66

4 sieve) in a porcelain dish with 30 c.c. NH2OH, sp.g. 0.910, and pass through a No. 3 sieve into a flask containing 270 c.c. of CHCl3, heating the mixture in the water-bath for 1 hour under a reflux. Cool, strain and press till the liquid measures to 185 c.c. Repeat the treatment twice with 220 c.c. and 200 c.c. portions of CHCl,, recovering 180 and 160 c.c., respectively, of the CHCl, extracts. Filter the combined liquids (525 c.c.) and concentrate by distillation to 100 c.c., then shake in a separatory funnel with four 30 c.c. portions of 1 per cent. AcOH. To the combined acetous extracts add a trace of blankit" or SO2, filter and shake out in separatory with 50 c.c. of Et2O to remove impurities. Add NH2OH in excess and shake out with Et2O until a test with Mayer's reagent shows that no more alkaloid is extracted. Wash the combined Et2O extracts once with water, dry over anhydrous Na,SO,, filter and distil to a very small volume, finally transferring the residue quantitatively to a 30 c.c. Erlenmeyer flask, then evaporate the Et,O completely. Dissolve the residue in 3 c.c. absolute EtOH and add 4 drops HCl (sp.g. 1·19). Let stand overnight, transfer to a tared 5 cm. filter, washing the last crystal remnants on to the filter with a mixture of EtOH and Et,0 (sp.g. 0-720) 3:1. Finally wash with Et,O, dry and weigh as yohimbine hydrochloride. (See also Y.B., 1922, 208.)

ANIMAL PRODUCTS

Adrenaline, Chemical Method for the Detection of. B. Stuber, A. Russmann, and E. A. Proebsting. (Z. ges. exp. Medizin, 1923, 32, 448, through J. Soc. Chem. Ind., 1923, 42, 739A.) One drop of cold saturated HgCl, solution acidified by the addition of 1 per cent. of N/200 H2SO4 3 drops of cold saturated sulphanilic acid solution, and 1 drop of N/20 potassium hydrogen iodate solution are added to about 4 c.c. of neutral or feebly acid adrenalin solution, which is then boiled for 1 minute. A yellowish-brown, reddish-yellow, or pale yellow coloration is produced according to the concentration of the solution; the colour gradually deepens until after several hours it remains of constant depth and can be used for colorimetric estimation. Urea, uric acid, creatine, phenylalanine, leucine, tyrosine, tryptophane, cystine, bilirubin, lipochrome, cholesterol, and hypophysine give no coloration on application

of this test, while methylaminocatechol gives a yellow coloration in concentrations higher than 1: 100,000. The final decomposition products of adrenalin have no effect on the reaction, but the coloration is apparently produced in the presence of intermediate decomposition products which can cause vascular contraction. Previous to applying the test to animal fluids, these are mixed with HCl and toluene and dialyzed against distilled water for 24-36 hours. The test is capable of detecting adrenaline in dilutions up to 1 in 50-100 millions.

Adrenalin Content of Suprarenal Capsules, Physiological and Chemical Determination of. A. Richaud. (Comptes rend. Soc. biol., 1922, 86, 26, through Bull. Sci. Pharm., 1922, 29, 284.) The physiological method of Cushny applied to the determination of adrenalin in powdered suprarenal capsules, invariably gives higher results than any of the chemical methods at present in use. This indicates either that there is present in the capsules some other constituent, besides adrenalin, which exerts a hypertensive action; or, as is more probable, that the methods at present used for the chemical extraction of adrenalin are faulty and fail to yield the whole of that active principle contained in the organs. (See also Y.B., 1921, 25; 1922, 210.)

Ambergris, Testing of. H. I. Cole. (Philippine J. Sci., 1922, 20, 105, through Perfum. Record, 1922, 13, 300.) Genuine ambergris has a waxy texture, an earthy odour, and when burnt gives off the odour of burning fæces. It melts at 65°, and the Et2O extract, ambrein, at 82° to 88° C. Sp.g. 0-908 to 1-5028; saponification of Et2O extract 17 to 35; I value of Et2O extract, 78; ash, traces to 6 per cent. A "supposed ambergris," occasionally stated to be picked up at sea in the localities where true ambergris is found, closely resembles the genuine product in physical characters but has the texture of chewing gum, and in odour recalls that of brown sugar and fresh sawdust. When burnt, it gives the odour of burning rubber. It softens at 100° C., and melts at 118° C. The Et2O extract softens at 100° C. and melts at 120° C., and has the saponification value 109.4 to 121-4, and the I value 52.9 to 78.54. The original substance has a variable sp.g., both over and below 1000; ash 3-6 per cent. It yielded 21.06 per cent. of gutta by the method for rubber analysis. This supposed ambergris has a very close resemblance to the dried latex of Artocarpus elastica, which

gives figures closely approaching the above when put through the same scheme of analysis. This leaves no doubt that the "supposed ambergris" is of vegetable origin, and is probably derived from a tree closely related to Artocarpus. Microscopical examination is often more useful than chemical tests for the identification of true ambergris. This is of a waxy nature, brown, with minute specks of white distributed through it, and there are also embedded in it many fragments of the chitinous part of the internal shell or gladius of a cuttlefish. Other chitinous fragments, in the form of a parrot's beak, and the remains of the mandibles of the cuttlefish, were also found. This chitinous material is often the horny gills of a cuttlefish species. These fragments appear as thin, dark brown, opaque, finely striated pieces of chitin varying in thickness from 0.04 to 0.1 millimeter. No moss, bark, or other vegetable material should be found in the sample. (See also Y.B., 1921, 27.)

