MEMCAIL SCHOOL Exchange Detroit Public Library iedical Science Department r Laboratory Guide IN Experimental Pharmacology DIRECTIONS FOR THE COURSE GIVEN IN THE University of Michigan BY CHARLES W. EDMUNDS, A. B., M. D. j5^ INSTRUCTOR IN PHARMACOLOGY IN THE UNIVERSITY OF MICHIGAN AND ARTHUR R. CUSHNY, A. M., M. D. PROFESSOR OF PHARMACOLOGY IN UNIVERSITY COLLEGE, LONDON; AND LATE PROFESSOR OF MATERIA MEDICA AND THERAPEUTICS IN THE UNIVERSITY OF MICHIGAN GEORGE WAHR PUBLISHER AND BOOKSELLER ANN ARBOR, MICH. COPYRIGHT 1905 BY GEORGE WAHR v, PREFACE. The following course has been gradually developed in the pharmacological laboratory of the University of Michigan, and it has been suggested that if put in a more permanent form than hitherto it might be helpful to other teachers of this subject. The practical course has hitherto been given as an intro- duction to the didactic lectures and it has been found advis- able to limit the number of drugs examined to those possess- ing the most typical action. The object has been to train the student entering on the subject in the method of work and to impress on him that the study of drugs is to be approached in the same objective way as other branches of medicine. The order in which the experiments are arranged is a purely arbitrary one, necessitated by the conditions of the laboratory, and can doubtless be modified with advantage to suit other classes. A few days are devoted to purely pharmaceutical work, but the chief weight is laid on the experimental part of the course. The few hours devoted to pharmacy, however, train the student in observing the characters of the much larger number of drugs met in the lecture course, and have been found to be of considerable value. A beginning may be 3 4 PREFACE. made in prescribing as occasion offers, but this, as well as the therapeutic applications of the results obtained in the phar- macological experiments, is better left to the teacher than expounded in the laboratory directions. ARTHUR R. CUSHNY. CHARTS W. EDMUNDS. Ann Arbor, March i8th, 1905. We wish to express our thanks to Mr. Arthur J. Jones for his excellent drawings of the turtle, the dog's head and the frog with its heart exposed. CONTENTS. PAGE. The Pharmacopoeia 7 General directions for anaesthetics and operations n Chemistry of drugs : 24 Physiology of frog's nervous system 44 Soporifics or Hypnotics 5 1 Percolation 59 Nux vomica 64 Thujon 79 Opium 80 Curara 88 Nicotine 92 Veratrine 95 Caffeine 96 Cocaine : 100 Cinchona 103 Emulsions 108 Pills 112 Digitalis series 119 Aconite 128 Drugs on the frog's heart 132 Drugs on the rabbit's heart 135 Drugs on the turtle's heart 139 Drugs on the blood pressure 147 Drugs on the dog's heart 151 Analysis of tracings 160 Perfusion of the kidney 167 Diuresis 168 Belladonna series 179 Drugs on the sympathetic system 187 5 6 CONTENTS. PAGE. Salivary secretion 191 The sphygmomanometer 199 Anaesthetics 204 Pilocarpus ....'. 208 Suprarenal extract 212 Nitrite series 216 Intestinal peristalsis 219 Vasomotor effects in the intestine 223 Vasomotor effects in the kidney 228 Drugs and reagents required 235 The Pharmacopoeia. When a physician prescribes a certain drug from time to time in his practice it is of greatest importance to him to know that his patients are always getting the same prepara- tion, containing the same active principles in uniform strength. These requirements could not be carried out if there were not some standard recognized by the manufac- turers and followed by them in the preparation of the various medicinal remedies. Most countries possess such a standard in their " Pharmacopoeias," which are generally published by the government. In this country, the government does not issue the pharmacopoeia, but it recognizes its au- thority. The book is revised every ten years by a Committee of Revision composed of members appointed or elected by a convention of various medical and pharmaceutical societies and colleges. It need hardly be remarked that in such a work only such remedies are included as have become well established. These are designated as "official" remedies, while drugs not mentioned are "non-official." The pharmacopoeia gives, first, the Latin title of the drug followed by the English name, and in the case of chemicals the formula and molecular weight are given. Any syno- nyms are mentioned and wherever it is necessary, a short, concise definition of the drug is added, telling its nature and its source. The book then gives tests by which the identity of the drug may be recognized, and its purity and strength 7 8 LABORATORY GUIDE IN tested and finally mentions in what ways and in what doses it may be administered. Official preparations are therefore made according to the directions given in the pharmacopoeias and they should possess uniform strength and properties; such an' ideal, however, is hard to attain, as some of the crude drugs, even when collected according to pharmacopceial directions, dif- fer in their content of the active constituents. EXPERIMENTAL PHARMACOLOGY. 10 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 11 General Directions. Before giving the specific directions for the experimental work a number of general directions are given for the com- moner operations which are carried out on animals some of which are usually necessary in subsequent manipulations. The various anaesthetics employed in experimental work are also described. Anaesthetics. No animal is to be operated on unless it is under complete anaesthesia. Ether and chloroform may be used for this pur- pose, but have the disadvantage of requiring constant atten- tion. They are therefore only used to supplement other anaesthetics which experience has shown can be used to give full anaesthesia in animals without the necessity of their being continually administered. For this purpose the following combinations of drugs usually give very good results, al- though in rare cases chloroform or ether may have to be given in addition. Anesthetic for Rabbits. Paraldehyde, 1.7 cc. per Kg. of body weight. While your assistant holds up the animal by all four legs and head, place a gag in the mouth and pass a stomach tube 1 through the opening in the gag, being very careful not to pass it into the lungs. Draw the paraldehyde into a pipette *A stomach tube for rabbits and cats may be made from a semi-elastic catheter (No. 7 or No. 8) by cutting off the ivory tip and fitting in its place a short piece of rubber tubing. If preferred, the soft rubber catheter may be used. 12 LABORATORY GUIDE IN and place the point of the pipette in the opening of the stomach tube and blow the drug into the stomach, and then withdraw the tube. Anesthetic for Cats. Morphine sulphate, 0.040 to 0.060 G. Chloretone, 0.3 G. per Kg. of body weight. Place the animal in a cat box, 1 draw the head forward and with a hypodermic syringe inject the morphine solution under the skin of the neck. Dissolve the chloretone in as small a volume of alcohol as possible and add about an equal volume of water. Keep the animal in the box and introduce this solution through the stomach tube in the way described under "Anaesthetic for Rabbits." Anesthetic for a Dog. Morphine sulphate, 0.2 G. to 0.25 G. Chloretone, 3 G. to 40. Three or four hours before the animal is needed inject the morphine solution under the skin by means of a hypodermic syringe. Just before operation, dissolve the chloretone in alcohol and add about an equal amount of water and give it to the dog by means of a stomach tube 2 passed through a wooden gag. a A cat box is of the greatest service as a means of protection to the operator whenever this animal is used. It consists of an ordinary wooden box about 30 or 35 cm. long, 18 cm. wide and 15 or 18 cm. deep. It is provided with a sliding cover, which has a V-shaped cut in the middle of one end. In the corresponding end of the box a second V cut is made. When the lid is shut these two cuts leave enough room for the animal's neck. The lid is kept closed tightly by means of a nail passed through the opposite end of the cover into the end of the box. 2 The average stomach tube, such as is used in clinical medicine, is about the right size for a dog. It should be about 75 cm. long. EXPERIMENTAL PHARMACOLOGY. 13 14 . LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 15 Operations on Frogs. Pithing. Before exposing the heart of a frog or carrying out any other operation upon the animal its brain is destroyed (pithed) in the following manner: Hold the frog in the left hand with its back upward, tip the head forward with the left index finger, and feel for the prominent angle made at the junction of the skull and spinal column. Just back of this angle make a cut with small scissors through the skin in the median line. Now push the sharpened end of a match forward through the foramen magnum into the skull, rotat- ing it in the cranial cavity to destroy the brain. Injecting into a Lymph Sac. In the intact animal, drugs to be tested are usually injected into the anterior lymph sac. This is carried out in the fol- lowing manner : Lay the animal back downwards in the palm of the left hand. Hold one of its forelegs firmly between the thumb and index finger and the other foreleg between the middle and ring fingers. Draw its hind legs downward and hold them against the palmar surface of the hand by means of the little finger. Having the drug in the glass injecting pipette, 1 which is held in the right hand, force the animal's mouth open with the point. Pass the pipette into the mouth avoiding the tongue, which is attached anteriorly and direct the point to- 1 To make a glass injecting pipette take a piece of glass tubing about 7 mm. in diameter and about 20 or 30 cm. long and heat the middle point in the Bun- sen flame, rotating the tube constantly, and when it is red hot remove it from the flame and draw it out to a capillary size; cut it in the middle so as to leave a fine tube about 8 cm. long. Heat the large end of the pipette to smooth it off so it will not cut the tongue or lips. 2-p 16 LABORATORY GUIDE IN ward the floor of the mouth which with a little pressure it will pierce, entering the lymph sac. As it is pushed down the sac the point can be seen beneath the skin of the abdominal wall. The finger is now removed from the upper end of the pipette and the drug allowed to flow into the sac, or if neces- sary blown in. FIG. No. i. To Expose the Heart. Pith the frog and tie it on its back on the frog board. With a pair of forceps raise the skin in the median line of EXPERIMENTAL PHARMACOLOGY. 