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Jan.

20, 1960 ALKALINEDEGRADATION


OF A L G I N A T E S 457

(6) 2,3,4,6-Tetra-O-methyl-~-mannose.-This chromato- aniline derivative of 2,3,4,6-tetra-0-methyl-~-glucose,


m.p.
graphically pure component (75 mg.) was obtained as a 137-138,32 it was concluded that only 2,3,4,6-tetra-O-
sirup [ c z ] ~ ~ +29.2'
D in water (c 3.9). Although chromato- methyl-D-mannose was originally present in this component.
graphically pure, the possibility existed that this component Acknowledgment.-The authors wish to express
could be a mixture of 2,3,4,6-tetra-0-methyl-~-glucose and
2,3,4,6-tetra-O-methyl-~-mannose since it has been shown their thanks to Mr. J. E. Jeffery under whose super-
that a variety of chromatographic solvents failed to separate vision the intrinsic viscosity determinations, sugar
these two rompounds.6 and methoxyl analyses were carried out. We also
The aniline derivative of this component was prepared wish to acknowledge the capable assistance of Mr.
and the crude crystals were separated from the adhering
sirup by use of a porous tile. These crystals were fraction- R. G. Rogerson. The authors are also indebted to
ally crystallized from ether-petroleum ether mixture into Dr. F. Smith, University of Minnesota, for authen-
four fractions, all of whirh had melting points and mixed tic specimens of crystalline 3,6-di-O-methyl-~-
melting points of 143-146'. The four fractions were com- glucose, 2,G-di-O-methyl-~-glucose and 2,3-di-0-
bined and had [ c Y ] ~-8.4'~ D in methanol (c 1.5) (lite3'm.p.
144-145", [ C Y ] ~ ~-7.5'
D (in methanol)). From the preced- methyl-D-mannonic acid.
ing data, and from the values given in the literature for the (32) M. L. Wolfrom and W. L. Lewis, THISJOURNAL,
60, 837
(1928).
(31) W. N. Haworth, R. L. Heath and S. Peat, J. Chem. Soc., 833
( 1941). WASH.
SHELTON,

[CONTRIBUTIONFROM THE DEPARTMENT


OF BIOCHEMISTRY,
PURDUE UNIVERSITY]

Alkaline Degradation of Alginates'


B Y ROY L. WHISTLERAND J. N. BEMILLER
JUNE 1, 1959
RECEIVED

Alginates are readily degraded by alkaline solutions a t 80-120' with the formation of new &carbon, dicarboxylic sac-
charinates, 3-deoxy-2-C-hydroxymethyl-pentarates.

