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diff --git a/34510.txt b/34510.txt new file mode 100644 index 0000000..ff6adda --- /dev/null +++ b/34510.txt @@ -0,0 +1,1941 @@ +The Project Gutenberg EBook of Some Constituents of the Poison Ivy Plant: +(Rhus Toxicodendron), by William Anderson Syme + +This eBook is for the use of anyone anywhere at no cost and with +almost no restrictions whatsoever. You may copy it, give it away or +re-use it under the terms of the Project Gutenberg License included +with this eBook or online at www.gutenberg.org + + +Title: Some Constituents of the Poison Ivy Plant: (Rhus Toxicodendron) + +Author: William Anderson Syme + +Release Date: November 30, 2010 [EBook #34510] + +Language: English + +Character set encoding: ASCII + +*** START OF THIS PROJECT GUTENBERG EBOOK SOME CONSTITUENTS--POISON IVY PLANT *** + + + + +Produced by Bryan Ness, Josephine Paolucci and the Online +Distributed Proofreading Team at https://www.pgdp.net. (This +book was produced from scanned images of public domain +material from the Google Print project.) + + + + + + + + +SOME CONSTITUENTS OF THE POISON IVY PLANT (RHUS TOXICODENDRON) + +DISSERTATION + +SUBMITTED TO THE BOARD OF UNIVERSITY STUDIES OF THE JOHNS HOPKINS +UNIVERSITY IN CONFORMITY WITH THE REQUIREMENTS FOR THE DEGREE OF DOCTOR +OF PHILOSOPHY + +BY WILLIAM ANDERSON SYME + +1906 + +1906 +THE SUN JOB PRINTING OFFICE +BALTIMORE + +Transcriber's Note: Underscores around words indicates italics while an +underscore and curly brackets in an equation indicates a subscript. + + + + +CONTENTS. + + +Acknowledgments 4 + +Literature 5 + +Introduction 7 + +Work of Khittel 11 + +Work of Maisch 12 + +Work of Pfaff 13 + +Experimental 14 + + Gallic Acid 18 + + Fisetin 20 + + Rhamnose 23 + + The Poison 28 + + Potassium Permanganate as a Remedy for Rhus Poisoning 35 + +Summary 37 + +Biography 38 + + + + +ACKNOWLEDGMENTS. + + +The author desires to avail himself of this opportunity to tender his +thanks to those under whose guidance he has worked while a student at +the Johns Hopkins University, namely to Professors Remsen, Morse, Jones, +and Andrews, and to Doctors Acree and Tingle for instruction in lecture +room and laboratory. + +He is especially indebted to Dr. S. F. Acree, at whose suggestion this +research work was undertaken, for counsel and assistance in its +prosecution. + +He would also thank Messrs. Parke, Davis and Co., of Detroit, Mich., for +the preparation of the crude material used in this investigation, and +the U. S. Department of Agriculture, Washington, D. C., for electrotypes +of figures 17, 18, and 19 in Bulletin No. 20, Division of Botany. + + + + +LITERATURE. + + +Acides Gummiques, Garros (Dissertation) 1895. + +American Chemical Journal. + +American Journal of the Medical Sciences. + +American Journal of Pharmacy. + +Annalen der Chemie und der Pharmacie (Liebig). + +Annales de Chimie et de Physique. + +Berichte der deutschen chemischen Gesellschaft. + +Biochemie der Pflanzen (Czapek) 1905. + +Brooklyn Medical Journal. + +Bulletin de la Societe Chimique. + +Bulletins 20 and 26 U. S. Department of Agriculture, Division of Botany. + +Chemie der Zuckerarten, Von Lippmann, 1904. + +Chemiker-Zeitung. + +Comptes rendus. + +Industries of Japan, J. J. Rein. + +Journal of the Chemical Society. + +Journal of Experimental Medicine. + +Les Sucres, Maquenne, 1900. + +Manual of Botany, 6th Edition, Gray. + +Medical and Surgical Reporter. + +New York Medical Record. + +Proceedings of the American Pharmaceutical Association. + +Treatise on Chemistry, Roscoe and Schorlemmer. + +Ueber Mategerbstoff, Reuchlin (Dissertation) 1904. + + + + +SOME CONSTITUENTS OF THE POISON IVY PLANT. + +(RHUS TOXICODENDRON) + + + + +INTRODUCTION. + + +Plants belonging to the natural order Anacardiaciae (Cashew family or +Sumach family) are found in all the temperate climates of the world and +quite frequently in semi-tropical climates. Many of these plants play +important parts in economic botany, yielding dye-stuffs, tanning +material, wax, varnish, and drugs. Several species are poisonous. At +least three poisonous species of the genus _Rhus_ are found in the +United States. These three are all common and well-known plants, but +confusion frequently arises concerning them on account of the different +names by which they are known in different localities. For example, +poison ivy (_Rhus toxicodendron_ or _Rhus radicans_) probably the best +known poisonous plant in America, being found in all the States except +those in the extreme West, is often confounded with and popularly called +"poison oak." The true poison oak is the _Rhus diversiloba_ of the +Western States.[1] The third and most poisonous species of this plant is +_Rhus venenata_ or _Rhus vernix_; it is the _Rhus vernicifera_ of Japan, +from which Japanese lac is obtained. It is popularly known in the United +States as "poison sumach," "poison dogwood" and "poison elder." It grows +in swamps from Canada to Florida. + +As the poison ivy is by far the most common of these plants in the +Eastern States, a brief description of it is given here:[2] A shrub +climbing by rootlets over rocks, etc., or ascending trees, or sometimes +low and erect; leaflets 3, rhombic-ovate, mostly pointed, and rather +downy beneath, variously notched, sinuate, or cut-lobed; high climbing +plants (_R. radicans_) having usually more entire leaves. It is found in +thickets, low grounds, etc. Greenish flowers appear in June. + +[Illustration: Fig. 1.--Poison ivy (_Rhus radicans_ or _Rhus +toxicodendron_). _a_, spray showing aerial rootlets and leaves; _b_, +fruit--both one-fourth natural size. + +(Chesnut, Bulletin No. 20, Division of Botany, U. S. Department of +Agriculture.)] + +[Illustration: Fig. 2.--Poison oak (_Rhus diversiloba_) showing leaves, +flowers, and fruit, one-third natural size. + +(Chesnut, Bulletin No. 20, Division of Botany, U. S. Department of +Agriculture.)] + +In the general description of the order Anacardiaciae, Gray[3] says: +"Juice or exhalations often poisonous." Whether it is contact with some +part of the plant, or with the exhalation from the plant, that causes +the well-known skin eruption has been a topic for discussion ever since +its source was known. On account of its intangible nature there has been +more speculation than experimental evidence bearing on this question, +although a few investigations have been made with the object of +isolating the poison. It is most generally believed that the exhalations +are poisonous. Dr. J. H. Hunt[4] states that the exhalations have been +collected in a jar and found to be capable of inflaming and blistering +the skin of an arm plunged into it. + +[Illustration: Fig. 3--Poison sumach (_Rhus vernix_), showing leaves, +fruit, and leaf-scars, one-fourth natural size. + +(Chesnut, Bulletin No. 20, Division of Botany, U. S. Department of +Agriculture.)] + +Prof. J. J. Rein,[5] in his treatise on Lacquer Work, describes the +poison of the Japanese lac tree, _Rhus vernicifera_, as being volatile, +as do also the Japanese chemist Yoshida[6] and the French chemist +Bertrand.[7] Recent work by Prof. A. B. Stevens,[8] however, seems to +show that this poison is not volatile, and is similar to, if not +identical with that obtained by Pfaff[9] from _Rhus toxicodendron_ and +_Rhus venenata_. + +Not many cases of internal poisoning by _Rhus toxicodendron_ are on +record in medical literature. Two cases of poisoning from eating the +fruit of this plant have been described.[10] The subjects of these cases +were two children who had eaten nearly a pint of the fruit. The symptoms +are described in detail, being in general, similar to those of +alkaloidal poisoning. Warm water was given to promote emesis; afterwards +large quantities of carbonate of soda were given in solution under the +belief that it was an antidote to the poison. Otherwise they were +treated on general principles. Both children recovered. + +Another case of internal poisoning is the following:[11] Three children +drank an infusion of the root of poison ivy thinking it was sassafras +tea. The first of these cases was diagnosed as measles, but on the +appearance of similar symptoms in the sisters of the first patient, the +cause of the trouble was found. All recovered. + +Dr. Pfaff[12] explains the few fatal cases that have followed Rhus +poisoning on the assumption that enough of the poison was absorbed +through the skin to cause renal complications in persons having chronic +kidney trouble. He showed that the poison, when given internally, +produces a marked effect on the kidneys, causing nephritis and fatty +degeneration of this organ. + +The irritating action of poison ivy has been attributed at different +times to the "exhalation," to a volatile alkaloid, to a volatile acid, +and to a non-volatile oil. Pfaff,[13] who made the most recent +investigation of this poison, obtained from the plant a non-volatile oil +having the same action on the skin as the plant itself. He found this +oil in all parts of the plant and concluded that it was the active +principle, and that one could be poisoned only by actual contact with +some part of the plant. He assumed minute quantities of pollen dust to +be in the air to account for the cases of "action at a distance" so +frequently quoted. Pfaff says: "In my opinion, it is more than doubtful +if ever a case of ivy poisoning has occurred without direct contact with +the plant or with some article that has been in contact with the plant. +The long latent period of the eruption in some cases may obviously +render mistakes extremely easy as to the occasion when contact with the +plant really occurred." Granting, however, that the active principle is +practically non-volatile when isolated from the plant, we cannot say +positively that it is not volatile in the juices of the plant, or under +the influence of vital forces. It is quite conceivable that the water +transpired by the leaves of the plant may carry with it a quantity of +the poison sufficient to produce the dermatitis on a person very +susceptible to its action. It is also conceivable that a volatile poison +manufactured by a living plant could become non-volatile by changes in +it consequent upon the death of the plant. + +Up to the present time, only three important chemical investigations of +the active principle of _Rhus toxicodendron_ have appeared in medical +and chemical literature, these being the researches of Dr. J. Khittel, +J. M. Maisch, a pharmacist, and Dr. Franz Pfaff, of the Harvard +University Medical School, to whose work reference has been frequently +made. The chemical work of these investigators and their conclusions are +given here in some detail for the sake of completeness. + +FOOTNOTES: + +[1] Chesnut. Bull. No. 20, U. S. Dept. of Agr., Div. of Botany. + +[2] Man. of Bot., p. 119. + +[3] Man. of Bot., p. 119. + +[4] Brook. Med. Jour., June, 1897. + +[5] Rein, The Ind. of Jap., p. 338, et seq. + +[6] H. Yoshida on Urushi Lacquer, Jour. Chem. Soc., 1883, p. 472. + +[7] Ann. de Chem. et de Phys., Series VII, Vol. 12, p. 125, 1897. + +[8] Amer. Jour. Pharm. 