1 I > A REVISION OF THE PROXIMATE COMPOSITION OF RHAMNUS PURSHIANA BY FRED WARD ALWOOD B. S. University of Illinois, 1920 THESIS Submitted in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE IN CHEMISTRY IN THE GRADUATE SCHOOL OF THE UNIVERSITY OF ILLINOIS 1921 UNIVERSITY OF ILLINOIS THE GRADUATE SCHOOL JUNE 4 192JL I HEREBY RECOMMEND THAT THE THESIS PREPARED UNDER MY SUPERVISION BY FRED V/ARD AL^VOOD C ENTITLED . /I LEVI! o: PURSHIANA BE ACCEPTED AS FULFILLING THIS PART OF THE REQUIREMENTS FOR Recommendation concurred in* Committee on Final Examination* *Required for doctor’s degree but not for master’s 'v >^3 - /^iq^mjjr ho YTiaHavmu ;•. V *« - s? I f 3 J 5 TA.ua AJBiu* 3 m ^ - ri: eidJHr nrr t.uit My ^ ' - ^ - .. ... .? ;: •% A ,c#:^c*^T Ya^yfn^RjKwj? ' 7 /;.;l^ . I.Ti;'’".- '"',* f' 'A": > .- », - - - » ,-i:* - ‘ iJfes^iau i !>iT Jo ^11 rr i^v.jiiti.rfct >/. M la * '• .:t 'KJs r.^-'-i viVU. vK.aispaff'im- «j<| i 1. * \ , .. : >■ : . .fo a Muu? '* ' H? * -I* ; _> ' • ' N *•. K? ? '. .‘ ;/« 52 ^ ' * T' * ■ ' /''’^>,r_I^ . . = a;^ ' *tti i:f^^uwJo r u)cbft3/nnvt>-->y .U . J ’ • , . . *■ i< j» iSihAt* *4 *»' io4 kfXfpif. e*iocw>f> »i| ■ AUC^ ACKiroV/LaDGUdElHT I wish to express my appreciation to Dr. George D. Beal under whose direction and ad'vice this work has "been carried out. The success and results are due largely to his helpfulness and suggestions given in carrying out the experimental Y/ork of this paper. J Digitized by the Internet Archive in 2015 https://archive.org/details/revisionofproximOOalwo TABLil 0? GOITTailTS Page I. IIIIHODUCIIOH 3 II. EISTOHICAL 4 III. EXPERILSETAL 8 A. Water Soluble l-iatter 12 B. Hot Water Extract 15 IV. SUMiAHY 17 V. BIBLIOGRAPHY 19 3 llITRQDUCglOH The work undertaken was a continuance of work done last year hy the author. At that time more attention was given to the insoluble resin and its products. The \7ater soluble matter was also examined, but during evaporation of the solution it caramelized, making identification of the sugars impossible. From the results obtained it v/as concluded that a portion at least of the anthraquinone derivatives are present as gluco- sides, or in some other combined form. The work on the water soluble matter was continued this year with the results which follow in the experimental part of the investigation. A f. ■ r ill 'i#^'* Tin ■**“ ^ -i T- ■- vr ' ^ '>/' Vk ^919 * V:. ' : V'fv'b ,™ ■", ■ ■j'vt ’■ % p-f .i- ■ ' ■' • • ‘"'^ ‘VA’^'i ' ‘ ' « : W> '^.>^ y ' '■' ■■ ' ^KrJifcJiA ^.’■'ii'viBB-. , •f ^ :, ^''". ■ ':‘t : V' - ■( -1 it'" ' . '^S- '.'■'! r UA .*V‘ Ha. .-'Su • ; I* ■ .•..i^‘tiu!» r^|||jj^,..,.,V:,,-i'|^}^ , ■ j.‘" Lsi ^ • ' -I- ,■■- '■ ,' '"'t®- . ■ . 'C --IE’' ■^>^*''’<’ ■/ -i t < ' *1 - . '® «; '''Vf '' , i!n|}.r^ .A • " '' " ■ r • » »# ■ . 1 . Ii* »Tl-» ■ ' ■* \ _ ;'.,, 4 , i.ff "-■^'- 'WWm . '> “Ih ', 1' - - r-,si£' ‘ ,'-’■ 'AV»Jow*«M. ' : Mwjhki l 5 r^y'. '<(♦’* 1 ' * .:-A '\. ' / ::^i^ ^ I' ' ',iia '(} ./W; ;■ » '‘■;i...... ..-T 5 v' :• : ■■ vScf » ‘' ^ 'M '■- m M 1^ &t'. t^«.« f , t. iWPH l Nii> W».i < > ■ »i| i i i t)» <^ i» [ ^,M%| M ;i I ’.,- &;■ . . ' ‘^.uf ' p V ’>‘V . ' ■ 'S . i*/.'. I i 1 j»IfV- ,(*^L ^S' ■# 4 •; . V'-.'^' \": 4 IiISTORIC.'lL Since the latter part of the eighteenth century, there have been described various naturally occurring dyestuffs which are yellow in the free state and give red colors with alkalies. These dyestuffs, as prepared, by early workers, were not chemical individ- uals . ( 1 ) In 1858 De La Rue and Miller isolated pure emodin, and in 1875 Schmidt (2) distilled aloes ?