I N D U S T R I A L A N D ENGINEERING CHEMISTRY
October, 1924
physiological action is entirely counterbalanced by the more important, more powerful substance, A . This substance A constitutes the truly important principle of the posterior lobe and it will be referred to as the hormone of this lobe. I n using this term we take it for granted that it does pass into the blood, although we cannot a t the moment give a conclusive proof for this statement in the case of the higher animals. SEPAR.4TION O F THE P R I N C I P L E
Horn does one separate a chemical principle of this sort from the many other substances contained in an extract of the posterior lobe? At the slaughterhouse the posterior lobe is separated from the anterior lobe of the chilled glands by trained operators. A great many of these small lobes have to be shelled out much as a small acorn might be removed from its woody cup, before enough material to work with is secured. The freed posterior lobe is then finely ground to a very fine paste and each 100 grams of paste is thoroughly mixed with 0.35 per cent hydrochloric acid containing 4 grams of mercuric chloride. There can be no decay or putrefaction as no bacteria or spores can develop in the presence of an excess of corrosivt: sublimate. When the material reaches the laboratory finely powdered corrosive sublimate is added in excess. The flask is then fastened in a shaking machine and the material is kept in agitation for a couple of hours. At the end of this time it is found that the protein material has coagulated and that it quickly settles to the bottom of the flask. We have now at the bottom of the flask what we may call the mercuric chlorideprotein cake with a clear, supernatant fluid. This top fluid contains all of the B and C depressor substances as well as almost all of the soluble crystalloid material contained in the posterior lobe, while the A substance is adsorbed by the surfaces of the mercuric chloride-protein precipitate. At one stroke, 1herefpre, we have almost completely separated from our active A principle, or hormone, the other constituents of the posterior lobe. The next step is to separate the mercuric chloride cake by means of pressure filtration, disintegrate it, and wash with cold, sal urated corrosive sublimate solution. This washing process is repeated and the cake is then disintegrated, almost neutralized to litmus paper with sodium hydroxide, treated with hydrogen sulfide in excess, then with a certain amount of a saturated sodium chloride solution (in order to flocculate the precipitiite of mercuric sulfide), and filtered. The filtrate is aerated to remove the excess of hydrogen sulfide, and is then completdy precipitated with phosphotungstic acid. The phosphotungstate precipitate is then decomposed according to a method previously describedG2 The next step consists in preparing a tannate which, after washing, is converted into a tartrate. The tartrate, after appropriate treatment, is subjected to a purification process by means of picrolonic acid. After this treatment the tartrate is carried through a number of stages of purification by the appropriate and consecutive use of various organic solvents, such as pyridine, absolute ethyl alcohol, n-butyl alcohol, and precipitating agents in which the tartrate is insoluble, such as ether and acetone. By such processes the tartrate is finally obtained in the form of a dry, white precipitate which is extremely soluble in water and also in 93 to 94 per cent alcohol. The final product then is a highly active tartrate. This has not yet been obtained in crystalline form, but it is hoped that this mill soon be accomplished. PHYSIOLOGICAL PROPERTIES S o w what are the physiological properties of this tartrate? In the first place, it is an extraordinarily powerful stimulant 2
J Pharmacol , 22. 290 (1923).
1033
for all smooth muscle tissues, as is well illustrated by its power to induce the virgin guinea pig’s uterus to contract. Here we must compare its action with that of the well-known amine, &imidoazolylethglamine, more generally known as histamine (the C constituent of pituitary extracts), which up to the present time has been the most powerful oxytocic or uteruscontracting agent known to medical science. The best and final tartrates are found to be from 1000 to 1250 times more powerful in their action than the acid phosphate of histamine. The tartrate has many other physiological properties. It acts upon the entire cardio-vascular apparatus, constricting the arterioles and capillaries and causing a significant and prolonged rise of the blood pressure. It acts in a remarkable way upon the respiration when the dose is more than a mere minimum, causing the periodic respiration known as CheyneStokes resbiration. It acts upon the kidneys in two ways. I n rabbits, for example, which have been fed for a week on cabbages or other succulent vegetables, it causes after each injection of a minute amount only a sharp but temporary flow of urine. Under other conditions it can cause a cessation or diminution of the urinary flow. This latter antidiuretic action is seen at its best in human beings in the disease known as diabetes i n s i p i d u s . Numerous trials in hospitals have shown that when our tartrate is administered subcutaneously to patients suffering from this form of diabetes the relief is secured as promptly as when a well-prepared aqueous extract of the posterior lobe is administered.3 This tartrate exhibits every known specific physiological property of a good extract of the posterior lobe; for example, it even darkens the skin of tadpoles that have been changed into pale tadpoles by removal of the posterior lobe of the hypophysis. There is further proof of a chemical nature that all the physiological properties described belong to one substance only. In other words, the tartrate, while still perhaps contaminated with some physiologically inactive material, consist3, not, of several active substances, but of only one physiologically active substance, or hormone. The chemical properlies of the tartrate cannot be described here in the proper detail with the proper qualifying statements. The method of isolation rests largely on adsorption methods, and it is interesting to observe that the final product can no longer be precipitated by such agents as mercuric chloride, picric acid, and tannic acid (in the presence of sodium chloride), and only imperfectly by phosphotungstic acid, while all these agents precipitate it in a practically quantitative manner from aqueous extracts of the posterior lobe. So, too, the behavior of the isolated substance towards solvents differs greatly from that observed when it isstill contaminated with other constituents of the gland. Naturally there are many other points relating to the physiological action, the clinical uses and chemical properties of this extraordinary substance which space does not permit considering. 3
J Pharmacol., 22, 317 (1923).
B u s t of P a s t e u r Dedicated O n t h e afternoon of Tuesday, September 16, ceremonies were held a t the American Institute 01 Eaking, Chicago, Ill., in connection with t h e dedication of a portrait bust in bronze of Louis Pasteur. H . E. Earnard, director OF t h e institute, made t h e opening address. The presentation 02 behalf of t h e donors of the bust was ma5e by Max Hexius, president of t h e Wahl-Henius Institute, and Raymond K. Stritzinger, president of t h e American Baker’s Association, made the acceptarce speech. S. P. L. Sorensen, director of t h e Carlsberg Laboratories, Copenhagen, Denmark, delivered an address, taking as his subject, “Louis Pasteur.” T h e Danish and French Consuls located in Chicago also spoke and during t h e ceremonies t h e national anthems for Denmark, France, and t h e United States were played. A lunch followed a t which members of the institute and specially invited guests were present.
