140 QOODSON CONSTITUENTS OF THE of the Leaves of Helinus ovatus. By JOHN AUGUSTUS GOODSON. Helinus ovatus E. Meyer (Nat. Ord. Rhamnaceae) is a climbing shrub indigenous to South Africa where it is found growing on the borders of woods and thickets. No species of this genus which, according to Oliver (“Flora of Tropical Africa”) is confined to Africa and India appears to have been examined previously, although the constituents of the genus Rhamnus from which the order is named are fairly well known including as they do such well-known drugs as cascara sagrada m d buckthorn. The resulh of the present investigation show that H . ovatus contains aconitic acid quercetin a saponin and scyllitol. The last-mentioned substance was first isolated from certain plagio-stomous fishes including the spur dogfish but has since then bee LEAVES OF HELINUS OVATUS.141 found in a number of plants such as the acorns of the common oak and the leaves of Cocos plumosa and C nucifera. The occurrence of scyllitol the second of the two known meso-inositols in these plants is of considerable biological interest in view of the suggestion made by Winterstein Contardi and others (compare Posternak Compt. rend. 1919 169 37) that phyt*in, which is believed to be the usual organic phosphorus reserve con-stituent of plants is a mesoinositol hexaphosphate. The material came from Komgha Cape Province and was sup-pliqd by Mr. I. B. Pole Evans Chief of the Division of Botany, Union of South Africa who stated that it is used medicinally by the natives and is known locally as “soap-plant,” since the leaves have the property of yielding a lather when rubbed in the hands with water.Preliminary Examination. The leaves contained 9.5 per cent. of moisture and 9.2 per cent. of ash of which 20.2 per cent. was potash (K,O) equivalent to 1.8 per cent. in the leaves. No alkaloid or cyanogenetic glucoside could be detected by the usual reagents. The finely ground leaves gave the following percentages of extract on exhaustion in a Soxhlet apparatus with solvents in the order named petroleum (b. p. 35-60°) 2.2 ; ether 2.3 ; chloro-form 2.2; alcohol 23.0. Isolation of Ceryl Alcohol. The petroleum extract consisted of brown waxy matter of which about one-third remained undissolved when digested with ether. This was boiled with alcoholic potassium hydroxide solution to remove traces of oil and wax.The residue left after removal of the alcohol was crystallised from ethyl acetate and then melted at 7 8 O ; a specimen of ceryl alcohol melted at 81O in the same bath, and a mixture of the two at 79O. (Found C=82*0; H=14*0. Ceryl alcohol C&H,,O [Henriques Ber. 1897 30 14151 requires C=81*6; H=14.2 per cent.) The remaining extracts were systematically examined with results which showed that the quantity of plant available (650 grams) could best be dealt with by extraction with chloroform to remove wax and resinous matter and then in succession with alcohol and water 142 GOODSON CONSTITUENTS OF THE Examination of the Alcoholic Extract. The buik of Ghe alcohol was removed and the1 rwulting syrup set aside for some days when it deposited a considerable quantity of potassium chloride The filtrate was poured into about four times its volume of water and treated succewively with lead acetate and basic lead acetate.The lead was removed from the two pre-cipitates and from the filtrate by hydrogen sulphide in the usual manner. Isolation of Quercetin. The aqueous solution of the material recovered from the lead acetate precipitate contained a considerable amount of tannin. Extraction with ether removed a small quantity of a yellow sub-stance probably quercetin (see below). The1 liquor was then acidified with hydrochloric acid boiled to hydrolyse glumsides, cooled and again extracted with ether the extract yielding a yellow substance crystallising in rosettes of needles.This was recrystallised from a mixture of alcohol and chloroform and then melted a t 309O. On acetylation i t formed matted colourless needles melting at 1 9 5 O and this melting point was not depressed when the substance was mixed with penta-acetylquerceth. The yellow colouring matter is therefore quercetin. The aqueous solution of the material recovered from the basic lsad acetate precipitate also contained