/ith zinc dust, obtaining methyl anthracene. I'his was the first real evidence of the struc- so ture of these compounds, and remained until 1911, when Fischer and Sapper (3), and Fischer, Falro and Gross were able to prove that the hydrocarbon obtained from chrysophanic acid was B methyl an- thracene . Oesterle has shov;n (5) that emodin monomethyl ether is usually associated with chrysophanic acid. It was due to his v;ork (6) that chrysophanic acid, aloe emiodin and rhein v;ere shown to be different stages in the oxidation of the same hydroxymethyl anthra- quinone, while emodin is a tri-hydroxy-methyl anthraquinone . The structure of aloe-emodin was nroved to be o This structure being proved, the structures of rhein and chryso- phanic acid were established as follows:- Ctti-OV\ 0^ ° OW ^ ah 0 Vik of emodin hy the condensation of emodin with triacetyl from rhamnose in the presence of alkali. The product, while impure, as shown hy an indefinite melting point, behaved in a manner similar to frangu- lin, and v;hen mixed with the supposed frangulin from frangula hark, gave only a very slight lowering of the melting point of the mix- ture . Jowett and Potter made a detailed examination of the consti- tution of the hark, special attention being directed to the results already obtained hy other investigators. Where definite substances were isolated, their purity was established, if possible, by chem- ical methods. The details of the experimental work are somewhat extended, so only a summary of the results will be given. In addition to emodin, an isomeric substance was isolated melting at 183°C, insol- uble in ammonia, and its acetyl derivative melted at 168°C. Glu- cose was proved to occur in the bark. IIo evidence could be obtained of the existence of chryso- phanic acid or chrysarobin in the bark, or of glucosides yielding on hydrolysis emodin or chrysophanic acid. It was found that . ( ' s-V r^i ^ ' -h 1 { ' ‘ -IH"' • • n- y • ■• * ■ I \ ^ :> T 1'. ’ ■ . ^ ^ ' 'tf .'v' ' .^v IC o. :to n .H T ri .1 ."jJ'f-t/’i' t> r ri- rf rs«T^, ?“ rr'iwv'f »rT o" ''if' -r o ff-I pi^ rf i. f %'■ , t*- o^f r «v f( t. »»'■ •» A <• r A "1 “f Pi r> «' ^ i> r 114 IV. r 0 ^ r 5= V X o # {- ' f f.O r ^'- r— r* r C t» fi ^ I:^ A y f '' . f) r ^ ’ ' 1.^' B . V f f' M f,o*T >^i'' , P A r t. f V r^ t 7 f 4 :! qn *Vo i f'’'" T* r»A r>rf <_. • / •-■■'.x!'o-c O^H ‘■*' ’ '•*' ■•* '«► ' - -■'. . — • - 1 ^ ^ ^ ^ v/% i-» .. W. 4 l- 4 A-> A* .w «■ . - - j ^ • ih. - - ■*• n '*. .''- ‘V i V ».' i:i .'■ '.'..C i, .1,-1 ■.«. JU ' Va • « < •* - • - ; ^* .'• f>rT J,-. t " ::■ r * , W 4 *- v' ■' ■ ■ - . 0 «A Vt 1 iV'i *4 ' • t ^ ‘ l'^'' ‘ ^ V A ^ J V .w M #<«< 7 emodin, although insoluble in water, was soluble in the aqueous extract of the bark and that it was extracted from such a solution only slowly and with difficiilty by shaking with the imiscible sol- vents. After treatment v/ith acids, the water soluble substances were decomposed with formation of insolube resins and the emodin vjas very readily extracted from such a mixture. The bark contained about two per cent of fatty or waxy material, which consisted of rhamnol arachidate, free arachidic acid, and substances yielding on hydrolysis linolic acid, and myris- tic acid. The name rhamnol has been assigned to the alcohol melting at 135^-156°, which is combined with arachidic acid in cas- cara. I I . .• : ‘ rt ‘'":\. . . i v po^A'^orfo 'trej f).'!