June 5 , 1956
2489
P E R I O D A T E OXIDATION OF C Y C L I C 1,3-DIKETOP;ES
standard. However, chondroitinsulfuric acid and hyaluronic acid did not exhibit a precipitation reaction with concanavalin-A. Upon desulfation and concomitant acetylation of sodium heparinatelo a desulfated heparin acetate is obtained which gives a precipitation reaction with concanavalin-A. This positive reaction persists when the above material is deacetylated with barium methoxide to give the N-acetyl desulfated heparin." It would appear therefore that, as in the case of glycogen, the precipitation reaction reflects the over-all molecular architecture of the whole heparin molecule or part of it rather than the presence of sulfate ester groups. It is interesting to note that desulfated mucoitin acetatelo also gives a precipitation reaction with concanavalin-A which lends support to the postulation of Jorpes12 that heparin is a polysulfuric ester of mucoitin. The galactogenI3 which is obtained together with (11) hI. L. Wolfrom, R. Montgomery, J. V. Karabinos a n d P. Rathgeb, THISJ O U R N A L , 72, 5796 (1950). (12) E. Jorpes, "Heparin," Oxford University Press, London, 1939; E. Jorpes. Biochem. J., 36, 203 (1942). (13) M. L. T;Volfrom, G. Sutherland and M. Schlamowitz, THIS JOURSAL, 74, 4883 (1952).
[CONTRIBUTION FROM
THE
heparin from beef lung does not give a precipitation reaction with concanavalin-A. It is suggested that the purity of a heparin preparation may be determined by the concanavalin-A precipitation reaction. Experimental Concanavalin-A Solution.-The concanavalin-A solution was prepared b y extracting jack bean meal" with 2% saline as described p r e v i ~ u s l y . ~ Concanavalin-A-Polysaccharide Precipitation Reaction .T o a n aqueous solution of the polysaccharide (1 ml. containing about 1 mg. of material) is added the concanavalin solution (9 tnl.). T h e two solutions are well mixed and then allowed. to stand for 10 minutes after which time the absorbance of the turbidity produced is determined in a n Evelyn colorimeter using a KO. 420 filter. A blank is prepared by using water in place of the polysaccharide solution. By interpolating the absorbance produced by the precipitation reaction on a standard curve prepared in the same way using purified human liver glycogen as a standard, the ratio of the absorbancies produced by equal weights of polysaccharide and the standard glycogen is calculated. (14) A product of the Arlington Chemical Co , Yonkers, DEPARTMENT OF AGRICCLTLJAL BIOCHEMISTRY UNIVERSITY OF MINNESOTA ST.PAUL1, MINNESOTA
DEPARTMEST O F CHEMISTRY O F THEOHIO
?rT.
Y.
STATE ENIVERSITY]
Periodate Oxidation of Cyclic 1,3-Diketones BY M. L. WOLFROX AND J. M. BOBBITT~ RECEIVEDNOVEMBER 2, 1955 The selective oxidizing ability of aqueous periodate ion is shown to encompass cyclic 1,3-diketones. Five- or six-membered cyclic 1,3-diketones, unsubstituted on carbon-2, reduce four molar equivalents of oxidant t o yield one equivalent of carbon dioxide and one molar equivalent of a dibasic acid. Carbon-2 substituted, cyclic, six-membered 1,3-diketones reduce three molar equivalents of oxidant to yield one molar equivalent of a monobasic acid and a like amount of a dibasic acid. Postulated reaction intermediates oxidize more rapidly than the initial diketone and give the same products. Satisfactory reaction occurs between bH 3 and 8 with a rate maximum between pH 5 and 6. The kinetics were second order a t low concentrations.