78, p. 53, Feb., 1906. + +[9] An account of Pfaff's work will be found in another part of this +paper. + +[10] Amer. Jour. Med. Sci. 51 (1866), p. 560. + +[11] Med. and Surg. Rep. 17, Nov., 1867. + +[12] Jour. Exp. Med. 2 (1897), p. 181. + +[13] Ibid. + + + + +KHITTEL'S INVESTIGATION. + + +The first attempt to find the poisonous constituent of this plant was +made by Khittel in 1857. His work was published in _Wittstein's +Vierteljahrresschrift fuer praktische Pharmacie_, VII, 348-359.[14] +Khittel obtained 37-1/2 ounces of fresh leaves of poison ivy from the +botanical garden in Munich, dried them, and got a residue of 9-1/2 +ounces which he analyzed. Not detecting anything to which the poisonous +qualities of the plant could be attributed, he made another series of +experiments which, as he thought, showed that a volatile alkaloid is the +poisonous constituent. It was obtained by the following process: "3 +ounces of the powdered leaves were infused with hot distilled water, +after three days strained, expressed, the liquid evaporated to 3 ounces, +and with the addition of potassa, carefully distilled to one-half. The +clear, colorless distillate had an alkaline reaction, and an odor +resembling henbane or hemlock. It was saturated with sulphuric acid, +evaporated, and treated with a mixture of equal quantities of alcohol +and ether which left sulphate of ammonia behind, the solution was +evaporated spontaneously, distilled with potassa, the alkaline +distillate neutralized with hydrochloric acid, and a precipitate could +now be obtained with chloride of platinum. Want of material prevented +further experiments." + +The editor of the _American Journal of Pharmacy_ inserts the following +note: "It would have been more satisfactory if the author had given some +physiological evidence of the poisonous nature of the alkaloid +substance obtained. It is quite interesting to hear that the hitherto +intangible venom of this plant has at last been detected." + +FOOTNOTES: + +[14] A free translation of this paper is given in Amer. Jour. Pharm. for +1858, p. 542. + + + + +WORK OF MAISCH.[15] + + +The next investigation of this plant was made by Maisch in 1864. He +criticizes Khittel's experiments as follows: "It is well known that the +_exhalations_ of _Rhus toxicodendron_ exert a poisonous influence on the +human body; the poisonous principle must, therefore, be volatile and, at +the same time, be naturally in such a loose state of combination as to +be continually eliminated and separated with the usual products of +vegetable exhalations. It is natural to suppose that during the process +of drying, the greatest portion of the poisonous principle should be +lost. The loss must be still greater if the dried leaves are powdered, a +hot infusion prepared from them, and this infusion evaporated down to +the original weight of the dried leaves. It is obvious that Dr. Khittel +could not have selected a better method for obtaining the least possible +quantity of the poisonous principle, if, indeed, it could be obtained by +this process at all." + +Maisch then worked up 8-3/4 ounces of the leaves of the plant in a way +to get the alkaloid, making some improvements on Khittel's method, but +failed to find it. Believing that the poison was a volatile acid, he +enclosed some fresh leaves of the plant in a tin box with several test +papers. The blue litmus paper became red showing the presence of an +acid. He concluded from this experiment that the exhalations of the +leaves contained a volatile organic acid which he thought was the +poisonous substance. To determine this point, he prepared the acid in +larger quantity by macerating the leaves with water, expressing and +distilling the expressed juice. He was poisoned in doing this work +although he had not been affected by handling the living plant and had +considered himself immune. He obtained an acid which investigation +showed to be somewhat like formic acid, more like acetic acid, but +having some reactions different from both. "Taking all the reactions +together, it is unquestionably a new organic acid for which I propose +the name of _Toxicodendric Acid_," writes Maisch. He further says: "That +it is the principle to which poison oak owes its effects on the human +system was proved to my entire satisfaction by the copious eruption and +formation of numerous vesicles on the back of my hand, on the fingers, +wrists, and bare arms while I was distilling and operating with it. +Several persons coming into the room while I was engaged with it were +more or less poisoned by the vapours diffused in the room; and I even +transferred the poisonous effects to some persons, merely by shaking +hands with them. + +"The diluted acid, as obtained by me, and stronger solutions of its +salts, were applied to several persons, and eruptions were produced in +several instances, probably by the former, though not always, which was +most likely owing to the dilute state of the acid. Whenever this was +boiled, I always felt the same itching sensation in the face, and on the +bare arms, which I experience on continual exposure of my hands to the +juice of the plant." + +Toxicodendric acid was thought to be the active principle from the time +of Maisch's work until the investigation by Pfaff in 1895. + +FOOTNOTES: + +[15] Proc. Amer. Pharm. Assn. 1865, p. 166, and Amer. Jour. Pharm. 1866, +p. 4. + + + + +PFAFF'S WORK. + + +By far the most valuable work on _Rhus toxicodendron_ is that of Pfaff. +From a clinical study of Rhus poisoning, Pfaff came to the conclusion +that the poison must be a non-volatile skin irritant. The more volatile +the irritant, the quicker is its action on the skin. Formic acid acts +very quickly; acetic acid, less volatile than formic, acts more slowly, +but still much more quickly than poison ivy, the latent period of which +is usually from two to five days. Pfaff thought that the volatile acid +obtained by Maisch might have contained some of the poisonous principle +as an impurity, but that it would not produce the dermatitis if prepared +in a pure state. He therefore prepared a quantity of the acid by +distilling the finely divided fresh plant with steam. The yield was +increased by acidulating the mixture with sulphuric acid before the +distillation. The acid distillate so obtained was freed from a +non-poisonous oily substance by shaking the solution with ether. Barium +and sodium salts were made by neutralizing the acid, and were purified +by crystallization. Analysis showed them to be salts of acetic acid, and +they gave the characteristic tests for this acid. The toxicodendric acid +of Maisch was thus shown to be acetic acid, and was therefore not the +poisonous principle of the plant. + +Pfaff obtained the active principle by the following process: The plant +was extracted with alcohol, the alcohol was distilled off, and the +residue was taken up in ether. The ether solution was washed with water +and dilute sodium carbonate solution, and the ether was evaporated. An +oily, black, poisonous substance partly soluble in alcohol was obtained. +To get the active principle in a pure state, this residue was extracted +with alcohol and filtered and the filtrate was precipitated fractionally +by lead acetate. The final precipitates consisted of the lead compound +of the poison in a pure state. On decomposing the lead compounds with +ammonium sulphide, shaking out with ether, and letting the ether +evaporate spontaneously, a non-volatile oil was obtained which gave the +characteristic skin eruptions. The pure lead compounds made in different +preparations were analyzed and assigned the formula C_{21}H_{30}O_{4}Pb. +The oil itself was not analyzed. Pfaff proposed the name _Toxicodendrol_ +for the oil. He found that it was not volatile, was decomposed by heat, +was soluble in alcohol, ether, chloroform, benzene, etc., but insoluble +in water. Its effects upon the human skin were studied in many +experiments upon himself and others. It was shown that an exceedingly +minute quantity of the poison will produce the dermatitis, even 1/1000 +milligram applied in olive oil being active. The oil was given +internally to rabbits, its effects being most marked on the kidneys. + +The oil obtained by Pfaff from _Rhus venenata_ seemed to be identical +with that from _Rhus toxicodendron_. + + + + +EXPERIMENTAL. + + +The writer's investigation was undertaken with the object of attempting +to throw more light on the chemical nature of the poisonous substance +found in _Rhus toxicodendron_. Soon after commencing work, however, it +became apparent that the poison could be more intelligently studied if +the substances associated with it in the plant were first identified; +the scope of the