** t *" r "ff'o rf»r Af*v^rr o"*(r *fllrOjA C i 8 I^LXP^HIIvliilllTAL The author's attention V7as nov/ directed towards the products of the hydrolysis of rhamnus purshiana, the water soluble extract being studied. When the bark was extracted in a percolator of the ordinary type, the extract being concentrated by almost continuous heating on the steam bath. An excellent yield of emodin was ob- tained on extracting the dried resin with ether. When another, and much larger, portion of the drug was extracted in the Lloyd apparatus, in which the extract after having been rapidly concen- trated was immediately chilled and kept cold, the yield of emodin obtained from the resin by direct extraction was very much smaller. It was thot that the difference in yield might be explained by assuming the presence in the drug of an emodin containing gluco- side, or other derivative, which was extracted in either instance by the alcohol, but in the one case was broken do?;n by continuous heating, yielding emodin which was easily extracted by ether. The purpose of this investigation was to study the water soluble extract and to attempt to establish the presence of a glu- coside in the bark. The material used for the examination was cascara bark, ob- tained through Fuller, liorrison and Company, Wholesale Druggists, of Chicago. The drug as received \vas in the form of quills, and was ground to about number twenty powder in a Kance Mill. The drug was moistened with alcohol, placed in the percolator of a Lloyd ilxtraction Apparatus, covered with 95 /^> alcohol, and allowed to macenate twenty-four hours, after which percolation was begun, and continued until the dinig v;as completely exhausted. • I V' '•* I I i. o JjU - : t - ' \ ■>- , . V. , . V W -'-, • »■< • ^ 1 r, f. ft ■''• r- ■ .>*.■. T:' T .•.».•'■ f •,•*»■ ^/- *or >r- r t "’ f* '*!"*' *'r^ "* . ■^”^1**1*’ *^,'1 If I .r.r u '■f^> ■■" r«'‘*‘r Pjfifrtrf ^ i^n-:r •\*^j*>'*r* '.' "tr, V” 7^■^'7 'r/7 'to.ktO'^lctX'' yrf t£ -o*!; nr f: ’^^■^ f!•^d’c. acT tri'; jEi.,etv -rr oorkO*s:o'^^I Mx.t t.ifft 'Tl tj'. . 'lifi^.AOO lo v>it j-ti 3’ .TT>eiv JfOr ,, .. 1 " . -dv C3»;^*. Itl f/aj'OfiJt. Xv3 .i.)£:iv7 , ■/■' T',i> f^Jtj '. > j7i. :£ctoo (uT j’ L-..'3'v I 10 bjit .1- IM 7-C . , V J. , I ' ’a j - Jl l'< 'J ^ ; -'.-u '4 X. .if.-}* '' • ' . , • • 'i*’/ It - aiAa jJ ■J i..u » i , . «» ./ S ’ f • r i» J • .v^ * • -:t "■ '*r'' r .-f>- . ,r h. « ' 0 V", l‘C:!*f e- Z ■}■ „ . u ' >•' / . O". .f 7 ■ i Ar '••r i) 9 Concentration of the percolate uas carried out in the con- centrator of the extraction apparatus, and was continuous during the extraction. This concentrator is of a type making use of the principle of surface e'vaporation, while the tendency is for the heavier concentrate to he carried hy gravity to the lov;er portion of the concentrator, thus removing it from the heated zone. The extract is never exposed to heat for any period longer than one minute, and is in addition rapidly cooled hy a water Jacket sur- rounding the apparatus immediately helow the heater. This short period of heating, coupled with rapid cooling, and the fact that the extract is at no time brought in contact v/ith air, leads one to believe that the extract, as thus obtained, must represent most closely the actual nature of the extractives as they are present in the tissues before extraction. The extract obtained, representing the total alcohol soluble material of the bark, was reddish brown in color, thick and syrupy. This was mixed with enough alcohol to make a liquid having the vis- cosity of a rather thin syrup, and sufficient "dT/o hydrochloric acid added to give an actual acid concentration of one percent. The flasK was placed on a steam bath, with a large funnel in the neck of the flask to restrict circulation of air and loss of solvent, and heated at such a rate that the alcohol simmered gently. The heat- ing was continued for twenty-four hours, twice replenishing the al- cohol lost by evaporation. At the end of that period the alcohol remaining was distilled off until a thick viscious extract remained, and this, while still warm, ?;as poured into a large volume of water contained in a stone Jar, the water being vigorously stirred by 10 means of a "blast of air. I'he precipitate