17 18 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 19 the body just below the point of the sternum and make a nick in it with a pair of scissors. Starting, at this point cut up and outward on each side as far as the pectoral girdle and turn this V-shaped piece of skin upward toward the head. Raise the sternum by placing the forceps beneath the point and with scissors cut it through its entire length avoiding in- jury to the underlying heart. Tighten the strings on the fore legs so as to pull the sides of the sternum well apart. The heart is now seen enclosed in the pericardium, which may be carefully opened and cut away. Directions for Operations on Mammals. The following are the most common operations necessary in experimental pharmacology. To Insert a Venous Cannula. This is to allow of the intravenous injection of drugs. FIG. No. 2. Two sizes of glass venous cannulas (with rubber tubing attached) are shown in Fig. No. 2. The larger is best suited for a dog, while the smaller is used in a rabbit or cat. Vari- ous sizes should be provided. Operation. When the anaesthesia is complete make an in- 20 LABORATORY GUIDE IN cision through the skin over the site of the vessel and deepen the cut through the fascia until the v.ein is exposed. Avoid- ing injury to the vessel from forceps or tearing, clear about two inches of the vein, removing all the fascia surrounding it so as to leave it entirely free. Pass two threads about a foot long under the vein and place "bull-dog" forceps on it toward the cardiac end of the cleared portion. This allows distention of the vessel by the blood. Tie one of the threads tightly around the upper end of the cleared portion. Hold the end of this ligature in the left hand and place the left index finger under the vessel, thus making it tense. With sharp scissors make a cut in the distended vein and place the tip of the cannula in it, and tie it in place with the loose thread previously placed around the vessel. Fill the cannula and vein with salt solution (0.8 %), taking great care that all air is expelled. PRECAUTION. When drugs are injected intravenously it is of the utmost importance to see that no air is allowed to enter the veins. A small bubble of air is almost invariably fatal to rabbits. Be sure the cannula and syringe have all the air expelled. To Insert an Arterial Cannula. 1 This is to allow records of arterial blood pressure to be taken. Operation. Expose and clear the artery, usually the carotid, in the manner described above for the vein. Pass two ligatures under the vessel and tie the thread which is distal to the heart. Now clamp the artery with the "bull- dog" forceps, placing them on the "heart end" of the cleared 'Arterial cannulas are exactly like venous cannulas. Fig. No. 2. EXPERIMENTAL PHARMACOLOGY. 21 22 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 23 portion. Place the index finger under the vessel as before and make a cut in it and tie a cannula in place with the loose ligature. This cannula is to be rilled with sodium sulphate solution (page 147) to prevent clotting of the blood. To Insert a Tracked Cannula. FIG. No. 3. Glass trachea! cannula with rubber tubing attached. A tracheal cannula is inserted to allow of artificial respira- tion being carried on in case the chest is to be opened or if the anaesthesia is so deep that the respiratory center is seriously depressed or paralyzed. A convenient form of tracheal tube is shown in Fig. 3. They can be made of various sizes of glass tubing so as to fit the trachea of any animal. When in use the piece of rubber tubing shown in the cut is partially constricted to give the proper degree of inflation to the lungs. Operation. Make a median incision in the neck through the skin and fascia beginning just below the thyroid cartil- age. Expose the trachea by separating the muscles with the fingers or blunt hooks and dissect away all structures from around it. Pass a strong thread under it and then make a transverse cut with the scalpel between two rings about two- thirds of the way through. Into this opening pass the tracheal tube and tie it in place with the loose ligature. 24 LABORATORY GUIDE IN Chemistry of Drugs. Alkaloids. Among the most important of the organic compounds em- ployed in medicine is the group of vegetable bases known as Alkaloids. They are found in almost all parts of plants, but in greatest abundance in the seeds and roots. They consti- tute in almost all cases the active constituents of the plants in which they are found and some of them are among the most powerful poisons known. The alkaloids contain carbon, hydrogen, nitrogen, and usually oxygen, but in a few instances the latter is not pres- ent. In their chemical properties they resemble ammonia very closely, having an alkaline reaction and in uniting with acids without the elimination of hydrogen to form salts. These salts differ from the alkaloids in their solubility, and upon this fact is based one method of isolating them in a pure form from the plants in which they are found. One of the most familiar alkaloids is quinine, which is de- rived from various species of cinchona bark, and it may be taken as a type of the group, as the reactions given by it are characteristic of most of the other alkaloids. Taste quinine and quinine sulphate. I. Test the solubility of a few crystals of quinine (alkaloid) and of quinine sulphate (alkaloidal salt) in 10 cc. of each of the following: EXPERIMENTAL PHARMACOLOGY. 25 26 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 27 Solvent. Alkaloid solubility. Alkaloidal salt solubility. Sts^fjjrf a Water. *~7/^ b Alcohol. *f~ *// se^-^Cci c Ether. ^J^^c^y - d Chloroform. gu fc^ < --^ EXPERIMENTAL PHARMACOLOGY. 67 solve this extract in the 35 cc. saved and make the total bulk up to 50 cc. with diluted alcohol. Define a fluid extract. III. Reserve 5 cc. of the fluid extract made in Exp. II. and from the remaining 45 cc. isolate the strychnine in the following manner : Dilute the fluid extract with about its own volume of dis- tilled water and pour it into a separating funnel and render it alkaline with ammonia. Add about 25 cc. of a mixture of chloroform and ether in the proportion of 2: I and shake gently for three minutes and set the funnel aside to allow the chloroformic solution to separate. When separation has taken place, draw the chloroform and ether off into a beaker and repeat the agitation of the watery extract, using a fresh supply of chloroform and ether. Draw this off as before into the beaker with the first solution. The watery extract can now be thrown away. Pour the chloroform and ether into the separating funnel, which has been washed, and add about 25 cc. of water and enough dilute H.,SO 4 to make the solution distinctly acid, shake gently for a few minutes and then separate the water and chloroform as before, placing the acidulated water in a flask. Repeat the agitation of the chloroform and ether with fresh acidulated water, adding the latter when separation has taken place, to the first lot in the flask. (The waste chloroform is to be poured into a large bottle provided for it and later redistilled.) If the acidulated watery solution is much colored it should be shaken in the flask with charcoal and filtered. Render the filtrate dis- tinctly alkaline with ammonia and shake 'it out twice as be- fore in the separating funnel, with small quantities of the mixture of chloroform and ether. When this has been done, pour this chloroformic solution into a glass evaporating dish, 68 LABORATORY GUIDE IN add to it 2 or 3 drops of concentrated hydrochloric acid ,and stir it well with a glass rod or put the mixture in a small flask and shake it well. The solution becomes milky from the formation of strychnine hydrochloride, which is thrown out of solution and may be separated from the chloroform by allowing the latter to evaporate spontaneously or by warming it gently over a water bath. CAUTION. In this process two things are of prime im- portance in determining the success of the manipulations : First, to agitate sufficiently the chloroform and ammonia or sulphuric acid solutions to allow thorough intermingling of the solutions. Three to five minutes should be devoted to each shaking. Second, make sure that the correct reaction is present. Test with litmus paper each time and after shaking for a minute or two, test again. (See Alkaloids, Exp. No. II.) The reaction must be distinct each time. Explain why each step in the manipulation was carried out. (Compare Alkaloids, Exp. Nos. I, III, VI.) The crystallized strychnine hydrochloride is to be used in tests given below. (Exp. Nos. IX, X, XII.) IV. To a few cubic centimeters of the infusion made in Exp. I add a few drops of ferric chloride solution. The dark color of the mixture is due to the tannic acid present in the nux vomica bean uniting with the iron to form ferric tannate. (To a dilute solution of tannic acid add ferric chloride.) V. Boil a small amount of Fehling solution and add 3 or 4 cc. of the infusion and see if there is any reduction. NOTE. There is a reducing substance found in the nux vomica bean. VI. Inject I cc. of the infusion into the lymph sac of a frog, and replacing the animal under a bell jar, observe the EXPERIMENTAL PHARMACOLOGY. 69 70 LABORATORY GUIDE IN /xX /? jp s^^+*-^r~&-~^' (Ze^Z^^ v /^.45 ^^ H^/-^^ ^1/^^^x^ 4^*^ /t/ U * r ^ >^-+ sy^r&^f- b c Picric acid. ' Iodine in potassium iodide. ^tx^-W~ 1-20,000 1-100,000 d Mercury-potassium iodide. /- ^1/2^6; 1-150,000 e Phosphotungstic acid. + */^ 1-200,000 XIII. With a hypodermic syringe inject subcutaneously into a rabbit I mg. of strychnine sulphate. Watch the effect on reflexes, etc. If convulsions come on control them with a few drops of chloroform on cotton held close to the ani- mal's nose. Compare the convulsions in this animal with those seen in the frog. Note the efficiency of chloroform in controlling the convulsions of strychnine. Therapeutic On account of their intensely bitter taste, preparations of nux vomica are employed in cases of loss of appetite and malnutrition. By their action on the taste organs they probably increase reflexly the secretion of the gastric juice and thus aid digestion. Strychnine is largely used as a stimulant to the central nervous system, being useful in various conditions in which depression of the brain or cord is found. It is employed in shock or collapse, and in failure of the respiratory or of the vaso-motor center. By stimulating the activity of the spinal cord to a slight extent it increases the "tone" of the muscles and of the blood vessels whose muscular coats are EXPERIMENTAL PHARMACOLOGY. 77 - f s&^tffi&i**-^^. . <^l. .jt^^J / # I ^ Cf^L<^r7>t^^^ EXPERIMENTAL PHARMACOLOGY. 