodium benzoate o? Slight white ppt. Gray ppt. on heating. VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee o Brownish ppt. Tea o Brownish ppt. Cocoa o Brownish ppt. Milk o Urine + Beef tea ... . Meat extract Physiological saline o Saliva + Liver extract Pancreas extract Blood o? Usually a very slight reduction. Soap solution o Inhibition due to alkalinity. Test-Barfoed: on 0.4 per cent, glucose solution. 36 Table VI. Substance added. Result. Remarks. I. NEUTRAL INORGANIC SALTS. Ammonium sulphate Barium chloride . + Calcium chloride . + Calcium phosphate . + Potassium chloride . + Potassium iodide . + Potassium nitrite . ol Lemon yellow solution-no ppt. Magnesium chloride . + Magnesium sulphate . + Manganese sulphate . + Sodium chloride . + Sodium nitrate . + Slight yellowish coloration. Sodium sulphate . + Sodium sulphite Zinc sulphate . + II. NEUTRAL ORGANIC COMPOUNDS. Chloroform . + Pale coloration. Ethyl alcohol . + Glycerol . + Formaldehyde . o?2 Heavy pink ppt. in a pink liquid. Chloral hydrate . + Acetone . + Glucose . + Sucrose . + Egg albumin . 4- Toluol . + Thymol . + Phenol . + Picric acid Slight yellow ppt. Urea . + III. ACIDS. Boric acid . + Hydrochloric acid Nitric acid . + Slight yellowish cast-no ppt. Sulphuric acid . + Phosphoric acid . + Acetic acid . + Butyric acid . + Benzoic acid . + Citric acid . + Formic acid . + Lactic acid . + Oxalic acid . + Salicylic acid . + Test-Seliwanoff: on levulose-0.1 per cent, solution. 1 This inhibition is due to the formation of nitrous acid. Resorcin 4- KNO2 = water colored solution. HC1 4- KNO, = a yellow liquid, due to presence of nitrous acid. HC1 4- KNO, + levulose = a yellow liquid, due to presence of nitrous acid. Resorcin 4- levulose = a water colored solu- tion. ' Resorcin 4- formaldehyde = clear solution. HC1 4- formaldehyde = clear solution. Seliwanoff's 4- formaldehyde = pink coloration - Seliwanoff reaction. Solution of resorcin added to HC1 4- formaldehyde while hot = no change, but if they are then boiled the Seliwanoff reaction appears. A solution of resorcin added to boiling H2SO4 (i : 2) 4- formaldehyde gave the same result as above with HC1. 37 TablE VI (Continued). Substance added. Result. Remarks. Tannic acid + Tannic acid 4- Seliwanoff's re- agent yields a white ppt. Tartaric acid . + IV. ACID SALTS. Copper sulphate .. o3 A yellow solution becoming red only after standing one hour. Dihydrogen sodium phosphate Ferric chloride . + On cooling there is a slight yellow- ish coloration due to Fe(OH)r Lead acetate . + Mercuric chloride . + Acid potassium oxalate . + Sodium acid tartrate . 4- V. ALKALIES. Ammonium hydroxide. . 4- Calcium hydroxide . + Sodium hydroxide . 4- VI. ALKALINE SALTS. Alum 4- Borax . 4- Disodium hydrogen phosphate 4- Potassium thiocyanate o4 Clear solution. Potassium ferrocyanide . 4- Sodium carbonate ...4- Sodium arsenate. . 4- Sodium sulphide . 4- Seliwanoff's reagent 4- sodium sulphide give a white ppt. Lead acetate (basic) . 4- Sodium acetate . 4- Sodium benzoate . 4- VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee . 4- Tea . 4- Cocoa : 4- Milk . 4- Urine . 4- Beef tea . + Meat extract . 4- Physiological saline . 4- Saliva. . + Liver extract . 4- Pancreas extract . 4- Blood . 4- Soap solution • + Test-Seliwanoff: on levulose-o.i per cent, solution. 3 CuSO4 4- HC1 - a green solution due, to formation of copper chloride. CuSO4 + HC1 4- heat = yellow solution. CuSO4 -I- resorcin = blue solu- tion. HC1 4- resorcin = a water colored solution. The inhibition of the Seliwanoff reaction seems to be due to the CuCl2. 4 KCNS 4- HC1 = a colorless solution. KCNS 4- resorcin = a water colored solution. KCNS 4- resorcin 4- levulose = a water colored solution. KCNS 4- resorcin 4- HC1 = a water colored solution. KCNS 4- Seli- wanoff's reagent 4- levulose - a water colored solution which turns red only after standing a long while. Table VII. (i) egg-albumin-(egg-white) sol. i : 100. (2) Witte's peptone-0.1 per cent. sol. (3) acid-albumin-approximately 1 : 50 of the wet metaprotein. (4) edestin-0.1 per cent. sol. (approxi- mately). (5) ovomucoid-less than 0.1 per cent, solution. (6) nucleoprotein (yeast)-0.1 per cent. sol. (approximately). Test-Biuret: reagent (5 drops) on Substance added. Result. Remarks. I. NEUTRAL INORGANIC SALTS. Ammonium sulphate . + Barium chloride . + White ppt. probably barium sul- phate. Calcium chloride . + White ppt. probably calcium sul- phate. Calcium phosphate . + White ppt. Magnesium chloride . + White ppt. probably magnesium sulphate. Magnesium sulphate . + White ppt. of Mg(OH)2. Manganese sulphate . o Vide supplementary table of biuret interferences. Potassium chloride . + Potassium iodide . + Potassium nitrite . + Sodium chloride . + Sodium nitrate . + Sodium sulphate . + Sodium sulphite . + Zinc sulphate . + Heavy, and sometimes gelatinous ppt. probably zinc hydroxide. II. NEUTRAL ORGANIC COMPOUNDS. Chloroform Ethyl alcohol . + Glycerol . + Formaldehyde . + Chloral hydrate . + Acetone . + Glucose . + Sucrose . + Egg albumin Toluol . + Thymol. . : . + Phenol . + Picric acid . O Vide supplementary table of biuret interferences. Urea . + III. ACIDS. Boric acid . + Hydrochloric acid . + Nitric acid . + Phosphoric acid . + Sulphuric acid . + Acetic acid . + Butyric acid . + Benzoic acid . + Citric acid . + Formic acid . + Lactic acid . + Oxalic acid . + Table VII {Continued). (i) egg-albumin-(egg-white) sol. i : 100. (2) Witte's peptone-0.1 per cent. sol. (3) acid-albumin-approximately 1 : 50 of the wet meta protein. (4) edestin-0.1 per cent. sol. (approxi- mately). (5) ovomucoid-less than 0.1 per cent, solution. (6) nucleoprotein (yeast)-0.1 percent, sol. (approximately). Test-Biuret: reagent (5 drops) on Substance added. Result. Remarks. IV. VI. Salicylic acid Tannic acid Tartaric acid ACID SALTS. Copper sulphate Dihydrogen sodium phosphate Ferric chloride Lead acetate Mercuric chloride Acid potassium oxalate Sodium acid tartrate ALKALIES. Ammonium hydroxide....... Calcium hydroxide Sodium hydroxide ALKALINE SALTS. Alum Borax Disodium hydrogen phosphate Potassium thiocyanate Potassium ferrocyanide Sodium carbonate Sodium arsenate Sodium sulphide Lead acetate (basic) Sodium acetate Sodium benzoate + 0 . + 0 . + . 0 . + . 0 . + . 4- . + . + . + . 4- . 4- + . 4- 0 . + . + . + . 4- . 4- Vide supplementary table of biuret interferences. Vide supplementary table of biuret interferences. Vide supplementary table of biuret interferences. Heavy white ppt. of lead sul- phate. Vide supplementary table of biuret interferences. Vide supplementary table of biuret interferences. Heavy white ppt. of lead sul- phate. VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee o Vide supplementary table of biuret interferences. Tea o Vide supplementary table of biuret interferences. Cocoa + Milk + Urine o Vide supplementary table of biuret interferences. Beef tea 4- Meat extract Physiological saline 4- Saliva + Liver extract 4- Pancreas extract 4- Blood 4- Soap solution 4- Table VIII.-Supplementary Table of Interferences with the Biuret Reaction. Egg Witte's Acid Substance added. albumin. peptone. albumin. Edestin. Ovomucoid. N ucleoprotein. CuSO/ o O o o o Blue- Blue and Bluish Blue and Blue and Bluish white dark ppt. PPt- dark ppt. black black ppt. PPt- PPt- MnSO/ o o o o o o Brown- Browni s h Browni s h Yellow to Yellow- Heavy yellow PPt- PPt- brown brown brown Ppt- PPt. PPt- PPt- Picric acid3 o o o o o o Clear sol. Clear sol. Clear yel- Clear yel- Clear sol. Clear sol. yellow yellow low sol. low sol. yellow yellow Tannic acid4.... o o o o o o Brownish Brownish Brownish Red and Red and Slight solution solution solution yellow yellow dark ppt. sol. sol. HeCl3 o o o o o o Yellow Yellow Yellow Yellow Yellow Yellow ppt- Ppt- PPt- Ppt- PPt- Ppt- Fe2Cl(>• o o o o o o Reddish Reddish Reddish Reddish Reddish R e d di s h Ppt- PPt- Ppt- PPt- PPt. PPt- Tea and coffee'.. o o o o o o Brownish Brownish Brownish Brownish Brownish Brownish solution solution solution solution solution solution K4Fe(CN)68 o o o o o o Clear yel- Clear yel- Clear yel- Clear yel- Clear yel- Clear yel- low sol. low sol. low sol. low sol. low sol. low sol. o + o + + Pale No ppt. No ppt. biuret Brown Brown sol. sol. Note.-The o indicates a negative result of the test. 1 CuSO« + proteins = a bluish white ppt. of copper albuminate, copper edestinate, etc., each of which fails to give the biuret reaction. 2 This inhibition is due to the union of KOH and MnSO4. A whitish ppt. is formed in large amounts. This is Mn(OH)a. This oxidizes rapidly to a brownish color. 3 Picric acid + proteins = a yellowish ppt. which is an insoluble salt that does not give the biuret reaction. This salt dissolves on adding just a little of the biuret reagent, leaving a clear yellow solution. 4 Tannic acid + proteins = a ppt. which is an insoluble salt that fails to give the biuret reaction. This salt usually dissolves on adding the biuret re- agent, leaving a solution which is usually yellow with a narrow zone of brown- ish or reddish liquid above it. B Mercuric chloride + proteins = a white ppt. which is an albuminate. This fails to give the biuret reaction. Mercuric chloride + biuret reagent = a yellow ppt., mercuric hydrate which further precludes the formation of biuret. 6 Ferric chloride + proteins = a brownish ppt. which is an albuminate. This fails to give the biuret reaction. Ferric chloride + biuret reagent = a reddish ppt., ferric hydroxide which further precludes the formation of biuret. ' Coffee and tea inhibit the biuret reaction because of the tannic acid present. Unless used in concentrated solution they do not produce the sharply defined effects that tannic acid does. 8 This inhibition is due to the formation of cupric ferrocyanide, a brownish red ppt. which forms in large amounts with very dilute CuSO4 and one drop of potassium ferrocyanide. In the biuret test this ppt. dissolves, leaving a yellowish solution. Occasionally a small amount of biuret may form. Table IX. Test-Millon: reagent (5 drops) added to 5 cc. of the typical proteins indicated. Result. 3 1; a 33 o « >> sol. 250 £ b* 6 a ci au a° a k .9.2 be ao a 2 0) a o a_. a a aS 0 s aS aj o a TJ 0 6 o & a*o O.i 3 g U a •• bl O Witte's p cent, so T* d'O .Sa <n ■S® vomu< mately sol. o g " 3 « s a bl 3 d g- 3 Substance added. < w O I. NEUTRAL INORGANIC SALTS. Ammonium sulphate . . . + + 3 + + 1 + 2 + Barium chloride • • + + 3a + + + + Calcium chloride . . + + 3 + + + + Calcium phosphate . . + + + + + + Magnesium chloride + 8 + + + + Magnesium sulphate .. + ?7 + + ?8 + ' o' o?19 Manganese sulphate .. + ?7 + + ?8 + ' +' o?19 Potassium chloride . . + 4 + 3 + + ?3 + ?3 + Potassium iodide5 . . O O O o o o Potassium nitrite .. + + + + + + Sodium chloride .. + + ' + 11 + ?3 + ?3 + Sodium nitrate ... + + + + + + Sodium sulphate .. +?7 + + ?3 + ' o?13 o?13 Sodium sulphite .. +?7 + 12 + ?8 + ' o?13 o?13 Zinc sulphate .. +?7 + + ?3 + ' + 14 o15 II. NEUTRAL ORGANIC COMPOUNDS. Chloroform .. + + + + + + Ethyl alcohol .. + + + + + + Glycerol ... + + + + + + Formaldehyde ... + + + + + + Chloral hydrate ... + + + + + + Acetone ... + + + + + + Glucose ... + + + + + + Sucrose ... + + + + + + Toluol Thymol .. + + + + + + Phenol Picric acid18 . . . O o o o o o 1 Millon reaction rather weak. 2 Yellow ppt. 3 No red coloration unless double the amount of Millon reagent used. 3a When barium hydrate is substituted for the chloride there is no inter- ference. 4 The red color appears only on standing. 5 Total inhibition owing to a reddish mercuric iodide pptd. in large amounts. 8 Clear yellow liquid unless double the amount of Millon reagent is used. 7 Red color comes only on doubling the amount of Millon's and after standing i hour. 8 Yellow ppt. No red coloration unless amount of Millon's used is doubled. 9 Yellow ppt., in some cases heavy. 10 A yellow ppt. usually inhibits the test. 11 Yellowish tinge to the liquid. 13 A yellow ppt. (heavy with nucleoprotein) usually inhibits the test. 12 Yellow sulphite tends to mask the reaction. 14 Yellow ppt. Red color appears slowly if at all. 15 Heavy yellow ppt. and no Millon reaction even with double the usual amount of Millon reagent. 18 With all the proteins a yellowish or brown ppt. was formed, except in the case of ovomucoid which yielded an amber colored solution and no ppt. after the Millon's was added. Table IX (Continued). Test-Millon: reagent (5 drops) added to 5 cc. of the typical proteins indicated. Resul t. 3 o ft If T sol. 250 * U) d d P. ci 8 . K'S O co 0 o Q <U a0. .9 « o >> is o ft .S I6 a v ci <j ucoid a ;ely 0.2 QU O 2. " S H ft o .8^ 2 w aj c . & cj-: M ° « Cl S w • w Q £ ■2^ •q W w 15 a «« o a o > w w Substance added w W O Urea .. o17 o" o17 + ?18 o?1' o17 III. ACIDS. Boric acid .. + + + + + + Hydrochloric acid .. o?20 o?20 o?20 + ?21 O22 O?23 Nitric acid ' Sulphuric acid .. + + + + + + Phosphoric acid .. + + + + + + Acetic acid .. + + + + + + Butyric acid .. + + + + + + Benzoic acid .. + + + + + + Citric acid .. + + + + + + Formic acid . . O24 + ?25 + p26 27 l 28 1 28 Lactic acid .. + + + + + 4- Oxalic acid .. + ?3" + 31 -p3l + + + Salicylic acid .. + + + + + + Tannic acid32 . . o o o o O o Tartaric acid .. + + + + + + IV. ACID SALTS. Copper sulphate . . . . | 33 + 33 _p 33 + + + Dihydrogen sodium phosphate. . . .. + + + + + + Ferric chloride .. +?34 + ?34 O35 + ?3« O?37 -1- ?3« 17 Whitish to yellowish ppts. were formed. The test is hindered by the formation of nitrates of urea and mercury. 18 The red coloration appears only after using double the usual amount of Millon's. The reaction is very slow. 19 Considerable froth (CO2) accompanies the white ppt. Sometimes the red color appears after an interval of about ten minutes. 20 Total inhibition and a clear yellow solution, even with 25 drops (five times the usual amount) of Millon reagent providing heat is applied. If no heat is applied a faint red eventually appears. 21 Light yellow ppt. No red color unless the amount of Millon reagent is doubled. 22 Yellow solution only. 23 If heated, even with 25 drops of Millon reagent there is total inhibi- tion. If kept cool, 10 drops gives a reddish color in one hour. 24 Heavy gray ppt. of mercury or mercuric formate inhibits the test. 2S Red coloration fails to appear unless double the usual amount of Millon reagent is used. 28 The quantity of Millon reagent used must be doubled. 27 Small amount of gray ppt. 28 After standing half an hour the red color vanishes and a gray ppt. forms. 29 The red color does not appear immediately. A heavy gray ppt. is formed. 30 A heavy white ppt. of oxalate inhibits the test unless an excess of Millon reagent is used. 31 A heavy white ppt. of oxalates makes the reaction faint. 32 Total inhibition due to the formation of characteristic ppts. with the various proteins. These vary from yellow to brown. 33 Pale blue ppt. is formed. 34 Slight brown or yellowish ppt., and no red color unless double the usual amount of Millon reagent is used. 35 Clear yellow liquid and usually no ppt. 36 Red coloration appears only when a double quantity of Millon reagent is used and 900 of heat attained carefully. 37 Amber-red liquid and no ppt. Test-Millon: reagent (5 drops) added to 5 cc. of the typical proteins indicated. Table IX (Continued). Result. " 3 .X 2*85 * - m ° " . 8 a aS 2 -s"? g2 &u a ■• »o 2 >. 2 4J *5*85 01 2 a a u tn o s »> ■g a o S » •? " '3 « « Substance added. W > Lead acetate 4- 4- 4- Mercuric chloride + + + Acid potassium oxalate 4- 4- 4- Sodium acid tartrate + + 4- V. ALKALIES. Ammonium hydroxide38 + ? 4- ? 4- ? Calcium hydroxide + 4- 4- Sodium hydroxide38 + ? + ? + ? VI. ALKALINE SALTS. Alum + + + Borax 4- 4- + Disodium hydrogen phosphate 4- + 4- Potassium thiocyanate 4- + ?40 + ?41 Potassium ferrocyanide o42 o42 o43 Sodium carbonate48 4- ? 4- ? + ? Sodium arsenate + + 4- Sodium sulphide + 4- 4- Lead acetate (basic) 4- 4- 4- Sodium acetate 4- 4- 4- Sodium benzoate 4- 4- 4- VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee o32 4- o32 Tea o 4- o32 Cocoa 4- 4- 4- Milk 4- 4- 4- Urine 4- 4- 4- Beef tea 4- 4- 4- Meat extract 4- 4- 4- Physiological saline 4- 4- 4- Saliva 4- 4- 4- Liver extract 4- 4- 4* Pancreas extract Blood . 4- 4- 4- Soap solution 4- 4- 4- 4- + : 4-4-4-4-4-4-4-4-4-4- + + + + + + °£ + + + + + + + + + + + ceR^L^ . _ Ovomucoid ap pr oxi - 4- +: ++++++++++ + + + + + + FL + + + + 4-4-4- 4- 4- 4- 4- mately 0.2 per cent, sol. °IOS uwiojdoapnu isnaA + 4-4-4- + + + + + + +"0+ + + + + + *© "0 + + + + + + + + '• + + 38 Heavy white persistent ppt. inhibits the test unless an excess of Millon reagent used. 39 Heavy yellow ppt. (oxide of mercury) inhibits the color unless an excess of Millon reagent is used. 40 Red coloration only slight after standing one hour without heating. Light ppt. 41 White flocculent ppt. No red coloration unless the amount of Millon reagent is doubled. 42 Dark green ppt. which vanishes on heating; then a light yellow and a drab ppt. 43 Heavy dark blue ppt. inhibits the reaction. 44 Total inhibition. A dark blue and a yellow ppt. formed. 45 Bluish yellow and green ppt., finally changing to a drab ppt. in a clear solution. 46 Heavy brown ppt. (mercu ric carbonate) and no red coloration unless an excess of Millon reagent is used. 44 Table X.-Conspectus of Interferences with the Millon Reaction. Note.-Five drops of Millon reagent were used except where an asterisk accom- panies the + or o signs. The asterisk is used with these signs to indicate that the amount of Millon's used was from io drops (slight excess) to 25 drops. As usual, the + means reaction without definite interference; o, a negative result. Substance added. c § a M at W Witte's pep- tone. Acid a 1 b u- min. •0 6 Q. 8 g •ax a .5 (A C •d a 0 > 0 BaClj,1 CaCl2,' MgCl/... + + * + + + + NaCl and KC12 + + * + +* +* + urt ( heated HU 1 cold3 o* O* 0* +* 0* 0* . + *20-60 min. + * 20-60 + min. * 20-60 min. +* 0* + * 20-60 min. FeaCV KCNS5 + * + * 0* +* 0* + * + + 1 hr. + * + + + Na2SO3 and Na2SO4°. . . + * cold, 1 hr. + + * + 0 + * Color faint. MgSO/ . cold, +* 1 hr. + + * + 0 + ?* MnSO/ . cold, +* 1 hr. + + * + + ? + ?* ZnSO46 . cold, +* 1 hr. + + * + + 0* Potassium iodide7 0 0 0 0 0 0 Picric acid8 0 0 0 0 0 0 Formic acid9 0 + * + * + + + Tannic acid10 . 0 0 0 0 0 0 Oxalic acid11 + * + + + + + NaOH and Na2CO3 . These bases inhibit in all proteins unless excess Millon. Tea and coffee12 0 + ? 0 + 0 0 K4Fe(CN)613 0 0 0 0 0 0 Urea14 0 0 0 +* 0? 0 1 and 2 The reaction of the nitrous and nitric acids with these chlorides may be one of the reasons for the difficulty here. 3 The interference here may be due to the formation of chlorides of mercury which remain in solution. There is no ppt. here to interfere. The chloride would form more freely with heat, hence the inhibition is more pronounced in hot than in cold solutions as the table shows. 