tannin and a small amount, of yellow colouring matter which could not be obtained in a crystalline condition. The filtrate after removal of the lead as sulphide was concen-trated and extracted with butyl alcohol which removed a saponin. The latter was purified by solution in water and precipitation with basic lead acetate the lead precipitate being decomposed with hydrogen sulphide in the usual manner and the filtrate evaporated to dryness under diminished premure.The quantity of saponin obtained was so small that no further purification could be effected. The material frothed strongly in aqueous solution gave no com-pound with cholesterol in alcoholic solution did not reduce Fehling’s solution and was not hzemolytic. It was hydrolyseld by boiling with dilute hydrochloric acid the resulting solution yield-ing a small amount of apparently crystalline sapogenin on extrac-tion with ether. The residual aquelous solution reduced Fehling’s solution strongly but did not give a crystalline phenylosazone. The aqueous liquid after extraction with butyl alcohol was con-centrated under diminished pressure and set aside when a furthe LEAVES OF HELINUS OVATUS.143 quantity of potassium chloride separated. d-phenylglucosazone on treatment with phenylhydrazine. The filtrate yielded Examination of the A pueozcs Extract. The aqueous extract was treated successively with lead acetate and basic lead acetat,e and the two 'precipitates were collected. The lead acetate precipitate was suspended in water and decom-posed by hydrogen sulphide. The filtrate was concentrated under diminished pressure and extracted with ether which removed 12.6 grams of a crystalline substance corresponding with 1.9 per cent,. in the leaves. This on recrystallisation from water formed minute prisms melt-ing a t 19l0 and gave all the reactions of aconitic acid including that described by Taylor (T.1919 115 886). (Found C=41.3, 41.4; H = 3.9 3-6. Acon-itic acid C,H,O, requires C =41-4 ; H = 3 - 5 per cent.) Isolation of Scyllitol. The basic lead acetab precipitate was decomposed in the usual manner and t h s filtrate concentrated under diminished pressure, wheln slightly brown crystals separated. Two crops of the crude substance amounting to 3 grams and corresponding wit.h 0-46 per ceat. in the leaves were obtained. The product was purified by recrystallisation from hot water from which it separated in anhydrous monoclinic rhombs. (Found C = 39.7 40.1 ; H = 6.8, 6.9. Scyllitol C,H,,O, requires C =40.0 ; H = 6.7 per cent.) The properties of the substance agreed closely with those recorded for scyllitol (J.Muller Ber. 1907 40 1821 and H. Muller T., 1907 91 1767; 1912 101 2383). When recrystallised slowly from cold water it separated in transparent hexagonal prisms, which on removal from the solvent became opaque and friable owing to loss of water of crystallisation. Crystals freed as rapidly as possible from adhering mother liquor lostl on exposure to air, 24.9 per cent. of water. C,€€,,0,,3H20 requires H,O=23*1 per cent. H. Muller noted this change in crystal habit and transparency, but did not establish the fact that it is due t o loss of water of cryst<allisation (T. 1907 91 1772) 144 HENDERSON AND SMEATON CONTRIBUTIONS TO THE When heated the scyllitol obtained from H . ovatus leaves coloured slightly a t 300° darkened considerably a t 320° and melted and effervmced at 353O as recorded by a mercury thermometer. J. Muller (Zoc. cit.) gave the solubility of scyllitol in water as about 1 gram in 100 C.C. a t 1 8 O ; the author finds a solubility of 1-03 grams in 100 grams a t 18O for his specimen whereas H. Muller (Zoc. c i t . ) gave it as 1.7 grams in 100 C.C. a t 1 5 O . Its identity with scyllitol was confirmed by the preparation of the hexa-acetyl derivative which melted a t 291O. H. Muller (Zoc. c i t . ) gives 290-291O (corr.). (Found C=50-1; H=5*6. Hexa-acetylscyllitol C,H,(C,H,O),O requires C = 50.0 ; H = 5.6 per cent.) The filtrate from the lead precipitates after removal of the lead, contained merely inorganic salts. I n conclusion the author desires to express his warmest thanks to Dr. Henry for his advice and criticism throughout the course of the work. WELL~OME CHEMICAL RESEARCH LABORATORIES, LONDON E.C. 1. [Received January ZFith 1920.