work was therefore extended to an investigation of the +other constituents of the plant, and it was hoped that a knowledge of +the properties of these constituents would suggest a more economical way +of getting the poison than the method of fractional precipitation. + +The crude material for this work was prepared by Messrs. Parke, Davis & +Co., of Detroit, Mich., according to special instructions submitted to +them: 67-1/2 pounds of fresh leaves and flowers of poison ivy were +collected near Detroit and carefully inspected by a competent botanist. +This material was thoroughly macerated and put into ten-liter bottles +with ether. The mass was thoroughly shaken, water being added to make it +more mobile. The ether was then separated off and the extraction was +repeated three times in the same way to insure complete removal of the +toxicodendrol. The ether extracts were combined, thoroughly dried with +anhydrous sodium sulphate, and the ether was distilled off, the +temperature being kept below 40 deg. C. during the entire distillation. +The residue after the removal of the ether was a thick, black, tar-like +mass, weighing 3 pounds 11 ounces. In extracting the plant, about +twenty-four gallons of ether were used. It is a significant fact in +regard to the volatility of the poison that during the process of +preparing this material none of the employees engaged in the work were +in any way affected, since proper precautions were taken and the +utensils were handled with rubber gloves. + +The crude ether extract, which will be designated as the "original +material," was shipped to Baltimore in August and was kept in a cool +place until November when the investigation was begun. When the bottle +was opened, there seemed to be an escape of a vapor, and a nauseating +odor suggesting crushed green leaves pervaded the atmosphere. Some days +later, irregular red patches appeared on the face though a mask of +cotton cloth was worn during the work, and the hands were protected by +rubber gloves. + +Assuming from Pfaff's work that this original material contained the +non-volatile oil toxicodendrol, the first experiment was to try to +distil it out under diminished pressure. For this purpose, an Anschuetze +distilling bulb containing ten grams of the tar was connected with a +vacuum pump. After a pressure of 2 mm. had been established the bulb was +gradually heated in a bath of Wood's metal. Nothing distilled over. The +material began to carbonize at a temperature of 140 deg. to 150 deg. + +It was then thought that perhaps the oil could be converted into an +ester which might be more volatile and could be distilled out. 20 grams +of the original material were dissolved in 100 cc. of absolute alcohol +containing 3 grains of hydrochloric acid gas, and the mixture was heated +10 hours on a water-bath under a return condenser. After the heating, +the mixture had a delightful ethereal odor. The flask was corked and +left standing several weeks while other work was in progress. The ester +solution was then put in a vacuum desiccator over sulphuric acid and the +alcohol evaporated. A black, tarry, solid mass was left having the ester +odor. It was extracted with warm water and filtered from insoluble tar. +The filtrate had a green color and the ethereal odor. It was shaken out +with ether; the ether layer had a blood-red color while the water layer +was deep green. The extraction with ether was continued until the water +layer was no longer green. The combined ether extracts were evaporated +in a desiccator without heat. A black tar-like solid was left very much +like the original material, but it had the ester odor. It was partly +soluble in water and readily soluble in alcohol. The alcoholic solution +was tested on the skin and found to be not poisonous. The ester, or +mixture of esters, was not investigated further in this connection, but +was later shown to give the reactions for gallic acid and methyl +furfurol. These reactions will be referred to in connection with other +experiments. + +After a few other preliminary experiments, it became evident that the +original material was a complex mixture of substances and that it would +have to be fractionated by some means and the fractions studied +separately. + +A portion of the original substance was treated with 50 per cent. +alcohol and was found to be partly soluble in this medium. The solution +was filtered from insoluble tar. A portion of the yellow filtrate gave a +reddish yellow precipitate with lead acetate. The alcoholic solution was +distilled in an Anschuetze flask under diminished pressure; a yellow +liquid condensed in the arm of the flask while most of the alcohol was +collected in a bottle connected with the arm. The yellow liquid was acid +to litmus. Water was added, the solution was shaken out with ether and +the ether was evaporated. When the small residue was completely dry, it +was a yellow solid soluble in dilute alcohol and acid to litmus. The +substance was not volatile enough to justify the use of this method for +getting it. + +Chlorophyll could not be removed from the original substance because the +solvents for chlorophyll such as alcohol, ether, fats, petroleum, and +carbon bisulphide dissolve large quantities of the mixture. + +A precipitate obtained by adding lead acetate to a filtered solution of +the original substance in 50 per cent. alcohol was suspended in water, +decomposed by hydrogen sulphide, shaken out with ether and the ether +evaporated. The residue appeared at first to be a yellow oil, but on +complete evaporation of the ether in a desiccator, a yellow solid was +obtained--apparently the same as that obtained by vacuum distillation. + +A solution of the original material in 50 per cent. alcohol was filtered +through bone-black and the filtrate was colorless. Examination showed +that everything had been removed by the bone-black and the filtrate was +apparently pure alcohol and water. + +In precipitating an alcoholic solution of the crude material with a +solution of lead acetate, it was noticed that at least two kinds of +precipitates were formed. The part that went down first was darker in +color than that thrown down later. Pfaff used the last fractions in +obtaining his oil and stated that these precipitates consisted of the +lead compound of the oil in a pure state. It was found by experiment +that the darker part was soluble in ether while the lighter part was +not. This indicated that the darker part consisted of tarry matter which +was brought down mechanically or separated out when the alcoholic +solution was diluted by the lead acetate solution, or was perhaps a lead +compound soluble in ether. To test this point an experiment was made as +follows: Some of the crude material was thoroughly extracted with 50 per +cent. alcohol. The tar insoluble in 50 per cent. alcohol was then +treated with 95 per cent. alcohol; most of it dissolved; the solution +was filtered and lead acetate in 50 per cent. alcohol was added. A +greenish colored precipitate was formed which was filtered off and found +to be completely soluble in ether and soluble to a considerable extent +in strong alcohol. These experiments suggested that the light colored +lead compound which was thought to contain the poison could be purified +by extraction with ether in a Soxhlet apparatus more conveniently than +by the tedious process of fractional precipitation. Further preliminary +experiments showed that 50 per cent. alcohol extracted from the original +material all of the substance or substances which gave the light colored +precipitate and dissolved only a small amount of the tar. + +Two hundred and eighty-eight grams of the crude material were then +extracted several times with 50 per cent. alcohol and filtered; the +insoluble tar was washed and saved for examination. To the filtrate was +added an excess of a solution of lead acetate in 50 per cent. alcohol. +The large precipitate, which will be designated as "precipitate A," was +filtered and drained by suction in a Buechner funnel. The alcoholic +"filtrate A" was saved. Precipitate A was extracted with ether in +Soxhlet extractors until the ether came over practically colorless, the +operation being interrupted from time to time to stir up the precipitate +in the thimble. The green colored ether solution was saved for future +examination. The lead precipitate, after extraction with ether and +drying, weighed about 116 grams. A portion of this lead compound was +decomposed by hydrogen sulphide in a mixture of water and ether which +was well shaken during the operation. The ether was separated, filtered, +and evaporated under diminished pressure without heat, and there +remained a yellow oily looking residue having a pleasant odor. When the +ether and water were completely removed in a vacuum desiccator, a hard, +brittle, yellow resin weighing about 16 grams was obtained. It was +soluble in alcohol, had a strong acid reaction and was free from +nitrogen[16] and sulphur. The nitrogen tests were made by the Lassaign +and soda lime methods,[17] and the sulphur test was made with sodium +nitroprusside after fusing the residue with sodium. The main portion of +the lead compound was decomposed under alcohol by hydrogen sulphide, +filtered, and the alcoholic filtrate evaporated in vacuo. The same +yellow acid resin was obtained. Experiments continuing through several +weeks were made in applying solutions of this resin to rats, rabbits and +guinea pigs. Finding it to be without effect upon these animals it was +tried on the writer and found to be not poisonous.