obtained was a dense, sticky mass, dark brown in color. This was filtered from an orange yellow liquid, washed with water, and examined in 1919-EO, with the following results. The work of this year is carried on as a continuance of that of last year, in which the hydrolysed resin was studied. The bark was extracted with alcohol, the alcoholic extract hydrolyzed for twenty-four hours with 1^ hydrochloric acid and poured into water. The resin separating out was studied with the results which follow. The precipitated resin, after being air-dried, was broken into small fragments and placed in the inner tube of a large Soxh- let extraction apparatus, where it was extracted with ether. The resin during this extraction, possibly due to the presence of a small amount of water, gradually cohered into a gummy mass, only slightly permeable to the ether. When this stage was reached, the resin -/as removed, dried and powdered again, then returned to the extraction apparatus and the extraction with ether completed. The resin, after the ether failed to dissolve more material, was dis- solved in alcohol, hydrolyzed a second time with one percent hydro- chloric acid, and precipitated and extracted with ether as before. Since the anthraquinone derivatives present in the ether ex- tract would differ in acidity by reason of variation in the number of hydroxyl a«d carbonyl groups present, the ether solution was shaken with solutions of progressively increasing basicity in order to effect a partial separation. Accordingly the ether extract was shaken in turn with solutions of ammonium carbonate, sodium carbon- ate, and sodium hydroxide. • 4«V*1 *''■ f ^ 7 ‘* c • 'J 1 J. I. Otv.'i « V .t - -rr, . , *Ti-_,t,-.'.- ‘ ' '■ J. '‘4 , ; r,Vl‘ ’ ) . ■ ; ' I ’V •' • •■•■» .i' '■* , ' k • ■ ■ 0- !. v‘ . *♦ ;>P,. • ' ' ■' ... ^ ■ '. 1 y ’In ■; , •' ■ - 1 . 1 ' .1, ; ‘ - : <• * ^ . ' - ■ ■ • ' ■ • ; ' *1, .14 V yt. lEt^i - f 4 ^ f «7, . t,'‘‘AAJ -i' •• - . ^ .,>.£ • * ' rf ^ * _ ww “ - .1 ■ ■ . ' .. . . ^ aUt; « k * * • ' * / ,i:Ia ■ , . ., J ■ '. , f ' ■ I, ' .'t . ) • « . ' :.v. «v 1 •< * ■fv • , .’fr’ .1 ! . t . :. *. 'vTf: t^. . i . ' ’■ • T, .‘ ; : • ;. ■ . * ^ ^ ^ ' 0 W- i - - ' : 'l l iy i . » . .-lir • Sm « 1 V. • ' ■t.i ■'V , , V. ftv ^ . .■ 1 •„ w " ■r"C;;T2. \ ' * * ‘ i. - ’i s. ■* \ ' ./ i> 1 4Uo.3{‘ . f.. ... ‘ 3 • - » »,'• J . . < 4 ♦ . .* V i . • t ■ i . — • I A 11 Ammonium Carlonate ij^xtract. Shaking the ether solution vjith b^o ammonium carbonate yielded a deep-red soliition. This extraction was repeated until the carbonate solution became only slightly colored. The extracts thus obtained were combined and acidified with hydrochloric acid, forming a yellow precipitate, rather small in 'volume. The precipi- tate thus obtained was recrystallized in part from alcohol and part- ly from glacial acetic acid. The crystals were dried in air, and melted at 248°C. uncorrected. From the melting point of the com- pound this was found to be emodin. A mixture vjith an equal quan- tity of emodin from other sources showed, no lowering of the melting point . An acetyl derivative was prepared by refluxing with acetic anhydride and fused sodium acetate for one hour. On cooling the solution, lemon yellow needles appeared. These were recrystallized from glacial acetic acid, which after drying melted at 197*^, ■on- corrected. This corresponds to the melting point and appearance of triacetyl emodin as given in the literature, and as previously obtained in this laboratory. Sodium Carbonate iixtract . The ether extract was then shaken out with b^o sodium carbon- ate giving a deep-red solution. The extraction was repeated until the sodium carbonate solution was only slightly colored. The ex- tracts were combined and acidified with hydrochloric acid forming an orange yellow precipitate, large in volume. The precipitate was filtered and recrystallized from glacial acetic acid. The melting ik r < ' I ' V- «• tr -4. i»r hr •!