4 Inhibition probably due to formation of mercurous chloride and ferric nitrate. 6 Interference due to the fact that KCNS tends to ppt. mercuric thiocyanate. 8 The sulphates tend to ppt. the mercury. The sulphates and sulphites vary from white to yellow and then to red when heated. For further discussion of this point see the comments on the Millon reaction. 7 The mercury is pptd. in such large amounts that red fails to be seen. 8 The protein is pptd. 9 There is a tendency to ppt. the mercury here. 10 The protein is pptd. In addition mercurous tannate, a greenish compound produces further complications. 11 The red color is always slow in appearing when oxalates are present. These tend to ppt. the mercury. 12 The dark color and the tannic acid cause interference. 13 Inhibition of the test here is due to the formation of mercuric ferrocyanide which is bluish gray, and very insoluble. 14 Urea is split by nitrous acid and the abundant white ppt. of urea nitrate tends to inhibit the Millon test. Sometimes an excess of the Millon reagent will tend to overcome this, but even then the red color is very slow in appearing. Test-Hopkins-Cole: on the typical proteins indicated: Table XI. Result. a 1 Ui a I? * a o-o o W a 5 3 o X> u, * a " H C -. hl J 0 73 X o * u* Substance added. M £ < I. NEUTRAL INORGANIC SALTS. Ammonium sulphate + + + Barium chloride +1 + l + l Calcium chloride +1 + 1 + 1 Calcium phosphate +l + 1 + 1 Magnesium chloride + + + Magnesium sulphate + + + Manganese sulphate + + + Potassium chloride + + + Potassium iodide + 2 + 2 + 2 Potassium nitrite + ?3 + ?3 + ?' Sodium chloride + + + Sodium nitrate o4 o4 o4 Sodium sulphate + + + Sodium sulphite + + + Zinc sulphate + + + II. NEUTRAL ORGANIC COMPOUNDS. Chloroform + + + Ethyl alcohol +' + + Glycerol + + + Formaldehyde o5 o?5a o?5& Chloral hydrate + + + Acetone + + + Glucose + + + Sucrose +6 + ' +6 Egg albumin + + + Toluol + + + Thymol + + + Phenol + ?7 + ?7 + ?7 Picric acid + + + Urea + + + III. ACIDS. Boric acid + + + Hydrochloric acid + + Nitric acid +?8 + ?' + ?8 Sulphuric acid Phosphoric acid + + + Acetic acid + + + Butyric acid + + + Benzoic acid + + + Citric acid + + + Formic acid + + + 1 White ppt. 2 Brown ppt. 3 Yellowish ppt. Very faint violet coloration. No purple ring. 4 Slight pale yellowish ppt. 5 No purple ring. A slight white to yellowish ppt. Faintest trace of violet in the solution. 5a No purple ring. Yellow zone of liquid. Faintest trace of violet in liquid. &b White ppt. Brown ring. Only slight trace of violet in the liquid below. 6 Sucrose causes discoloration and obscures the Hopkins-Cole test if the percentage is at all high. 7 Slight whitish ppt. No ring. Very faint violet coloration. This inter- ference is due to the formation of orthophenolsulphonic acid. 8 Yellowish ppt. No ring. Faint violet coloration. 9 Very faint violet coloration. No ring. Yellow liquid above. The interference of the nitric acid may be due to its reaction with the sodium of the reagent. Table XI (Continued). Test-Hopkins-Cole: on the typical proteins indicated: Result. albumin' per cent. e's p e p - le 0.1 per it. sol. albumin per cent. 3 ® 8 tj d s 2 g Substance added. W £ Lactic acid . + + + Oxalic acid . + + + Salicylic acid * . + + Tannic acid + ?10 + ?12 + ?11 Tartaric acid . + + + IV. ACID SALTS. Copper sulphate . + + + Dihydrogen sodium phosphate . + + + Ferric chloride . o?13 o14? o?13 Lead acetate . +15 + 16 + 15 Mercuric chloride.. . ; . +18 +16 + 18 Acid potassium oxalate . + + + Sodium acid tartrate V. ALKALIES. . + -f- + Ammonium hydroxide . + + + Calcium hydroxide . + + + Sodium hydroxide . + + + VI. ALKALINE SALTS. Alum . + + + Borax . + + + Disodium hydrogen phosphate . + + + Potassium thiocyanate . + 17 +17 + 17 Potassium ferrocyanide . + 18 + 18 + 18 Sodium carbonate . + + + Sodium arsenate . + + + Sodium sulphide . + 19 + 19 +19 Lead acetate (basic) | 20 | 20 | 20 Sodium acetate . + + + Sodium benzoate . + + + VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee . + + + Tea . + + + Cocoa . + + + Milk . + + + Urine . + + + Beef tea . + + + Meat extract . + + + Physiological saline . + + + Saliva . + + + Liver extract . + + + Pancreas extract . + + + Blood . + + + Soap solution . + + + 10 White ppt. violet color faint. 11 Dark green ring surmounted by a violet ring. White ppt. 12 White ppt. Violet color faint. 13 Slight yellow ppt. The faintest trace of violet coloration in the liquid below. No ring. 14 Clear yellow solution. No ring. Very faint trace of violet in the sub- natant liquid. The interference of the iron is probably due to the formation of ferrous oxalate which is a yellowish compound. 15 White ppt. formed. 19 Slight white ppt. formed. 17 The violet color is very faint. 18 A white and blue ppt. No sharp ring, but a violet zone below. This color fades on standing. 19 A white ppt. Violet color often faint. 20 White ppt. 48 Test-Xanthoproteic: on the typical proteins indicated; Table XII. Result. C o O tn 8 a . a a $ . a a 23 a Substance added. Egg albumin (1 egg-w h i t e 1 : 100). Witte's p e p t 0.1 per cent. - o *® 73 U •a ° Edestin 1 per solution. Ovomucoid 1 cent, solutio o u •aS aa§ w o s-H I. NEUTRAL INORGANIC SALTS. Ammonium sulphate ... + + + + + + Barium chloride . . . + + + + + + Calcium chloride . . . + + + + + + Calcium phosphate . . . + + + + + + Magnesium chloride . . + + + + + + Magnesium sulphate ... + + + + + + Manganese sulphate ... + + + + + + Potassium chloride .. . + + + + + + Potassium iodide . . . +1 + l + 1 + 1 + 1 + 1 Potassium nitrite . . . + + + + + + Sodium chloride . . . + + + + + + Sodium nitrate . . . + + + + + + Sodium sulphate .. . + + + + + + Sodium sulphite ... + + + + + + Zinc sulphate ... + + + + + + II. NEUTRAL ORGANIC COMPOUNDS. Chloroform ... + + + + + + Ethyl alcohol . . + + + + + + Glycerol . . + + + + + + Formaldehyde . . + + + + + + Chloral hydrate . . + + + + + + Acetone . . + + + + + + Glucose . . + + + + + + Sucrose . . + + + + + + Egg albumin + + + + + Toluol . . + '+ + + + + Thymol . . + + + + + + Phenol . . +2 + 2 + 2 + 2 + 2 + 2 Picric acid . . . + + + + + + Urea .. +2 + 2 + 2 + 2 + 2 +2 III. ACIDS. Boric acid ... . . + + + + + + Hydrochloric acid . . + + + + + + Sulphuric acid . . + + + + + + Phosphoric acid . . + + + + + + Acetic acid . . + + + + + + Butyric acid . . + + + + + + Benzoic acid . . . . + + + + + + Citric acid . . + + + + + + Formic acid . . + + + + + + Lactic acid . . + + + + + + Oxalic acid . . + + + + + + Salicylic acid . . + + + + + + Tannic acid . . +2 + 2 + 2 + 2 + 2 + 2 Tartaric acid . . + + + + + 1 A slight tendency toward interference is readily overcome by a good excess of the acid and the base. 2 A slight tendency to interfere overcome by an excess of the acid and base. 49 TabliS XII {Continued). Result. 43^-3 w o £ o'® o O a Q 1 per- >n. £3 . § - o' 1^2 a o II •fi w 2* V o . fl o S3 '5 fl S3 O'© o w 1 fl •q & 5Qd -.2 ■p fl fl Substance added. w •tid £ o w O 3 « ® IV. ACID SALTS. Copper sulphate .. +?3 o4 o4 o4 o4 o4 Dihydrogen sodium phosphate. .. .. + + + + + + Ferric chloride o® o5 o® o® o® Lead acetate .. + + + + + + Mercuric chloride .. +8 +8 +8 +8 +8 +8 Acid potassium oxalate .. + + + + + + Sodium acid tartrate .. + + + + + + V. ALKALIES. Calcium hydroxide .. + + + + + + VI. ALKALINE SALTS. Alum .. + + + + + + Borax ■ .. + + + + + + Disodium hydrogen phosphate. . . .. + + + + + + Potassium thiocyanate .. + + +7 +7 +7 +7 Potassium ferrocyanide .. + + +7 +7 +7 +7 Sodium carbonate .. + + + + + + Sodium arsenate .. + + + + + + Sodium sulphide .. + + + + + + Lead acetate (basic) .. + + + + + + Sodium acetate .. + + + + + + Sodium benzoate .. + + + + + + VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee .. + + + + + + Tea .. + + + + + + Cocoa .. + + + + + + Milk .. + + + + + + Urine8 .. + + + + + + Beef tea .. + + + + + + Meat extract .. + + + + + + Physiological saline .. + + + + + + Saliva .. + + + + + + Liver extract .. + + + + + + Pancreas extract .. + + + + + + Blood .. + + + + + + Soap solution .. + + + + + + Test-Xanthoproteic: on the typical proteins indicated. 3 With an excess of the acid and base only a trace of orange color. 4 Total inhibition of the test because of the formation of copper hydroxide, a bluish ppt. which dissolves in an excess of NaOH, leaving an almost clear solution. Usually there is no orange ppt. or coloration at all. 6 Total inhibition due to the formation of Fe(OH)3 which is itself orange and is freely formed when a strong base is added. 6 Slight white ppt. Tendency to interfere overcome by excess of acid and base. 7 Slight tendency to interfere overcome by a liberal excess of the acid and base. 8 Urines in the small percentages used, 5 drops in 2-5 cc. of protein solution cause no interference with the xanthoproteic test. 50 VII. Supplementary Studies of the Millon Test. In the compilation of the tables of Millon interferences the greatest difficulty was experienced in attempting to verify and confirm a large proportion of the observations. Many experiments were repeated from six to twelve times with vary- ing results. This merely bears out the common experience as to the rather unsatisfactory character of the Millon reac- tion. In those places in the table where a question mark follows the + or o no definite conclusion could be positively stated for the reason that with the same substances both posi- tive and negative results were observed. The effects of chlor- ides and sulphates were peculiarly difficult to establish. E. g., there was often no interference so long as the liquids were not heated, but heat however carefully applied frequently caused the red color to vanish. Sometimes there were two distinct precipitates, one protein and the other non-protein, and no clue appeared to explain interferences. In view of these things, it was decided to carry on some further experiments to determine if possible the influence of tempera- ture, amount of Millon reagent used, neutralizing agents, and the interrelations among these. I. Two sets of test tubes were prepared as follows and heated respectively to 400 and 500 C. in a water bath: Tubes 1 and 7 = egg albumin and HC1. Tubes 2 and 8 = egg albumin and urea. Tubes 3 and 9 = edestin and HC1. Tubes 4 and 10 = edestin and urea. Tubes 5 and 11= peptone and HC1. Tubes 6 and 12 = peptone and urea. Tubes 13 and 14 = acid albumin + HC1. Tubes 15 and 16 = acid albumin + urea. To each of these tubes the minimum amount of Millon reagent used in the regular tests was added, viz., 5 drops. The results were as follows: 40° tubes. 50° tubes. i = good reaction. 2 = faint reaction. 3 = good reaction. 4 = no reaction. 7 = good reaction. 8 = no reaction. 9 = no reaction. io = no reaction. 51 5 = no reaction. 6 = no reaction. 13 = no reaction. 15 = no reaction. 40° tubes. 11 = no reaction. 12 = faint tinge of red. 14 = no reaction. 16 = no reaction. 50° tubes. Conclusion.-No general conclusion is possible except that the reaction varies with temperature. The data show merely individual facts-e. g., that taking urea as an interfering factor with peptones the higher temperature was favorable. Both sets of tubes were treated after being prepared a sec- ond time with 10 drops of Millon's, with the following results: 1 = negative. 2 = good reaction. 3 = negative. 4 = good reaction. 5 = negative. 6 = slight reaction. 40° tubes. 7 = negative. 8 = faint reaction. 9 = negative. io = slight red coloration. ii = negative. 12 = very slight reaction. 50° tubes. Conclusion.-With HC1 as an interfering factor the Millon reaction is inhibited at both temperatures-400 and 500 C. Urea seemed to have less of an inhibitory effect at the lower temperature. II. Influence of heat where sulphates are present Two sets of tubes were prepared as follows: Five cc. of egg albumin, Witte's peptone, acid albumin, and edestin were put into test tubes. To one set were added in each case 5 drops of Na2SO4 and ZnSO4, respectively. Five drops of Millon reagent were used in every tube. One set of tubes was heated to 400 and the other to 500 C. The results were as follows at the end of 30 minutes: 40° tubes. Reaction. 50° tubes. Reaction. Egg albumin + Na2SO4 good good Egg albumin + ZnSO4 good good Edestin + Na2SO4 faint good Edestin + ZnSO4 faint good Peptone + Na2SO4 good ■very good Peptone + ZnSO4 very good exceptionally good Acid albumin + Na2SO4 fair good Acid albumin + ZnSO4 fair good 52 All those reactions underscored were better than the corre- sponding ones at 40°. Conclusion.-500 C. is more favorable to the Millon reac- tion where sulphates are present than the lower temperatures. The general table of Millon interferences show certain excep- tions to this generalization. This experiment confirms the previous ones in demonstrating that the Millon reaction varies decidedly with even moderate variations in temperature. III. Relation of temperature to the Millon reaction when formic acid is present as an interfering factor. Peptone + formic acid (5 drops) + 10 drops (slight excess) Millon's put into each of three tubes; one kept cool, one kept at 400 and one at 500 C. All these showed a reaction in 10 minutes but the tube at 500 had by far the best color. Conclusion.-With formic acid present, a temperature of 500 is far better for the Millon test than a lower temperature. IV. Relation of temperature to the Millon reaction when no interfering factors are present. Six tubes were prepared, each containing 5 cc. of peptone (0.2%) + 10 drops of Millon reagent. Number 1 was kept cool; 2 was heated to 40°; 3 to 500; 4 to 750; 5 to 8o°; and 6 to 920 C. Results.-All the tubes showed a Millon reaction with an intensity directly proportional to the degree of heat up to 92 °. A temperature of 96° to boiling was applied to many tubes previously exhibiting good Millon reactions, with the result that decolorization often occurred. Conclusions.-1. 900 C. is the optimum temperature for the Millon reaction when no interfering factors are present. This is also true if formic acid is present, and probably true for most other interfering factors. 2. Temperatures of 95° to ioo° C. very often cause decolorization of the Millon reaction. 3. Boil- ing is not so likely to completely destroy the Millon red if a heat of 500 to 900 has been used to produce the Millon reaction. The influence of varying temperatures on the Millon reac- tion was studied in the same way in connection with acid albu- min, edestin, and nucleoprotein. The results on these confirmed the three conclusions just formulated. V. Further studies on the amount of Millon reagent neces- sary to produce a good reaction. 1. Egg Albumin.-Experiments with the sulphite, sulphates, 53 and urea were repeated but io drops of Millon reagent were used with the 5 cc. of albumin instead of 5 drops. Conclusion.-In cold solutions this slight excess of Millon reagent is usually effective in from 1 to 1% hours. In boiled solutions even this slight excess is ineffective except in the cases of MgSO4 and MnSO4. 2. Edestin.-Experiments similar to the above proved that usually 10 drops will give a reaction where 5 drops fail to do so. Similar results were noted in connection with NaCl, KC1 and HC1 as interfering factors. In the case of urea the test was in- hibited where the amount of Millon reagent was under 20 drops (good excess). In this case the reaction appeared after the lapse of 30 to 45 minutes. 3. Ovomucoid.-Experiments with the sulphite, sulphates, chlorides and HC1 were repeated, using 10 and 15 drops of Millon reagent. With the 10 drops no inhibition occurred where chlorides were present, but with the other compounds even this amount of Millon reagent failed to give a reaction. But 5 cc. of ovomucoid + 5 drops HC1 + 10 drops of Millon reagent at 400 C. gave a good Millon reaction at the end of 10 minutes. 4. Acid Albumin.-Experiments with the sulphates, chlor- ides, HC1 and urea were repeated, using 10 drops or more of Millon reagent. With this extra amount there was no inhibition except in the case of HC1 and urea. But if the solution contain- ing the HC1 was allowed to stand without being heated a good reaction occurred in about an hour. Tests made with 10, 15, 20 and 25 drops of Millon reagent, respectively, showed that HC1 inhibits the Millon reaction more effectively when boiled, but urea inhibits it best when left cold, or at a temperature below 500 C. 5. Witte's Peptone.-The experiments noted above were re- peated on Witte's peptone with approximately similar results. 6. Nucleoprotein.-With slight variations (noted in the general table of Millon reactions) the results noted above were confirmed on the yeast nucleoprotein. Conclusion.-1. In spite of minor exceptions and complica- tions it is to be noted that often where chlorides or sul- phates cause inhibition, the Millon reaction "occurs when a slight excess of the reagent is used. 2. The amount of heat 54 and the manner of its application are active factors in influ- encing the reaction as well as the amount of the reagent, espe- cially in certain cases of interference. 7. Witte's Peptone.-Another test was made by introducing formic acid as an interfering factor, using 5, 10, and 15 drops of Millon reagent in the usual tests. Even at varying temper- atures 10 drops (slight excess) was found to give the best red coloration. VI. Relation of the Millon reaction to various neutralizing agents when a base inhibits the reaction. Tubes were prepared as follows: 1. Witte's peptone + 5 drops Na2CO3 + 5 drops H2SO4 + Millon (5 drops). 2. Witte's peptone + 5 drops Na2CO3 + 5 drops HNO3 + Millon (5 drops). 3. Witte's peptone + 5 drops Na2CO3 + 5 drops acetic acid + Millon (5 drops). 4. Witte's peptone + 5 drops Na2CO3 + 5 drops Millon + Millon (5 drops). Results.-All gave the Millon reaction, but 4 was by far the best. Number 1 was next best. Conclusion.-An excess of Millon reagent is best for over- coming the otherwise inhibitory tendency of sodium carbonate. The above experiment was repeated, substituting NaOH for Na2CO3. Results.-A Millon reaction appeared only in the tube con- taining sulphuric acid. Conclusion.-Sulphuric acid is exceptionally effective in neutralizing the inhibitory effect of sodium hydroxide in the Millon reaction. Another set of tubes was prepared as follows: 5 drops of 5%. 5 drops. i. Edestin 4- Na2CO3 4- H2SO4 4- Millon = good reaction. 2. Edestin 4- Na2CO3 4- HNO3 4- Millon = fair reaction. 3. Edestin + Na2CO3 4- Acetic acid + Millon = no reaction. 4. Edestin + Na2CO3 4- Millon 4- Millon = good reaction. Conclusion.