79 controlled by the spinal centers. Perhaps more than any other drug it deserves the name "tonic." Thujon. Thujon is the poisonous principle derived from the volatile oils of sage, tansy and absinthe. It is an isomere of camphor (C 10 H 16 O) and may be taken as an example of that series, as it acts in the body in a manner closely resembling that t drug. I. Place a cat in a cat-box (page 12) and pass a tube through the mouth gag into the stomach and inject by means of a pipette about i cc. of thujon. The end of the pipette containing the thujon is inserted into the opening of the stomach tube and the drug blown in and afterward washed down with some water. The stomach tube and gag are now removed and the animal is freed and placed in a cage. Compare the symptoms with those caused by strychnine in the rabbit. It will be seen that the convulsions caused by thujon are of a distinctly different type from those following a poisonous dose of strychnine. Those caused by the latter drug are tonic in type and are due to an action on the spinal cord. On the other hand, the thujon (and camphor) convulsions are "clonic" and are caused by stimulation of some unknown part of the cerebrum or lower part of the brain. Therapeutic uses** Thujon is not employed in therapeutics. Its pharmacolog- ical action would indicate that it would be of value as a stimulant to the central nervous system and especially to 6-p 80 LABORATORY GUIDE IN the respiratory center. As such it would be of value in shock and collapse and perhaps in narcotic poisoning. Camphor has long been used in shock and collapse. Opium. Opium is the inspissated juice obtained from the fruit- capsules of the white poppy (Papaver somniferum). The capsules are incised while yet unripe and the milky juice allowed to exude and dry by spontaneous evaporation until it becomes a brownish gummy mass. The poppy is grown in Turkey, Asia Minor, Persia, India and China, but that obtained from Smyrna and Turkey is by far the most valuable. Persian and Indian opium is im- ported mainly as sources of the opium alkaloids and the Chinese opium is consumed entirely in that country. Opium contains about twenty alkaloids of which Morphine and Codeine are the most important, and, indeed, it may be said that the value of opium depends upon the per cent of morphine which it contains. The U. S. Pharmacopoeia specifies that the crude opium when moist shall contain not less than 9 per cent of crystallizable morphine, while the dried product must assay from 13 to 15 per cent. Opium from the different localities varies exceedingly in its mor- phine content. Morphine was discovered in 1817 by Seitiirner, of Eim- beck, Germany, and was named by him morphium. It exists in the opium principally in combination with meconic acid, although some may be present as a lactate or a sulphate, both of these acids being found in the crude drug. Meconic acid is of most importance as it is strictly characteristic of EXPERIMENTAL PHARMACOLOGY. 81 82 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 83 opium and its preparations, and its positive detection is proof of the presence of some opium preparation. In their solubilities morphine and codeine differ widely from the other alkaloids, for morphine is almost insoluble in chloroform and only very slightly soluble in ether (1-6100); codeine, on the other hand, is more soluble in water than any other alkaloid. Morphine Reactions. I. Put a few crystals of morphine on a porcelain cover and add a drop or two of pure concentrated sulphuric acid and mix the two with a clean glass rod. No color should be formed if the morphine is absolutely pure, but occasionally a faint pink tint is seen. Heat the mixture very gently. At about 150 a dirty green or rose-red color is developed, and on still further heating, the solution becomes almost black. Allow it to cool and add some water, when a greenish blue color appears which changes to green when ammonia is added in excess. / II. Put a few crystals of morphine on a cover and mix them with a drop or two of concentrated sulphuric acid as in Exp. I. To this add a drop of distilled water which will heat the mixture. Now treat with a drop of concentrated nitric acid, which should give a rose-red color, changing to brown. (This reaction for morphine is very delicate, being well shown by o.oi milligram of the alkaloid.) III. To a drop of concentrated nitric acid on a porcelain cover add a minute amount of morphine. An orange-red color is obtained, which changes to yellow on heating. IV. (a) Mix a small amount of morphine with about twice its weight of cane sugar. By means of a glass rod add /^ 84 LABORATORY GUIDE IN a drop of concentrated sulphuric acid. This should produce a purple color, changing gradually to blood-red and brown- ish-red and becoming an olive-brown on dilution with water. (b) The above test can be modified as follows: To a solution of morphine add cane sugar until the solution is saturated. Now pour this solution carefully down the side of a second test tube in the bottom of which is concentrated sulphuric acid. By inclining the second tube the solution can be poured in so that no mixture of the two fluids takes place and a purple or rose-red ring will form at their junc- tion. This ring becomes more intense on standing. V. Dissolve some morphine sulphate in a little water and add it to a solution of iodic acid. What color does it turn ? Add to the resulting mixture some thin starch paste. What was the first color due to? NOTE. In order tp color the starch paste blue morphine must be present in the strength of i to 1000. If no blue is obtained mix the solution well and pour down the side of the test tube some very dilute ammonia. A double ring will be seea at the junction of the two fluids; it will be blue below and brown above. VI. To a few cubic centimeters of water add some mor- phine sulphate and to this solution add a few drops of ferric chloride solution. A deep blue color is obtained. I. Meconic acid reaction. To a cubic centimeter of tmcture of opium diluted with a little water add two drops of ferric chloride solution. What color does the solution become? If the color is obscured by a precipitate, filter the solution. VIII. Inject into the anterior lymph sac of a frog 25 mg. of morphine sulphate. Replace the animal under the bell jar and observe. Examine it from time to time, comparing the effects with those caused by chloral or strychnine. EXPERIMENTAL PHARMACOLOGY. 8S ^-Z^z- - p/^ - ~/3^^5 Xi^^O ^ ^c* ,t?, ^ ^^^_^^C^^, .- ^^/^L^fe ^-i * s&^ -^<.^^-<^Y/ 86 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOWGY. 87 NOTE. This experiment (VIII) should be begun early in the laboratory ses- sion as it sometimes takes two or three hours for the full effects to appear. Among the alkaloids found in opium which are of less importance than those studied is thebaine, which resembles strychnine very closely in its pharmacological action. In fact, many of the alkaloids of opium may be arranged in a series with the depressant, morphine, at one end of the series and thebaine at the opposite end. IX. Inject subcutaneously into a dog 50 mg. of morphine sulphate and observe the effects. X. Inject 30 to 40 mg. of morphine sulphate subcutane- ously into a cat. Put the animal in a wire cage and compare the effects with those seen in the dog. The peculiar action of morphine on the cat is not confined to that animal, but is found in all the members of the cat tribe, and also in the horse and ass. Examine a. crude opium, b. powdered opium, c. extract of opium, d., tincture of opium (Laudanum), e. camphorated tincture of opium (Paregoric), f. powdered opium and ipecac (Dover's powder) . g. and codeine. Note the odor of all the opium preparations and taste d. e, and f. Therapeutics. The symptoms induced by opium in man resemble more closely those in the dog than those in the other animals. It depresses the brain, and for this reason is used in cases of sleeplessness when this condition results from pain, because in depressing the cerebrum it acts on the pain centers, for 88 LABORATORY GUIDE IN which it seems to have a special affinity. In the dog the respiration was slowed in the later stages of the action from depression of the respiratory center, and opium prepara- tions are often employed in cough mixtures to allay exces- sive irritability of this part of the medulla. The -numerous other purposes for which opium and morphine are used cannot be illustrated experimentally. Curara. Curara, the South American arrow poison, varies in com- position with the different localities from which it is ob- tained. The best known product comes from Guiana and is prepared from the bark of Strychnos toxifera, a tree native to that country. Curara as obtained in commerce consists of dark brown, shining, brittle masses which contain vegetable extracts in addition to two alkaloids, tubocurarine and curine. The characteristic action of the drug is dependent principally upon the first alkaloid mentioned, the latter hav- ing quite a different action and being much weaker. Not only do solutions of curara 1 deteriorate on keeping, but, what is of greater importance, the crude drug, as ob- tained in the market, varies greatly in strength, many of the preparations being entirely inert. There is no chemical method by which the activity of curara can be estimated; the only test is pharmacological. Each solution to be used has to be "standardized" by tests on animals and the dose necessary to produce characteristic effects ascertained. I. Inject into a frog a poisonous dose of a standardized solution of curara. Study the effect on spontaneous move- 1 The curara solution should be made up in physiological salt solution to ,'hich some thymol has been added. EXPERIMENTAL PHARMACOLOGY. 89 x*^rz 90 LABORATORY GUIDE IN x-***^r* r^ (7. ^xr /tL^J} t^v EXPERIMENTAL PHARMACOLOGY. 91 ments, reflexes, ability to turn over when placed on its back, and length of time leg muscles will remain contracted when stimulated by applying the tetanizing current to the lumbar spine. If paralysis should occur see if it is due to central or peripheral action (see Soporifics, Exp. Ill, page 55). This animal is to be replaced under the bell jar and kept moist, as it will probably recover in a few days. II. Pith a frog, tie it on the frog board and expose the sciatic nerve and artery along the back of the thigh by a short longitudinal incision through the skin, and careful separation of the muscles with the dissecting needle. Sepa- rate the sciatic nerve from the surrounding tissues along a short distance of its course and then pass a thread under the nerve and tie it tightly around the rest of the thigh as high up as possible. Free the animal, inject curara as in I, and from time to time compare the reflexes in the hind legs. Stimulate the lumbar cord with the electric current and note any differences between the two sides. See whether the reflexes remain equal in the two legs and whether cross reflexes can be obtained. If the experiment is successful the operated leg should not be affected by the poison and should respond to electrical stimulation. In such a case cut off the leg just above the knee, leaving attached to it the sciatic nerve which has been carefully separated from the thigh muscles and cut toward the upper part of its course. Holding the leg preparation by the severed end of the femur lay the hanging sciatic nerve for a few moments in the curara solution. Remove it and stimulate it with the electric current. Does curara affect the nerve trunk? Dip the muscle in the solution and after a few moments stimulate the sciatic nerve again. Is there 9: LABORATORY GUIDE IN any change? If so, test the activity of the muscles by direct stimulation. Where does curara act ? Examine the heart of the animal in Exp. II. Examine Curara. Therapeutics. The main importance of curara is in physiological experi- ments. It has been suggested, and in some few cases used, to allay the convulsions due to tetanus or to strychnine poisoning. It is not recommended for such conditions as its action is too uncertain and too dangerous. Nicotine. The alkaloid, nicotine, is obtained from the dried leaf of the tobacco plant (Nicotianum Tabacum) in which it is found to the extent of from 2 to 8 per cent. The pure alka- loid is a colorless, oily fluid, which, on exposure to air, be- comes yellowish or brown. It has a strong, unpleasant odor resembling somewhat that of tobacco. When tobacco is smoked, only a small amount of the alkaloid appears in the smoke, the larger part is decomposed in the process and it may be that some of the effects of smoking are due to the absorption of these decomposition products rather than to the nicotine itself. In an aqueous solution the alkaloid gives a strong alkaline reaction and it unites with various acids to form salts. I. Inject i cc. of nicotine chloride solution 1 into the anterior lymph sac of a frog. Observe the effects, noting especially the posture of the animal in the early stages of 1 The nicotine chloride solution is formed by neutralizing a small quantity ot nicotine with 1-20 normal hydrochloric acid. Each cubic centimeter will then contain about 4 mgs. of nicotine. EXPERIMENTAL PHARMACOLOGY. 