> I>t fc* *V w W. ^ •' •-«>. ^ L' o... , . . i 7 , . '/., V I'" u' t"--- '■ '-K f i . ^ I 'bin -Bii J' 0 1 a.P> rr o ^ ^•V,.i;,,r^r,,4 ; 1.' Jt ii ■■' ! < iX' . • 1 'w' J. ti f ,r ■ r- :;,j,'.vj. If*' r 12 point of this precipitate was 250-255*^0. This was shown to he emodin by the method previously described. The triacetyl deriva- 0 tive prepared as before gave the yellow needles melting at 197 , corresponding to triacetyl emodin. taodium Hydroxide iilxtract. The ether extract was shaken out with a 5^ solution of sodium hydroxide until fresh portions of alkali added took on only a slight color. The extracts were combined and acidified with sulphuric acid, forming a dark brovm precipitate, very small in volume. The precipitate was recrystallized from glacial acetic acid. On attempting to determine the melting point of this material, it darkened greatly and apparently decomposed, making it impossible to observe the true melting point, The value lies above 260OG. The water soluble extract was treated and clai^ified for examination last year, but on evaporating the solution caramelized making any further identification impossible. It was due to this fact that the following work was carried out. The orange yellow filtrate, on standing in a closed con- tainer, gradually deposited a lemon yellow precipitate. This was filtered off, and the filtrate evaporated to a small volume under a diminished pressure. A. Water Soluble Hatter. The filtrate from the hydrolyzed resin was filtered and neutralized with barium carbonate, and filtered again. The fil- trate was evaporated, under diminished pressure, to a small vol- ; -'v’ ' ' ■' I - ; ; v^- — V .y ^ - .^.v.,%^- ,. ^ .. ! TM v'">- '■ ■ -c • - • ’tf ■f ■ i «i ' --f ':•; ■•;■ Ao'i«r.i>jif^,|ji; “A r ..ir.-, ■ '-' V » I J'l •■ h'll :V' 'i; 'll 1 , - ■' '• ^ fu . ' , ... ■: "•■-■■'I ■!' '*v t~!jf f'lrt.^v’.l’.-'*- '’r./ ),;. : . ■’ •.»' -u.;- . ‘ ,‘ .: .ru; tl w' . -t ‘ ’•■ v.>, ,^.. •yi s V- 't'/.>S , -n. , .i,;, \ L 4 F^'^/v..W-C ■ '■'■ - -. 1 ;^’^' n_^ 4 i<'ij ■!, . • .wtl:- ■*■ ' '*• •-- -“^ .Jwoiii.i-Jiiw. ..o.^yt '.'. 'ctL^. r i.y'iy I'ut.- till . v-:»T ''t ft, ^ jiiiy, .Ik m « . t^jLi 1 all t.a 'i'Jj OU ;vfti X flv, ^ ! ' * ''. j I ' , ' *, i’j V ,'. " •i. - , . . \ ** ' w ■» t • \ ‘ %f*X . ' (,-i .j 6 v:i ."ig'fc'XJ} AGS jl , .. AX'! f •J" I (i ■ a. >«> U> J 4 > ^ 4 > - . iALwij au-Is; enJ tvi ‘1300 03 ui0-i'...f5c rr'/:- V .y , 4;:: •K •:^^ ' '“^1 .>♦ V . , . -fif/ATV ‘ • L uc'H yy >f

r- ■fk'itLl'- i . . ' < •V >. . 1 ^ ' ' ■ •: ' xr /• ji A* . S3'«’^xg ' : lo 1C w'c.i!i£v; nccif l v.rr«fin i-o <1 ' ■ ■ ?! .vii'ii. cv i.iB.ire 2 oj '‘'OJ'j? £ 3 -,oq/jTe Gil-ffivtXi'q elf,^ f’i'i .=t- 0 I .■ " t-raa ' • ' M ( ■ • ' ''i ‘>''' '^'•^.i n/.l K;, ?* I '>i. 1': i .Kh .-''3 ^ 'i«Ni I ' 1. 1. L a' 'i : ■ ■ '.V:V;'^;<\x. •, ii ,1! 1 f J. . \ ,& 4 'a:rucii 3 G .iX'j . ' A: *: C.'!! .'.coilj^ ' A: n., " -x .' X. ftrt I- V ■ / «. V . -i < f . .i,*' u. ;■■ 13 ume and filtered. The precipitates obtained were dried and united, and ex- tracted in a Soxhlet apparatus with ethyl ether. The ether ex- tract was shaken out siiccessively with 5^ solutions of ammonium carbonate, sodium carbonate and sodium hydroxide. T'rom the ammonium carbonate extract, after acidifying, only a small amount of amorphous red resin was obtained. The sodium carbonate ex- tract yielded emodin v/hich was recrystallized from glacial acetic acid and was identical with the emodin described in the litera- ture. The sodium hydroxide extract did not yield anything. The filtrate was