-An excess of Millon reagent and a very small amount of sulphuric acid are equally effective in overcoming the interference of sodium carbonate in the Millon test on edestin. 55 The above experiment was repeated, substituting NaOH for Na2CO3. Conclusion.-An excess of Millon reagent is the best for neutral- izing the otherwise inhibitory effect of NaOH on the Millon test on edestin. Similarly, sets of tubes were prepared for the purpose of studying these relations in egg albumin and in yeast nucleopro- tein. The conclusions reached as a result of all these experiments are shown below: Table of Relation of Millon Reaction to Neutralizing Agents. Note: + denotes that the reagent mentioned helps the Millon test; o denotes that the reagent mentioned fails to overcome a negative result with the Millon test; ! denotes a perfect Millon reaction. Interfering factor I. Na2CO3 2. NaOH i. Na2CO3 2. NaOH i. Na2CO3 2. NaOH i. Na2CO3 2. NaOH 5 drops of 5 per cent. sol. 5 drops of Millon. + o + + + + + + !■ Peptone Edestin Nucleoprotein Egg albumin Io - Leo + + + + + + + + HNO3 Acetic ac. (very weak). + + o o o o + O o o o o + o o o Conclusions.-From the above tables and experiments the following general conclusions covering the various types of pro- teins may be deduced: i. An organic acid like acetic is relatively useless in over- coming the inhibitory effect of a base on the Millon reaction. 2. Five drops of 5 per cent, sulphuric acid was useful in every instance in overcoming such inhibitory effects. 3. An excess of Millon's was effective in every instance but one (peptone with NaOH) in overcoming the inhibition due to bases. 4. Nitric acid of strength similar to that in the case of sul- phuric is comparatively useless in this connection. 5. With peptone + sodium hydroxide it is extremely easy to get the Millon reaction if sulphuric acid is present; otherwise impossible, even with 5 drops excess of Millon reagent. VII. Filtration in relation to the Millon reaction. The con- tents of various test-tubes used in the above experiments were 56 filtered to determine whether the Millon coloration appeared in the filtrates. Conclusion.-In the case of the tubes containing peptone a good Millon reaction appears in the filtrate. In the case of the other less soluble proteins none of the red compound goes through the filter. VIII. Comment on the Tables of Interference. The various interfering factors detected in this test were assembled and applied to all the carbohydrates studied in such a way as to admit of synchronous observations. In this way certain discrepancies were reconciled and errors which had crept in were eliminated. In point of time the first set of ob- servations was compiled about six months previous to the date of the second set. Furthermore, wherever possible, new lots of both carbohydrates and interfering substances were used, as well as new ingredients for the Molisch test. The interference of nitrites, nitrates, and nitric acid in this test is so strong that one drop of any of them caused a total inhibition. In addition to these, although there were many substances which formed heavy precipitates only two showed distinct properties of interference. These were ferric chloride and formaldehyde. For comparison with the latter acetaldehyde and benzaldehyde were substituted for it but they failed to in- hibit the violet coloration. The nature of the formaldehyde interference is not clear. COMMENT ON THE RESULTS WITH THE MOLISCH TEST. COMMENT ON THE RESULTS WITH THE IODINE TEST. All of the hydroxides form with iodine both soluble iodates and iodides in direct proportion to their dissociability. It is for this reason that for every drop of hydroxide added, from 2 to 50 drops of iodine solution must be added in order to pro- vide an excess of iodine available for combining with the starch. Many other compounds form precipitates with iodine, e. g., lead, but either because of their being white in color, or because these precipitates are not formed in large amounts, they fail to interfere. Some of the neutral organic compounds like thymol form with 57 iodine a compound which attracts iodine from mixtures of these substances plus starch plus iodine faster than the starch can attract iodine from the same mixture. Hence there is a pale- ness in the coloration or even inhibition at first. In the tables this fact does not appear for the reason that such an interfer- ence, being merely ephemeral, cannot be considered an inter- ference in the sense in which the term is commonly used and is used in this paper. Peptone, acid albumin, edestin and ovomu- coid decolorize the starch iodide mixture just as egg albumin does.1 In general, it may be said that the iodine reaction is subject to many disturbing factors because of the numerous affinities of this element. The successful application of the test then resolves itself into a question of making sure that these affinities are satisfied by using a generous excess of the reagent-in some cases 30 to 50 times the amount that would ordinarily be re- quired. This having been done, there remain only a few com- pounds that cause further disturbance. Mercuric chloride inhibits because of the heavy dark character of the mercuric iodide precipitate. Potassium ferrocyanide causes the blue color to disappear very rapidly whenever it is produced. The cause of this phenomenon is not clear. Potassium thiocyanate has a somewhat similar effect, though in a degree much less marked unless the amount of the interfering substance is in- creased. Saliva of course causes total inhibition by acting upon the starch. Finally soap solution, because of its basic property, has a strong interfering effect. Obviously the numerous disturbing compounds (other than those just mentioned) which are noted in the tables become much more serious when the percentage of such materials associated with the starch is increased. COMMENTS ON THE RESULTS WITH THE FEHLING-BENEDICT TEST. It will be noticed at once on referring to the table (No. 4) that only one neutral salt inhibits the Fehling-Benedict test, viz., ammonium sulphate. Many tests were made to determine the nature of the difficulty here. The following compounds were substituted in the usual way as interfering factors to re- place ammonium sulphate in the test: (NH4)2HPO4; (NHJ2- 1 Cf. Puchot: loc. cit. 58 CrO4; (NH4)2C2O4; (NH4)2CO3; NH4C1; NH4SCN; (NH4)2A12SO4; NH4C2H3O2; and (NH4)2C6H6O7. Every one of these nine com- pounds caused inhibition except the double salt with aluminium. In this instance there was initial inhibition, but reduction occurred on cooling. Then ammonium benzoate, ammonium magnesium phosphate and an excess of sodium citrate were tried and in all cases good reduction occurred. The inference is that the inhibition is due to the ammonium ion and depends upon the proportion and dissociability of this in the particular compound used. The inhibition seems to be due to the splitting of the ammonium salt by the sodium carbonate of the reagent, with the subsequent production of compounds of copper solu- ble in the resultant solutions. A few tests made seemed to indicate that the introduction of another sulphate with the ammonium sulphate tended to minimize or overcome the inhibition discussed above. Most of the acids used had no interfering effect because the percentage was too low. The Fehling-Benedict test is comparatively free from inter- ferences, but the following are effective inhibitory agents even in the small percentages used: ammonium sulphate, and other ammonium compounds, salts of heavy metals, tannic acid and potassium ferrocyanide. The mineral acids and the stronger organic acids merely retard reduction when present in these small amounts. COMMENT ON THE RESULTS WITH THE BARFOED TEST. Of all the tests included in the present investigation the Bar- foed test for monosaccharides proved to be the most uncertain and the most easily affected. The table shows that compounds from every one of the various groups cause trouble. Even the neutral organic compounds which rarely show any such influence have one representative in the class of inhibitory substances, viz., chloral hydrate. All of the neutral salts except nitrates and nitrites affect the test. Even the basic and acid sodium phosphates which usually are inert show strong inhibitory tendencies. The significance of all these facts is best seen by noting that of some 75 compounds and substances studied only about 20 show no interfering effect on the Barfoed test. 59 COMMENT ON THE RESULTS WITH SELIWANOFF TEST. This reaction for the detection of levulose is one of the easiest dealt with because of the fact that usually the coloration is so vivid that judgment upon it is easy and definite. The tabu- lated results show an extremely low coefficient of interference- only four out of 77 factors studied exhibiting inhibitory effects. These four are nitrites, copper sulphate, potassium thiocyanate and formaldehyde. The last-named interferes only in the sense that it will itself produce a red coloration with the Sali- wanoff reagent very similar to that resulting with levulose. Nitric acid in sufficient amounts will of course show a similar relation. Seliwanoff reagent then must be ranked as one of the most efficient from the standpoint of freedom from disturbing fac- tors. COMMENT ON THE RESULTS WITH THE BIURET TEST. The conclusions presented in the tabular summary (Table No. 8) were reached after repeated experimentation and review of doubtful points. The proteins selected were representa- tives of the main groups. All of the protein material was freshly prepared except the edestin and ovomucoid. The latter were made up from dry products of guaranteed quality. The test was applied after the well-known method of Gies,1 using the biuret reagent made up under his direction from the best materials obtainable. The results of these experiments showed a gratifying uni- formity, and the fact that less than ten active interfering fac- tors manifested themselves is a further proof of the great value of the biuret reagent. I found nothing in my results which one might not look for on a priori grounds except possibly the influence of manganese sulphate in producing a precipitate in amounts sufficient to cause trouble. The proteins used in connection with this test were the same as those mentioned in the comments on the biuret tests. How- ever, in many experiments I resorted to solutions of dry prepara- tions of these proteins in order to expedite the work. This COMMENT ON THE RESULT WITH THE MILLON TEST. 1 Gies: Jour. Biol Chem., VII., p. 60, 1910. 60 was on account of the necessity of repeating experiments so often. The results of the studies on the Millon test are in direct contrast to those on the biuret in several respects-in the number of interfering factors disclosed, in the non-uniformity of results on the various proteins, in the difficulty of forming definite conclusions and even specific judgments upon colora- tion, and in the amount of evidence accumulated to show that Millon reagent is, to state the case mildly, rather unreliable. Some 25 interfering factors as shown in the table (No. 10) were found. Since some of the sulphates and sulphites produce when combined with mercury, compounds that may take on a reddish tinge there is added difficulty in forming judgments about the relation of such to the Millon test. In the supplementary studies where reference is made to the sulphates the conclusions based on the appearance of a red coloration may be misleading if used in any wide generalization. They are stated for what they are worth-merely as deductions based on limited experimentation. The only broad conclusion pos- sible in the premises is that sulphates introduce a factor into the Millon test which is very uncertain. COMMENT ON THE RESULTS WITH THE HOPKINS-COLE TEST. The work with this test demonstrated the existence of quite a number of partial interferences, i. e., interferences which are absolute when the percentage of the interfering factor is raised. KNO2, NaNO3, formaldehyde, HNO3, tannic acid, Fe2Cl6, all affect the test in an unfavorable manner. In the case of sodium or potassium nitrate a very slight in- crease in the amount of these present causes total inhibition of the violet coloration. Usually the amount of nitrate used, 5 drops in 2 cc. of protein solution plus 3 cc. of Hopkins-Cole re- agent plus about 5 cc. of sulphuric acid, was sufficient to cause inhibition. The same is true of ferric chloride. Formaldehyde showed a strong tendency to interfere but there were often traces of violet coloration without the formation of a definite ring. The same is true of nitric acid. On the whole, this test must be ranked as one of the most reliable that we have for proteins. 61 COMMENT ON THE RESULTS WITH THE XANTHOPROTEIC TEST. This test is one which lends itself readily to such a study as the present one. The orange coloration is so very definite that even in the presence of all sorts of extraneous substances there is never any doubt as the facts in regard to whether a substance does or does not interfere. The proteins used were the same as those used in the Millon tests. Only two out of 75 substances investigated showed strong inference. These are copper sulphate and ferric chloride. Many other substances, tannic acid, potassium thiocyanate, potassium ferrocyanide, potassium iodide, mercuric chloride, phenol, urea, urine, tea and coffee, showed a tendency to in- hibit the test but in every case this was readily overcome by a liberal excess of the acid and base. Urines in small amounts, 5 drops in 2-5 cc. of the dissolved proteins, are ineffective as interfering factors in this test. This test therefore has a very low coefficient of interference. COMMENT ON THE RESULTS WITH THE SUDAN III TEST. This test was applied to olive oil, lard, and butter in the manner described previously in this work. Negative results (i. e., a total lack of interference), were obtained with every interfering substance used. For this reason no tables of the Sudan III tests are included with the tables of the other nine tests. In view of the lack of alcohol-soluble materials among the interfering substances used in this work the results are quite in accord with what one would expect. 62 Table XIII. IX. A General Summary of Results Deduced from Tables I tO 12. (Note.-As usual the zero, o, means that the result was negative, i. e., that there was interference.) Blanks indicate normal reactions. Substances added. .d Q cn o 3 al o t>«U - o a 43 u vm & 13 o s m Seliwanoff. i PQ a 5 g s m a 44 a o oo M Xantho- proteic. a a a co I. NEUTRAL INORGANIC SALTS. Ammonium sulphate - - o o - - - - - - Barium chloride - - - o - - ? - - Calcium chloride - - - o - - - - Calcium phosphate - - - o - - - - - - Magnesium chloride - - - o - - ? - - - Magnesium sulphate - - - o - - o - - - Manganese sulphate - - - O - o O - - - Potassium chloride - - - o - - ? - - Potassium iodide - - - o - - o - - - Potassium nitrite o - - - o - - - - - Sodium chloride - - - O - - ? - - - Sodium nitrate o - - - - - -. O - Sodium sulphate - - - o - - o - - Sodium sulphite - - - o - - o - - - Zinc sulphate - - - o - - o - - - II. NEUTRAL ORGANIC COMPOUNDS. Chloroform - - - - - - - - - Ethyl alcohol - - - - - - - - - - Glycerol - - - - - - - - -. - Formaldehyde o - - - o - o o - Chloral hydrate - - - o - - - - - -. Acetone - - - - -. - - - - - Glucose - - - - - - - - - - Sucrose - - - - - - - - - - Egg albumin Toluol Thymol - - - - - - - - - - Phenol - - - - - - - - - - Picric acid - - - - - o o - - - Urea - - - - - - o - - - III. ACIDS. Boric acid - - - O - - - - - - Hydrochloric acid - - - o - - O - - - Nitric acid o - - o - - - - - - Phosphoric acid - - - O - - - - - - Sulphuric acid - - - o - - - - - - Acetic acid - - - O -. - - - - - Butyric acid - - - o - - - - - - Benzoic acid Citric acid O - Formic acid - - - o - - o - - - 63 IX. A General Summary of Results Deduced from Tables I tO 12. Table XIII (Continued). (Note-As usual the zero, o, means that the result was negative, i. e., that there was interference.) Blanks indicate normal reactions. Substances added. Lactic acid / Oxalic acid Salicylic acid Tannic acid Tartaric acid "6 Wo c 8 2 •U o® ® ft - O *0 0 PQ O O o o ia o c d g to 3 o 1 O 1 O 1 Millon. W a 0.0 oU W 0 . •a .3 a o " a a re 13 a to IV. ACID SALTS. Copper sulphate - - - O o o - - - Dihydrogen sodium phosphate. .. O Ferric chloride . . O - O o - o o o o - Lead acetate - - - - - - - - - - Mercuric chloride - O o o - o - - - - Acid potassium oxalate - - - o - - - - - - Sodium acid tartrate - - - - - - - - - - V. ALKALIES. Ammonium hydroxide - - o? O - Calcium hydroxide - - - - - - - - - - Sodium hydroxide - - O - VII. ALKALINE SALTS. Alum - - _ o - - - - - - Borax - - - O? - - - - - Disodium hydrogen phosphate. . . o - Potassium thiocyanate - O? - O o - ? - - - Potassium ferrocyanide - o? o O - o o - - - Sodium carbonate - - - - - - - - Sodium arsenate - _ - O - - - - - Sodium sulphide - - - o - - - - - - Lead acetate (basic) - - - - - - - - - - Sodium acetate - - - - - - - Sodium benzoate . . - - - O? - - - - - - VII. BIOLOGICAL MIXTURES AND MISCELLANEOUS. Coffee - - o o o o Tea - - o o - o o - - - Cocoa - - - o - - - - - - Milk - - - o - - - - - - Urine - - - - - o - - - Beef tea - - - - - - - Meat extract - - - - - - - Physiological saline - - - o - - - - - Saliva - o - - - - - - - Liver extract - - - - - - - - - Pancreas extract - - - - - - - - Blood : - - - o? - - - Soap solution O? - o - - - - - - 64 In the foregoing table (No. 13) the zero, o, as usual means that in the small percentages present there was decided inter- ference. Wherever the space is left blank the conclusion in- dicated is, of course, that there was no interference under the conditions of the test. Taking the individual interfering substances, the following general conclusions may be deduced from the table: 1. Ferric chloride is the most active interfering compound, affecting 70 per cent, of all the tests. 2. Potassium ferrocyanide is the next most active, affecting 50 per cent, of all the tests. 3. Cupric sulphate, mercuric chloride, potassium thiocyanate, tannic acid, tea, coffee, and formaldehyde, rank next in this respect, showing in each instance about 40 per cent. 4. Manganese sulphate inhibits 30 per cent, of the tests. 5. Fifteen other compounds actively interfere in 20 per cent, of the tests. 