[18] In the meantime +the resin was being studied in the laboratory. + + +GALLIC ACID. + +An alcoholic solution of the resin was just neutralized with potassium +hydroxide. During the titration, the solution rapidly became dark brown. +After neutralization it was shaken with ether; the water solution +remained brown while the ether layer was nearly colorless and contained +practically no dissolved substance. A portion of the water solution of +the potassium salt on being acidified with sulphuric acid and standing +over night, deposited a slight precipitate. The solution of the +potassium salt gave a heavy precipitate with lead acetate somewhat +similar to the original lead precipitate A, and also slight precipitates +with salts of zinc, mercury, copper, and silver (with reduction). It +gave a bluish-black color with impure ferrous sulphate and a dark color +with ferric chloride. It reduced ammoniacal silver nitrate and Fehling +solution. These experiments indicated the presence of a tannin compound. +An alcoholic solution of the resin gave the same color reactions with +iron salts as did the potassium salt. To determine which one of the +tannin compounds was present was a matter of some difficulty since the +di- and tri-hydroxybenzoic acids have in general the same color +reactions. The presence of other plant substances in the solution also +interferes with the color tests, and finally, a substance which gives a +blue color with iron salts and one giving a green color may be found +together in the same plant.[19] Further tests with a solution of the +resin in dilute alcohol, and with a water solution of the acid +precipitated by adding sulphuric acid to a solution of the resin in +potassium hydroxide, led to the conclusion that the acid is gallic acid. +These tests were the following: + +(1) Boiling with an excess of potassium hydroxide gave a black substance +(tauromelanic acid). + +(2) The acid was not precipitated by gelatin. + +(3) On addition of potassium cyanide a transitory red color appeared +which reappeared on shaking with air. + +Gallic acid is distinguished from tannic acid by tests (2) and (3). At +later stages in the work the potassium, barium, and sodium salts of +gallic acid were obtained, and finally the pure acid was made by +decomposing the sodium salt with sulphuric acid and crystallizing from +water. A portion of the acid so obtained was further purified by +dissolving in absolute alcohol and pouring into absolute ether.[20] The +melting point behavior of the acid corresponds with that of gallic acid; +it melted with decomposition at about 230 deg. For further identification, +some of the acid was converted into an ester by the following process: +it was dissolved in 80 per cent. alcohol, hydrochloric acid gas was +passed in, and the solution was heated an hour on the water bath. It was +then evaporated to a small bulk, neutralized with barium carbonate and +extracted with ether. The ether, on evaporation, left the ester which +was crystallized from water and dried in a desiccator over sulphuric +acid. The anhydrous ester agreed in melting point (156 deg. to 159 deg.) and +other properties with the ester of gallic acid described by Grimaux.[21] +For the sake of comparison, an ester was made from gallic acid obtained +from another source and the two agreed in properties. A mixture of the +two esters melted within the limits given for the ester of gallic acid. + +While the tests leading to the identification of gallic acid were being +made, another series of experiments was in progress. Eleven and one-half +grams of the resin obtained from lead precipitate A by decomposition +with hydrogen sulphide were treated with 0.1 n. potassium hydroxide +added from a burette until the acid was exactly neutralized. All went +into solution. On shaking with ether, some of the potassium salt +separated out and was saved for examination. The solution became brown +on exposure to air and got darker as the work proceeded. The acid in +solution as a potassium salt was precipitated out in four fractions by +adding for each fraction one-fourth the amount of 0.1 n. sulphuric acid +required to neutralize the potassium hydroxide used. The precipitates +were filtered off and examined. The first was small in amount, gummy and +hard to filter. The solution was shaken with ether after each +precipitate had been filtered off. The succeeding precipitates were in +better condition, but were not pure. All appeared to be impure gallic +acid which had become brown by absorption of oxygen. They were saved, +however, to be tested for poison. After the last fraction had separated, +the filtrate was shaken several times with ether and saved for further +examination, which will be described under "Rhamnose." This filtrate is +designated as B. + +At this stage of the work a portion of the resin obtained from lead +precipitate A was tested and found to be not poisonous as already +mentioned. By this test, all the substances contained in the lead +precipitate A after its extraction with ether in the Soxhlet apparatus, +were eliminated from the possible poisonous substances. The poison must +therefore have been extracted by the ether. + +A fresh portion of the original poisonous material was treated with 50 +per cent. alcohol and filtered from insoluble tar. The filtrate was +precipitated in six fractions by lead acetate. The last fractions were +lighter in color and apparently much purer than the first. The sixth +lead precipitate was decomposed by hydrogen sulphide, the light-yellow +water solution was tested and found to be not poisonous. It gave the +characteristic reactions for gallic acid. The poison, if precipitated at +all by lead acetate, must have gone down in one of the preceding +fractions. Later experiments showed that it is brought down partly +mechanically and partly as a lead compound in the first precipitates. + + +FISETIN. + +Having identified gallic acid, and not finding any other phenol +derivative in the lead precipitate, some of the original material was +extracted with hot water to remove gallic acid and filtered from tar +while hot. The filtrate had a deep yellow color. On cooling over night, +an olive green precipitate separated out which was dried and found to be +a light powder. It was practically insoluble in cold water, soluble +with great difficulty in boiling water from which it separated in yellow +flakes, slightly soluble in ether and in acetic acid, but readily +soluble in alcohol. The solutions were not acid to litmus, gave a dark +color with ferric chloride, an orange-red precipitate with lead acetate +which was easily soluble in acetic acid, and an orange-yellow +precipitate with stannous chloride. These properties and reactions +indicated that the substance was the dye-stuff fisetin and that it +occurs in the free state in this plant though it is usually found as a +glucoside of fisetin combined with tannic acid. A compound of this kind +was found in _Rhus cotinus_ and named "fustin-tannide" by Schmid[22]. He +showed that the fustin-tannide could be decomposed by acetic acid into +tannic acid and a glucoside, fustin C_{46}H_{42}O_{21}. Fustin, on +heating with dilute sulphuric acid, gave fisetin and a sugar supposed to +be rhamnose. Fisetin was also found as a glucoside compound in _Rhus +rhodanthema_ by Perkin.[23] + +The yellow substance which separated from the boiling water solution was +further purified by dissolving in a small quantity of hot alcohol and +adding hot water. On cooling, the yellow substance separated out in a +flocculent condition. Examined under the microscope, the flakes appeared +to be made up of masses of fine crystals. + +An alcoholic solution of the substance gave a black color with ammonia +which became red on addition of more ammonia. Concentrated acids +intensified the yellow color of the alcoholic solution. Fehling solution +and ammoniacal silver nitrate were reduced by it. Potassium hydroxide +added to an alcoholic solution gave at first a deep red color +accompanied by a green fluorescence which disappeared, leaving a yellow +liquid. With an excess of caustic potash, the red color returned and was +permanent. These reactions are characteristic for fisetin.[24] + +Furthermore, fisetin should give protocatechuic acid and phloroglucinol +by fusion with caustic potash under proper conditions.[25] The +experiment was carried out as follows: 2 grams of fisetin were gently +heated in a nickel crucible with 6 grams of caustic potash dissolved in +6 cc. water. An inflammable gas, apparently hydrogen, was evolved during +the fusion. The pasty mass was dissolved in water, acidified with +sulphuric acid, and filtered. The filtrate was shaken out with ether +containing one-fourth its volume of alcohol. The ether was evaporated +and the residue was extracted with warm water and filtered. Lead acetate +was added to the filtrate to precipitate protocatechuic acid, while +phloroglucinol remained in the filtrate from this precipitate. The lead +precipitate was suspended in water, decomposed by hydrogen sulphide, +filtered, and evaporated to obtain protocatechuic acid. That the +substance obtained was protocatechuic acid was shown by the following +characteristic tests: + +(1) It gave a greenish brown color with ferric chloride; on addition of +one drop of a dilute solution of sodium carbonate, the color became dark +blue; on adding more sodium carbonate the color became red. + +(2) A violet color was obtained when a solution of the acid was treated +with a drop of sodium carbonate solution and then with a drop of ferrous +sulphate. + +(3) It reduced ammoniacal silver nitrate. + +(4) It did not reduce Fehling solution. + +The filtrate supposed to contain phloroglucinol was treated with +hydrogen sulphide to remove lead, filtered, and shaken with ether. The +residue left on evaporating the ether was taken up in water. This +solution gave the following reactions characteristic for phloroglucinol: + +(1) It reduced both silver nitrate and Fehling solution. + +(2) It colored pine wood moistened with hydrochloric acid red. + +(3) It gave a red color with vanillin and hydrochloric acid, and + +(4) A deeper red color with oil of cloves and hydrochloric acid, +becoming purple on standing. + +(5) It