treated with lead acetate and a precipi- tate, large in volume, was obtained. The filtrate was treated with hydrogen sulphide to remove the excess lead, and the lead sulphide filtered off. This solution was evaporated under diminished pressure, passing carbon dioxide through the solution during the evaporation. A portion of this solution, after re- moval of the excess hydrogen sulphide, was tested to determine the amount of sodium hydroxide necessary to neutralize the acid present. Then, to the remainder of the solutions, a little less than the amount needed to neutralize was added. This solution was evaporated under diminished pressure, watching closely for any signs of caramelization or formation of a product. A dark brown resinous substance separated out during the evaporation. This was filtered and dried, but no melting point could be obtained. The filtrate was diluted with five volumes of absolute alcohol, but no product crystallized out. This solution was allowed to evaporate spontaneously, but nothing separated out ex- 1 ' ' i-'i r ‘ H. V zf^ I r . A , 6(j^j r '■'wp f *■ '^'T ■jT*'' T t T r ! i' .' ' '• 3 f t'^Tf 0t»X*rO 3'^'V .T > ?• f- • '■'' > •'•■'■•f', , t i' l03 Bri\j’ u"("XrO‘X£f;t rr>CXO.r» j’’OCf*ri ^ nrrr^^orr - I ' ■ t H ■ -■ *-£ 3 ^X 13 .ftoituloa serf* lO 'fOl:tlonf /, . '^0 t.tf»7.0., 7 ;C'n inH'J ^\L. %'i.Jij^i u,i (’•*!, df-iori , m* .ii» 3 :./ ct:ij\T ■' . ^ . . 1 .;, ■ ■' . ■ ••:’'•'■ ••.‘3 # :« • ^ Tv L ■; . .: r ; f,vA'; rv; . 0 hit..,- V ■■;'■ ;j-' 5 r....o> i jjcj.. .ift.r. •-: ■;< j/ ,r. .; re *•; •' • 1 . 1 n ■1 rs-acfe-. 14 cept more of the dark brown resin. A portion of the solution was evaporated to dryness, and. some crystals separated out, but on examination it was found to be sodium acetate. The solution was tested for the presence of sugars by the Molisch napthol test. A few drops of the solution were diluted v;ith water, and to this was added napthol in chloroform solution; concentrated sulfuric acid was poured down the side and a ring was formed, which is characteristic of sugar solutions. An attempt was then made to make an osazone by boiling some of the solution with phenyl hydrazine hydrochloride and sodium acetate, llo product of osazone could be obtained. A brown precipitate separated out, but on trying to recrystallize from 50/O alcohol nothing vms obtained. A portion of the solution was then tested with I’ehling's solution to see if there vjas any reduction. A precipitate of red cupious oxide was obtained showing that sugars might possibly be present. The reduction of the Fehling's solution, however, may be due to some other constituent in the clarified solution. The precipitate formed, on the standing of the water soluble extract of the hydrolyzed resin, was dissolved out of the con- tainers with 95% alcohol. The alcohol v/as then distilled off, after extraction with ether, and recovered. The ether extract was in turn shaken out with ammonium carbonate, sodium carbonate, and sodium hydroxide. The ammonium carbonate and sodium hydroxide solutions on acidifying yielded nothing. The sodium carbonate on acidifying gave a brown precipitate. This precipitate was re- crystallized from alcohol, most of the precipitate going into 15 solution leaving a small amount of yellow substance, which did not go into solution. This v;as filtered off, dried and. identified, as emodin. B. Hot Y/ater iixtract. A hot water extraction of the ground hark was made and ex- amined. The extract was dark brown in color and was evaporated under dimiinished pressure to a small volume and clarified by add- ing lead acetate and sodium phosphate, the phosphate being added to remove the excess lead, thus doing away with the use of hydro- gen sulphide and subsequent evaporation of the excess hydrogen sulphide. This solution was evaporated, further to see if anything ?