93 94 LABORATORY GUIDE IN , v ^ _ HXPHRIMHNTAL PHARMACOLOGY. 95 poisoning as this is very characteristic of nicotine action. In addition, watch for any twitching of the muscles, and if paralysis should come on see whether it is due to central or peripheral action. The action of nicotine upon the sympathetic nervous sys- tem is studied later (pages 187, 195). Therapeutics. Nicotine is not employed in therapeutics. Veratrine. Veratrine is an alkaloid prepared from the seeds of Asa- groea officinalis. It is a grayish powder which causes very marked irritation when even a minute quantity is inhaled. The alkaloid is not of great medical importance but is of pharmacological interest, owing to a peculiar action it possesses which can be best studied in cold-blood ani- mals, although the same effect may be elicited in mammals. I. Inject i mg. of veratrine sulphate (i cc. of o.i % sol.) into a frog. Watch and describe the effects. Observe the awkwardness of its movements which appears in a few min- utes. When the clumsiness is well developed locate the point at which the drug acts by removing the cerebrum, optic lobes, and finally destroying the spinal cord if the symptoms do not disappear earlier. Stimulate the sciatic plexus with single shocks from the induction coil, comparing the results with those seen in a normal frog. Finally, stimu- late the muscle directly, using single shocks. At what point does the drug act? 7-p 96 LABORATORY GUIDE IN Therapeutics. The phenomena seen in this experiment give no hint as to the use of the drug in therapeutics; it is employed for ex- ternal application, and internally to a limited extent for its effects on the circulation. Caffeine. Caffeine differs from the other alkaloids in the fact that it is contained in a number of plants belonging to different species. It is obtained from the berry of the coffee (Coffea Arabica) ; from the leaves of tea (Thea Chinensis), besides being present in the kola nut of Central Africa; in the Paraguay tea and Guarana paste of South America and in the Apalache tea of Virginia and Carolina. In some of these plants it is associated with two closely related alkaloids, Theobromine and Theophyllin, which re- semble caffeine very much in some of their actions. All three alkaloids are derivatives of xanthine, which in turn is closely related to uric acid. Caffeine is a very feeble base and forms salts only with difficulty and these salts, as a rule, are very unstable, being easily decomposed, even by water. I. Test the solubility of caffeine in Reagent. Solubility. / J^c-tx^ a Water, cold. + rf A A / tQ V /_ // b c d hot. Alcohol. Chloroform. + If' ^ (ah e^u ^^ EXPERIMENTAL PHARMACOLOGY <0~ /? 9 8 LABORATORY GUIDti IN EXPERIMENTAL PHARMACOLOGY. 99 II. Apply the various alkaloidal reagents to a i% solu- of caffeine. Reagent. Precipitation? a Tannic acid. ^s5n!^ b Picric acid. c Iodine in potassium iodide. - d Mercury-potassium iodide. e Phosphotungstic acid. + / -. ^r EXPERIMENTAL PHARMACOLOGY. 119 The Digitalis Series. This series embraces a large number of drugs which re- semble each other in their pharmacological action. While they act on various organs in the body, the cardiac action is characteristic of the group and distinguishes it from all others. The members of the series are derived from plants widely distributed in nature and as widely separated in their botanical relationships. The most important is digitalis, which is obtained from Digitalis purpurea (purple fox glove) ; strophanthus from Strophanthus hispidus; and squills from Scilla maritima. I. Prepare the tincture of digitalis according to the U. S. P., using 50 G. of the powdered drug and the proportionate amount of dilute alcohol. NOTE. For the strength of "dilute alcohol" consult the U. S. P. under "Alcohol Dilutum." Define a tincture. This tincture is to be used in the following experiments: II. To 5 cc. of the tincture add 15 cc. of distilled water. Is a precipitate formed? Cork and set aside the mixture and examine it after a few days. III. Examine the tincture for the presence of tannic acid by adding ferric chloride. IV. To 5 cc. of the tincture add 3 cc. of tannic acid solu- tion and let it stand for a time. Is a precipitate formed ? X x, ' * (Digitalis contains glucosides some of which are precipi- tated by tannic acid.) V. To 5 cc: of the tincture add a few drops of mercury- 120 LABORATORY GUIDE IN potassium iodide. Is a precipitate formed? What does, the ^ result indicate? VI. Evaporate over a water bath 10 cc. of the tincture to about one-half its volume and then make up the concentrated tincture to the original bulk by adding physiological salt solution. Now, pith a frog and tie it on a board. Expose the heart and inject into a lymph sac I cc. of the modified tincture. Record all changes in the heart. Describe any change in the rate of the organ and also any differences in the size of the various chambers or in their manner of contracting or dilating. VII. Pharmacological assay of tincture 1 of digitalis. Digitalis preparations vary considerably in their strength and it is impossible to standardize them by any chemical means such as can be employed in the cases of those drugs whose activity depends upon alkaloids which may be isolated in pure form and weighed. It is very hard to isolate the glucosides in pure form as they are likely to break up, making the results obtained very unreliable. For this reason a pharmacological estimation of the strength of a preparation is employed as follows : Evaporate 10 cc. of the tincture over a water bath to about half volume and then make it up to the original volume by the addition of physiological salt solution. Select 3 frogs of about the same size, weigh them carefully and mark them with string so as to be able to identify them. Into the an- terior lymph sac of one of the frogs inject by means of a glass pipette 0.3 cc. of the modified tincture diluted with x The assay of the tincture may be performed by one-half the class while the infusion of digitalis (page 124) is assayed by the remainder. EXPERIMENTAL PHARMACOLOGY. 121 122 LABORATORY GUIDE IN 3 * \A^^^C^i^^ ~a^<_ f r^ / -3 3 o*-*#- /& sCSttf /^ s {'**.---& 3.33 - .fj, 3 . - / - /. ;/ 170 LABORATORY GUIDE IN /7'o /? S / $ J * EXPERIMENTAL PHARMACOLOGY. 171 ranged according to the directions given under "Blood Pres- sure," page 147. To insert a cannula in the bladder to collect the urine, make an incision about an inch and a half long in the median line of the abdomen, beginning the incision at the symphysis and extending it upward the distance named. Deepen the cut until the peritoneal cavity is opened. The bladder is usually found partially distended and lying amid folds of fat. Draw it out through the opening made and empty it' by gentle pressure, catching the urine in a dish as it flows from the urethra. Tie a knot loosely in a piece of thread and lay it on the abdomen so that the knotted thread will encircle the bladder and be ready to tie the cannula in place when the latter shall have been inserted. After locating the position of the ureters, so as to avoid including them in the ligature, take hold of the upper sur- face of the bladder on the two sides with two pairs of forceps and draw them gently up and outwards so as to exert a slight tension on the organ. While one operator holds the bladder in this way the other operator makes a cut with a sharp pair of scissors midway between the forceps in the fundus of the bladder. The cut will vary with the size of the cannula flange (a, Fig. 10) and will probably average about two centimeters in length. Into this incision now place the cannula so that its flange shall be entirely sur- rounded by bladder tissue and tie it in place with the loose ligature already mentioned. Examine it after it is tied to see that the ureters are not included. Fill the bladder and cannula with the urine which was ob- tained earlier or with warm physiological salt solution and 172 LABORATORY GUIDE IN connect the end of the cannula with the piece of bent tubing (b, Fig. 10) which can now rest on a support so as to ex- tend over the end of the operating board where small weighed dishes are to be placed to catch the urine. As soon as the urine begins to drop from the tube a record of the time is to be made, and thereafter to the end of the experiment, every five minutes, the dish with the urine is to be replaced by a clean receptacle and the first one weighed, and the weight of urine re- corded. The blood pressure tracing is to be taken on a very slow drum upon which the five minute intervals are to be marked under the corresponding trac- ing, and the amount of urine for that period is also to be marked within the intervals indicated. By keeping the record in this way the drum will show the drug used, the time, the amount of urine excreted in the different intervals and the height of the blood pressure during the same periods. FIG. No. 10. Bladder Take two or three readings until a C T ula M ^' Fla " get bc fair regularity is obtained, and after tied m bladder. B. Bent J delivery tube. this normal is ascertained inject into the vein 40 mg. of caffeine dissolved in hot water. After this injection the urine probably will be increased and the next drug must not be injected until the flow has re- turned to about the normal. EXPERIMENTAL PHARMACOLOGY. 