6. The following substances, in the percentages used, failed to cause interference under the conditions noted in the various tables: Chloroform, alcohol, glycerol, acetone, egg albumin, toluol, thymol, phenol, benzoic acid, salicylic acid, plumbic acetate, sodium acid tartrate, calcium hydroxide, sodium car- bonate, basic lead acetate, sodium acetate, beef tea, meat ex- tract, liver extract, pancreas extract. Taking the various groups of interfering factors, the follow- ing general conclusions may be deduced from the table: 1. The acid salts are the most active of all the groups, af- fecting as they do over 24 per cent, of all the tests. 2. The activity of the other groups is as follows: X. A Summary of General Conclusions. Per cent. Neutral salts 16.5 Alkaline salts 13-5 Acids Biological mixtures, etc 12.0 Alkalies 10.0 Neutral organic compounds 5-7 (This series is offered only for what it is worth; the percent- ages apply merely to the limited but representative list of compounds used.) 65 3. The neutral organic compounds are the most inert of all the groups in causing interference. 4. There is a grand average for all these interfering groups of about 13.5 per cent., or, in other words, these substances ex- hibit interference phenomena in connection with 13.5 per cent, of the various tests included in this work. 5. By multiplying the number of substances in each group by ten (the number of tests studied) and then dividing this number into the actual number of recorded interferences we can get a comparative percentage table for all the tests. The following table represents the conclusions arrived at by this method, the ranking being according to freedom from inter- ference : I. Test. Sudan III Approximate per cent, of interference. 0.0 2. Xanthoproteic 4-0 3- Hopkins-Cole 4° 4- Seliwanoff 5° 5- Molisch 6.5 6. Iodine 6.5 7- Fehling-Benedict 10.0 8 Biuret 130 9- Millon... 22.0 IO. Barfoed 60.0 The above table is in its way an "efficiency table" for the tests mentioned and under the conditions incident to this research. 66 XI. Literature Cited. G. Colasanti, Gazetta Chim., (1891) XX., pp. 299-305. B. Reinbold, Pfliiger's Archiv., (1904) CHI., pp. 581-617. Molisch Test. Iodine Test. *E. DuClaux, Ann. Chim. Phys., (1872) [4] XXV., pp. 264-284. *Fr. Goppelsroeder, Poggend. Annalen, (1863) CIX., p. 57. P. Grelot, Jour. Pharm. Chim., (1906) [6] XXIV., pp. 154-161. ' H. Labbe and J. Chabriez, Compt. rend. soc. biol., (1907) LXIIL, pp. 32-34. C. Lonnes, Zeit. Anal. Chem., (1894) XXXV., pp. 409-436. C. Meinecke, Chem.-Ztg., (1894) XVIII., pp. 157-160. Guerbet Marcel, J. Pharm. Chem., (1903) [6] XVII., pp. 313-314. E. Puchot, Compt. rend., (1877) LXXXIIL, pp. 225-226. E. Riegler, Z. Anal. Chem., (1907) XLVL, pp. 315-318. Charlotte F. Roberts, Amer. Jour. Sci., (1894) [3] XLVII., pp. 422-429. *A. D. Tschirikow, PRu., (1891) XXX., pp. 802-807. Fehling-Benedict Test. S. Benedict, Jour. Biol Chem., (1909) V., pp. 485-487. E. Boivin and D. Loiseau, Compt. rend., (1875) LXIX., p. 1263. F. Repiton, Ann. de le Chim. analyt. appl., (1908) XIII., pp. 269-270. O. Rosenheim and P. Schidrowitz, Chem. News, (1897) LXXVI., pp. 318- 319; and Ibid., LXXVII., p. 97. W. Muller, Pfliiger's Archiv., (1878) XVI., p. 562; and W. Muller and J. Hagen, Ibid., (1878) XVI., 567. W. H. Welker, Proc. Amer. Soc. Biol. Chem., (1908) pp. 33-34. Barfoed Test. R. and O. Adler, Pfliiger's Archiv., (1905) CVI., pp. 323-328. W. O. deConinck, Bull. soc. chim., (1910) V., p. 569; Compt. rend. Soc. biol., (1910) LXVI., p. 509. H. Malfatti, Ztschr. phys. Chem., (1908) LVIII., p. 544-546. J. Pieraerts, Bull. soc. chim., (1909) [4] V., pp. 248-250. H. Rosin, Ztsch. phys. Chem., (1903) XXXVIII., pp. 555-556. Seliwanoff Test. *E. Feder, Arch, der Phar., (1905) CCXLH., pp. 680-704. W. J. Gies, Proc. Soc. Biol. Chem., Jour. Biol. Chem., (1910) VII., p. 60. H. B. J. Stockvis, Z. Biolog., (1898) XXXIV., pp. 466-470. *L. A. Tschugaeff, Jour. Russ. Phys. Chem. Soc., (1906) VII., pp. 1083-1085. K. H. Van Norman, Biochem. Jour., (1909) IV., pp. 127-135. Biuret Test. Millon Test. O. Nasse, Pfliiger's Archiv., (1901) LXXXIIL, pp. 361-368. F. Repiton, Compt. rend. Soc. biol., (1907) LXIL, 339-340. BIOGRAPHICAL. Chester Arthur Mathewson was born in Cincinnati, Ohio, December n, 1878. He was graduated from the Technical School of Cincinnati in 1896. During the two succeeding years he was engaged in business pursuits. From 1898 to 1903 he was a teacher in the Technical School of Cincinnati, studying during the same period at the University of Chicago, Columbia University, and the University of Cincinnati. In the year I9O3~'O4 he was a student in Yale University. During the field season of 1904 he was connected with the United States Forest Service. In 1905 he received the degree of B.S. from Columbia University and was then appointed assist- ant in biology and nature-study at Teachers College. In 1906 he received the degree of Master of Arts, together with the Master's Diploma in Education from Columbia University. After spending a year at Plainfield (N. J.) in establishing a department of biology in the high school, he was appointed in September, 1907, as a teacher of biology in the New York High School of Commerce. This position he resigned in Feb- ruary, 1910, in order to accept his present post in the science department of the Brooklyn Training School for Teachers. In September, 1907, he matriculated as a candidate for the degree of Doctor of Philosophy in the Faculty of Pure Science, Columbia University. PUBLICATION. The Behavior of the Pollen Tube in Houstonia coerulia. Bull. di Torrey Botanical Club, (1906) xxxiii, pp. 487-493. ARCHIVES OF PEDIATRICS ADVERTISER xvli MEAD'S DEXTRI-MALTOSE (MALT SUGAR) A substance resulting from the action of diastase upon starch. A pure form of carbohydrate, free from proteid, cellulose and fat. Wholly soluble in milk or water. Advantages in Infant Feeding Mead's Dextri-Maltose may be taken by the infant in larger quantities than cane or milk sugar, without causing diarrhoea and with greatly decreased chances of sugar fermentation. Being more readily assimil- able and oxidizable, it increases weight and energy to a greater extent than other sugars. Its Assimilation Limit About 100% Higher Than Either Cane or Milk Sugar The assimilation limits of the sugars as demonstrated by Reuss, Grosz, et. al., are: Malt sugar 7.7 grams per kilogram of body weight. Either milk or cane sugar, 3.1 grams per kilogram of body weight. Mead's Dextri-Maltose is made and marketed by MEAD, JOHNSON & CO. Jersey City, N. J. oflnasarcm Oxydendron Arboreum (Sour Wood) Sambucus Canadensis (Elder) and Urginea Scilla (Squill) Eliminates Effused Serum Controls heart action and restores physiological balance between arterial and venous systems. Thousands of Physicians are continually using t^nasarcin with most gratifying results in all dropsical effusions whether caused by disease of the heart, liver or kidneys. TRY dlnasarcin in any obstinate case of Dropsy - then judge its merits. Clinical results prove Therapeutics. Sre1onUrequtestarod 'K'™' 7^ ANASARCIN CHEMICAL CO. physicians only. WINCHESTER, TENN. Messrs. Thomas Christy & Co., London Agents. ■Rotes and Rews. Motherhood and Infancy.-This is the most unique, at- tractive and instructive little treatise yet published on this subject. You should give a copy to every patron who is raising a family. They are very beneficial to both patient and physician. Address the author, Dr. E. S. Harris, Box 722, Blue Springs, Mo., enclosing 10 cents, and he will send you a copy. They are fine. Sleeplessness.-Insomnia is a condition which is met with al- most daily, yet it is one which is treated only too frequently in some routing fashion, and often enough baffles treatment altogether. In the case of persons sleepless owing to troubles, says Dr. C. Willet Cunningham, writing in "The Hospital," words of wisdom will be more valuable than even the most ingenious prescription. There re- main, however, a number of patients who have nothing on their minds and yet cannot sleep. In some there is a family tendency, in others a habit has grown perhaps from small beginnings. These are exceedingly difficult to manage successfully. In this class of sleepless- ness one should have the bedroom absolutely dark and quiet at night. Ventilation is also important. Cold feet will keep other people awake. In habit cases it may help to advise a change of bedroom or even of house for a time. The evening meal is an important matter, but no general rule can be laid down. The late dinner of the full-blooded, over-nourished man needs curtailing, while the thin, dyspeptic, nervous woman requires more nourishment. For them some easily digested milk food, such as Benger's Food, the last thing at night, is useful. Tea and coffee are best avoided, especially in the latter part of the day. A "nightcap" has its obvious objections. Extra pillows are useful. The patient should spend as much time as possible in the open. Above all, Benger's is the Food for restful nights. It is so easily digested and so soothing and agreeable, that, while giving full nourishment to the system, it really promotes sound, healthy sleep. Benger's Food is mixed with new milk when prepared. It forms a dainty and delicious cream, entirely free from rough and indigest- ible particles. Infants thrive on it and aged people enjoy it. The composition is known to, and approved of, by medical men. Consumption.-The indifference of phthisical patients towTard the maintenance of sanitary conditions is proverbial. That the en- vironment of all such patients should be absolutely aseptic, both for the good of the patient and for the welfare of those who are brought into contact with them, is a well-established fact. The odor of the ordinary antiseptic is exceedingly disagreeable to the average person, and intensely so to the average consumptive, hence the strong disin- clination to the constant use of them. In Glyco-Thymoline we have an antiseptic which, while mild and soothing in its effect on mucous and other surfaces, is still a powerful agent for promoting asepsis, and a potent factor in the maintenance of sanitary environment in the sick room. Inhaled from a vaporizer or a fine spray atomizer, it will loosen the mucus in a marvelous manner and in a wonderfully short time shorten the paroxysms of coughing to a marked degree, at the same time reducing the danger of contagion to a minimum. ARCHIVES OF PEDIATRICS ADVERTISER xix Opium in the fullness of its therapeutic excellence Opium freed from all inert, useless stuff, exhibiting the total alka- loids of the drug as a unit, in their naturally-occurring proportions, in a water-soluble, non-irritating, injectable, diffusible form, ' n generally without constipating or other by-effects. > Provided in Powder, Tablets (for hypo. Brilliant results in U or ora' use) and Ampoules. Internal Medicine, II IR , _ .-. Sample and literature Gynecology and Ob- I lllliMIc-^ on reQuest- stetncs, Anesthesia L I and Psychiatry are re- corded in the clinical re- THE HoffmannLaRoche P°rts of Sahli, Ewald, Chemical Works Grafenberg, Leipoldt and other well- (TV-?V> 65 Fulton Street, New York known writers. (pain) " The physician has no higher nor nobler service to perform than to secure for his suffering patients prompt surcease of pain." In accomplishing effectual analgesia, however, the painstaking practitioner will at the same time always aim to cause the least possible embarrassment to physiologic processes. Of all anodynes, therefore, PAPINE is the most acceptable, because it affords not only the most gratifying relief from pain but with none of the narcotic or toxic effects common to other opiates. Papine presents the anodyne principle of opium with the narcotic, nauseat- ing and constipating elements removed. In consequence it does not suppress the secretions, cause cerebral excitation, nor show habit-forming tendencies. To the result-seeking, conscientious physician, Papine cannot fail to appeal as "the ideal analgesic.'' BATTLE & CO. LONDON ST. LOUIS PARIS XX notes and news. Neurotic Anorexia.-While loss of appetite and nausea are usually symptoms of a host of diverse pathological conditions, they sometimes constitute a disease in themselves-a kind of neurosis. In these cases the physician will find Gray's Glycerine Tonic Comp, of almost specific value for restoring the impaired appetite. It is not only agreeable to take, but produces its benefits at once in such a natu- ral way that before the patient realizes it the normal amount of food is being taken. Its efficacy in these neurotic cases makes Gray's Glycerine Tonic Comp, exceedingly useful in relieving the severe nausea that often occurs in early pregnancy. I On .returning home on a cold and wintry night, per- haps from a case sorely taxing his mental and physical powers, the physician can find no better solace than in a cup of hot, appetizing and refreshing bouillon. With a box of "Steero" Bouillon Cubes the material for quickly preparing such a delectable beverage is al- ways at hand. One of these cubes, dropped in a cup with boiling water added, gives a deliciously flavored drink that is grateful to the stomach and agreeably stimulating. If taken with a biscuit it forms a digestible, light repast before retiring that, unlike coffee or tea, allays wakefulness from over-tired nerves and promotes sleep. The "Steero" Cubes are prepared from the purest materials, and the great- est care exercised in obtaining just the right degree of seasoning. Write Schieffelin & Co., New York, for samples. Moore on Physiological Standardization.- Benjamin Moore, D.Sc., M.R.C.S., Professor of Bio-Chemistry, University of Liverpool, criticises physiological standardization as a test of "killing power." Underlying the test of killing is the assumption that the same body which kills produces the favorable therapeutic effect. However sound such argumentation might be in the case of a single active body, it loses all application when dealing with at least four bodies so very different in action and toxicity as are the glucosides of digitalis leaves. The test would only hold good if there were the same ratio between maximum therapeutic action and minimal lethal dose for each of these bodies, and experience of both pharmacologist and clinician demonstrate that this does not hold true. He further emphasizes that it is not maximum toxicity or brute force that should be aimed at. The active agent to pick out is one which possesses what might be described as the greatest factor of safety, that is, the highest ratio between the minimal lethal dose and the dose giving a good therapeutic effect. Some attention should also be paid to the possibility of accurate dosage, and here a body which can be com- pletely isolated holds an advantage because the physician knows exactly how much he has given to his patient. Moore, from a series of experiments with digitoxin amorphous (Digalen), concludes that "the results indicate that in the use of ordinary pharmacopoeial prepa- rations of digitalis the tonic effects and toxic effects are produced in varying degree by the different glucosides, but that soluble digitoxin (Digalen) is one which possesses more valuable therapeutic properties, with a large factor of safety, than the mixture as extracted from the tincture, and that the safe dose is treble that of the latter." He also states that its low toxicity as compared with the mixed glucosides in the tincture of digitalis renders intravenous injection safe, at the same time "a full therapeutic effect is obtained on the heart."-British Medical Journal, January 13, 1912. ARCHIVES OF PEDIATRICS ADVERTISER . xxi "while the paramount serviceability of a remedy is its therapeutic value, its adapt- ability is an item of no small import. This is particularly true in the application of hot moist heat, the generally accepted treatment for inflammatory conditions, where its continued application is so es- sential for results. To subject a patient to frequent and un- necessary dressings and exposure, as is the case where poultices, moist packs, etc., are used, not only retards the progress of treat- ment, but disturbs and annoys the patient and is not in keeping with advanced therapeutics. The serviceability of antiphlogistine as a therapeutic agent is best attested by the preference accorded it by the medical profession in the treatment of Congested or Inflammatory conditions where hot moist heat is called for. The adaptability of antiphlogistine indi- cates it as the best medium for the employ- ment of hot moist heat, as it is easy to apply, holds its heat for hours and thus does not disturb or annoy the patient." xxil , ARCHIVES OF PEDIATRICS ADVERTISER NATIONAL ASSOCIATION FOR THE STUDY AND EDUCATION OF EXCEPTIONAL CHILDREN An Incorporated Society, not for Pecuniary Profit, and Devoted to the Promotion of a National Movement for the Solution of the Problem of the Exceptional Child. It Maintains (a) Herbart Hall, Institute for Atypical Children. An educational laboratory and school for the care and treat- ment of truly atypical boys and girls. (Feeble-minded, insane, epileptic, or otherwise clearly abnormal chil- dren are not accepted.) (b) Observation Clinic. For the care and study of borderland cases where positive prognosis of restoration to normality or permanent defectiveness can only be made upon the basis of extended observation, (c) Adolescent Department. For ineffective boys between the ages of sixteen and twenty-five. (The so-called "after- care" cases.) (d) Practical Teachers' Course. For regular teachers specializing to organize or take up work in ungraded classes of public schools, (e) Physiological and Psychological Labora- tory. Devoted to research on the broad aspects of the problem of the exceptional child. ( f) Publicity Bureau. For the distribution of society proceedings, scientific monographs, reports, etc., and the arrangement of public lecture courses, round tables, etc. For particulars on the general scope of the Association or any of its activities, address, SECRETARY N. A. S. E. E. C. " WATCHUNG CREST," PLAINFIELD, N. J. BOOKS ON COSMETICS HAND BOOK OF LECTURES ON Cosmetics Cosmetic Treatment By DR. MAX JOSEPH, Berlin, Authorized By DR. EDMUND SAALFELD, Berlin, Au- English Translation, Revised from the Third thorized English Translation from Second German Edition German Edition 12mo, 151 Recipes, Cloth, Prepaid, $1.00 202 Pp., 131 Recipes, Ulus., Prepaid, $1.75 E. B. TREAT & COMPANY, 241-243 West 23d Street, New York . • Jr- yes'. ARCHIVES OF PEDIATRICS ADVERTISER xxlii Looking Into The "Coffee Question" ww necessarily involves a consid- ■ fT eration of the now well-known f and well-established cereal ■ Ip. beverage • POSTUM With modern advances in pharmacy-the discover}- and isolation of alkaloids and a knowledge of their physiological action on the human system-has come a more intimate knowledge of the Uses and Abuses of Caffeine -the alkaloid in Coffee and Tea. As in some other instances, people have learned that something in Coffee causes various ailments. It remained for science to dis- cover and define that "something." It is now known to be Caffeine. But, to induce, mankind to forego Coffee, even though it contain a drug, was no easy task for the physician or nurse. Postum, however, has made it practical for the physician to obtain the cooperation of patients, the successful treatment of whose cases demand that Coffee be abandoned. When Postum is properly prepared (see directions on package) it is a most palatable beverage. With cream, it resembles, in flavor, high-grade Java; yet possesses a "tang" peculiar to itself which many come to like the more they use it. Made of clean, hard wheat and a small per cent, of New Orleans molasses, Postum is free from Caffeine or any other harmful substance. The"Clinical Record" for the physician's bedside use will be sent prepaid to any physician or nurse who has not already received one. Also a box of samples of Postum, Grape-Nuts and Post Toasties. Postum Cereal Co., Ltd., Battle Creek, Mich., U. S. A. xxiv ARCHIVES OF PEDIATRICS ADVERTISER NOW READY THIRD EDITION DISEASES OF THE Stomach and Intestines By BOARDMAN REED, M.D. No more positive proof of the value of this work could be afforded than its augmenting sales and this early demand for a new edition, which is more than an "edition," in the ordinary meaning of the term, as the entire text has been thoroughly revised and largely reset. It is rather a new book. The author is clear and sound in his teachings, simplifies conditions as far as pos- sible, and gives a good practical working knowledge, such as he has gathered from a large experience in his special field. There was need of such a book by a practical observer, who gives to the reader the easiest and most useful methods to be employed in general practice.