gave a violet color with ferric chloride. + +The substance is then, without doubt, fisetin. The formula[26] of +fisetin is supposed to be C_{15}H_{10}O_{6}. + + +RHAMNOSE. + +It was stated above that Schmid obtained a sugar solution by the +decomposition of a fisetin-glucoside from _Rhus cotinus_, and Perkin +obtained the same from a glucoside in _Rhus rhodanthema_. These +investigators thought that the sugar was isodulcite or rhamnose, but +they did not isolate it on account of the small quantities of material +at their disposal. Moreover, the sugar is very hard to crystallize in +the presence of other soluble substances and is not found in large +quantity in plants. Maquenne[27] could obtain only 15 to 20 gm. of +rhamnose by working up 1 kilogram of the berries of _Rhamnus +infectorius_. Assuming that the free fisetin found in poison ivy leaves +had its origin in the decomposition of a fisetin-glucoside by natural +processes, it was reasonable to suppose that the sugar would also be +found in the free state, although, according to Roscoe and +Schorlemmer:[28] "Isodulcite does not occur in the free state in nature, +but is found as a peculiar ethereal salt belonging to the class of +glucosides. On boiling with dilute sulphuric acid, this splits up into +isodulcite and other bodies...." The more recent works on the sugars and +on plant chemistry[29] mention the occurrence of rhamnose only in the +glucoside form, with one possible exception. The exception referred to +is the occurrence of a free sugar, supposed to be rhamnose, in a certain +palm-wine.[30] Czapek says:[31] "The well-known methyl pentoses do not +occur in the free state in plant organisms so far as we know." + +Since rhamnose forms a lead compound, the sugar, if present, should be +found in the first lead precipitate, A, and also in filtrate A in case +it is not completely precipitated in the presence of acetic acid and +alcohol. + +The filtrate A (about two liters) was examined first. It had a light +yellow color, contained an excess of lead acetate, and was acid from the +acetic acid liberated in the precipitation of the lead compound A.[32] +This filtrate was evaporated to dryness under diminished pressure to +remove alcohol, water, and acetic acid. The clear distillate had a +peculiar odor suggesting both tea and amyl formate. It was saved for +examination and was found to be not poisonous. The residue in the dish +after evaporation was a tough reddish brown, gummy mass which could be +drawn out into fine threads. It had a pleasant sweet odor. It was +extracted several times with hot water, each portion being filtered. A +brownish yellow powder remained undissolved and was saved. The combined +filtrates deposited more of the yellow solid on standing. This powder +will be referred to later as "P." The filtered liquid was freed from +lead by hydrogen sulphide. The solution then had a lemon yellow color, a +sweet odor and was acid from acetic acid. On concentrating the solution +by evaporation and making a small portion of it alkaline with sodium +hydroxide, the yellow color came out very intense[33]. The alkaline +solution reduced Fehling solution and ammoniacal silver nitrate, +indicating the presence of a sugar. Another portion of the solution gave +a slight precipitate with phenyl hydrazine in the cold. The remainder of +the solution was evaporated to dryness, extracted with water, filtered, +and again evaporated. A dark sticky syrup was left which was only partly +soluble in water. This was treated with water, filtered, and the +filtrate was evaporated, the water being replaced from time to time to +remove acetic acid. Finally the liquid gave the following tests for +rhamnose, besides those already mentioned: + +(1) With alpha-naphthol[34] and sulphuric acid, a purple violet +color. + +(2) With thymol[35] and sulphuric acid, a red color. + +(3) With resorcinol[36] and sulphuric acid, red color. + +(4) With orcinol[37] and hydrochloric acid, red color. + +(5) With ammonium picrate and sodium picrate, yellowish red color. + +(6) With phloroglucinol and hydrochloric acid, red color. + +(7) It decolorized an alkaline solution of potassium ferricyanide. + +(8) It gave a white precipitate with lead acetate. + +The filtrate B (p. 20) from which gallic acid was precipitated by +sulphuric acid in four fractions was saved to examine for sugar. To +remove gallic acid completely, and other vegetable matter, it was shaken +out several times with ether, and was kept at a low temperature with +salt and ice for a long time. It was left standing for several weeks, +during which time more brown matter separated out and was filtered off. +The filtrate was evaporated to a small bulk, cooled, and filtered from +crystals of potassium sulphate. The filtrate was evaporated to dryness, +the residue taken up in water and filtered through bone-black. Addition +of alcohol caused complete precipitation of potassium sulphate. The +solution then gave the above mentioned characteristic tests for +rhamnose. + +All attempts to get the osazone of the sugar by the method of +Fischer[38] failed, probably on account of the small quantity of the +sugar present. The plant, it will be remembered, was originally +extracted with ether in which rhamnose is practically insoluble. The +above described tests, however, can leave no doubt as to the identity of +the sugar. + +Additional evidence that the sugar is rhamnose was obtained by a method +described by Maquenne[39] as follows: + + "The production of methyl furfurol in the dehydration of + isodulcite furnishes a very simple means of characterizing + this sugar in mixtures which contain it; it is sufficient, + for example, to distil 50 gm. of quercitron wood with as + much sulphuric acid and about 150 gm. of water, then to + rectify the liquid obtained in order to get several drops of + the crude furfurol, which on addition of alcohol and + concentrated sulphuric acid gives immediately the green + coloration characteristic of methyl furfurol. This procedure + is applicable to extracts as well as to entire plants, and + has the advantage that it does not require the separation of + isodulcite, the crystallization of which is often very slow + and at times impossible when it is mixed with other very + soluble substances." + +The experiment was tried with the crude ether extract of the plant +according to the directions of Maquenne, and the green color with +alcohol and sulphuric acid was obtained from the thicker oily portion of +the distillate. This test can be made with hydrochloric acid[40] as well +as with sulphuric. Therefore the color test was tried with the ester +mixture prepared in one of the early experiments by boiling the original +plant material with hydrochloric acid and alcohol. Methyl furfurol was +found here also, this method indeed giving better results than that of +Maquenne. + +The presence of free rhamnose has thus been shown in the original +material, in the first precipitate by lead acetate, and in the filtrate +from this precipitate. Experiments to be described under "The Poison" +showed that the ether extract from the Soxhlet apparatus contained a +substance which yielded rhamnose when hydrolyzed by dilute sulphuric +acid. + +The presence of free gallic acid, fisetin, and rhamnose in the plant can +be readily explained by a series of assumptions for which there is a +considerable amount of experimental evidence. There is reason to believe +that tannin-like bodies are formed at the expense of chlorophyll,[41] +that complex tannin bodies can be broken down by acetic acid (also found +in _Rhus toxicodendron_) into a tannic acid and a glucoside (for +example, the "fustin-tannide" mentioned above yields tannic acid and +fisetin-glucoside); and finally that the glucoside can be hydrolyzed by +acids or enzymes giving, in the sumach plants, fisetin and rhamnose. + +Nitrogenous ferments which can effect the hydrolysis of glucosides and +give rise to sugars are frequently found in plants, for example, emulsin +in almonds, myrosin in mustard, and erythrozym in madder. Acree and +Hinkins[42] found that diastase, pancreatin, and a number of other +enzymes cause hydrolysis of triacetyl glucose with the formation of +glucose and acetic acid. Stevens[43] obtained a nitrogenous oxidizing +enzyme from _Rhus vernicifera_. The close relationship between the +poisonous species of _Rhus_ would lead us to suppose that the same +soluble ferment exists in poison ivy, though it was not detected in the +original material used in these experiments, probably because the plant +was extracted with ether in which the enzyme is insoluble. The existence +of such a soluble ferment would explain the presence of free sugar and +free fisetin. + + +EVIDENCE OF THE PRESENCE OF A FATTY ACID IN FILTRATE A. + +The brown substance P, obtained from filtrate A by evaporation and +extracting the residue with hot water, was suspended in warm water and +dilute sulphuric was added. A white precipitate was formed and a strong +fatty acid odor was developed. After the mixture had been heated for +some hours on the water bath a small portion was made alkaline and it +reduced Fehling solution. The main solution was filtered and the +precipitate supposed to be a fatty acid was saved. The filtrate was +neutralized with barium carbonate, filtered, evaporated, freed from +caramel, and the solution then gave the tests mentioned above for +rhamnose. + +A portion of the precipitate supposed to be a fatty acid was ignited in +a porcelain spoon. It fused, carbonized, and burned. The remainder was +heated with alcoholic potash and reprecipitated with hydrochloric acid. +The precipitate was washed and heated with alcohol. Part of it +dissolved. The insoluble part was found to be a lead compound. On +boiling it with hydrochloric acid and cooling, lead chloride +crystallized out. This was confirmed by dissolving the lead chloride in +hot water and precipitating as lead sulphide. These experiments were not +carried farther on account of the small quantity of material, but they +show that the gummy substance obtained from filtrate A contained +rhamnose (either as a lead compound of free sugar or as a lead compound +of a rhamnoside), and also, most probably, the lead compound of an +organic acid.