173 174 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 175 When the amount of urine has thus fallen, inject 0.250 G. (5 cc. of a 5% solution) of sodium nitrate and collect the urine as before. When normal is reached once more, inject 0.250 G. (5 cc. of 5 % solution) of sodium chloride and compare the results with the nitrate. NOTE. If time allows, the diuretic effect of digitalis may be studied in the same way. When the experiment is concluded fix the tracings and later study them, not only as regards the drug injected, but also in relation to the changes in blood pressure. Caffeine, digitalis and sodium nitrate are taken as repre- sentatives of the large class of drugs known as Diuretics from their action in increasing the amount of urine. They may act either on the heart or on the kidney, and 1;hey are known as cardiac or renal diuretics according to the point of action. Digitalis is an example of the former class, as it causes diuresis indirectly through its effects on the cir- culation. Caffeine probably acts directly on the renal epi- thelium, while various salts, of which sodium nitrate is an example, induce diuresis by their salt action (saline diuretic) in addition to which there may be a stimulant action on the kidney epithelium. Therapeutic uses of the Diuretics. See also under Caffeine and Digitalis, Therapeutic uses, (pages 100 and 128). These drugs or their allies are chiefly employed in diseased conditions in which the amount of urine excreted is below the normal, a condition which may result from disease of the kidneys or of the heart. In conditions of oedema or ascites, 12-p 176 LABORATORY GUIDE IN there is an abnormal collection of fluid in the tissues ; its removal may be aided by the use of the diuretics. In poison- ing with an irritant drug which is eliminated by the kidneys, the latter may be protected from the action of the toxic substance by increasing the volume of urine through the ad- ministration of these drugs. EXPERIMENTAL PHARMACOLOGY. 177 LABORATORY GUIDE IN f) t<_ -** i ^U^ ~~b\J(j L v/ EXPERIMENTAL PHARMACOLOGY. 179 The Belladonna Series. Several plants belonging to the Solanacese furnish alka- loids which are very closely related, not only chemically, but also in their pharmacological effects. Chief among these alkaloids is Atropine which is contained in the leaves and roots of the deadly nightshade (Atropa Belladonna). Next in importance to atropine are Hyoscyamine and Hyoscine. These alkaloids are all obtained from other sources than that named, viz.; henbane (Hyoscyamus niger), the thorn- apple (Datura Stramonium) and other plants of less im- portance. They are very hard to isolate in absolutely pure form because hyoscyamine is very prone to change to atro- pine during the manipulations. As stated above, they resemble one another closely in their properties and actions, so that atropine alone will be studied. Examine Belladonna leaves and roots. I. Prepare the alcoholic extract of belladonna leaves ac- cording to the U. S. P., using 50 G. of the powdered drug. Define an "extract" of a drug. II. Take some of the extract, an amount about the size of a pea and rub it up in 5 cc. of salt solution and put a drop of the watery extract in a cat's eye and watch for any changes comparing it with the normal eye. The cat is best put in a cat box (footnote, page 12) with the head exposed during the experiment; drop the solution near the outer canthu?oi the eye so that it will have to cross the eyeball before escap- i ing\by the nasal duct. *This action of the atropine series upon the pupil is known as mydriasis and the drugs, producing it are mydriatics. It 180 LABORATORY GUIDE, IN is the most delicate test known for the group, although a few other drugs, of which cocaine is the most important, act as mydriatics, though to a less marked degree. III. Isolate the atropine from the extract of belladonna by the "shaking-out" process as described below. Observe the precautions mentioned under the isolation .of strychnine, (page 68). Mix the extract thoroughly with about 100 cc. of distilled water and render it alkaline with ammonia. Put the mix- ture in a separating funnel and add 20 cc. of chloroform. Shake gently for 3 to 5 minutes and set the funnel aside to allow the chloroform to separate, drawing it off into a flask. Add 15 cc. more chloroform to the watery extract and shake as before, and draw off the chloroform into the flask with the first lot. The watery extract can now be thrown away. Put the chloroform containing the alkaloid in the separa- ting funnel and add about 50 cc. of water rendered distinctly acid with sulphuric acid. Agitate the mixture several min- utes as before, drawing off the chloroform from the water when separation has taken place. Repeat the process with the chloroform and 50 cc. of fresh acidulated water. (The waste chloroform is to be placed in a jar on the side table for redistillation.) If the watery solution is colored, shake it in a flask with powdered charcoal and filter it. The fil- trate containing the atropine is now rendered alkaline with ammonia and shaken out again with 20 cc. of chloroform, which is drawn off into an evaporating dish. Repeat the process with 10 cc. of chloroform and add it to the 20 cc. in the dish and set the whole amount aside, protected from the dust to allow the chloroform to evaporate and the atropine to crystallize. IV. Place a few crystals of atropine (from Exp. Ill) EXPERIMENTAL PHARMACOLOGY. 181 182 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 183 on a porcelain cover and add to them a drop of concentrated nitric acid and evaporate it to dryness over the water bath. Allow the cover to cool and then touch the residue with a glass rod which has been dipped in an alcoholic solution of caustic potash. A rich violet color is produced, changing to a dark red, which finally fades away, but can be reproduced by the addition of fresh alcoholic potash. This reaction (known as Vitali's test) is almost peculiar to the atropine series and is said to be given by o.oooi mg. of the alkaloid. V. Inject into a frog 15 mg. of atropine sulphate and de- scribe all the symptoms which are observed following the in- jection of the drug. Keep the animal for several days until complete recovery takes place. Compare the effects with those induced by any other drugs used earlier in the course. VI. Dissolve the remainder of the atropine in about 20 cc. of distilled water which has been rendered acid with sul- phuric or hydrocholoric acid. Divide the solution into five parts, put it in test tubes and add the alkaloidal precip- itants as follows: Reagent. Precipitate? a Tannic acid. b Picric acid. ^~ c Iodine in potassium iodide. + d Mercury-potassium iodide. /- e Phosphotungstic acid. -/- VII. Take o.ooi G. of atropine sulphate (which will be furnished you) and describe all the symptoms experienced. Count the rate of your heart before and after taking the drug and see if there is any change. The action of atropine on the innervation of the heart was studied on the turtle (page 144) ; and on the rabbit (page 139)- 184 LABORATORY GUIDE IN Its action on secretions will be studied on the salivary gland (page 196). Its mydriatic effect when applied locally was shown in Exp. II, and the same effect from internal administration is studied under the heading "Cervical Sympathetic Nerves," (page 188). Examine Atropa Belladonna, Hyoscyamus niger, Datura Stramonium, the alcoholic extract of belladonna leaves, tincture of belladonna leaves and the extract and tincture of hyoscyamus. Therapeutic Uses. Owing to the fact that atropine has such a wide range of activities, its uses in medicine are very numerous. Among these a few may be mentioned as being indicated by the va- rious experiments in which the drug is employed. It is used as a stimulant in the depressed state of the central nervous system occufring in shock, collapse and in narcotic poison- ing. (Compare strychnine and caffeine.) For its peri- pheral action it is employed for many purposes. In cases of extreme slowness of the heart ( Brady cardia) due to over- activity of the vagus, atropine may be employed. It is used to dilate the pupil of the eye to allow of ophthalmoscopic examination, or as a therapeutic agent in the treatment of many eye diseases. In excessive secretion of the saliva such as occurs in poisoning with mercury, and in the night sweats of tuber- culosis, atropine is very valuable. Finally it is used in cases of poisoning by pilocarpine or muscarine, the latter being an alkaloid resembling pilocar- pine in many of its actions. EXPERIMENTAL PHARMACOLOGY. 185 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 187 Action of Drugs on the Cervical Sympathetic Nerves. Anaesthetize a rabbit (an albino if possible) with paralde- hyde (page n) and when anaesthesia is complete tie the animal on the operating board and remove all the hair from the neck region. Expose the jugular vein on one side and in- sert a cannula. Expose the carotid sheath on the other side, open it, laying bare the vessels and nerves. The largest nerve seen is the vagus and the smallest is the depressor nerve, while the third is the cervical sympathetic trunk, which is to be separated very carefully from all connective tissues and a fine thread passed under it. Make sure you have the correct nerve by gently raising it and stimulating it with a weak tetanizing current, observing meantime the effect on the pupil on the corresponding side ; it should dilate during the stimulation. If the correct nerve has been isolated tie the ligature around it as low down in the neck as convenient and cut the nerve trunk below the ligature. Gently hold the animal's ears up against the light so that the vessels may be clearly seen, and observe the difference in their size on the two sides ; note also the difference in the temperature of the two ears. Compare the size of the pupils on the two sides. How would you explain these changes? Raise the trunk of the cut nerve carefully by the ligature and stimulate it with the electric current as before. During this stimulation look especially for any changes in the calibre of the ear vessels on that side; and for any changes in the position of the eyeball, or of the eyelids, or in the size of the pupil. After the changes have been seen and described, in- ject into the jugular vein 3 mg. of nicotine chloride. Im- 188 LABORATORY GUIDE IN mediately after the injection notice the effect of the drug on the animal's respiration, and also look for any movement in the animal's whiskers. Again stimulate the cut cervical sympathetic comparing the results with those obtained at the first stimulation. (Should they be the same as before, it will be necessary to inject more nicotine.) Now using very careful dissection trace the trunk of the sympathetic toward the head until the superior cervical ganglion is reached. It will be recognized as a small whitish enlargement on the trunk of the nerve. When it is found, stimulate the trunk of the sympathetic be- yond the ganglion, that is, between the ganglion and the head of the animal. Compare the results of stimulation at this point with those obtained below the ganglion. NOTE. In addition to the nerve fibres which run to the eye by way of the cervical sympathetic and which are distributed to the radial muscle fibres of the iris there are nerves which control the circular fibres. These come from the motor oculi to the ciliary ganglia and . run from that point to the iris. The size of the pupil can be changed by drugs acting along the course of these fibres also. Inject into the jugular vein 5 mg. of pilocarpine (i cc. of 0.5% solution) and observe any change in the pupil. (Note any effect on the heart rate.) Finally inject I mg. atropine sulphate (i cc. of 0.1% solution). What effect has it upon the pupil ? What effect has nicotine upon the sympathetic ganglia? Cocaine dilates the pupil by stimulating the terminations of the nerves from the superior cervical ganglia which end on the radial fibres. Physostigmine (Eserine) acts on the same point and in the same manner as pilocarpine, i. e., on the nerve termina- tions on the circular muscular fibres; atropine acts at the same place, but instead of stimulating the nerve endings, its action is to paralyze them, removing the pilocarpine effects. EXPERIMENTAL PHARMACOLOGY. 