-Medical Record, New York. Third Edition, thoroughly revised and largely re-written Octavo, 1037 pages, fully illustrated, cloth, prepaid, $5.00 net Send for large descriptive circular with contents E. B. TREAT & CO., Medical Publishers, 241-243 West 23d Street, NEW YORK a.. Mcv jA** ■ Why Take a Chance? Pills tablets and gelatine capsules are often insoluble. Powders "in Konseals" give sure and quick results, for Konseals are always soluble and tasteless. The U. S. Dispensatory (p. 1118) says: For most powders of disagreeable taste, cachets afford the best method of administration, - Konseals are the best cachets. - If you write for Konseals see that you get Konseals; if you don't, investigate, for substitutors in little things are not de- pendable when it comes to the more expensive medicaments. For literature and samples, write J. M. Grosvenor & Co., Boston, Mass. ARCHIVES OF PEDIATRICS ADVERTISER xxv BacterinS Bacterins (bacterial vaccines) are killed bacteria suspended in physiological salt solution. They stimulate the production of protective substances (antibodies) and are used to prevent or overcome bacterial infections. Each bacterin is indicated for the infection caused by its corresponding bacterium; for instance, a staphylococcic bacterin is used for staphylococcic infection. Accurate diagnosis is therefore necessary. For the general practitioner the use of stock bacterins is advisable be- cause valuable time is thereby saved. It is well recognized that mixed infections are usually present in infectious diseases. "Mixed" and "polyvalent" (many different strains) bacterins are therefore becoming deservedly popular. As regards their use, Polak states: " The mixed vaccines of reliable laboratories have given better results than when a single variety was used. This has been shown repeatedly in the blood picture when an auto- genous vaccine of a single strain used in large doses up to 500,000,000 has failed to increase the leucocyte-count or diminish the polynuclear percentage, the mixed vaccines of several strains have promptly produced a marked leucocytosis. Even colon bacillus infections, such as the infection of a pelvic hematocele by the colon bacillus, have yielded more promptly to mixed vaccines of polyvalent strains than when a single autogenous germ has been used." -(Journal American Medical Association, November 25, 1911, p. 1738.) The prophylactic value of bacterins is proved beyond question in typhoid fever, and preventive medicine suggests immunization against streptococcic, colon, staphylococ- cic, pneumococcic and tubercular infections by the use of their corresponding bacterins. The results following the general use in the U. S. Army of typho-bacterin in protective vaccination against typhoid fever are little short of marvelous. "During the past three years 60,000 men completed the three inoculations; but twelve cases of typhoid fever developed during this time and no death occurred." (Phalen and Callison, Medical Record, December 9, 191 1, p. 1203.) We prepare the following: Acne-Bacterin (Acne Vaccine) Pulmonary-Bacterin Mixed Cholera-Bacterin (Cholera Vaccine) Pyocyano-Bacterin (Pyocyaneus Vaccine) Coli-Bacterin (B. Coli Vaccine) Scarlatina-Bacterin (Scarlet Fever Vaccine) Influenza-Bacterin Mixed (Influenza Vaccine Mixed) Staphylo-Bacterin (Staphylococcic Vaccine) Friedlaender-Bacterin (Friedlaender Vaccine) Staphylo-Bacterin Mixed (Mixed Staphylococcic Vaccine) Neisser-Bacterin (Gonococcic Vaccine) Staphylo-Acne-Bacterin (Staphylo-Acne-Vaccine) Neisser-Bacterin Mixed (Gonococcic Vaccine Mixed) Staphylo-Albus-Bacterin (Staphylo-Albus-Vaccine) Neoformans-Bacterin (Neoformans Vaccine) Staphylo-Aurens-Bacterin (Staphylo-Aureus-Vaccine) Pneumo-Bacterin (Pneumococcic Vaccine) Strepto-Bacterin (Streptococcic Vaccine) Pneumo-Bacterin Mixed (Pneumococcic Vaccine Mixed) Typho-Bacterin (Typhoid Vaccine) Copies of Mulford's Working Bulletins mailed free upon request. State bacterins in which specially interested. H. K. Mulford Company New York P h I1 eld QI P h I3 Chicago St. Louis Kansas City Minneapolis San Francisco Atlanta Seattle Toronto xxvi ARCHIVES OF PEDIATRICS ADVERTISER JUST PUBLISHED THE DISEASES OF INFANTS AND CHILDREN By EDMUND CAUTLEY, M.D., Cantab., F.R.C.P. London Senior Physician to the Belgrave Hospital for Children ; Physician to the Metropolitan Hospital, London Journal of the A. M. A., says:-"We recommend this as a text-book on diseases of children, both to the practicing physician and to the student. It is a marvel of complete- ness and thoroughness; the style is clear and forcible; the arrangement is excellent and the matter is strictly up-to-date." Medical World, says:-"It is a complete, authoritative, and absolutely dependable work for regular consulting, and is in every way one of the best books we have on the subject." London Lancet, says:-" For its size this is the best English text-book on the sub- ject of the diseases of children with which we are acquainted." Medical Record, says:-' ' It would be extremely difficult to name a work on pediatrics which, within the limits of one volume, covers the ground so completely as does this treatise on diseases of children. Written for the practitioner rather than for the student, it aims to present this subject with such fulness as to render reference to other works unneces- sary. A careful examination of the book will reveal that hardly anything of importance in the modern developments of pediatrics has escaped the vigilance of the author." Send for Pamphlet Giving Full Contents and Specimen Pages Large Octavo, 1058 Pages. Substantially Bound in Cloth, Prepaid, $7.00 net E. B. TREAT & COMPANY, 241-243 West 23d Street, New York Two New Works 1 By Prof. DR. CARL von NOORDEN Professor of the First Medical Clinic, Vienna, in his series on the Pathology and Therapy of the Disorders of METABOLISM AND NUTRITION Edited and Translated under the Supervision of Alfred C. Croftan, M.D., Chicago X VIII. Inanition and Fattening Cures $1.50 The author's criticism of innumerable dietetic fads and fallacies is of the greatest practical use, as he provides something of value in their stead IX. Technique of Reduction Cures and Gout I lumber $1.50 The practical dietetics and general treatment of the subjects are given With great exactness and make very interesting and instructive reading OTHER MONOGRAPHS IN THE SERIES I. Obesity $0.50 V. Saline Therapy $0.75 II. Nephritis 1.00 VI. Drink Restriction 75 III. Colitis 50 VII. Diabetes Mellitus 1.50 IV. The Acid Autointoxications 50 Others in Preparation Complete set (9 volumes) $8.50, prepaid. Also sold singly E. B. TREAT & CO., Medical Publishers, 241-243 West 23d Street, NEW YORK ARCHIVES OF PEDIATRICS ADVERTISER xxvii "Just to be sure!" Thus spoke a careful physician when asked why he used our diphtheria antitoxin exclusively. Our antitoxin justifies his confidence. It is evolved in the blood of healthy, vigorous horses-horses that are carefully selected; that are under the watchful eyes of expert veterinarians at t our model biological farm. Z Z • II It is perfected in laboratories that afford = 7 II unequaled facilities for serum manufacture Z * A / il - laboratories in which it is possible to Lr Ui observe, at every step of the process, the Zf The most satisfactory I | vital principles of asepsis. V syringe container ( ® It is exactingly standardized, bacterio- / ever devised. VJB logically and physiologically. / " It is guaranteed as to purity, potency and uniformity. • CONCENTRATED ANTIDIPHTHER1C SERUM (GLOBULIN.) Containers of 500, 1000, 2000, 3000, 4000 and 5000 units. Specify "P. D. & Co." on your orders. Parke, Davis & Co. Write it plainly in the prescription. There is just one preparation of Cascara Sagrada that affords in agreeable form the full laxative effect of this val- uable drug. That preparation is CASCARA EVACUANT. This product is prepared from the well-seasoned bark of is=====ia true Rhamnus Purshiana. Its marked therapeutic activity, its agreeable flavor, and the beneficent effect which it exerts upon the stomach, bowels and liver, commend it as one of the I most eligible laxatives available to the medical profession. fesSEsitfi CAUTION.-There are numerous so-called aromatic casca- • ras-some of them of doubtful therapeutic worth. To pre- Bu vent confusion with them, write plainly in your prescription, " CASCARA EVACUANT, P. D. & CO." Pint, %-pint, J4-pint, 5-pint and gallon bottles. Home Offices and Laboratories, Parke. Davis & Co. Detroit, Michigan. * xxviii ARCHIVES OF PEDIATRICS ADVERTISER 1 ' B T7PII T7PQV § VnKUlN i-L'l^i 1 Qqpn THRTG no remedy gives such uniformly sat- $ XJxxO I 1x1 I 10 isfactory results as B 5 with its train of hepatic and intesti- S KlLrkV<V7V<lX O nal complications, rarely fails to B BROMIDES ^nd^oncetothebe^^ | This is due, first to the happy /^TTTAAXTT A H ! combination of several different bro- > VjlTlvliNllX I mide salts; second, to the remark- 5 , B | able chemical purity of each salt; . Although exerting a pronounced .1 S and finally, to the pharmaceutical 5 influence on the secretory structures l|.| H ■ I character of the composite product. < of the stomach and liver, Chionia has |I * g . It is easy to understand, there- ; the special advantage of increasing L S |l I fore, with their purity and potency the physiologic activity of the gastro- H i B .1 so marked, how Peacock's Bromides < intestinal and hepatic functions hn have been able to accomplish such $ without producing severe or undue B ||| pronounced therapeutic effects in > catharsis. It is in consequence, the I J epilepsy and kindred ailments, even 5 ideal remedy in all forms of hepatic || when administered over long periods. $ torpidity. : PEACOCK CHEMICAL CO.. St. Louis, Mo.L I Free from P R I । Al 0 I D Produce W Cathartic | l\v/ »Vz| Permanent Iniquities ■ S Benefits JW OVERCOME CONSTIPATION r/» $ Free from CACTIMA Sustain and fob* Cumulative Dll I Regulate Effect r I L L t I J the Heart TONE THE CARDIAC MUSCLE A Promotes LK Digestive f " Natural III Tonic Processes Ayim W INCREASES FUNCTIONAL ACTIVITY pr -ii 1-SULTAN DRUG CO., 5t. Louis. Mo.-1 fcffl rwaWM ARCHIVES OF PEDIATRICS ADVERTISER xxix A crystal, liquid Antiseptic, slightly Alkaline. UNEXCELLED in the treatment of Catarrh, Eczema, Pruritus and itching so frequently affecting the a aged, trial bottle by mail, on request. NEURILL A FOR NERVE DISORDERS NEURILLA If Patient suffers FromTHE BLUES (Nerve Exhaustion), Nervous Insomnia,Nervous Headache,Irritability or ■■■■BUK1 General Nervousness,£ive four times a day one BRBK998HHBHBHI teaspoonful NEU RI LLA In nervous fretfulness of teething Children g'iva five to twenty drops, DAD CHEMICAL COMPANY. NEW YORK PARIS^ FRESH AIR WITHOUT A DRAFT Patented March 1, 1910 AN abundant supply of fresh air without the least possibility of a draft is supplied by means of a GEM VENTILATOR. Adjustable to windows of different widths, equipped with a fine wire screening, having controlling features. The above features make it the most adaptable window ventilator on the market. A necessity in every room occupied by human beings. Thousands of users report perfect satisfaction. GEM VENTILATOR COMPANY, BOSTON, M.». The BLUES Agents Wanted /c i l • m ,i • x to introduce our line of new (oplanchmc Neurasthenia) 1 , i i j- i l i and standard medical books By ALBERT ABRAMS, M.D. Exclusive Territory Quick Sales The object of this volume is to direct atten- tion to a form of nerve weakness, popularly Good Profits known as "Ike blues." , . , . Write for full particulars From the author's vast experience, he ... . . e , , . . . F R TREAT & CO Medical Publishers knows of no variety of neurasthenia which r»D. ik a u , , . , , , . TT- ,, j 241-243 West 23d Street : New York is more amenable to treatment. His methods are described in detail and may be easily g~a executed. CLINTON In meeting the demand for a new edition the Pion*n* AnTIl/E author has taken opportunity to thoroughly vAOvAnA Hull VC revise the text and to add a most important FOR CHRONIC CONSTIPATION chapter on "Augmenting the Tone of the does Not Gripe Splanchnic Circulation." A palatable and highly active prepara- J tion of CASCARA SAGRADA. ( FOURTH EDITION Revised and Enlarged Each fluidounce imperial represents one avoirdupois ounce of selected drug. A PALATABLE AND 8vo. 304 Pages. Illustrated. Cloth, $1.50 Sure and Safe Laxative for j ACT|vcpRtPARAnoN Children and Adults. E. B. TREAT & CO., Medical Publishers WRITE FOR FREE SAMPLE. i^^OKLYN-NEW YW I 241-243 West 23d Street - NEW YORK BRISTOL-MYERS CO.JMjM'1 - - ---1 BROOKLYN • NEW Y0»". ''fcdJtJ®- XXX ARCHIVES OF PEDIATRICS ADVERTISER The Great Combination "The Practical" with "The Necessary" Archives of Pediatrics, $3.00 Review of Reviews, $3.00 ■ DnTU h I II | THE AMERICAN UU 1 11 I REVIEW^; $ 4 50 REVIEWS^ : • S edited by ALBERT SHAW \ 1 0 gel 1 1 \JI\l v S Subscriptions must * ' NECESSARY be paid m advance $ -niAxi nrrn but may be new THAN EVER or renewal. Mag- I ! azines may be sent I | j f | I to tne same or -1, different addresses. Archives of Pediatrics is the practical Review of Reviews is most necessary in medical journal because the disorders the year preceding presidential election, of infants and children occupy the larger It not only reviews every important arti- part of the general practitioner's time. cle in all the other leading magazines. It is the representative journal of its but, in addition to this, has each month specialty and is recognized as an author- contributed articles of its own on every ity among pediatrists the world over. conceivable important topic. Other Magazine Combinations You may also secure the following $1.50 magazines: AMERICAN PEARSON'S SHORT STORIES COSMOPOLITAN RED BOOK SUNSET McCLURE'S EVERYBODY'S DELINEATOR WOMAN'S HOME COMPANION GOOD HOUSEKEEPING by adding, at the rate of $1.15 for each, to the price of Archives of Pediatrics ($3.00) or the price of "The Great Combination" ($4.50). Subscriptions may commence any time. Sample "Archives'' upon request. ADDRESS ARCHIVES OF PEDIATRICS . . . 241-243 West 23d Street NEW YORK ARCHIVES OF PEDIATRICS ADVERTISER ttyI r.f m' r- । > rui ' j/ Ml wlklLM £----- - CREOSOTE is an old remedy, It's value is jOfeWww 1 recognized by all, yet it is little used because wAmlSiK of the difficulties encountered in its adminis- tration. We have perfected a creosote that overcomes all difficulties. This perfected ■ i j Al W 1 product is a chemical combination of creosote A *41 11 with calcium. It is a reddish-brown granular powder and contains approximately per pure beechwood creosote. It might properlv be | called SOLUBLE CREOSOTE since it mixes ' in all proportions with water. S It is a remarkable remedy, not only in tuber- K1A S culosis and bronchitis, but in coughs and colds, KB pneumonia, typhoid fever, indigestion, diar- ' ® rhoea, &c. || Published reports of results obtained from |L. this product seem exaggerated until you have opportunity to observe them, then the skeptic pb)sx S becomes an enthusiast. price list. j J CALCIUM CREOSOTE POWDER, (MALTBIE). Pre- | ' > pared by simply adding water. Per pound, sufficient p ,/L for one gallon solution. $1 50. <Bv express prepaid. $1.75) K CALCIUM CREOSOTE TABLETS, (MALTBIE , « chocolate coated representing one teaspoonful solu- tion, per 1000, $2.00. (By express prepaid, $2.25.) SjMMsMWiiW 1====;=====^^ Piihw^^ % bp it '0 ; 3 ' TIiLY British Journal of Children's Diseases MONTHLY, 2/- net Annual Subscription, 20/- net, $5, post free ADLARD & SON, Bartholomew Close, LONDON, ENGLAND ft A I limiTIAl is especially valuable THE. BIG BEN BINDER &AL HEPATIGA is simple, practical, satisfactory ing from organic derangement of the liver and kidneys. It is the best agent M .. L' ri i ah ESsl -JR for the relief of that form of costive- Ino punching or holes necessary. All ness that is ushered in by an attack of that is required is a slight slit between c?llc and '"digestion, and not only iii- IL I cIears away the effete and irritating the pages to accommodate the clip ! 