[44] + + +THE FRAGRANT DISTILLATE. + +Several times in the course of this work, extracts of the original plant +material in alcohol and in water were distilled under diminished +pressure for the purpose of concentrating the solutions. The distillate, +in every case, had an ethereal odor suggesting amyl formate in very +dilute solution, but was more fragrant. The distillate from a water +extract was examined. It was a clear liquid, a little darker than pure +water, was not poisonous, was neutral to litmus paper, gave no color +with ferric chloride, reduced ammoniacal silver nitrate, but not Fehling +solution, and gave a faint red color with dilute ammonium hydroxide and +with sodium carbonate. + +A small quantity of a finely divided black precipitate separated out +from the water distillate on standing. + +The substance with the fragrant odor was extracted by shaking the +distillate with ether and letting the ether evaporate spontaneously. A +very small quantity of a yellow solid was deposited on the sides of the +dish. This substance had a strong and persistent odor, so sweet as to be +almost nauseating. Not enough was obtained for examination or analysis. +This fragrant residue was difficultly soluble in water and the solution +reduced silver nitrate in ammonia. A steam distillate of the original +plant material had the same fragrant odor as the distillate from a water +extract. + + +THE POISON. + +288 grams of the original poisonous material were extracted with 50 per +cent. alcohol, and this alcoholic solution was precipitated with lead +acetate in the manner already described (p. 17). The lead precipitate so +obtained was extracted with ether in Soxhlet extractors and after the +extraction was found by test to be free from poison. Therefore the +poison, if precipitated by the lead acetate, must have been extracted by +the ether. This ether solution had a dark green color, and was acid from +acetic acid brought down in the lead precipitate. The ether was +evaporated in a vacuum desiccator without heat and there remained a +small quantity of an acid mixture of water and a soft tar; the watery +part was colored green, showing that the tar was soluble to some extent +in dilute acetic acid. The mixture had the peculiar odor of the original +material. A small drop of the green watery part was applied to the +wrist, allowed to remain a few minutes and was then removed by absorbent +paper, but the spot was not washed. Itching and reddening of the skin +commenced within twenty-four hours. At the end of forty-eight hours, +there was a well developed case of poisoning. How this was cured will be +described in another place. + +A small portion of the poisonous mixture was dissolved in alcohol, and +this solution was divided into three parts. The first part was treated +with ferric chloride, but it gave no color reaction. Another portion of +the alcoholic solution was diluted with water. It became turbid. The +third portion gave a dirty-green precipitate with lead acetate, which +seemed to come down more readily when the solution was diluted with +water. The main portion of the poisonous mixture was then dissolved in +95 per cent. alcohol and lead acetate in 50 per cent. alcohol was added. +The precipitate was filtered, washed, and decomposed by hydrogen +sulphide in a mixture of water and ether. The ether solution was +filtered and evaporated. The residue was a tar which, on standing in a +desiccator for some time, became dry enough to break into sticky lumps. +An alcoholic solution of this substance gave a dark color with ferric +chloride and a light colored precipitate with lead acetate. + +To get more of the poisonous tar for study, 233 grams of original +material were extracted with 95 per cent. alcohol. Strong alcohol was +used in order to dissolve as much of the tar as possible. The solution +had a dark greenish color, but was somewhat yellow in thin layers. The +undissolved tar was filtered off and extracted twice again in the same +way. The tar left after the third extraction was only slightly soluble +in alcohol, and its solution was not poisonous. The three filtrates from +these three extractions were precipitated separately by lead acetate in +50 per cent. alcohol. The first precipitate was largest, darkest in +color, and carried down more tarry matter. The second was light green, +and the third was quite small, black, and was not a lead compound at +all, but some of the tar which separated out on diluting the strong +alcohol with the weaker grade containing lead acetate. It was soluble in +ether and less soluble in alcohol. The alcoholic solution of this third +lot gave no precipitate with hydrogen sulphide. The first and second +lead precipitates were filtered by suction and washed with water. They +were kept a day or two in a desiccator over sulphuric acid, but did not +become completely dry. The weight of these two moist precipitates +together was 172 grams. They were combined and extracted with ether in +Soxhlet extractors which were kept in operation during work hours for +three days. + +In the meantime the alcoholic filtrates from these lead precipitates +were combined and concentrated on the water bath by distilling off two +liters of alcohol. The alcohol obtained had the peculiar odor of the +original material, but was not poisonous. + +After a long extraction of the lead precipitate in the Soxhlet +extractors, the green ether solutions were combined and washed by +shaking them with water to remove lead acetate and acetic acid in case +any should have been held in the lead precipitate. The ether was +distilled off at a low temperature and there remained a soft tar, a +portion of which was not completely soluble in 95 per cent. alcohol. The +alcoholic solution had a greenish yellow color and was poisonous. The +tar was also partly soluble in acetic acid, and this solution was found +to contain lead. Thinking that the lead acetate had not been completely +washed out, the main part of the tar was dissolved in ether and shaken +with water. The wash water continued to give a test for lead as long as +the washing was continued. This indicated probably the hydrolysis of an +unstable lead compound. Hydrogen sulphide was passed into the ether +solution mixed with water to remove the lead. Lead sulphide was filtered +off, and the ether was evaporated. A small portion of the tar residue in +alcoholic solution gave a color reaction with ferric chloride. As this +may have been due to traces of lead gallate dissolved in the extraction +with ether and afterwards decomposed by hydrogen sulphide, the main +portion of the tar was redissolved in ether and shaken with water until +it no longer reacted with ferric chloride. The ether was then evaporated +and a soft, black, poisonous tar or gum of uniform consistency was left +which was shown by tests to be free from gallic acid and lead. These +experiments showed that some of the poison was precipitated as a lead +compound soluble in ether and some was brought down mechanically in the +free state. To see if the extraction with ether in the Soxhlet apparatus +was complete, the residue in the thimbles was decomposed by hydrogen +sulphide and shaken with ether. The dark colored ether solution was +freed from gallic acid by shaking with water and dilute sodium carbonate +solution, and was evaporated. A small quantity of tar was obtained which +was added to the main portion. + +A solution of the poisonous tar in 95 per cent. alcohol did not reduce +Fehling solution and did not give a precipitate with lead acetate except +the separation of a small quantity of tar, which was not a lead +compound. The lead compound of the poison was apparently soluble in 95 +per cent. alcohol as well as in ether, for it would not precipitate in +this medium, although it was found in the original precipitate by lead +acetate. The alcoholic solution of the tar became turbid on diluting +with water. + +In order to see if the poison is volatile with vapor of acetic acid, +since this acid is found in the plant and it is thought by some that the +poison is volatile, a portion of the tar was distilled under diminished +pressure with acetic acid. It was soluble to some extent in the acid. +The temperature did not go higher than 55 deg. during the distillation. A +tube containing cotton wet with sweet oil was placed between the +receiver and the water suction so that the uncondensed vapors would have +to pass through the cotton. This cotton was rubbed on the skin and was +not poisonous. The yellow distillate collected in the receiver was also +tested and was not poisonous. + + +HYDROLYSIS OF THE POISON. + +About 5 grams of the poisonous tar free from gallic acid and sugar was +dissolved in alcohol, and dilute (2 per cent.) sulphuric acid was added. +Some of the tar separated out on diluting the alcohol with the acid. The +mixture was heated on a water bath during work hours for four days. A +purple and green fluorescent solution was formed, though much tar was +left apparently unchanged. The alcohol was evaporated off and the +solution was filtered from tar. The fluorescent filtrate was shaken +with ether, by which the green substance was removed, leaving the +solution purple. The ether left, on evaporation, a small quantity of a +green substance having a pleasant ester odor. It was not further +examined. A portion of the purple solution was exactly neutralized with +sodium carbonate. This solution gave a blue-black color with ferric +chloride which became red on addition of another drop of sodium +carbonate, indicating gallic acid. It