Oxidations of carbon compounds with aqueous and a-keto-acids,8 since in each case the reduction periodate ion a t room temperature or below can con- of one equivalent of oxidant is accompanied by the veniently be divided into two types : (1) those pro- cleavage of one carbon-carbon bond. The theory ducing carbon-carbon bond cleavage and ( 2 ) those of this type of oxidation has been extensively studwhere no such cleavage takes place. The second ied. The second type of periodate oxidation was noted reaction type occurs when a hydrogen attached to a and acetoacecarbon flanked by carbonyl groups is transformed to take place in malic,'O malonics*lO.ll into an hydroxyl function. This gives rise to an hy- tic acids" and their derivatives, and, indirectly, in droxy ketone, thus making the compound suscepti- malonaldehyde." Other examples of this type of oxidation have appeared in the sugar series.12 Abble to the first type of oxidation. The first type of periodate oxidation is the well (8) D. B. Sprinson and E. Chargaff, ibid., 164, 433 (1946). (9) R. Criegee, Sitzber. Ges. B e f 6 r d e v . ges. X a f t ~ r w M . n r b u r R , 69, known a-glycol cleavage reaction discovered by 25 (1934); C. A , , 29, 6820 (1935); C. C. Price and H. Kroll, THIS Malaprade3and initially developed by Fleury4and JOURNAL,60, 2726 (1938); C . C . Price and M. Knell, i b i d . , 64, 552 by H u d ~ o nand , ~ their associates. The same type of (1942); L. J. Heidt, E . K. Gladding and C. B. Purves, Paper T r a d e J., oxidation probably prevails in the oxidation of a- 121, h-0. 9, 35 (1945); F. R. Duke, THISJOURNAL,69, 3054 (1947); E. Taylor, i b i d . , 7 6 , 3912 (1953); F. R. Duke and V. C. Bulgrin, hydroxyketones,6 a-diketones,6 2-aminoalcohol~~J. i b i d . , 76, 3803 (1954); G. J. Buist, C. A. Bunton and V. J . Shiner, ( 1 ) Reported in part in A b s i i a c l s P a p e r s A m . Chem. Soc., 127, 3 1 N (1955).
(2) Allied Chemical and Dye Fellow, 1954-1965. (3) L. Malaprade, Compt. r e n d . , 186, 382 (1928); BuU. soc. chim., [41 43, 683 (1928); [SI 1, 833 (1934). (4) P. Fleury and J. Lange, Compt. r e n d . , 196, 1396 (1932); J. pharm. chim.,17, 107, 196 (1933). (5) E . L. Jackson and C . S . Hudson, THIS JOCRNAL, 68, 378 (1936); 69, 994 (1937); 61, 959 (1030). ( 6 ) P. W. Clutterbuck and I:. Keuter, J . Chrm. Soc., 1407 (1935). ( 7 ) B. H . Nicolet and L. A. Shinn, THISJ O U R N A L , 61, 1615 (1939); I.. H. Shinn and R. €I. Nicolet, J . Bioi. C/ztwz., 138, 91 (1941).
Research, 6 , 45 (1963); G. J. Buist and C. A. Bunton, J . Chem. S o c . , 1406 (1954). (10) P. Fleury and J. Courtois, Bull. soc. chim.,151 14, 368 (1947); [ 5 ] 16, 190 (1948). (11) C . F. Huebner, S. R. Ames and E. C. Bubl, THISJOURNAL, 68, 1621 (1946). (12) C. Niemann and J. T . Hays, ibid., 67, 1302 (1945); T. G. Hallsall, E. L. Hirst and J. K. N. Jones, J. Chem. Soc., 1427 (1947); R. K. Ness, H. G. Fletcher, Jr., and C. S. Hudson, THISJ O U R H A L , 73, 3742 (1961); M. L. Wolfrom, A. Thompson, A. N. O'Seill and T. T. Galkowski, ;bid., 74, 1062 (1952); E. L. Jackson and C. S.Hudson, ibid., 7 5 , 3000 (1953).
11.L. WOLFROM AND J. M. BOBBITT
2490
sence of reaction was noted in esters of malonic and acetoacetic acids and in the only 1,3-diketone investigated, the acyclic diketone 2,4-pentanedione.l 1 I n the light of this work, it was somewhat surprising when the cyclic 1,3-diketone 5,5-dimethyl-1,3-cyclohexanedione, was found to reduce periodate select i ~ e 1 y . l ~The reaction appears to be general for cyclic five- or six-membered lj3-diketones although an explanation for the reactivity of the cyclic over the acyclic structures can only be conjectured. Eleven diketones have been investigated in the work reported herein. They can be divided on the basis of structure into three groups: (1) cyclic 1,3diketones unsubstituted on C-2, ( 2 ) cyclic 1,3-diketones substituted on C-2 and (3) acyclic 1,3-diketones. Five cyclic 1,3-diketones unsubstituted on C-2 were subjected t o the action of sodium metaperiodate. These were 1,3-cyclohexanedione (I), 5methyl-1,3-cyclohexanedione (11), 5,5-dimethyl1,3-~yclohexanedione (III), 1,3-cyclopentanedione (IV) and 1,3-indandione (V).
5-
RI-IvCOJ~
+ coz
Rz
0
IV
0
il
Substrate,
M
NaIO,,
M
Oxidant b Acid reduced, produced, moles per moles per mole of mole of substrate substrate
Temp.,a Time, "C. hr.