8gggrH<8SM I matter in the alimentary tract but elim- *1111 HlhTS if 'nates the semi-inspissated bile that too Each binder holds one volume of ' KV^^bU|J1 f I frequently induces the so-called "bil- ARCHIVES OF PEDIATRICS ious" condition; at the same time an AKLHIVL3 Ut rtUIA 1 KILD pr abundant secretion of normal bile is c- i d- J *1 nn -J !irJAUNLUwMnl t assured, thereby demonstrating its □ingle Binder - $I.Uu prepaid unit Arm* eniVFIIT I vaIue as a liver stimulant and true Special price on quantity KSmS cholagogue' „ „ - _ ~ BRISTOL-MYERS CO. B. B.Treat 6 Company 277-281 Greene Avenue, 241=243 W. 23d St., New York write for free sample. BROOKLYN, NEW YORK, U.S.A. xxxil ARCHIVES OF PEDIATRICS ADVERTISER Quinine is the sulphuric acid salt of the base quinine. Euquinine is the ethylcarbonic acid salt of the same base. Therapeutically, therefore, they are identical; but Euquinine is Children's quinine particularly, for the reasons that: It IS tasteless. It does not disturb the stomach. It is better than chocolate tablets. It is more reliable than pills or capsules. Samples and Literature on application to MERCK CO. New York "It's real Coffee" W7* yU Genuine Coffee from which about 90% of 1 J r V Ju jAL its Caffeine has been removed. __ - f Flavor and aroma unimpaired. Merck S Thls is the Coffee used for past two years "Caffeine-extracted" at Carlsbad, Marienbad. Bad Nauheim, &c Coffee Write for leaflet and details. MERCK & CO. N. Y. ARCHIVES OF PEDIATRICS ADVERTISER i JUST PUBLISHED The most complete review of the entire field of medicine. -Interstate Medical Journal It is one of the best books in English covering the progress made in all branches of medicine during the past year. -Bulletin of the Johns Hopkins' Hospital There is no single volume annual anywhere near its equal. -Medical Summary No other single volume at anything like the price will keep the practician so thoroughly abreast of all that is new in the various branches of medicine. " Medical World A comprehensive review of the year's work. -Journal of the A. M. A. There is no better compend of the year's progress. The arrange- ment is the acme of simplicity and convenience -Medicalstandard 1912 International Medical Annual 30th Year, Complete in One Volume Octavo, over 700 pages, fully illustrated by plain and colored plates, cloth, prepaid, net, $3.50 Send for Large Descriptive Contents Circular E. B. TREAT & Co., Medical Publishers, 241-243 West 23d Street, New York SAL LITHOFOS A Valuable Effervescent Saline Laxative Especially indicated in the treatment of Rheumatism, Rheu- matic Arthrica, Neuralgia and all Uric Acid Diseases SAL LITHOFOS is a preparation containing in an active state Lithia and Sodium Phosphates. It is of special service in the treatment of Chronic Rheumatic and Gouty conditions, their allied affectations and in many other disordered states. Expert knowledge and chemical skill of a high order were required to combine in this palatable preparation the necessary active constituents without it in any way producing the deterioration so often found in many advertised remedies. SAL LITHOFOS is of value in restoring the organism to a normal state in a very short time. Sal Lithofos by virtue of its saline aperient qualities is of distinct service in the treatment of cirrhosis of the liver and its attendant disorders. A three ounce bottle mailed on request. The WINGATE CHEMICAL COMPANY, Ltd. MANUFACTURING CHEMISTS Montreal, P. Q. Rouses Point, N. Y. ii ARCHIVES OF PEDIATRICS ADVERTISER | Maltose and Dextrine The continued endorsement and increasing use of the highly efficient carbohydrates, maltose and dex- fa trine, in infant feeding is good evidence that the strong claims made for Mellin's Food as a modifier of P milk are fully warranted. Study the following analy- sis, noting especially the carbohydrate content, that you may realize why Mellin's Food and milk makes pj such a satisfactory diet for infants. ANALYSIS OF MELLIN'S FOOD Fat 16 ftf Proteins 10.3S Maltose (58.88 fa Dextrines } S°luble Carbohydrates . . } W Salts 4.30 Water 5.62 fa 100.00 I The readjustment of cow's milk, by adding the proper amount of Mellin's Food, is easily and scien- tifically done with the result that the baby has a diet that furnishes all the elements for the structure of brain, fa nerve, muscle, bone and gland, as well as for the re- pair of waste tissue. W Formulas for Infant Feeding" (for physicians' use) sent upon request. fa MELLIN'S FOOD COMPANY, - - BOSTON, MASS. ARCHIVES OF PEDIATRICS ADVERTISER iii TO CONTRIBUTORS AND CORRESPONDENTS OF ARCHIVES OF PEDIATRICS Subscription $3.00 a Year, in Advance. (England, 14s.) Single Copy, 30 Cents Editorial Communications address to R. S. HAYNES, M.D., 267 West 79th Street, New York Business Communications address to E. B. TREAT & CO., 241-243 West 23d Street, New York ORIGINAL ARTICLES, brief reports of rare and interesting cases, or new modes of treatment are solicited and should be sent, together with all other editorial communications, direct to the editor. All articles must be typewritten; practical, not theoretical, and none will be con- sidered for publication unless with the distinct understanding that it is contributed exclusively to this journal. The editor and publishers will not be responsible for the views of contributors. ILLUSTRATIONS, as in the judgment of the editor are necessary, will be furnished free when black aud white drawings or photographs are supplied by the author. Temperature charts must be neat and in black ink on white paper. REPRINTS of original articles will be furnished gratis to authors making the request direct to the pziblishers, immediately upon receipt of galley proof of their article. Covers to these and extra reprints will be furnished at cost. Or, in lieu of reprints, the publishers will, if so de- sired, mail free to individual addresses furnished by the author, twenty-five magazines. COPYRIGHT.-Matter appearing in this journal is covered by copyright, but no objection will be made to its reproduction in reputable journals, if proper credit be given. CHANGE OF ADDRESS notice should give both the old and the new address. DISCONTINUANCES.-The publishers must be notified, by letter, when a subscriber wishes his journal stopped and all arrearages must be paid. Without such notification it is assumed that a continuance is desired. Journals returned are not notice of discontinuance. REMITTANCES should be made by check, bank draft, money or express order. If currency is sent, the letter should be registered. ^CONTENTS^ EDITORIAL. The Final Report of the Royal Tuberculosis Commission 241 Popular Lectures on Hygiene 243 ORIGINAL COMMUNICATIONS. Is there any Valid Objection to Vaccination? By Charles W. Banks, M.D. 244 Contact Infection in Contagious Diseases. By B. Van D. Hedges, M.D 250 The Prevention of the Commoner Contagious Diseases of Infancy and Childhood. By Royal Storrs Haynes, M.D 257 The Duty of the Community to its Backward and Defective Children. By Isa- belle Thompson Smart, M.D 268 Outdoor Life for City Children. By Walter Lester Carr, M.D 281 On the Diagnosis of Atypical Scarlet Fever. By D. J. Milton Miller, M.D. 289 Rumpell-Leede Phenomenon of Scarlet Fever. By May Michael, M.D 298 What New Jersey is Doing for the Epileptic. By David F. Weeks, M.D 303 CURRENT LITERATURE. DISEASES OF THE EYE. LeFever, C. W.: The Management of Squint in Children 312 Tibone, P.: Ophthalmia Neonatorum 313 DISEASES OF EAR, NOSE AND THROAT. Hopkins, F. E.: A Case of Multiple Foreign Bodies in Smaller Bronchi 313 Wagner, Henry Lewis: Contribution to Passive Immunization in Diphtheria of the Upper Respiratory Organs 314 Contents continued on page v. iv ARCHIVES OF PEDIATRICS ADVERTISER ESSENTIAL BLOOD ELEMENTS Which all convalescents lack, have been found by thousands of the leading physicians for their patients in BOVININE BOVININE suppl es all this as no Beef Extract can. It • raises the Opsonic Index to normal standard and prevents chronic invalidism. BOVININE s not only a perfect nutritive tonic in itself, but being rich in elementary iron and all essential elements neces- sary for complete cell reconstruction and nutrition, it re-establishes completely normal metabolism, thus assuring a quick recovery from all wasting diseases. Write for Sample, also for one of our new Glass (sterilizable) Tongue Depressors / THE BOVININE COMPANY \ \ 75 West Houston St.. New York City A NEW BOOK ON DISEASES OF CHILDREN Case Histories in Pediatrics Containing One Hundred Histories of A dual Patients, seleded to illustrate the Diagnosis, Prognosis and Treatment of Diseases of Childhood By JOHN LOVETT MORSE, M.D. TABLE OF CONTENTS Section /-Diseases of the Newborn Section VII-Diseases of the Heart and Peri- Section II-Diseases of the Gastro-Enteric Tract cardium Section ///-Diseases of Nutrition Section of the Liver Section /K-Specific Infectious Diseases Section °f tEe K'dney and Section V Diseases of the Throat, Nose, Ears i and Larynx Section X -Diseases of the Blood Section P7-Diseases of the Bronchi, Lungs and Section ^/-Diseases of the Nervous System Pleura Section XII-Unclassified Diseases A thorough Index of Symptoms and Diagnoses makes the book a fairly complete reference book It has been the author's purpose to furnish for practitioners, by the discussion of these actual and selected case-histories, a Post-Graduate Clinical Course in Diseases of Children 8vo, 320 pages, with a few illustrations. Price, express prepaid, $3.00 W. M. LEONARD, Publisher, 101 Tremont Street, BOSTON ARCHIVES OF PEDIATRICS ADVERTISER V CONTENTS-Continued CURRENT LITERATURE-Continued SURGERY. Homans, J.: Osteomyelitis of Long Bones 314 Cecil, R. L., and Bulkley, K.: Lesions Produced in Appendix by Oxyuris Vermi- cularis and Trichocephalus Trichiura 3T5 Kirmisson, E.: Diverticulum of Meckel Opening into the Umbilicus. (Per- sistance du Diverticule de Meckel Ouvert a 1'Ombilic.) 315 Simmons, C.: Two Cases of Intracranial Cerebral Hemorrhage in the Newborn Relieved by Operation ... • 316 MEDICINE. Roleston, H. D., and Maloney: Purpura in Acute Infectious Diarrhea. A Re- port of the Meeting of the Royal Society of Medicine held November 24, 1911 316 Pisek and Coffen: The Polygraph as an Aid in Diagnosis of Cardiac Conditions in Children 316 Basch, Carl, and Rohn, Adolf: Physical Evidence of the Thymus 317 Smith, Eric Bellingham, M.D., Lond., M.R.C.P., Lond.; A Type of Nervous Vomiting in Childhood 317 Beuker, O. H.: The Tuberculin Reactions and Their Comparative Value as Diagnostic Aids 318 Contents continued on page vii. A Deficient or Scanty Milk Secretion may be brought to Normal and kept there by the regular use of Maltropon. Maltropon is a combination of the albumen Tropon (Finkler) with Malt and Lecythin. A trial is seriously recommended especially in multiparae, where previously difficulty was encountered. TROPON WORKS 81 Fulton St., New York V1 ARCHIVES OF PEDIATRICS ADVERT IS E R \ Xh-Z" . "X C, " ■ ' S -XX . X J . ' " ?\ \?J ' £ y \ A\ v , - j The formation of a rich, nutrient, circulating fluid: blood which shall contain an abundance of red corpuscles of the necessary structural and physio-chemical integrity. How to "build" such blood is an ever-present therapeutic problem for the physician to solve. i m a a <» jav j ; rang is a powerful blood-forming agent; it induces the generation of hemoglobin, the oxygen-carry- ing constituent of the blood; it is a genuine hemoglobinogenetic. It feeds the red cor- puscles with organic iron and manganese which are quickly and completely absorbed in cases of Anemia from any cause, Chlorosis, Amen- r orrhea, Dysmenorrhea, Chorea, Bright's >/ Disease, etc. In eleven ounce bottles only; never sold in bulk. 78 % MJBREITENBACH CO., NewySm Our Bacteriological Wall Chart or our Differential Diagnosis Chart will be sent to any Physician upon request ARCHIVES OF PEDIATRICS ADVERTISER VH CONTENTS-Continued CURRENT LITERATURE-Continued THERAPEUTICS. Wood, Horatio C., Jr.: The Effects of Caffein on the Circulatory and Muscular Systems 318 Rogers, Leonard: The Use of Hypertonic Salines Controlled by Estimation of the Specific Gravity of the Blood in Infantile Diarrhea 319 Schamberg, J. F., and Kolmer, J. A.: The Treatment of the Vaccination Site with Picric Acid Solutions 319 BOOK REVIEWS. The International Medical Annual, 1912 Issue. A Year-book of Treatment and Practitioner's Index 320 Forty-third Annual Report of the Children's Hospital, Huntington Avenue, Boston, for the Year 1911 320 ITEMS. The Congenital Mongolian Blue Spot 249 Treatment of Whooping-cough by Fluoroform 256 Epilepsy in Children and Its Treatment with Bromide 280 Streptococcus of Scarlatina and the Reaction of Fixation 288 Presence of Diphtheria Toxins Circulating in the Blood 297 Occurrence of Adenoids 302 Concerning the Period of Incubation of Serum Sickness 311 THE ORIGINAL AND ONLY GENUINE HORLICK'S MALTED MILK Endorsed by the medical profession and the public for over twenty-five years. Has become well and favorably known as a reliable substitute for mother's milk, and as a valuable dietetic adjunct in the treatment of all diseases involving, either directly or indirectly, the intestinal tract. Its value is based not only on chemical properties, but also on the possession of certain physical attributes, e.g., palatability, solubility, ease of digestion and assimilation. Always specify " HORLICK'S," Original-Genuine Samples sent on request HORLICK'S MALTED MILK COMPANY <A°n^"wisco^ I viii ARCHIVES OF PEDIATRICS ADVERTISER Hunyadi Janos NATURAL LAXATIVE WATER. Prescribed in small doses it acts by stimulating the liver cells and removing from the ducts, retained secretions. It is of decided value in ngaStro intestinal" ailments as well as in constipation of infants and children. The average dose for children is three to four tablespoonfuls in a cup of hot milk, taken in the morning-the milk serves to hide the bitter taSte and makes it palatable and the children like it that way. <J For adults a wine glass full before breakfast. I Cj In prescribing use the full name Hunyadi Janos to avoid substitution. The Nursing Mother whose milk is deficient is decidedly helped by taking thrice daily IMPERIAL GRANUM The Unsweetened Food It provides with least tax on the digestive organs that extra pure unmedicated nourishment necessary to augment the scanty milk supply and maintain the mother's strength while nursing. Samples and literature sent on request JOHN CARLE & SONS, 153 WATER ST., NEW YORK rSANMETTO GENITO URINARY DISEASES. | A Vitalizing Tonic to the Reproductive System. * Specially Valuable in Prostatic Troubles of Old Men-Irritable Bladder- i * Cystitis-Urethritis-Pre-Senility. SOOTHING -RELIEVING -RESTORING. DOSE:-One Teaspoonful Four Times a Day. OD CHEM. CO., NEW YORK. ' ^g^Beware of the so-called Elixir Compounds claiming to be "the same thing'' s Jr or "just as good" if you do not wish to be disappointed. ARCHIVES OF PEDIATRICS ADVERTISER Is ^K^^TYPHOID > FEVER B ^Rr Fhe nourishment of the patient being pl hl of primary importance, the question of diet la needs careful attention; where Bengerised ^1 CW Milk is used, from the inception of illness pl and through the various stages of convales- |l cence, both the nourishment of the patient $1 JSl and the question of diet are at once settled. ;|l JHr Bengerised Milk is readily retained; even tf in very weakened gastric conditions, it supplies a it nourishment in a pleasant and acceptable form l|l gg to the patient the fear of tympanites is les- M sened, while the waste products of combustion II gl are reduced to a minimum. If II Dr. C. Buchanan Ker, the authority on || pl fevers, writes as follows in his text IJ book on "Infectious Diseases." I M "I find Bengers a most useful prepa- ration, and employ it perhaps more pl frequently than any of the others.'' pl Samples and Literature from Wk Bengers Food Co., Ltd. 17^^' Dept. 2 Wvn 92 William Street ( Wk V\ NEW YORK CITY \ J|» i ARCHIVES OF PEDIATRICS ADVERTISER IMPORTANT NEW BOOK The MONTESSORI METHOD Of Scientific Pedagogy, as applied to Child Education in "The Children's Houses" By MARIA MONTESSORI, M.D. With important revisions and additions by the author. Translated by Anne E. George Introduction by Prof. HENRY W. HOLMES of Harvard University A complete, authorized translation of Dr. Montessori's famous book. It expounds a new method of child education so important and original that a knowledge of it is vital to every one interested in children. Professor Holmes calls the system "remarkable, novel and important'' and says "for years no educational document has been so eagerly expected by so large a public, and and not many have better merited general anticipation.'' An inspiring book that should be read by everyone interested in human progress. It contains chapters of great special interest to physicians Nothing like a conception of the method can be gained except from the book itself. Yet, it may be said that the system is the product of years of scientific experiment, that it is based not on abstract theories, but on a study of the nature of the individual child, and that its purpose is to develope self-dependence and to encourage the growth of strong, complete human beings, physically, mentally and morally. 8vo. 437 pages With many illustrations $1.75 net; postpaid $1.90 FREDERICK A. STOKES COMPANY. Publishers. New York Send orders to E. B. TREAT & CO., 241-3 West 23d Street, New York Loef land's Malt Soup {Keller's Malt Soup) The best dietetic remedy for infants suffering from Marasmus and Atrophy. Used with the greatest success in children's hospitals here and abroad. Loeflund's Malt Extract plain, and in various suitable combinations. For over 50 years standard therapeutic agents. Used by physicians all over the world. Loeflund's Food Maltose A food of excellence for children and invalids. Ail these products BEWARE OF IMITATIONS contain no Alcohol Literature Upon Request Sole Agents for U. S. Britt, Loeffler & Weil Physicians and Hospitals 255 Canal Street, New York ARCHIVES OF PEDIATRICS ADVERTISER xi FRESH~llfTHE"MEMORYn OF OUR OLD FRIENDS WHO HAVE TESTED IT AS A BLOOD-MAKER-A TISSUE REGENERATOR-A STRENGTH > BUILDER-IS THE PLEASING RECOLLECTION THAT HAS SERVED THEM WELL. As a reconstructive Cord. Ext. 01. Morrhuae Comp. (Hagee) has hdd high, favor for many years and bids fair to find among the coming generation of physicians the same sturdy friends its merit has always won for it. Free from grease and the taste OF nsn. «-^^Suyipliedin sixteen ouncebottiei on/g. -Mipenscd b# ill druggisis. Haitiarmon Chemical Co., SiAouis.mo. Il/ ITU IFM1A11 WILL BE FOUND A MOST I KATHARMON represents in combination Hydrastis I IK ATn ADMON "°™nc a^nt >n p^ws 11 FT1 l^"lUI\'>!^!