also reduced Fehling solution. + +Another portion of the purple solution was made alkaline with sodium +carbonate. A reddish-brown flocculent precipitate was formed and was +filtered off. The filtrate did not give any color with ferric chloride, +but it reduced Fehling solution. It also gave the test for rhamnose with +alpha-naphthol. + +The main portion of the purple solution was made alkaline with sodium +carbonate; the precipitate was filtered off and dissolved in acetic +acid. This solution was yellow and gave a reaction with ferric chloride +similar to that of gallic acid. The filtrate from the precipitate by +sodium carbonate was concentrated by evaporation until sodium sulphate +began to crystallize out. Alcohol was added to precipitate the sodium +sulphate completely, the mixture was heated and filtered. The alcoholic +filtrate was concentrated to a syrup which reduced Fehling solution and +gave the characteristic tests for rhamnose already described. By this +hydrolysis, the tar was split up into rhamnose and some form of gallic +acid which could be precipitated by sodium carbonate. This compound, +whose acetic acid solution was yellow, probably contained fisetin also. +The reason for this last statement will appear from the following +experiment: + + +DECOMPOSITION OF THE POISON WITH ACETIC ACID. + +A portion of the poisonous tar was heated in an open dish with strong +acetic acid. The tar seemed to be decomposed to some extent, giving a +yellow substance. Acetic acid was added from time to time as it +evaporated. After several evaporations, water was added, the mixture was +heated to boiling and filtered. This filtrate No. 1 will be mentioned +later. The residue in the dish consisted of undecomposed tar and an +olive-green flaky substance. This substance was heated with a fresh +portion of glacial acetic acid. Water was added, and the mixture was +boiled and filtered. The filtrate had a deep yellow color suggesting +fisetin. It was shaken out with ethyl acetate which became colored +yellow. A portion of the ethyl acetate solution gave an orange red +precipitate with lead acetate showing the presence of fisetin. The ethyl +acetate was removed from the remainder of the solution by evaporation +and the yellow residue was taken up in alcohol. This alcoholic solution +gave the characteristic reactions for fisetin with stannous chloride, +with potassium hydroxide, with ferric chloride and with Fehling +solution. + +Filtrate No. 1 obtained by heating the poisonous tar with acetic acid +and hot water as described above was investigated as follows: A portion +of it gave a reddish colored precipitate with sodium carbonate as in the +case when the tar was hydrolyzed with sulphuric acid. The remainder was +nearly neutralized with sodium carbonate and lead acetate was added in +excess to remove gallic acid. The excess of lead was removed by +sulphuric acid, and the sulphuric acid was removed by barium carbonate. +The solution on evaporation reduced Fehling solution to some extent, but +a white precipitate was also formed. + +In this experiment, gallic acid and fisetin and probably sugar were +formed by decomposition of the poisonous gum with acetic acid, the acid +found in the plant by Pfaff. The presence of free gallic acid, fisetin +and rhamnose in the plant can therefore be explained by the natural +hydrolysis of a complex gum or tar or a constituent thereof. The +poisonous property is lost in the general rearrangement which takes +place during hydrolysis. + +The poisonous tar was not hydrolyzed by boiling with a dilute solution +of sodium carbonate. + +It was found, as has been stated elsewhere, that the lead compound of +the poison could not be precipitated in 95 per cent. alcohol. Further +experiments, however, showed that on extracting the poisonous gum with +50 per cent. alcohol, a portion of it dissolved, and this solution gave +a precipitate with lead acetate which was a true lead compound. The +remainder of the purified tar (about 10 gm.) was treated with 50 per +cent. alcohol and filtered. Very little dissolved in alcohol of this +strength, but on addition of lead acetate in 50 per cent. alcohol to the +solution, a light colored precipitate was formed, which became dark on +standing. It was filtered off, washed free from lead acetate, decomposed +by hydrogen sulphide, and shaken out with ether. The ether left, on +evaporation, a yellow resinous substance having a faint odor like +garlic. By drying in a desiccator, a small quantity of a solid yellow +resin was obtained which was completely soluble in alcohol. A very small +drop of this solution applied to the skin on the end of a glass rod +which had been drawn out to a point caused an eruption in about +thirty-six hours. Following the nomenclature used by Maisch and Pfaff, +this substance will be designated as _Toxicodendrin_, the ending "in" +indicating its glucoside nature. + +The filtrate from the lead precipitate just described was freed from the +excess of lead acetate by hydrogen sulphide, was tested for poison, and +was found to be poisonous, showing that the precipitation by lead +acetate was not complete even in 50 per cent. alcohol. On spontaneous +evaporation of the solution, a yellow, sweet smelling resin was left. + +A portion of the alcoholic solution of the toxicodendrin gave a dark +coloration with ferric chloride, did not reduce Fehling solution and was +slightly acid to litmus. + +To see whether the toxicodendrin could be hydrolyzed, the remainder was +dissolved in alcohol and dilute sulphuric acid was added. A fine, white +precipitate was formed at once which rose to the surface on standing as +a light flocculent substance. The mixture was heated for several days on +a water bath, filtered from unhydrolyzed resin and the filtrate was +neutralized and concentrated in the way already described. The solution +obtained reduced Fehling solution. Not enough was obtained for further +sugar tests, but all the hydrolysis experiments point to the conclusion +that the poisonous substance is a rhamnoside, and is the source of the +sugar in the plant. + +The reaction with ferric chloride observed whenever a lead compound of +the poison is decomposed by hydrogen sulphide may be explained by the +formation of traces of gallic acid or fisetin through the action of the +weak acids present. + +The supply of purified poisonous tar having been exhausted in the +preceding experiments, further study of the active principle is +postponed until more can be prepared. It is highly desirable to +investigate the white precipitate formed by addition of sulphuric acid +to an alcoholic solution of the toxicodendrin. + + +OXIDATION OF THE PURIFIED TAR WITH NITRIC ACID. + +When the purified poisonous material (p. 32) was extracted with 50 per +cent. alcohol, only a small quantity was dissolved as was stated above. +The insoluble residue was treated with fuming nitric acid. Violent +reaction took place at once with copious evolution of red fumes and +heat. When the reaction was over, a sticky red gummy mass was left which +was slightly soluble in cold water and readily soluble in warm alcohol. +The water extract was yellow, and the alcoholic solution was red. That +the water extract contained picric acid was shown by the following +experiments: + + (1) A portion was gently warmed with a few drops of a strong + solution of potassium cyanide and two drops of sodium + hydroxide. The red color of potassium isopurpurate was + formed. + + (2) A portion of the water solution was heated with glucose + and a few drops of sodium hydroxide. The deep red color of + picraminic acid was produced. + + (3) A few drops of an ammoniacal solution of copper sulphate + was added to the water extract. A yellow-green precipitate + was formed. + + (4) The water extract dyed silk, but did not dye cotton + cloth. + + +DISTILLATION OF THE TAR WITH SODA LIME. + +About 25 gm. of the tar left after extracting the original material with +hot water was dissolved in ether and poured into a glass retort +containing soda lime. The ether was distilled out, leaving the tar +intimately mixed with the soda lime. The retort was then gradually +heated. Vapors and liquid were given off, both of which turned red +litmus blue and had a strong odor like tobacco smoke. No odor of ammonia +was detected.[45] At the high temperature of the triple burner, a +semi-solid, red, greasy substance collected in and closed the condenser +tube. This substance had the same powerful odor as the liquid portion of +the distillate. The clear, watery portion of the distillate was +separated from the thicker parts, and was found to contain pyrrol and +pyridine derivatives by the following characteristic tests: + + (1) Wood moistened with hydrochloric acid was turned red by + it. + + (2) Colorless fumes were formed when brought near + hydrochloric acid; mixed with hydrochloric acid, a red + insoluble substance was formed. + + (3) It precipitated the hydroxides of iron, gave a light + blue precipitate with copper sulphate, and a white + precipitate with mercuric chloride. + +The greasy, semi-solid mass was extracted with 10 per cent. hydrochloric +acid and filtered. On addition of a solution of mercuric chloride to the +red filtrate, a brown flocculent precipitate was formed. It was filtered +off and distilled with caustic soda, but the distillate did not contain +pyridine. + + +POTASSIUM PERMANGANATE AS A REMEDY FOR RHUS POISONING.