/ould crystallize out, but only a dark brown substance came down. The solution did not yield any precipitate on attempting to pre- pare an osazone, but did reduce Fahling's solution forming the characteristic red cuprous oxide. The precipitate formed on standing of the extract, and the precipitate from» the clarified solution was treated with acid and hydrolyzed for twenty-four hours, but no test for sugars was ob- tained on examination of the solution. An attempt ?/as made to isolate the glucoside as Dohme and Bnglehardt did in 1898. The solution was treated v/ith lead acetate and filtered. This precipitate was taken up in hot water and. hydrogen sulfide was passed in. The sulfides were filtered off and the filtrate treated v;ith lead subacetate. The precipi- tate was filtered off, taken up ?;ith hot water and hydrogen sul- fide passed in to decompose the precipitate. The filtrate from the sulfides was evaporated to dryness. A dark brownish-red sub- 16 stance was obtained. An attempt to determine the melting point was made, but could not be determined. 17 ^UluivIARY The follov/ing conclusions -were drawn from the preceding year’s Vi/ork on the resin; 1. Powdered cascara was extracted under such conditions that the extract coiilcl he concentrated with the minimum amount of exposure of the concentrated material to heat, thus avoiding decompositi on. 2« The extract so obtained was hydrolyzed in alcoholic solution by means of one percent hydrochloric acid. 3. The resin precipitated from this solution showed the pre- sence of a larger amount of emodin than the unhydrolyzed extract. 4. The presence of sugar in the water soluble extract from the hydrolyzed resin was shovm, but the sugar was not identified. 5. Prom the results obtained it is concluded that a portion, at least, of the anthraquinone derivatives are present as gluco- sid.es, or in some other combined form.. The following conclusions are dravm from the work of this year on the v;ater soluble extract of the hydrolyzed resin. 1. The material separating out in the containers of the v;ater soluble extract was examined and found to contain emiodin. 2. The filtrate from this material was neutralized, clari- fied, and evaporated under diminished, pressure, but only a dark brown resinous substance separated out. 3. The solution gave a test for sugars with Llolisch alpha duct naphthol reagent, but on testing for osazone formation, no pro- was obtained. 18 4. On lolling some of the solution with Fehling’s solution a large precipitate of cuprous oxide was obtained, showing the presence of sugars. 5. The hot water extract of the hark v/as clarified and test- ed for sugars, hut no osazone could he obtained. 6. On attempting to isolate the glucoside, as Dohme and iinglehardt did, only a dark hrovmish-red substance vms obtained. It did not correspond with the substance which they obtained. 7. ITrom these results it is concluded that sugars are present in the water soluble extract, but they could not be isolated. (O It does not seem probably that the glucoside can be isolated by methods which have been used. 19 . BIBLIOGHAiPHY (1) Am. J. Pharm. 30 (1858) 442-447 (2) Ber. 8 (1875) 1274 (3) J. Prakt. Chem. 83 (1911) 203 (4) Ibid. 83 (1911) 208 (5) Aroh de Pharm. 243 (1905) 434; 248 (1910) (6) Proc. Amer. Pharm. Assoc. 45 (1897) 193 (7) Trans. Chem. Soc. (1903) 1329 ( 8 ) Arch de Pharm. 246 (1908) 114 ( 9 ) Arch de Pharm. 252 (1915) 327 fic) J. Amer. Pharm. Assoc. 45 (1897) 198-202 (11) J. Chem. Soc. 61 (1891) 1. / V • \ *■ .. I '.M • ' ■ \ • ■ • : - f;,. \ \ V \ V F/‘' \ UNIVEHSmr OF ILLINOIS-URBANA* V 3 0112 108856722 >■? ' t ■ ■ ■ s I ■U.J" h