±J^!5^ ^rie&tlegA Reg. U. S. Pat. Off. (ENGLISH) "Roseberry" and Gabardine Cloths Reg. U. S. Pat. Ojff. The ideal cloths for Rain Coats, Auto Coats and Golfing, Riding, Driving, Yachting and Iramping Suits. They are of unusual strength and tailor into very graceful garments. THOROUGH RAIN-PROOF Contain no Rubber-will not overheat or cause perspiration-have no disagreeable odor. They come in a large variety of the latest English colorings in plain and fancy effects. For sale in " Ready to Wear" garments by the leading dealers in men's and women's cloth- ing-or your tailor can make you a garment to measure by securing the cloth from his woolen jobber. A postal to us will bring booklet. This circular registered This circular registered Trade Mark is stamped Trade Mark is stamped on the back of all Priestley the back of all Priestley "Cravenette" "Roseberry" "Cravenette" Gabardines. JVMVMMBLg Cloth -7\ * American ENGLAND NEW YORK xii ARCHIVES OF PEDIATRICS ADVERTISER HEALTHY CHILDREN Are the Most Valuable Asset of a Nation BORDEN'S EAGLE BRAND CONDENSED MILK PROVIDES SUITABLE INFANTILE NUTRITION FOR A STRONG AND ROBUST MATURITY Sample and Analysis Sent upon Receipt of Professional Card BORDEN'S CONDENSED MILK CO. Est. 1857 " Leaders of Quality" New York Are You Interested In PRACTICAL GYNECOLOGY? Asa progressive practitioner we know you are, and you will therefore be pleased with The MAY NUMBER of the INTERNATIONAL JOURNAL OF SURGERY The General Practitioner's Surgical Journal which will contain a Symposium on this important subject This issue alone will be worth the price of a whole year's subscription, which is only ONE DOLLAR NOW IS THE TIME TO SUBSCRIBE ADDRESS: INTERNATIONAL JOURNAL OF SURGERY COMPANY 100 William Street, New York City ARCHIVES OF PEDIATRICS ADVERTISER xiii A SELECTED LIST OF NEW AND ■ 1 Practical Medical Books jOh 1912 International Medical An- At W^BB&: JMu nual. A review of all Medical Aj vM ■ books and journals for the past year. 7<'>2 pages, fully illustrated $3.50 , W JsjlL llllj Spondy lotherapy. Abrams. ^7^ * Third Edition, 698 pages, and 146 illustrations 5.00 ^1018 Landmarks and Surface Mark- J. ings of the Human Body. N, > Rawling. Fifth Edition. Color- I7 ed plates 2.00 Diagnosis of Nervous Diseases. II Purves Stewart. Third Edi- tion, 500 pages, 225 illustrations 4.50 . ■ ' , ■ Diseases of the Stomach and In- testines. Boardman Ri ed, |M^EW|^^^^^^^B Third Edition, 1037 pages, fully || HV IM illustrated 5.00 I Surgical Diseases of Children. Kelley. 768 pages, 300 illus- IN ■ ■ N MkJ ill B 1^1 lM 5.00 Blood Pressure and the Nau- | heim Treatment. Bishop 3.00 Il flVA EE Bl * B >4 ■ W* B Bl Inanition and Fattening Cures. vonnoorden 1.50 M■! Handbook of Cosmetics. Joseph 1.00 |M Lectures on CosmeticTreatment. Saalfeld 1.75 The Blues (Nerve Exhaustion). Abrams. Fourth Edition 1.50 Books sent prepaid. Medical catalog, with full descriptions, on request " 7^ 't-i-r-r , . f/t E. B. TREAT & CO., Medical Publishers ||RBMHHBB 241-243 West 23d Street, New York KHMMIHiHBnHHBHHBHHHBHB xiv ARCHIVES OF PEDIATRICS ADVERTISER For Thirty Years Vaporized Cresolene £ has held its position as a valuable remedy for the bronchial diseases of childhood. It is particularly useful in the treatment of the very young. Cresolene is indicated in Whooping Cough, Croup, Bronchitis, Asthma, . gj riggnJH Coughs and the bronchial complications incident to Scarlet Fever and U , VoPjJHM Measles. Vaporized Cresolene is destructive to Diphtheria bacilli and may be Hl " advantageously used in connection with the treatment of this disease. Eg? £ 4 Let us send you our descriptive and test booklet which ^^H gives liberal sample offer. Tur xz a rnrcm rMf Streets NEW \ ORK THE V Ar O-CKEdOLEIN E CU., Leeming-Mlle* Building. Montreal, Canada IN WHOOPING COUGH GOMENOL SYRUP acts promptly and effectively in the great majority of cases. Any druggist can obtain from his wholesaler CHARLES R. BARD, Sole Agent 37 East Twenty-Eighth Street, New York The Control of Fever Quilted I^attYPSS Paas occurring in the course of colds, Grippe A VlttU attacks, malaria and similar affections is Tuner crnor j tcm vcadc • often one of the principal indications for 1 rirxmt. ^GUKE, and IrJN ILAKc is a .. treatment. For this purpose . long life, yet about one third of it is spent w in bed. Then why not make your own bed f-- ... ... , _ as comfortable as can be made? gHTHgnwni Quilted Mattress Pads gi will not only make it • comfortable, but as is exceptionally serviceable inasmuch as its fg/ they are spread over administration is followed by the most grati- gf ^//z\ \a \n\ the mattre« thev will fying results. A few doses suffice not only r//^Fn\ mattress, they will to promote a free perspiration promptly llxT'1''' I Protect '*• anc* w"' followed by a lowering of the temperature, I I keep your bed or but the headache, myalgia, pain and general \ \ / / babv's crib in a ner soreness are also markedly relieved. \ / ^aby s ,crlb m a Per' Clinical experience has proven beyond all \ zXf bl " / *ectsamtarycondition, doubt that Phenalgin is the ideal analgesic and antipyretic. Remarkably free from all Examine closely stitch- depressing tendencies, its use assures the .... . ,lL . ing on our pads and see relief of pain without producing constipa- IVora genuine without that sizes correspond with tion, upsetting the stomach or leading to I rade-Marii size on ^cket. Seconds, habituation. Countless physicians employ it damaged or "Just as good" pads are sold under other in preference to all other anodynes, because labels. Insiston it serves them best. EXCELSIOR QUILTED MATTRESS PADS Samples on request See that our trade-mark is on each piece of goods you buy THE ETNA CHEMICAL CO. rVfriCIAD mill TINf m 708 Washington St., New York. N. Y. tAttLSIOK QU1L11NG CO. \ J 18 Laight Street NEW YORK CITY ARCHIVES OF PEDIATRICS ADVERTISER xv ' LAIBOSE' is a Food Wherever there is trouble with the feeding, or where it is im- portant to avoid trouble, 'Laibose' is a promising resource for nutrition. 'Laibose' is a milk-and-wheat food; contains the solids of pure whole milk and the entire digestible substance of whole wheat-no fibre, no cellulose-the wheat converted into a soluble state by a physiological process without chemical action. 'Laibose' is one of the most complete and valuable of foods, a product which has only been possible by original scientific laboratory methods. It is a dry food made ready for use by simple dilution with water, the ration thus readily adapted to individual needs and the quantity of fluid in the diet restricted at will. 'Laibose' was originated and is made by Fairchild Bros. & Foster New York Monosodium-Diethyl-Barbituric Acid Brom-Isovaleric-Acid-Borneolester A freely soluble hypnotic for Combines the action of vale- use by mouth, by rectum and rian with that of bromine, but is subcutaneously. Being read- readily taken and well borne, ily absorbed and rapidly ex- causing no eructation or other creted, it is distinguished by untoward symptoms. Exhibits PROMPT AND RELIABLE VIGOROUS SEDATIVE AND SOPORIFIC EFFECT NERVINE EFFECT FREEDOM FROM CUMULA- INNOCUOUSNESS EVEN IN T1VE TOXIC ACTIONS LARGE DOSES Superior to the sparingly soluble Decidedly effective in neuras- diethyl-barbituric acid of Mering. thenic and hysterical conditions, Advantageously replaces chloral in obviating subjective difficulties- threatening delirium tremens; use- mental and physical fatigue, head- ful in the treatment of morphinism. ache, nervousness, insomnia, etc. Dose : 5 to 15 grains (1 V* 3 tablets) Dose : x to 3 pearls several times daily Literature from SCHERING & GLATZ - - New York xvi ARCHIVES OF PEDIATRICS ADVERTISER j ..I i is indicated in Rickets, Anemia, Chlorosis, Leciinoi Tuberculosis and nerve disorders. pi. . TT is an aid to digestion and a good Elixir 01 Enzymes veh icle for the iodids, brom ids, etc. r . . r n in iui is recommended in Extract ot Ked Bone Marrow , _ cases where an increase in hemoglobin and red corpuscles is desired. Thyroid, Thymus, Spleen, Suprarenal, Pituitary, Para- thyroid and other substances under our label are pure and are made from fresh material. armour accompany 3 GASTRIC DEBILITY"^- | MOST FORMS OF DYSPEPSIA ARE TRACEABLE TO FATIGUE AND WEAKNESS OF THE STOMACH MUSCLES. ■ THIS IS WHY I Eran's EliicErinETnniECnmp. p ACCOMPLISHES SO MUCH IN THIS CLASS OF CASES, FOR IT PROMPTLY INCREASES THE ACTIVITY OF THE GASTRO-INTESTINAL MUSCLES, AND THEREBY : NOT ONLY RESTORES DIGESTIVE FUNCTIONS, BUT REMOVES DISTRESSING SYMPTOMS. If TWO TO FOUR TEASPOONFULS IN WATER AFTER MEALS AUGMENTS THE DIGESTIVE CAPACITY TO A MARKED DEGREE. JF THE PURDUE FREDERICK CO. BROADWAY, NEW YORK.^B^I ARCHIVES OF PEDIATRICS ADVERTISER xvii MEAD'S DEXTRI-MALTOSE (MALT SUGAR) A substance resulting from the action of diastase upon starch. A pure form of carbohydrate, free from proteid, cellulose and fat. Wholly soluble in milk or water. Advantages in Infant Feeding Mead's Dextri-Maltose may be taken by the infant in larger quantities than cane or milk sugar, without causing diarrhoea and with greatly decreased chances of sugar fermentation. Being more readily assimil- able and oxidizable, it increases weight and energy to a greater extent than other sugars. Its Assimilation Limit About 100% Higher Than Either Cane or Milk Sugar The assimilation limits of the sugars as demonstrated by Reuss, Grosz, et. al., are: Malt sugar 7.7 grams per kilogram of body weight. Either milk or cane sugar, 3.1 grams per kilogram of body weight. Mead's Dextri-Maltose is made and marketed by MEAD, JOHNSON & CO. Jersey City, N. J. oHnasarcin Oxydendron Arboreum (Sour Wood) Sambucus Canadensis (Elder) and Urginea Scilla (Squill) Eliminates Effused Serum Controls heart action and restores physiological balance between arterial and venous systems. Thousands of Physicians are continually using (^nasarcin with most gratifying results in all dropsical effusions whether caused by disease of the heart, liver or kidneys. TRY offnosarcin in any obstinate case of Dropsy - then judge its merits. Clinical results prove Therapeutics. tawXS'to 7^ ANASARCIN CHEMICAL CO. physicians only. WINCHESTER, TENN. Messrs. Thomas Christy & Co., London Agents. Vlotes and iRewg. Motherhood and Infancy.-This is the most unique , at- tractive and instructive little treatise yet published on this subject. You should give a copy to every patron who is raising a family. They are very beneficial to both patient and physician. Address the author, Dr. E. S. Harris, Box 722, Blue Springs, Mo., enclosing 10 cents, and he will send you a copy. They are fine. Fulfills the Standard of " Hygienic Merit."-In the days of Queen Elizabeth the leading scientist of that time, a Dr. Priestley, made the first reference we know of to rubber, in a lecture he delivered on a new and curious substance that returning voyagers had brought to England from some unknown island, where they had found the natives using rubber for head coverings and as a ball in natives games. Experiments followed to discover uses for rubber, but it was not until the early part of the 18th Century that a man by the name of McIntosh discovered the rubber process that was developed in the garments bearing his name. These coats, while rainproof, were hot, smelly, uncomfortable and unhealthy, and so did not long hold the public's fancy. By a strange coincidence a descend- ant of that same Dr. Priestley discovered the Cravenette Proof, which makes all fabrics rainproof without the use of rubber, and so the Cravenette coat became at once a necessity for well-dressed men and women. The merits of a garment that is rainproof, yet not air- tight and which does not compel its wearer to undergo a Turkish Bath every time it is worn, have been so recognized that last year the "London Institute of Hygiene" awarded the certificate of merit to The Bradford Dyers' Association, Ltd., of Bradford, England, for fulfilling the standard of Hygienic Merit. So popular has the word Cravenette become that it is now given in the New Webster's Dictionary. After Scarlet Fever and Measles.-After the acute dis- eases of childhood there is no remedy that will do more to hasten con- valescence than Gray's Glycerine Tonic Comp. Children are par- ticularly responsive to the tonic effects of "Gray's" and it is always gratifying to see the prompt improvement in the appetite, digestion and general nutrition that follows its administration. The palatabil- ity and clean bitter taste of "Gray's" make it exceptionally acceptable to children. The Approach of Summer with its attendant gastro-in- testinal troubles brings with it the necessity of weaning and fortify- ing baby against the dangers of the hot weather. Weaning is most satisfactory when it is gradually accomplished. When age permits, now is the best time to begin by accustoming baby to one or two feed- ings daily of Imperial Granum. Milk which is modified by Imperial Granum, is the most efficient food. It makes the milk more digest- ible, more nutritious and provides the necessary salts and phosphates in their natural combinations. Practitioners wishing to test Imperial Granum for infant feeding or for the mother who is nursing, may obtain samples gratuitously by Addressing John Carle & Sons, Water Street and Maiden Lane, New York City. In Convalescence from Febrile Disorders. Seng imparts a desirable tonicity to the alimentary structures, coaxing back the vital functions that have been sadly exhausted and depressed. Digestion, absorption and assimilation are substantially promoted, and gratify- ing improvement in the whole bodily condition follows as a natural sequence. ARCHIVES OF PEDIATRICS ADVERTISER xix Opium in the fullness of its therapeutic excellence Opium freed from all inert, useless stuff, exhibiting the total alka- loids of the drug as a unit, in their naturally-occurring proportions, in a water-soluble, non-irritating, injectable, diffusible form f p generally without constipating or other by-effects. Provided in Powder, Tablets (for hypo. Brilliant results in U or ora* use) and Ampoules- Internal Medicine, U Sample and literature Gynecology and Ob- I on reQuest. stetrics, Anesthesia and Psychiatry are re- e»row^ corded in the clinical re- THE HOFFMANN-LAROCHE ports of Sahli, Ewald, - Chemical. Works Grafenberg, Leipoldt and other well- Of 65 Fulton Street, New York known writers. HEMATIC INSUFFICIENCY -■1 I 1 *~~ -not only in physical or corpuscular elements but in certain indefinable chemico-physio- logic properties-is now known to be the all-important factor in the development and pro- gress of countless human ills. Their treatment to be effective, therefore, must be able not only to increase the so-called blood count, but what is of far greater importance to raise the vital index or protective properties of the blood. Unquestionably it is the efficiency of ECTHOL in this direction that accounts for its remarkable therapeutic activity in many of the acute affections such as erysipelas, tonsillitis, puerperal fever, phlegmasia alba dolens, typhoid fever, adenitis, septicemia, scarlet fever and so on. Clinical experience shows that Ecthol-a combination of Echinacea Angustifolia and Thuja Occidentalis-possesses the power when introduced into the living organism of markedly augmenting the protective, restorative and reparative properties of the blood. It is broadly indicated, therefore, sn all forms of blood dyscrasia and wherever an anti- septic, anti-suppurative and anti-morbific remedy is required. The prompt benefits that attend its use furnish its most eloquent testimonial. BATTLE & CO. LONDON ST. LOUIS PARIS XX NOTES AND NEWS. Children should not be allowed to drink Tea or Coffee on account of the disturbing action on the nerves and heart of the alkaloid caffeine. Physicians well know this, but parents either do not or are neglectful of their children's best interest in this regard. Postum, the well-known food beverage, contains no coffee, only the wholesome elements of clean, hard wheat, roasted, and a small per cent, of pure molasses. Children may drink freely of Postum, which, with good cream, affords them an agreeable, warm drink, healthful and nourishing. Let the children have all they want of Postum while their young bodies are growing and their nervous sys- tems are getting "settled." Meningo-Bacterin.-Bacterin therapy is long past the experi- mental stage, and the immunizing effect of typho-bacterin, for instance, is thoroughly established, the results from its use being sufficient evidence of the worth of this method of controlling the spread of typhoid fever. Remarkable results likewise have followed the use of cholera-bacterin, and it is hoped that equally good results will follow the use of meningo-bacterin in controlling epidemics of cerebrospinal meningitis. While immunization with Meningo- Bacterin has thus far been used in relatively few cases it is entirely leasonable to believe that it will prove a most valuable aid in the suppression of epidemics of cerebrospinal meningitis. The usual site for inoculation is the arm at about the insertion of the deltoid muscle. The dose is given subcutaneously and not into the muscle nor into the skin. An area about the size of a five-cent piece is painted with tinc- ture of iodine. The syringe needle is plunged through this area. No after treatment is necessary. The complete immunization treatment consists of three doses given at intervals of from five to ten days. The first dose is 500 million, the second dose 1,000 million, and the third dose, 1,000 million. For children smaller doses should be used according to weight. It has been suggested that the unit of body- weight for a full dose be considered 150 pounds. The H. K. Mul- ford Company, who prepare Meningo-Bacterin can also supply Anti- Meningitis Serum, prepared after the method of Flexner and Jobling, and will mail, upon request to the Philadelphia Office, Mulford Working Bulletin No. 8, on Anti-Meningitis Serum, giving a de- tailed and impartial review of the literature. Pneumonia. -Last winter a little eighteen months old boy baby, suffering from pneumonia, was rapidly growing worse under the usual camphorated oil jacket treatment. Breathing continued fast and labored, temperature high, and the usual attending nervous symptoms were alarming. I changed to Antiphlogistine, and was surprised on returning the following morning to find a decided im- provement in the patient's condition. The nervous symptoms had abated, the temperature had fallen, and the patient's breathing was much more comfortable. This change was not due to crisis as the disease ended by lysis. In my opinion Antiphlogistine was the sheet anchor in the case, and the patient's life was saved by its use.-Case reported by E. Clinton Murray, M.D., Houston, Texas. An Important Merger.