[46] + +In the early stages of this work some experiments were made to see if +potassium permanganate could be used to purify the lead precipitate by +oxidizing the tar brought down in precipitation. It was found that the +permanganate attacked the lead precipitate as well as the other organic +matter in the vessel. This fact and the well-known value of permanganate +in treating skin diseases, its use as an antidote for some kinds of +alkaloid poisoning,[47] as an antidote given to cattle poisoned by +plants,[48] and as an antidote for snake bites,[49] suggested its use as +a remedy for Rhus poisoning. Maisch[50] mentioned that he had used it +with success, but it never came into general use, probably on account of +its staining the skin and clothing. In carrying out this work abundant +opportunities for testing its value as a remedy for the dermatitis +caused by poison ivy were afforded by many cases of accidental and +intentional poisoning. The best example of the latter was obtained with +the ether solution from the extraction of the lead precipitate in the +Soxhlet apparatus (page 28). After removing the ether, a small drop of +the residue was applied to the wrist as described. An itching red spot +about the size of a dime was noticed in thirty-six hours, and it +steadily increased in size. Nearly two days after the application of the +poison, a dilute solution of potassium permanganate containing a little +caustic potash was rubbed into the spot until the pimples were +destroyed. A little black spot was left wherever there had been a +pimple, showing that the permanganate had been reduced to oxide in the +skin. The place was washed and nothing more was thought of it until the +morning following, when it was noticed that the wrist had commenced to +swell during the night, and the characteristic watery secretion was +running from the poisoned spot. More permanganate solution was applied +without potash and the wrist was bandaged, thinking that this would +prevent the spreading of the eruption, but it really facilitated +spreading by becoming saturated with the poisonous fluid and keeping it +in contact with a larger surface of skin. In the meantime the swelling +and inflammation had extended nearly to the elbow. The arm now had the +appearance of having been bitten by a snake. To reduce the swelling it +was immersed in hot water. This seemed to bring out the eruption very +quickly and the blisters were treated with permanganate as fast as they +appeared. The swelling was reduced, but returned during the night. On +the evening following, the forearm was immersed in a bowl of hot +permanganate solution containing a little caustic potash. The solution +was kept as hot as could be borne for about half an hour. After this +bath, the poison was completely oxidized, for the swelling was reduced +and did not return, nor was there any fresh eruption. What appeared to +be a severe case of poisoning was thus cured very quickly. The use of +hot water not only reduces the swelling, but also helps to destroy the +poison. The action of permanganate is also more rapid at high +temperatures. + +The oxidizing power of permanganate, as is well known, is greater in +acid solution than in alkaline, five atoms of oxygen being available in +the former and three in the latter, according to these equations: + + 2 KMnO_{4} + 3 H_{2}SO_{4} = K_{2}SO_{4} + 2 MnSO_{4} + 3 H_{2}O + 5 O. + 2 KMnO_{4} + H_{2}O = 2 MnO_{2} + 2 KOH + 3 O. + +Permanganate was used as a remedy in some cases mixed with dilute +sulphuric acid, and in others, with zinc sulphate; also with lime water. +It was found to be satisfactory whether used alone or with any of the +substances mentioned, provided it was well rubbed into the skin. The +concentration of the solution used was varied according to the location +and condition of the eruption. Where the skin was thin or already +broken, dilute solutions (one per cent.) were used. In one case, the +eruption appeared in the palm of the hand where the skin was so thick +that it was necessary to open it before the remedies could reach the +poison. The difficulty of getting the remedy in contact with the poison +in the skin is the reason why the eruption is hard to cure. + +The remedy most commonly used for this eruption is an alcoholic solution +of lead acetate. This remedy is unsatisfactory for the reason that its +action consists in depositing an unstable lead compound of the poison in +the skin where the conditions of moisture and temperature are favorable +for its decomposition, liberating the poison with all its irritant +properties. Moreover, alcoholic preparations should not be used because +the alcohol dissolves the poison and, on evaporation, leaves it spread +over a larger surface like a varnish. Potassium permanganate, however, +oxidizes the poison completely. The only objection to the use of +permanganate of which the writer is aware is that it stains the skin. +The stain can be removed by vigorous scrubbing with soap, or it will +wear off gradually in a few days. It can be removed at once by certain +acids, but these should not be used by persons not familiar with their +action. + +With the knowledge of the facts mentioned, many solutions were tested +for poison by applying them to the skin, and when an eruption appeared, +it was cured quickly and permanently by rubbing in a permanganate +solution, usually mixed with dilute sulphuric acid. + +FOOTNOTES: + +[16] Nitrogen was found very readily by the soda lime test in the tar +left after extracting the original material with 50 per cent. alcohol, +but was not found by the Lassaign test. + +[17] Stevens. Amer. Jour. Pharm. 77, 255, June, 1905. + +[18] Whenever it is stated in this paper that a solution was poisonous +or not poisonous, the test was made by the writer upon himself. + +[19] Liebig's Annalen, CXI, p. 215. + +[20] Ueber Mategerbstoff, p. 20. + +[21] Bull. Soc. Chim. (II), Vol. 2, 95 (1864). + +[22] Berichte 19, 1735 (1886). + +[23] Jour. Chem. Soc. 71, 1194 (1897). + +[24] Berichte 19, 1740. + +[25] Ibid. 1747; Annalen, 112, 97. + +[26] Biochem. Pflan. II, 521. + +[27] Ann. de Chim. et de Phys., 6th Series, XXII, 76 (1891). + +[28] Treatise on Chem., Vol. III, Pt. III, 492. + +[29] Les Sucres; Chem. der Zuck.; Biochem. der Pflan. + +[30] Chem. Zeit. 23, Rep. 177. + +[31] Loc. cit. 1, 209. + +[32] On standing several weeks, a small quantity of tar separated out on +the walls of the vessel, also a brown precipitate which was filtered +off, suspended in water, and hydrogen sulphide was being passed in when +an accident occurred and it was lost. + +[33] "By warming with alkalies or barium hydroxide, rhamnose is colored +yellow." Chem. der Zuck. I, 177. + +[34] Ibid. 188. + +[35] Ibid. + +[36] Rayman, Sur L'Isodulcite, _Bull. Soc. Chim._ 47, 668 (1887). + +[37] Acides Gummiques. + +[38] Berichte XX, pp. 1089, 1091, 1188, 2566. + +[39] Ann. de Chim. et de Phys. (6) XXII, 93 (1891). + +[40] Biochem. der Pflan. I, 210. + +[41] Comptes rendus CXV, 892. + +[42] Amer. Chem. Jour. 28, 370. + +[43] Amer. Jour. Pharm. 77, 255 (June, 1905); 78, 53 (Feb., 1906). + +[44] A wax obtained from _Rhus succedanea_ was shown by Stahmer to +contain palmitic acid and glycerol in the form of glycerol palmitate. +_Annalen_ 43, 343, (1842). + +[45] See Amer. Jour. Pharm. 77, 256. + +[46] This section is added in the hope that it may be of use to others +who are subject to this form of poisoning. + +[47] Moor, N. Y. Med. Rec. 45 (1894), 200. + +[48] Bull. No. 26, U. S. Dept. Agr., Div. of Bot. 47. + +[49] Lacerda, Comptes rendus 93 (1881) 466-469. + +[50] Amer. Jour. Med. Sci. 52 (1866), 285. + + + + +SUMMARY. + + +Leaves and flowers of the poison ivy plant were extracted with ether and +the ether was removed by evaporation. In the residue, the following +substances were found and studied: gallic acid, fisetin, the sugar +rhamnose, and a poisonous tar, gum, or wax. + +The lead compound of the poison was soluble in ether; this fact gave a +means of separating the poisonous substance from the non-poisonous +matter in one operation. + +The poison was not volatile with vapor of acetic acid, or with vapor of +alcohol. + +The poisonous tar or wax was decomposed by acids and yielded gallic +acid, fisetin, and rhamnose, showing the probable source of these +compounds in the plant, and indicating that the poison is a complex +substance of a glucoside nature. + +It was found that a portion of the poisonous substance could be +precipitated by lead acetate from a solution of the purified tar in 50 +per cent. alcohol. + +All cases of poisoning developed on the writer were easily cured with +potassium permanganate. + +The following method is suggested for obtaining the poisonous substance +from the plant: Extract the plant with alcohol, filter, and precipitate +at once with lead acetate. Wash the precipitate, dry, and extract with +ether in Soxhlet extractors (loosely filled). Combine the ether +extracts, mix with water, and pass in hydrogen sulphide. Separate the +water and the ether solution, and filter the latter. Wash the ether +solution thoroughly by shaking with water, and then evaporate at a low +temperature. + + + + +BIOGRAPHY. + + +William Anderson Syme, the author of this dissertation, was born in +Raleigh, N. C., on July 11, 1879. He was prepared for college at the +Raleigh Male Academy, entered the North Carolina College of Agriculture +and Mechanic Arts in 1896, and was graduated in 1899 with the degree B. +S. He was an Instructor in Chemistry at the same College from January +1st, 1900, until June, 1903, when he received the degree M. S. for +graduate work. In October following, he entered Johns Hopkins University +as a graduate student in Chemistry, and was awarded one of the North +Carolina Scholarships. His minor subjects are Physical Chemistry and +Biology. + + + + + + +End of the Project Gutenberg EBook of Some Constituents of the Poison Ivy +Plant: (Rhus Toxicodendron), by William Anderson Syme + +*** END OF THIS PROJECT GUTENBERG EBOOK SOME CONSTITUENTS--POISON IVY PLANT *** + +***** This file should be named 34510.txt or 34510.zip ***** +This and all associated files of various formats will be found in: + https://www.gutenberg.org/3/4/5/1/34510/ + +Produced by Bryan Ness, Josephine Paolucci and the Online +Distributed Proofreading Team at https://www.pgdp.net. 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