Cantharidin, Microdetection of. G. Denigès. (Bull. Soc. Pharm. de Bordeaux, 1923, 61, 63.) Although there is no known reaction by which a small quantity of cantharidin can be easily identified, its distinctive micro-crystalline form renders it possible to identify it even in quantity of less than one-thousandth of 1 Mgm. When the material may be expected to yield about 1 Mgm. of cantharidin it may be treated direct, as follows. About 0-1 or 0.05 Mgm. of the material is crushed, with the drawn out and rounded end of a glass rod, in the centre of a micro-slide. A droplet of CHCI, is applied, and allowed to evaporate spontaneously. The residue, uncovered, is then examined under the microscope. In presence of cantharidin characteristic crystalline tablets and prisms either isolated or grouped in step-like formation will be seen. If the crystalline form is not very distinct, a droplet of pure C,H, is applied and allowed to evaporate. The crystals will then be sharply developed. The cantharidin may then be sublimed, as follows. Another microslip is placed over the deposit, supported by two clean pieces of match stems, so as to leave a clear space between the two slips. A drop of water is placed on the upper surface of the top slip, just over the object, to act as a condenser. The whole is placed on a small disc of asbestos tissue having a minute circular orifice of about 15 mm. diameter in the centre. This orifice should be placed exactly under the object between the double slips. A small chimney conveying heated air, made

from a cut test tube, is placed over a very small gas flame from a drawn out tube or minute burner not more than 2 or 3 cm. high. In from 5 to 15 minutes, according to the heat employed, a white deposit will appear due to the formation of the sublimate which is then examined under the microscope. The characteristic form of the crystals is illustrated. If the amount of cantharidin in the material is likely to be very minute, it must be extracted first with CHCl, or CH, and concentrated by evaporation on the water-bath. When only a few drops of residue are left this can be transferred gradually to the microslide, and treated as above. In this manner as little as 0.005 Mgm. of cantharidin can be detected.

Cantharidin, Native American Source of. Macrobasis albida, Say. A. Vie hoever and Ruth G. Ca pen. (J. Assoc. Offic. Agr. Chem., 1923, 6, 489, through Chem. Abstr., 1923, 17, 2472.) The beetle, Macrobasis albida, Say, abundant in Texas and Kansas, is found to contain both free (0.6 to 1 per cent.) and combined (4 per cent. and more) cantharidin, this material thus representing a possible domestic commercial source of the drug. The eggs contain large, the heads small, amounts of cantharidin; the wings contain none.

Cantharidin, Characterization of, New Method of Extraction. G. Marchiolo. (Boll. Chim. Farm., 1923, 62, 65, through J. Soc. Chem. Ind., 1923, 42, 421A.) In applying the test described below, cantharides powder requires no preliminary treatment, but a vesicatory product is treated first with boiling alcohol and then with petroleum ether to free it from waxy and resinous substances, etc. Any liquid suspected of containing cantharides, such as a hair lotion, is evaporated to dryness on a water-bath, the residue being treated with petroleum ether and the insoluble matter then tested; a solid product is treated with boiling water and filtered, the residue on the filter being washed with alcohol and petroleum ether and then tested. In the case of a post-mortem examination, since the food residues and the intestinal juices may interfere with the reaction, one portion is tested directly and another portion either treated with EtCHO, or mixed with ten times its weight of HCHO2, sp.g. 1.16, left for some hours, treated with a large volume of water, and stirred at intervals during 12 hours; in either case a little of the extract is evaporated on a microscope

slide. A small portion of the intestinal epithelium showing the characteristic vesications may also be tested. A little of the preparation obtained in any of the above ways is treated on a slide with a drop of strong H2SO4, which causes decolorization and clarification but no crystallization; if, however, a drop of boiled, distilled water is added after about 10 minutes, radiating square and hexagonal plates of cantharidin immediately appear, especially at the edges of the water drop. A blank test with the materials is always advisable. Traces of cantharidin may be detected by means of a reagent freshly prepared by adding 5 Gm. of boiled and filtered distilled water to a solution of 5 Gm. of crystalline CrO, in 10 Gm. of H2SO,. If a few crystals of cantharidin are moistened with a drop of this reagent, a green cloudiness appears after some hours, this giving way to a yellowish-green coloration after some days. To extract cantharidin or to estimate it in the fresh or dried insects, etc., 20 Gm. of the ground material is mixed in a separating funnel with 200 Gm. of HCHO2, sp.g. 1.16, the paste obtained being treated after half an hour with 200 Gm. of water and shaken occasionally for a day. It is then drained on a percolator and washed with a considerable volume of cold water, the filtrate and washings being evaporated to dryness in a porcelain dish on a water-bath and the residue treated with just sufficient strong H,SO, to dissolve the cantharidin and destroy the organic and colouring matters present. The cantharidin is precipitated from the acid solution by addition of water, collected on a tared filter, dried and weighed. The small amounts of impurity present may be eliminated either by treatment with boiling EtOH and then with petroleum ether, or by recrystallization from ethyl formate. (See also Y.B., 1920, 33, 34; 1922, 24.)

Cholesterol, Microidentification of. G. Denigès. (Gaz. hebdom. Sci. med. de Bordeaux, through Répertoire Pharm., 1923, 35, 225.) Advantage is taken of the fact that cholesterol crystallizes readily in two forms; hydrated, in rhombic plates or tablets with notched or serrated edges; and anhydrous in long needles. The first are obtained by crystallizing from a slightly hydrated medium; the second from an anhydrous mother liquor. A few particles of the substances, such as the scrapings from a calculus, are placed on a micro slide and a droplet of a mixture of CHCI, 2 and glacial HC2H ̧O, 1 is run on them by means of a drawn out glass rod. As the drop evaporates crystalline