-The American Practitioner, a new medical journal incorporating the American Practitioner and News, formerly published in Louisville, Ky., and the New England Medi- cal Monthly with Annals of Medical Practice, of Boston, Mass., is being issued from this city. It is edited by John W. Wainwright, M.D., 80 Washington Square, East, New York City. It is a month- ly, subscription price for which is $1 per year. ARCHIVES OF PEDIATRICS ADVERTISER xxl "in the treatment of muscular cramps, hot applications according to S. Solis Cohen, lessen the excitability and energy of the voluntary muscles and relieve the excru- ciating pains accompanying these attacks. In neuralgias as shown by Du Bois Raymond, where there is compression of the nerve trunk by the over filling of blood vessels in contiguous areas, hot applications relieve the congestion and pain by diver- sion of the blood to other parts. In the use of hot moist heat as a thera- peutic agent in the treatment of muscular spasms and neuralgias, antiphlogistine seems to be particularly indicated. It relieves muscular tension, stimulates capil- lary and arterial circulation, thus removing the congestion and its accompanying pain. Antiphlogistine is the most convenient, sanitary and satisfactory method of apply- ing hot moist heat. It retains its thermic value for hours, a feature of recognized importance in the treatment of inflamma- tory and congestive conditions." xxiv ARCHIVES OF PEDIATRICS ADVERTISER Two New Works By Prof. DR. CARL von NOORDEN Professor of the First Medical Clinic, Vienna, in his series on the Pathology and Therapy of the Disorders of METABOLISM AND NUTRITION Edited and Translated under the Supervision of Alfred C. Croftan, M.D., Chicago J VIII. Inanition and Fattening Cures N„°±er $1.50 The author's criticism of innumerable dietetic fads and fallacies is of the greatest practical use, as he provides something of value in their stead IX. Technique af Reduction Cures and Gout N„°ler $1.50 The practical dietetics and general treatment of the subjects are given With great exactness and make very interesting and instructive reading OTHER MONOGRAPHS IN THE SERIES I. Obesity $0.50 V. Saline Therapy $0.75 II. Nephritis 1.00 ✓!. Drink Restriction 75 III. Colitis 50 VII. Diabetes Mellitus 1.50 IV. The Acid Autointoxications .5° Others in Preparation Complete set (9 volumes) $8.50, prepaid. Also sold singly E. B. TREAT & CO., Medical Publishers, 241-243 West 23 d Street, NEW YORK lesiA £4^ wJ Br isliir ■ 4 Why Take a Chance? Pills tablets and gelatine capsules are often insoluble. Powders "in Konseals" give sure and quick results, for Konseals are always soluble and tasteless. The U. S. Dispensatory (p. 1118) says: For most powders of disagreeable taste, cachets afford the best method of administration, - Konseals are the best cachets. - If you write for Konseals see that you get Konseals; if you don't, investigate, for substitutors in little things are not de- pendable when it comes to the more expensive medicaments. For literature and samples, write J. M. Grosvenor & Co., Boston, Mass. ARCHIVES OF PEDIATRICS ADVERTISER xxv MENINGO-BACTERIN ANNOUNCEMENT w? are prepared to supply Meningo- Bacterin (Meningococcus Vaccine) for immunization against cerebrospinal menin- gitis. The immunization treatment consists of three doses given at intervals of from five to ten days. First dose contains 500 million killed bacteria Second dose contains 1000 million killed bacteria Third dose contains 1000 million killed bacteria Meningo-Bacterin is supplied as follows: Single immunizing package containing 3 doses (each dose is a separate aseptic glass syringe). No. 5, Hospital and Board of Health pack- age, containing 30 doses, each in a separate ampul, sufficient for 10 immunizations. We also prepare Anti-Meningitis Serum, after the method of Flexner and Jobling, for the treatment of cerebrospinal meningitis. Anti-Meningitis Serum is supplied in pack- ages containing 2 syringes of 15 c.c. each, with a special needle for intradural injection. Working Bulletins Mailed Free upon Request H. K. Mulford Company (chemists) New York Chicago Philadelphia St. Louis San Francisco Minneapolis Kansas City Atlanta Seattle Toronto Dallas Boston xxvi ARCHIVES OF PEDIATRICS ADVERTISER PUBLISHED APRIL 15 No Practitioner Can Afford to be Without It The International Medical Annual for 1912. 30th Year of Publication. Price, $3.50 The most complete review of the entire field of medicine. The reviews in the Annual are compact and exact, yet full enough to be of use without re- ferring to the original article, which, however, is credited and given as reference. This Annual has the great advantage over the ordinary year-book in that it covers the most valuable productions of the year without being too extensive for practical use.-Interstate Medical Journal, St. Louis. The merit of the book will sustain its long enduring popularity and the man who has fewest books needs some such work more than the man with a large and constantly leplenished library. No other single volume at anything like the price will keep the practitian so thoroughly abreast of all that is new in the various branches of medicine.-Medical World, Philadelphia. The articles are carefully prepared by physicians thoroughly acquainted with the subjects discussed by them. It is one of the best books in English covering the progress made in all branches of medicine during the past year.-Bulletin oj the Johns Hopkins Hospital, Baltimore. It is a valuable aid to the busy practitioner and even to the one who has kept closely in touch with current literature, it provides an admirable review and the means of connecting many scattered threads of information.-Medical Record. There is no single volume Annual published anywhere near its equal. We have been commending this most excellent work for years and while other Annuals have come and gone, this one has weathered the storm of twenty-nine seasons and it seems that each succeeding volume has been an improvement over the preceding one.-Medical Summary, Philadelphia. There is no better compend of the year's progress in medicine than the Annual. This we unhesitatingly affirm. The arrangement is the acme of sim- plicity and convenience. Its contributors are men of unquestioned authority and sane conservatism, and its contents is precisely what it purports to be.-Medical Standard, Chicago. ORDER FORM 79/ Please send me, as soon as ready, all charges prepaid, The International Medical Annual for 1912 30TH YEAR OF PUBLICATION Enclosed find $3.50 Send C.O.D. $3.50 Mame . To E. B. Treat &, Company Street 241-243 WEST 23D STREET NEW YORK CITY Town ARCHIVES OF PEDIATRICS ADVERTISER xxvii ! 111 ly BWi io i^i i A Preparation of the Active Principle of the Thyroid Gland. This product consists of certain proteids of normal thyroid glands, extracted, purified, assayed and adjusted to a standard content of 0.33 per cent, of iodine. It is diluted with the necessary amount of lactose to make two-grain tablets, which we supply in three denominations, containing, respectively, 1, 2 and 5 per cent, of Thyro- protein. The 1- and 2-per-cent. tablets are used in the treatment of various types of goitre, the average dose being one tablet three times daily. The 5-per-cent. tablets are intended for administration in metabolic disorders, as skin diseases, affections of the joints, myxedema and cretinism. Thyroprotein has been employed for upward of two years by a number of experi- enced clinicians and in a wide variety of pathologic conditions. It is offered to the medical profession with full confidence that it will satisfactorily displace the crude and more uncertain thyroid preparations heretofore available. Supplied in 2-grain tablets, bottles of 50. Three strengths: 1%, 2% and 5% of Thyroprotein. FULL DESCRIPTIVE LITERATURE ON REQUEST. Home Offices and Laboratories, DADV^C FA AX7TQ JP Detroit, Michigan. r /A1V1S.1L, DA V 13 Gt LU. IfBii i i 11 For the Treatment of Bacterial Infections. Acne Vaccine (Acne Bacterin).-For non-pustular acne due to the Bacillus acne. Colon Vaccine (Colon Bacterin).-For colon infections. Combined Vaccine (Van Cott).-For erysipelas, puerperal sepsis, etc. Furunculosis Vaccine.-For boils, carbuncles, pustular acne, etc. Gonococcus Vaccine (Gonococcus Bacterin).-For acute gonorrhea and its complications. Gonorrheal Vaccine, Combined (Gonorrheal Bacterin, Combined).-For gonorrheal infections complicated by the presence of staphylococci. Staphylococcus Vaccine (Albus) (Staphylococcus Albus Bacterin). Staphylococcus Vaccine (Aureus) (Staphylococcus Aureus Bacterin). Staphylococcus Vaccine (Citreus) Staphylococcus Citreus Bacterin). Staphylococcus Vaccine, Combined (Staphylococcus Bacterin, Combined). - For fu- runculosis and carbuncle, sycosis, suppurative acne, etc. Streptococcus Vaccine (Streptococcus Bacterin).-For erysipelas, puerperal septice- mia, cellulitis, etc. Typhoid Vaccine (Prophylactic).-For preventive inoculation only. Supplied in 1 Cc. rubber-stoppered glass bulbs and in 1 Cc. syringe containers. HANDSOMELY ILLUSTRATED BOOKLET FREZ. Home Offices and Laboratories, D A DI/T TA A X7IQ JP /^/'A Detroit, Michigan. A /AIXAlIIi, DA V IO Gt VW. xxviii ARCHIVES OF PEDIATRICS ADVERTISER fl l ^^In the treatment^f fl I EPILEPSY I CHRONIC I no remedy gives such uniformly sat- $ vT/\^ I IV I I I!% g isfactory results as $ g PFACOCK'S with its train of hepatic and intesti- S x o nal complications, rarely fails to BROMIDES resjwid^at once to the beneficial | This is due, first to the happy > y JT/'WTT A H B ' combination of several different bro- VjlllvUvlxX. S B j mide salts; second, to the remark- B B | able chemical purity of each salt; . Although exerting a pronounced B B and finally, to the pharmaceutical 5 influence on the secretory structures 11.| Eg = . I character of the composite product. < of thestomachand liver, Chionia has I n = I It is easy to understand, there- ; the special advantage of increasing I. S = 11 I fore, with their purity and potency < the physiologic activity of the gastro- H । = । I so marked, how Peacock's Bromides < intestinal and hepatic functions = lin have been able to accomplish such $ without producing severe or undue H = ||| pronounced therapeutic effects in > catharsis. It is in consequence, the E I | epilepsy and kindred ailments, even 5 ideal remedy in all forms of hepatic E || when administered over long periods. | torpidity. r [PEACOCK CHEMICAL CO.. St. Louss, Mo. L I i I MB Free from PRllAIAln^Produce W Cathartic | |\ L?/ I g LZ 1 Permanent Iniquities ■ Benefits JW Br OVERCOME CONSTIPATION . W |» - rtf J Free from CACTI/1A Sustain and Cumulative Dll I E* T" C Regulate Effect r I L !■ t I the Heart TONE THE CARDIAC MUSCLE w A Promotes IF Digestive ■■ g ■ Natural Tonic Processes Aynu W INCREASES FUNCTIONAL ACTIVITY fcr" Mg J-SULTAN DRUG CO.. 5T. Louis. Moji ARCHIVES OF PEDIATRICS ADVERTISER xxix A crystal, liquid Antiseptic, slightly Alkaline. UNEXCELLED in the treatment of Catarrh, Eczema, Pruritus and itching so frequently affecting the aged, trial bottle by mail, on request. _ _ NEURILLA FOR nerve disorders NEURILLA - If Patient suffers fromTHE BLUES (Nerve Exhaustion), Nervous Insomnia,Nervous Headache,Irritability or General Nervousness,£ive four times a day one *■■■■■■■■■■ teaspoonful NEURILLA i In nervous fretfulness of teething Children WMSSKHH^Ml give: five to twenty drops. ■ DAD CHEMICAL COMPANY, NEW YORK and PARIS.. BOOKS ON COSMETICS HAND BOOK OF LECTURES ON Cosmetics Cosmetic Treatment By DR. MAX JOSEPH, Berlin, Authorized By DR. EDMUND SAALFELD, Berlin, Au- English Translation, Revised from the Third thorized English Translation from Second German Edition German Edition 12mo, 151 Recipes, Cloth, Prepaid, $1.00 202 Pp., 131 Recipes, Illas., Prepaid, $1.75 E. B. TREAT & COMPANY, 241-243 West 23d Street, New Yorkf JUST PUBLISHED These trademark cqss-cross lines on eyery pacKage HI IIT&O <8 F0R THE DIAGNOSIS OF ULU I Ml rUJU|fDIABETICS yvpcabsQr 1\T FA. Kidney and Liver lAWilt^weumatism, Obesity INervous Diseases and ills a^^ng fryf^cce^^^Vric Acid Rich in Proteu^r Askp^sician^d-eading grocers. By Purves Stewart, M.D., Edin., F.R.C.P. < W fl R V Physician to Out-Patients, Westminster Hospital; FARWELL & RIHNES, 3 C OWH, . ., Physician to the West End Hospital for Nervous Diseases This volume approaches the subject GASTROGEN «S®! of diagnosis of nervous diseases from the clinical standpoint. The author' TABLETS has the happy gift of giving the gist of a matter in a few pregnant sentences A Neutralizing Digestive and of seeing clearly what is wanted, I and consequently of being singularly sample and formula lucid in expression. mailed to physicians x upon request. ! ANeutrauzingDigestive | Third Edition, Revised and Enlarged, 8vo, about 500 BRISTOL-MYERS CO ""''"tom's otR'°US pages, 225 illus. and colored plates. Prepaid, $4.50 ' rn^Wfr^TWl ' 277-281 Greene Avenue ™ YORK. ]|j E. B. TREAT & COMPANY Brooklyn - New York 241-243 West 23d Street, New York City U. S. A. - XXX ARCHIVES OF PEDIATRICS ADVERTISER JUST ISSUED THIRD EDITION Spondylotherapy Physio-Therapy of the Spine Based on a Study of Clinical Physiology By ALBERT ABRAMS, M.D. (Heidelberg), F.R.M.S. Consulting Physician to the Mount Zion and French Hospitals, San Francisco; formerly Professor of Pathology and Director of the Medical Clinic, Cooper Medical College, San Francisco The subject of spinal therapeutics has received less attention from the medical profession than it deserves, although others, less scientific but more astute, have determined empirically that manipulation of the spine does sometimes cure conditions that have failed of cure in the hands of experienced physicians. The author has endeavored to put spinal therapeutics in a place befitting its scientific importance, and to emphasize its great practical helpfulness in disease. When the first edition of this book was published, nearly two years ago, it was a pioneer effort and only the cognoscenti could correctly interpret its real significance, viz., that spondylotherapy was suggested by the study of human physiology on the principle that "The proper study of mankind is-man." After this manner, clinical physiology is made the basis of clinical pathology. To launch an innovation in medicine, with its surfeit of theories and theorists, is fraught with much risk to the innovator, and the author anticipated the usual fate accorded to the originator, viz., condemnation, discussion, and possibly acceptance. Neither fear of difficulty, nor adverse criticism, deterred him from regarding scepticism as an argument against the truth of his observations. But the favorable reception accorded to previous editions has induced him to under- take the enlargement of this work by the addition of seven chapters and forty-nine new illustrations.-From Author's Preface to the Third Edition. CHAPTERS ADDED TO THIRD ADDITION For synopsis of other chapters see reverse side of this circular CHAPTER XII. The Reflexes and the Peripheral Symptomatology of Visceral Disease. CHAPTER XIII Eleven Illustrations Tonus of the Vagus and Pharmacology of the Reflexes CHAPTER XIV Nine Illustrations Further Advances in the Diagnosis and Treatment of Diseases of the Circulatory System. Sixteen Illustrations CHAPTER XV. Further Advances in the Diagnosis of Diseases of the Digestive System. CHAPTER XVI Six Illustrations Physio-Therapy of Pulmonary Tuberculosis CHAPTER XVII Five Illustrations Treatment of Whooping Cough One illustration CHAPTER XVIII. Miscellaneous Data One Illustration Third Edition. Revised and Enlarged. Octavo, 698 Pages, / 46 Illustrations, Cloth, Prepaid, $5.00 E. B. TREAT & CO., Medical Publishers .... 241-243 West 23d Street, NEW YORK ARCHIVES OF PEDIATRICS ADVERTISER xxxi CREOSOTE is an old remedy, It's value is 111(111111^^^^ recognized by all, yet it is little used because of the difficulties encountered in its adminis- tration. We have perfected a creosote that overcomes all difficulties. This perfected ■ J Al W product is a chemical combination of creosote ■ with calcium. It is a reddish-brown granular powder and contains approximately 50 per pure beechwood creosote. It might properly be ' called SOLUBLE CREOSOTE since it mixes *1 fl * I in all proportions with water. W M It is a remarkable remedy, not only in tuber- culosis and bronchitis, but in coughs and colds, W I pneumonia, typhoid fever, indigestion, diar- p' B MBE®2E5iAjyi«l|£3£»yHH ■ rhoea, &c. - A I'ublLlied reports of result- obtained from h this product seem exaggerated until you have opportunity to observe them, then the skeptic 1M| becomes an enthusiast. । . PRICE LIST. gtO CALCIUM CREOSOTE POWDER, (MALTBIE). Pre- pared by simply adding- water. Per pound, sufficient ;; zwIbwI for one gallon solution. $1 50 (Bv express prepaid. 51.75) m? ' CALCIUM CREOSOTE TABLETS. (MALTBIE , z x chocolate coated representing one teaspoonful solu- tion, per 1000, $2.00. (By express prepaid, $2.25.) I 1 _ ■■■■■■■■■■I Wir1 hmf-u;ij1,;i[i, rj। British Journal of Children's Diseases MONTHLY, 2/- net Annual Subscription, 20/- net, $5, post free ADLARD & SON, Bartholomew Close, LONDON, ENGLAND Agents Wanted SAL HEPATICA We solicit the careful considera- !tn*~3^ 'W^- * --pH to introduce our line of new tion of the physicians to the merits j j j. i i i of Sal Hepatica in the treatment and standard medical books Of Rheumatism, in Constipation I! f 1 • T •. and Auto-intoxication, and to its T»l| iLT fl nW ExclllSlVe 1 emtory highly important property of '> I ' cleansing the entire alimentary 11 Quick Sales tract'thereby eliminatinsand pre- j; pure combination] venting the absorption of irritating i I lAXAT|vt«tUMlN*NT I I toxins and relieving the conditions I mTil.. ,_.... .IJT! I Good Profits arising from indiscretion in eating I 8*~*l 11',1S; UlUrj ■ and drinking. | • 'T I ii , , , „ , ,. , Write for free sample. m Write for full particulars (jnp*»ea<x»«««a^^ E. B. TREAT & CO., Medical Publishers BRISTOL-MYERS CO. 241-243 West 23d Street : New York Manufacturing Chemists ^_J 277-281 Greene Avenue, Brooklyn, New York, U.S.A. xxxii ARCHIVES OF PEDIATRICS ADVERTISER Quinine is the sulphuric acid salt of the base quinine. Euquinine is the ethylcarbonic acid salt of the same base. Therapeutically, therefore, they are identical; but ELviqvinine is Children's quinine particularly, for the reasons that: It IS tasteless. It does not disturb the stomach. It is better than chocolate tablets. It is more reliable than pills or capsules. Samples and Literature on application to MERCK CO. New-York Rheumatism ) K I >* I Diplosalsii Div. in Chart. No. viii. Sig.: One powder 4 to 6 times per day. The action of DIPLOSAL, compared with the other derivatives of Salicylic Acid, has been shown to be most prompt and efficient, and free from the usual after-effects of Salicylic Acid. Write for Clinical Reports. MERCK & CO.. N.Y.