189 *- ' 190 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 191 Therapeutic Uses. Physostigmine (eserine) and, less commonly, pilocarpine are used in ophthalmological practice to contract the pupil of the eye in certain diseased conditions, the most important of which is glaucoma. For this purpose a solution of l /\ to I % is dropped in the eye as often as necessary. Atropine is used to dilate the pupil to permit of ophthal- moscopic examinations, and as a therapeutic agent in many ocular diseases. Homatropine, an artificial alkaloid re- sembling atropine in its action on the eye, is largely used as a substitute for the latter, as its action is not nearly as pro- longed. Cocaine is also largely employed to dilate the pupil to allow of examination of the interior of the eye. Mydriatics are drugs which dilate the pupil ; myotics con- tract it. Salivary Secretion. Anaesthetize a large dog with morphine and chloretone (page 12) and if necessary supplement them by the use of chloroform. When anaesthesia is complete, expose a vein in the leg and insert a cannula. As a matter of precaution, the trachea may also be exposed and a cannula tied in it to permit of artificial respiration should it become necessary. The submaxillary duct and chorda tympani nerve are to be dissected out according to the following directions. (This dissection should have been previously carried out on the head of a dead animal (from Exp., page 159) so as to allow greater speed in operating on the living dog). "Make an incision 3 or 4 inches long through the skin and platysma muscle along the inner border of the lower jaw, be- 13-p 192 LABORATORY GUIDE IN ginning about the angle of the mouth and continuing back- ward towards the angle of the jaw. Legate doubly and divide any vessels that come in your way. Divide the digas- tric muscle about its anterior third and clear it from its at- tachment. The broad, thin mylo-hyoid will now be seen with its motor nerve lying on it. Divide the muscle across FIG. No. ii. Dog's head dissected to show Chorda Tympani. A. Chorda Tym- pani, B. Wharton's duct, C. Hypoglossal nerve, D. Mylo-hyoid muscle turned back, E. Lingual nerve, F. Digastric muscle cut, G. Ra- mus of jaw cut away to show nerve emerging from behind it. its middle and dissect it up carefully. The lingual nerve is seen emerging from under the ramus of the jaw, running transversely towards the middle line and then passing for- ward parallel to the larger hypoglossal nerve. In its trans- verse course it crosses the submaxillary and sublingual ducts. EXPERIMENTAL PHARMACOLOGY. 193 194 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 195 The chorda tympani leaves the lingual nerve and runs back- ward along the duct towards the gland. Put a glass cannula into the duct in the same way as you would put a cannula in a vein. Trace back the lingual and put a ligature around it and tie it as far back as possible, and then cut it centrally so as to permit of the chorda tympani being put on electrodes." (Stewart.) Connect the cannula with a long, horizontal tube on which a scale is marked. While the dissection of the duct is being carried on, a second operator is to dissect out the cervical sympathetic trunk on the same side. In the dog the cervical sympathetic is contained in the carotid sheath and closely connected with the vagus, from which it may be separated by careful dis- section, as the fibres run in distinct strands. For some dis- tance below the superior cervical ganglion the two nerve trunks are not connected. After the sympathetic is isolated tie a ligature around it and cut the nerve below it. If the experiment has been properly done, both nerves (viz., the chorda and the sympathetic) can be stimulated by placing them on electrodes, and any secretion from the gland will pass into the horizontal tube, where it can be measured on the scale. 1. Stimulate the chorda tympani with a weak induction current and watch the effect on the secretion. 2. When the effect of chorda stimulation has passed off stimulate the sympathetic trunk (central or head end) and observe the difference in the quantity and quality of the saliva when compared with the "chorda saliva." 3. 'Inject 30 mg. of nicotine chloride and observe closely the effect on the secretion. 4. Stimulate the chorda again and compare the results with those obtained in i. 196 LABORATORY GUIDE IN 5. Stimulate the sympathetic and compare with 2. 6. Trace the trunk of the sympathetic to the superior cervical ganglion and stimulate the nerve beyond, observing if secretion can be obtained. Inject the following drugs in the order named, observing their effects on the secretion: 7. Nicotine, 5 mg., compare^with 3. 8. Pilocarpine, 5 mg. 9. Atropine, i mg. 10. Pilocarpine, 5 mg., compare with 8. 11. Suprarenal extract, Adrenalin chloride, i-iooo sol., 5 drops. What effect has nicotine upon the sympathetic ganglia? Compare also the experiment on the sympathetic ganglia in the rabbit (page 187). As was shown in this experiment, the salivary gland is innervated from two sources, both of which are acted on by nicotine. Pilocarpine and atropine affect the salivary secre- tion by acting on the cranial nerve supply of the gland. The action of the suprarenal gland extract in causing secretion is not as well known, but it is believed to be upon the termina- tions of the sympathetic fibres in the gland cells. (It is not always possible to get secretion of saliva by the injection of suprarenal extract.) EXPERIMENTAL PHARMACOLOGY. 197 198 LABORATORY GUIDE IN HXPZRIMZNTAL PHARMACOLOGY. 199 The Sphygmomanometer. Among the pharmacological experiments which the stu- dent may carry out upon himself, there are very few which allow of absolutely accurate records being kept such as are obtained in animal experimental work. The recent develop- ment of instruments by which the human blood pressure may be determined opens up a new field in which the student may study upon himself the effects of drugs whose actions he has demonstrated upon the lower animals. Any of the newer sphygmomanometers, such as those de- vised by Brlanger, Stanton, Janeway or the Cook modifica- tion of the Riva-Rocci, may be employed for this purpose. The underlying principle is essentially the same in all, and for complete descriptions of these instruments reference must be made to special works on the subject. 1 The upper arm of the "patient" is encircled by a broad rubber bag (12-15 cm - wide) connected by rubber tubing with a mercury manometer and a rubber bulb. The pressure in the closed system of tubing is raised by means of the bulb until the pressure in the bag is sufficient to obliterate the radial pulse under the finger of the operator. The pressure is now lowered slowly until the pulse reappears and then the pressure may be raised to cause the pulse to disappear once more. In this way the systolic or maximum pressure may be obtained and recorded by observing the height of the mercury column. With some of the instruments the minimum or diastolic J The Clinical Estimation of Blood Pressure. Janeway, New York, 1904. 200 LABORATORY GUIDE IN pressure may also be estimated, but this hardly necessary in the following experiments. A few precautions are necessary and they may be given most concisely by saying that the subject of the experiment should as far as possible refrain from all mental or physical exertion. During the course of the experiment he should sit or lie quietly and take no interest whatever in his sur- roundings. It is usually most satisfactory to have three students work together, one to receive the drug, one to man- age the air bulb and take the readings, and the third to direct his attention entirely to the pulse. The frequency of the readings and the length of time over which they should be taken will depend upon the drug used. For instance, the effects of amyl nitrite may be said to be over in a very few minutes, but during that time the readings should be taken as frequently as possible. With the other drugs used, readings should be made every five minutes and if possible they should be continued for an hour or two or until the pressure has returned to the normal height again. In any case, two or three estimations of the normal pressure should be made before the drug is administered. The experiments mentioned below may be varied almost indefinitely by using the drugs in varying dosage or in com- bining two drugs in one experiment, administering the sec- ond after the effect of the first is plainly manifest. The pulse rate is to be counted and recorded together with the pressure readings on the blanks provided. The following drugs are suggested : Amyl nitrite, I to 3 drops given by inhalation. Strychnine sulphate, 0.002 G. to 0.005 G., by hypodermic or by mouth. UXPBRIMHNTAL PHARMACOLOGY. 201 202 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 203 Atropine sulphate, 0.0005 G. to o.ooi G., by hypodermic or by mouth. Sodium nitrite, o.i G. by mouth. Nitroglycerin, I to 3 drops by mouth. Digitalin, o.ooi G., by mouth or hypodermic. The effects of smoking upon the blood pressure may be tested upon a suitable individual; the most pronounced re- sults will probably be obtained from inhaling the smoke from a "strong" pipe or cigar. 204 LABORATORY GUIDE IN Anaesthetics. The most important members of the group of drugs used to produce general anaesthesia are. 'Chloroform and Ether. Their depressant action on the central nervous system has been observed in some of the experiments previously carried out in which they have been used to supplement the other drugs used as anaesthetics in animals (chloretone, etc.). The cardiac effects were studied in the frog, rabbit and dog. In these animals is was noticed that the heart dilated and became very weak before there was very much change in the rate. The importance of this in practical anaesthesia is that information as to the condition of the heart cannot be derived entirely from the pulse rate, as the heart may be seriously affected and yet show very little change in the nu'mber of its contractions per minute. The quality of the pulse must also be noted. The fact that chloroform acts very much more strongly on the heart than ether, was also demonstrated in the ex- periments mentioned above. Some of the early effects of the anaesthetics are shown in the following experiment : I. Place a rabbit on the table and hold it by placing your hand under its chest in such a way that you can count its heart and respiration. Pour some chloroform on a little ab- sorbent cotton and hold it to the animal's nose for a few moments and observe any change in the rhythm of the heart or respiration. Repeat the experiment using ether and afterwards am- monia. It will be seen that the same effects are produced by EXPERIMENTAL PHARMACOLOGY.. 