2.04 0 24 3.94 I 0.0178 0.1218 2.04 4.00 6.5 .I218 I1 ,0136 2.00 10 20 4.00 .1821 I11 ,0143 0 96 3.92 1.92 ,0609 IV .0048 2.08 48 3.64 ,1240 1' ,0068 3.08 24 2.92 ,1218 .0138 VI I 3.05 2.75 2.95 VI11 .005gd ,0609 24 3.04 3.04 IX .0 156d .I218 24 0.60 ,0486 X .00a7 24 .52 XI .0486 ,0073 24 .08 X1I .0486 ,0078 2.02 7 3.03 ,1251 XIV .00726 2.15 21 2.10 ,2437 ,0272 X 1' a Room temperature, 22-28", unless otherwise stated. Determined by arsenite m e t h ~ d . ~ J By ~ titration t o Solution of substrate was not phenolphthalein end-point. immediate. e Transient iodine appeared a t higher concentratioiis.
TABLE I1 ACID
PRODUCTS FROM THE PERIODATE OXIDATION OF DIKETONES COz,d moles/ mole Subxp.,c of strate, NaIOp, Acidsaib Yield, 'C., subM M obtained yo cor. strate
I 0.0110 0.1268 ,0431 ,2437 Glutaric I 0278 ,2437 3-Methylglutaric' I1 ,0301 111 .2483 3,3-Dimethylglutaric! ,2437 SuccinicQ .0100 IV ,0442 ,2437 V .0455 ,2500 Phthalonich VI Phthalic ,0176 ,2481 Acetic' VI1 Glutaric ,2437 Propionici ,0236 VI11 Glutaric ,2437 Phenylacetici IX ,0309 Glutaric ,0139 ,2437 Glutaric XIV 0312 ,2437 Glutaric XV
0
0
TABLE I ANALYTICAL DATAON PERIODATE OXIDATIONSOF DIKETONES
/COzH
4IOP-
Rr--/,y==O Rz
Vol. 78
0
0.98
86. Bk
90k
94-9Bm 87-89"
92k
101-1020
144-145"
1.05 1.02 1.16
0.61 18.3
18.5' 94 59k
195-1 96g 0.00
95-96
90
figk 85' 73k
81k 7 tik
95-96 75-77' 95-96 94-96 95-96
1.03 0.99
I' VI Unless otherwise noted, all final identifications were made by infrared spectral comparisons. * Unless otherwise Three cyclic 1,3-diketones substituted on C-2 noted, acids were removed from the reaction mixtures by were oxidized. These were 2-methyl- (VII), 2- continuous ether extraction and isolated by removal of ethyl- (VIII) and 2-benzyl-1,3-cyclohexanedione ether and recrystallization. Fisher- Johns melting point
(1x1. 0 li
/C02H
A-R L,
"
=o
-1
310,vC0zH
+ RCOlH
1'11, R = CH3 VIII, R = CzH6 IX, R = CsHsCH?
Three acyclic 1,3-diketones, 2,4-pentanedione (X), l-phenyl-1,3-butanedione(XI) and 1,3-di(13) This observation was made in this Laboratory in 1952 by Dr. C. S. Rooney incidental t o a n investigation of t h e reaction products formed from reducing sugars and amino acids (Quartermaster Food and Container Institute for t h e Armed Forces, Chicago, Ill., Contract U A - I 1-009-gm-1329-4, T h e Ohio S t a t e University Research Fountlatioil l'rc,ject 477)
block. By sweeping from the reaction mixture with a stream of nitrogen into barium hydroxide and gravimetric determination of the barium carbonate. e Neutral equivalent, 73.0; cyclic anhydride, m.p. 42-42.5' (accepted 41'). f Neutral equivalent, 80.00; cyclic anhydride, m,.p. 126127" (accepted 124'). g Mixed m.p. 144-145 , undcpressed. Isolated as aniline d e r i ~ a t i v e , ' m.p. ~ 162.5163.5' (accepted 165") from concentrated ether extract after filtration of phthalic acid. By steam distillation from concentrated ether extract using acetyl assay apparatus.I6 Positive identification by comparison of the infrared spectra of the sodium salts. 1 Obtained by extraction of reaction mixture with one portion of ether prior t o continuous extraction. After recrystallization from benzene; [ -4fter recrystallization from water. Repo;ted 95-96 , 97.5'. n dccepted 87". Accepted 103-104 p Acceped 1441-G'. Accepted 206-208'. .4ccepted 76.7 .
.
( 1 L) E . I,. Jackson. O r g . Reaclions, 2 , 311 (1944). ( l a ) I