205 v^^l 206 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 207 all three drugs. They are caused by a reflex stimulation of the vagus and respiratory centers from the action of the irri- tant fumes on the mucous membranes. Other drugs that are used as general anaesthetics are Nitrous oxide gas and Ethyl chloride. 14-p 208 LABORATORY GUIDE IN Pilocarpus. r * Pilocarpine is an alkaloid derived from the leaves of sev- eral species of Pilocarpus. -It has .been employed in certain of the earlier experiments, which may be summarized in order to give a more complete picture of the action of the drug. Describe the effects of pilocarpine on the frog's heart (Exp. page 135) ; the turtle's heart (Exp. page. 144) ; the rabbit's heart (Exp. page 136) ; the pupil of the eye (Exp. page 188) ; the salivary secretion (Exp. page 196) ; and on the intestinal peristalsis (Exp. page 220). How does atropine affect the activity of the drug? I. (These drugs will be furnished you.) Take 0.003 G. of pilocarpine hydrochloride and describe any symptoms noted. Should any of these become unpleasant take atropine sulphate 0.0005 G. and describe any changes. Other drugs which resemble pilocarpine in many of their actions are Muscarine, from one of the poisonous mush- rooms (Agaricus muscarius), and Physostigmine or Eserine, from the Calabar or Ordeal bean (Physostigma venenosum). Examine the leaves of Pilocarpus jaborandi, and the calabar bean. Therapeutic Uses. The employment of pilocarpine in diseases of the eye is discussed elsewhere (page 191). The drug is practically never used for its cardiac action. . EXPERIMENTAL PHARMACOLOGY. 209 (/^^(2#^^ ^ sfa^f^ ^^^^ <^-L^^ x^s ^ 210 LABORATORY GUIDE IN BXP&RIMMNTAL PHARMACOLOGY. 211 It is used to increase the activity of the sweat glands (diaphoretic action) in various diseases in which there is an abnormal collection of fluid in the body. (Compare Di- uretics, page 175). Through its action a very large amount of fluid may be excreted so that in some cases the body may lose from four to nine pounds in weight from a single dose of the drug. Pilocarpine is employed in some diseases of the kidneys in which the urine excretion is lessened, with the idea of se- curing the elimination of toxic materials by the sweat glands. 212 LABORATORY GUIDE IN Suprarenal Gland Extract. The suprarenal glands contain a very active substance, Epinephrine, which can be isolated, but with difficulty, as it is very unstable and easily decomposed. Various preparations of the gland are put on the market, of which Adrenalin (Takarnine) is one of the best known. For experimental work it may be used in two forms, viz.: A solution may be prepared by boiling a minute amount of the powder (adrenalin) in physi- ological salt solution until the fluid becomes a light pink. The objection to this method is that it does not allow of ex*act dosage. By using the commercial adrenalin chloride (i-iooo) this objection is overcome. From one to three drops of this preparation give good results in experimental work. Describe the effects of extract of the suprarenal gland upon the rabbit's heart (Exp. page 139) ; the blood pressure (Exp. page 151) ; the dog's heart (Exp. page 159) ; the salivary secretion (Exp. page 196) ; the kidney vessels (Exp. page 168) ; and upon intestinal vessels (Exp. page 227). I. Examine the local effects of the drug by putting one drop of adrenalin chloride (i-iooo) in the eye. Compare with the normal eye. How would this experiment aid you in explaining the results obtained in the experiment on blood pressure ? Therapeutic Uses. The powerful effects of the drug on the circulation are limited in their application to therapeutics by their transitory nature and by the fact that in order to act on the heart or blood vessels (except the local action), the drug has to be EXPERIMENTAL PHARMACOLOGY. 213 214 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 215 injected intravenously. In some cases of shock and collapse it has been given in this way, but it requires constant admin- istration as the effects pass- off so rapidly. It is of most value for local application in surgical operations to constrict the vessels and thus render the field of operation bloodless. 216 LABORATORY GUIDE, IN The Nitrite Series. This series comprises several drugs whose action in the body is confined almost exclusively to the blood vessels. The principal members are Amyl Nitrite, Nitro glycerin, Sodium and Potassium Nitrite. They have been employed in various experiments and the results are to be collected and summarized. Describe the effects of amyl nitrite on the rabbit's blood pressure (Exp. page 151) ; the vessels of the kidney (Exp. page 232) ; and on the vessels of the intestine (Exp. page 227). Describe the effects on the human blood pressure of amyl nitrite (Exp. page 200) ; nitroglycerin (Exp. page 203) ; sodium nitrite (Exp. page 203), and of sodium nitrite on the excised kidney vessels (Exp. page 167). Therapeutic Uses. The members of this series are employed in medicine to lower the blood pressure and to relieve vascular spasm. EXPERIMENTAL PHARMACOLOGY. 217 218 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 219 The following experiments are not suitable for the regu- lar laboratory course, but may be carried out by advanced students ; they will also serve very well for class demonstra- tions by the instructor in charge. Intestinal Peristalsis. The peristaltic movements of the intestines and the effects of various drugs upon them may be demonstrated very well by submerging an animal in a saline solution, opening the abdomen and keeping the intestines under the surface of the water by means of a pane of glass. For this experiment a metallic tank is needed which will be large enough to allow a rabbit to be placed in it when the animal is tied to an operating board. A tank measuring 70x30x30 cm. is large enough for most purposes and it should be mounted on legs long enough to allow a Bunsen burner to be placed under it. The operating board should be weighted with lead so that it will sink in the water. A pane of glass to cover the intestines and some small lead piping, which can be bent over the sides of the tank to act as a support for the glass, and a bath thermometer complete the special apparatus necessary for the experiment. Fill the tank about two-thirds full of water and add sodium chloride to make a physiological salt solution (8 G. per liter) . Light the Bunsen burner under the tank and heat the solution until it is of the body temperature. 220 LABORATORY GUIDE IN Operation. In the meantime anaesthetize a rabbit in the usual manner and when anaesthesia is complete insert a venous cannula in the jugular vein and also insert a tracheal cannula. Place short pieces of rubber tubing on both free ends of the tracheal cannula and close one by means of a clamp. The second piece of tubing is to extend above the surface of the water when the animal is submerged. Place the animal in the warm salt solution which should completely cover it. Take great care no water enters the tracheal tube, the open end of which is to be tied to a support above the tank. Make an incision through the skin and muscles in the median line of the abdomen from the end of the sternum to the symphysis. The abdominal walls may be drawn outward and secured to the operating board by means of tacks. The intestines will float to the top of the water and they must be held just under the surface by placing the pane of glass over them and supporting the latter in position by means of the lead tubing. The peristaltic movements may now be observed as well as the vascular changes which will follow the injection of the drugs named below ; the changes in color of the intestines indicate the alterations in the circulation of the blood. Inject into the jugular vein the following drugs in the order named : Nicotine chloride, 5 mg., Pilocarpine hydrochloride, 5 mg., Atropine sulphate, I mg., Veratrine sulphate, i mg., Adrenalin chloride (i-iooo sol.), 2 drops, Amyl nitrite, 2 or 3 drops given by inhalation. EXPERIMENTAL PHARMACOLOGY. 221 222 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 223 Vasomotor Changes in the Intestine. The special apparatus needed for this experiment is, first, some form of intestinal plethysmograph and second, an in- strument to record the intestinal vascular changes upon a blackened drum. The plethysmograph may be made very easily of plaster of Paris after the pattern devised by A. Edmunds. 1 With the plaster make a hemispherical basin ten centi- meters in diameter and four deep with the walls about seven millimeters thick and with an elliptical opening in the bottom of the basin three centimeters by one and a half. Make a small opening in one side of the apparatus and cement in it a short piece of glass tubing which connects with the interior of the basin and which extends outside three or four centi- meters to allow of rubber pressure tubing being connected with it. Smooth off the upper edge of the apparatus with sand paper so that a piece of glass will fit fairly tightly on it. A plethysmograph made in this way will answer all the pur- poses of the experiment. Its more complicated forms may be made as described in the article referred to. A glass plate which will close the top of the plethysmograph is needed as well as a piece of pressure tubing long enough to connect the plethysmograph with the recording apparatus, which may be either a tambour, a delicate piston recorder or a Brodie bellows recorder. Either of these should be provided with a side tube to allow of proper equalization of pressure on both sides of the piston or membrane of the recorder. Vaseline is needed as well as a petrolate which melts at r. of Physiol., Vol. XXII, 1898, page 380. 15-p 224 LABORATORY GUIDE IN a higher temperature than vaseline and which may be made by melting the latter with wax, so that the resulting mixture is fairly solid when cold. Operation. Anaesthetize a rabbit or a cat as usual, tie it on an operat- ing board and insert a venous cannula. If it is desired to take a tracing of the carotid blood pres- sure at the same time, the artery must be isolated, a cannula inserted and connected with the mercury manometer accord- ing to the directions given on page 148. By thus taking simultaneous tracings of the general blood pressure and of the vascular conditions in the intestinal area very instructive records may be obtained. It is best to warm the plethysmograph slightly before ap- plying it and to have plenty of hot salt solution ready for use as well as some absorbent cotton. Make an incision about 4 cm. long in the median line of the abdomen about midway between the sternum and symphysis and with the fingers carefully pull out of the abdominal cavity a short loop of the small intestine. Place the elliptical opening of the plethys- mograph over the incision and intestinal loop and support the instrument in place with an iron ring. Draw the loop of the intestine into the apparatus until a section from 20 to 30 cm. long is lying in the basin. Cover the intestine temporarily with absorbent cotton dipped in the hot salt solution. Ligate doubly each end of the section of intestine and cut across it between the ligatures, tearing down the mesentery as far as possible and ligating any bleeding vessels. Now return to the abdominal cavity the upper and lower attached- ends of the intestine, leaving the isolated loop in the plethysmograph EXPERIMENTAL PHARMACOLOGY. 225 226 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 227 and connected to the animal by its section of mesentery con- taining the vessels and nerves. Pack absorbent cotton rubbed up with the hard vaseline around the mesentery so as to completely close the elliptical opening, using the soft vaseline to make it air tight near the vessels ; great care must be taken not to constrict the vessels. Remove the cotton covering from the intestines ; vaseline the upper rim of the plethysmograph and close it in with the pane of glass which has been dipped in warm salt solution to prevent the condensation of moisture on its surface. Connect the instrument with the recorder, close the side tube on the latter and take a tracing, which, if the experi- ment has been properly carried out, will show both cardiac beats and the respiratory waves. Arrange the lever to write in line with the blood pressure pointer and the time marker which has been placed below. Take tracings to show the effects of the following drugs on the general arterial blood pressure and on the vessels of the splanchnic area. Adrenalin chloride (i-iooo), 3 drops in salt solution. Digitalis, tincture, I cc. in salt solution. Nicotine chloride, 5 mg. in salt solution. Amyl nitrite, 2 drops to be inhaled, on cotton. 228 LABORATORY GUIDE IN Vascular Changes in the Kidney. To record the changes taking place in the size of the kid- ney due to the differences in its blood content, an oncometer and a recording apparatus are needed. The former can be made from plaster of Paris in the same manner as the intes- tinal plethysmograph. Several sizes should be made to ac- commodate the kidneys of different sized animals. A cylin- drical-shaped oncometer with a flat base, having a diameter of about 5 cm. and with walls about 3.5 cm. high is large enough for the average sized cat or rabbit. One side should have a cut in it about 0.5 cm. wide and extending nearly to the bottom of the apparatus, where the cut should be widened to allow greater room for packing around the pedicle of the kidney. On the opposite side of the oncometer is a glass tube communicating with the interior of the apparatus and extending outward for the attachment of rubber pressure tubing. A piece of glass is needed to fit tightly the top of the instrument. Vaseline together with a mixture of vaseline and wax (as described on page 223) is also necessary. For recording the volume changes on the kymograph, a tambour, piston recorder, or a bellows recorder as mentioned under the experiment on the intestinal vessels (page 223) is required. It should be fitted as before with a side tube. Operation. Anaesthetize a rabbit or a cat and insert one cannula in the jugular vein and a second in the carotid artery. Connect the latter with the mercury manometer for blood pressure, which has been arranged according to the directions given EXPERIMENTAL PHARMACOLOGY. 229 230 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 231 on page 148. Either kidney may be employed in the experi- ment, the choice largely depending upon the arrangement of the apparatus and the light. If the left has been selected, make an incision in the abdomen well over in the left flank, extending from the costal margin downward, the length of the incision necessary depending upon the size of the onco- meter. Draw the side muscles outward, if necessary con- tinuing the cut in the muscles along the costal margin, and nail the reflected muscles to the operating board. The intes- tines and stomach should be packed back with absorbent cotton which has been dipped in hot salt solution and the abdominal cavity closed in as far as possible with a hot towel. This should leave the left kidney well exposed. Loosen it very carefully from all its fibrous attachments and remove the fat as far as possible, so that the kidney remains connected only by the structures entering the hilus. Place the organ in the warmed oncometer so that the pedicle passes through the widened end of the cut in the side of the apparatus. Pack around the pedicle with ab- sorbent cotton saturated with the hardened vaseline and completely close the opening in the side in the same way, employing the soft vaseline to make it entirely air tight. Of course it is essential to see that no undue pressure is exerted on the kidney vessels ; the circulation must remain perfectly free in them. Close in the top of the oncometer with the pane of glass which is made air tight with vaseline. Con- nect the outlet tube of the instrument with pressure tubing, the other end of which is connected with the recorder. Ar- range the writing point of the recorder so that it marks directly above the pointer of the blood pressure manometer and the time marker. Close the side tubes of the recorder 232 LABORATORY GUIDE IN and the lever should show the distinct cardiac pulsations as they are indicated by the changes in the kidney volume. If the pulsations do not show, either the kidney vessels are con- stricted or the apparatus is not air tight. When all the apparatus is working satisfactorily inject the following drugs into the jugular vein : Adrenalin chloride, i-iooo, I to 3 drops in salt solution. Tincture of digitalis, I cc. in salt solution. Nicotine chloride, 5 mg. in salt solution. Amyl nitrite, 3 drops, given by inhalation. EXPERIMENTAL PHARMACOLOGY. 233 234 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 235 List of Drugs and Reagents Required, Acacia, powdered. Acids : Hydrochloric, con. lodic, 5%. Nitric, con. Phosphotungstic, 2%. Picric, saturated. Sulphuric, con. Sulphuric, 10%. Tannic, 10%. Aconite, tincture. Adrenalin ( Takamine ) . Adrenalin chloride, i-iooo. Almonds, bitter. Alcohol, 95%. Ammonium hydrate, 10%. Amygdalin. Amyl nitrite. Asafoetida. Atropine sulphate, 0.1% solution. Atropine sulphate, for hypoderm. use. Belladonna, No. 60 powder. Caffeine, cryst. Caffeine, 2% solution. Calcium chloride. Charcoal. Chloral. Chloretone. 236 LABORATORY GUIDE IN Chlorine water. Chloroform. Cocaine hydrochloride, 4% sol. Curara, standardized sol. (page 88). Digitalis leaves, No. 60 powder. Digitalis leaves, bruised. Ether. Fehling's solution. U. S. P. Ferric chloride. Iodine in potassium iodide. U. S. P. Lycopodium. Manganese dioxide. Mercury-potassium iodide. U. S. P. Milk sugar. Morphine sulphate. Nicotine chloride. (See note, page 92). Nux vomica, No. 60 powder. Oils: Castor. Cotton seed. Turpentine. Opium. Paraldehyde. Pilocarpine hydrochloride, 0.5% sol. Pilocarpine hydrochloride, 0.02 G. tablets. Potassium bichromate. Carbonate, 10%. chloride, hydrate, 10%. " hydrate, 10%, in absol. alcohol. Quinine. EXPERIMENTAL PHARMACOLOGY. 237 238 LABORATORY GUIDE IN EXPERIMENTAL PHARMACOLOGY. 239 Quinine hydrochloride. " sulphate. Sodium chloride, cryst. nitrate, $% sol. " nitrite, cryst. " sulphate, sat. sol. Starch. Strychnine sulphate, 0.1% sol. sulphate, 0.002 G. tablets. Sugar. Sulphonal. Veratrine sulphate, 0.1% sol. 16-p I INDEX. PAGE. Absinthe 79 Aconite I2 ^ Aconitine I2 Acrole'in 4 Adrenalin 2I2 Alkaloids 2 4 Alkaloidal precipitants 3* Amygdalin 3 2 Amyl nitrite 216 Anaesthetics 204 Anaesthetic for cat 12 for dog 12 for rabbit 1 1 Analysis of tracings 160 Apalache tea 96 Aqua 4 Assay of tincture of digitalis 120 of the infusion of digitalis 124 Atropine 179 isolation of 180 Balsams 43 Belladonna series 179 Berberine 32 Bladder cannula, to insert, 171 Blood pressure in man, to estimate 199 drugs on 200 Blood pressure in animals, drugs on 147 Brncine 64 Caffeine 96 Camphor 79 Cat box 12 Cannulas, arterial 20 tracheal 23 venous 19 241 242 INDEX. PAGE. Chemistry of drugs 24 Chloral 51 Chloralose 55 Chloretone . .- 55 Chloroform 204 Chorda tympani nerve, to find 191 Cinchona 103 Clonic convulsion 79 Cocaine 100 Cocaine on pupil 188 Codeine 80 Coffee 96 Coniine 32 Continental method of emulsification HI Curara 88 Curine 88 Deadly nightshade 179 Diaphoretic 208 Digitalin 36 Digitalis 119 Digitalis on kidney 172 Digitonin 124 Digitoxin 36 Diuresis' 168 Diuretics 175 Diuretic, cardiac 128 Dog's heart, drugs on 151 Dover's powder 87 Drugs and reagents required 235 Emulsifiers 108 Emulsin 35 Emulsions 108 Epinephrine 212 Eserine 208 Eserine on pupil 188 Essential oils 36 Ether 204 Ethereal oils 36 Ethyl chloride 207 Excipient 115 Extract 179 INDEX. 243 PAGE. Fehling's reaction 31 Fixed oil emulsion 1 1 1 Fluid extract 64 Frog, operations on. See Operations. Frog's heart, drugs on 132 Glucosides 32 Guarana paste 96 Gums 43 Gum resins 43 Gum resin emulsion m Henbane 179 Homatropine , 191 Hyoscine 179 Hyoscyamine 179 Hypnotics 51 Infusion of digitalis . . 124 Isolation of strychnine 67 of atropine 180 Intestinal peristalsis 219 Kola nut 96 Laudanum 87 Mammals, operations on. See Operations. Meconic acid 80 Meconic acid reaction 84 Monk's hood 128 Morphine 80 Murexoin test 99 Muscarine 208 Mydriasis 179 Mydriatics 179, 191 Myocardiograph for turtle 144 for dog 155 Myotics 191 Nicotine 92 Nicotine chloride footnote 92 Nicotine on sympathetic nerves 187 244 INDEX. PAGE. Nitrite series 216 Nitrite, amyl 216 sodium 216 potassium 216 Nitroglycerin 216 Nitrous oxide 207 ''Non-official" drugs 7 Nux vomica 64 ''Official" drugs 7 Oils, fatty 36 fixed 36 volatile 36 Oil of bitter almonds 35 Oleoresins 43 Oncometer for kidney 228 Opium 80 Operations on frogs, pithing 15 to inj ect in lymph sac 15 to expose the heart 16 Operations on mammals, to insert an arterial cannula 20 to insert a tracheal cannula 23 to insert a venous cannula 19 Paraguay tea 96 Paraldehyde 51 Paregoric 87 Percolation .- 59 Perfusion of kidney 167 Pharmacopoeia 7 Physiology of frog's nervous system 44 Physostigmine 208, 188 Pills 112 Pill mass 115 Pilocarpine on pupil 188 Pilocarpus 208 Pipette, glass injecting 15 Pithing frogs 15 Plethysmograph, intestinal 223 Quinine 103, 24 Quinine sulphate 24 INDEX. 245 PAGE. Rabbit's heart, drugs on 135 Reagents and drugs required 235 Resins 43 Sage 79 Salicin 36 Saline diuretic ". 175 Salivary secretion 191 Salivary secretion, drugs on 195 Seed emulsion 108 Shaking-out process 31 Soporifics 51 Sodium chloride, on kidney 175 Sodium nitrate, on kidney 172 Sodium nitrite, on kidney vessels 168 Sphygmomanometer 199 Spiritus 40 Squills 119 Stomach tube for cats 1 1 dogs 12 rabbits 1 1 Strophanthus 119 Strychnine 64 Submaxilliary duct, to find 191 Sulphonal 51 Suprarenal gland extract 212 Sympathetic system, drugs on 187 Syrupus 40 Takamine 212 Tansy 79 Tea 96 Tetronal 55 Theobromine 96 Theophyllin 96 Thornapple 179 Thebaine 87 Thujon 79 Tincture IJ 9 Tobacco 92 Tonic 79 Tonic convulsion 7 1 Trional . 55 246 INDEX. PAGE. 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