Vollmert reported that methyl alginate is would be the isosaccharinate typeg with a carbox-
depolymerized in alkaline solutions while sodium ylate group a t the C5 position (C6 of the original
alginate is relatively more stable. However, uronic acid unit). Because of the presence of both
both esterified and ionic alginates are degraded in D- and L-hexuronic acid units, this new acid, 3-
alkaline solutions and form salts of saccharinic- deoxy-2-C-hydroxymethyl-pentaric acid, would be
type acids by the ordinary process of sequential produced in both D- and L-isomeric forms. Experi-
elimination of the reducing end unit and its sub- ence has shown that the process of alkaline degrada-
sequent transformation into a saccharinate (Fig. tion forms a saccharinic-type acid with predomi-
l ) . 3 The alkaline degradation reaction is more nantly one configuration of C2.
pronounced in high concentrations of alkalies The isomeric, 6-carbon, dicarboxylic saccharinic
and a t high temperatures. acids have been isolated in the course of the
Alginic acids are heteroglycuronoglycans of D- present work. Structures are assigned to these
mannopyranosyluronic acid units and L-guluronic acids on the basis of the currently accepted mech-
acid units. Alginic acid from Macrocystis pyrifera anism for the formation of products from (1+4)-
contains mainly D-mannopyranosyluronic acid units linked D-hexo- and D-pentoglycans. Supporting
and 20-40% of L-guluronic acid units.(O The D- evidence for the proposed structures is afforded by
mannopyranosyluronic acid units are connected the equivalent weight (detd. 100, calcd. 97), the
with P-(l+-4)-linkage~.~-'The position on the production of formaldehyde upon periodate oxida-
L-guluronic acid units to which the glycosidic tion (0.5 mole per acid equivalent), and by com-
linkages are attached is not known, but periodate parison of the nitric acid oxidation products with
oxidations gives no indication of linkages a t either those of a-D-isosaccharinic acid. Nitric acid oxida-
C2 or C3. If all uronic acid units are connected tion of a-D-isosaccharinic acid (I) produces a
with (1+4)-linkages, the saccharinates formed carboxyl group from the primary alcohol group
(1) Journal Paper No. 1438 of the Purdue Agricultural Experiment
(C2') so that the asymmetry of C2 is destroyed.
Station, Lafayette, Ind. A tricarboxylic acid, 2-C-carboxy-3-deoxy-~-glycero-
(2) B. Vollmert, A n g e v . t h e m . , 61, 329 (1949); Makromol. Chem., pentaric acid (111), is formed in which only the
5, 110 (1050). penultimate carbon atom is asymmetric. Nitric
(3) For a general discussion of saccharinic acid formation see:
R. L. Whistler and J. N. BeMiller, Advances in Carbohydvale Chem., 13, acid oxidation of the two 3-deoxy-2-C-hydroxy-
289 (1958). methyl-pentaric acids produces two tricarboxylic
(4) (a) F. G. Fischer and H. D6rfe1, Hoppe-Seylev's Z . physiol. chem., acids, one which has the same rotation as that of
302, 186 (1955); (b) D. W. Drummond, E. L. Hirst and Elizabeth the known 2-C-carboxy-3-deoxy-~-glycero-pentaric
Percival, Chemistry &' I n d u s f r y , 1088 (1958); (c) R. L. Whistler and
K. W. Kirby, Hoppe-SerIev's Z . physiol. chem., 314, 46 (1959).
acid and one which has an opposite rotation. The
(5) G. Lunde, E. Heen and E. c)y, K o l l o i d - Z . , 88, 196 (1938); E. isolated 3-deoxy-2-C-hydroxymethyl-pentarate
Heen, T i d s . K j e m i Bergeresen, 17, 127 (1937); C. A . , 3 2 , 5792 (1938). whose nitric acid oxidation product has the same
( 8 ) E. L. Hirst, J. K. N. Jones and W. 0. Jones, Nature, 143, 857 rotation as that of the known 2-C-carboxy-3-
(1039); J. Chem. Soc., 1880 (1939).
(7) S. K. Chanda, E. L. Hirst, E. G. V. Percival and A. G. Ross, (9) For the action of alkali on 4-0-substituted sugars see: W. M.
rhzd , 1833 (1952). Corbett and J. Kenner, J . Chem. SOL.,2245 (1953); 1780 (1954);
(8) H. J. Lucas and W. T.Stewart, TEISJOURNAL, 62, 1782 (1940). J. Kenner and G. N. Richards, ibid., 1810 (1955).
458 ROYL. WHISTLERAND J. N. BEMILLER Vol. 82

-0 & coo-

H H
o >- -0 &cn20H
coo-

H O
action. illkali-treated pectin and pectic acid
show similar b e h a v i ~ r . ~ItJ ~is~ likely
~ ~ that 3-
deoxy-2-C-hydroxymethyl-pentaratescould also
be produced by alkaline treatment of dinitrogen
tetroxide oxidized starch or cellulose. Although
the alkaline degradation of glycuronoglycans is
I
W slow a t temperatures of 20-40, it can be completed
in several hours at 120'.
yoo-
7(OH)(CH20H) COOH COOH COOH
I 1
y 2 C(OI)(CH?OH) HOACOOH C(OH)(CH20H)
H-C-OH
-
HSOl 1 HN03 I
+ CH? -E+---
~

I CH, CH2
coo- 011 0
I I HAOH
Fig. l.-The formation of 3-deoxy-2-C-hydroxymethyl-n- HYOH HCOH
erythro(or n-three)-pentaric acid from 4-O-substituted-n- I
CHzOH COOH COOH
mannuronate.
I IT1 I1
deoxy-D-glycero-pentaric acid has been designated
D-erythro(or D-threo) . The other whose oxidation Experimental
product has an opposite rotation has been desig- Chromatography.-Chromatographic separations were
done a t 26" using the following irrigants: .A, l-butanol-
nated L-erythro(or L-threo). ethanol-3 N ammonium hydroxide ( 4 : 1 : 5 v . / v . ) ; B, ethyl
The combined evidence presented here that the acetateacetic acid-formic acid-water (18: 3: 1:4 v./v.);
two saccharinic-type acids which have been C, ethyl acetate-pyridine-water (10:4:3 v./v.); D , 1-
isolated are D- and L-isomers of 3-deoxy-2-C-hy- butanol-ethanol-water (40: 11: 19 v./v.). The compo-
droxymethyl-pentaric acid is an indication that both nents were detected with silver nitrate aiid sodium hydrox-
ide.14
the D-mannuronic acid units and the L-guluronic Degradation.-Several methods of degradation nere
acid units are linked through the 1- and 4-positions. used; four typical experiments are given here. Sodium
It has been shown that P-(l-t4)-linked D-man- alginate (10 9.) was dispersed in 500 ml. of oxygen-free
noglycans form 8-D-isosaccharinates when treated sodium hydroxide solution, and the dispersion mas kept a t
37' for 28 days. After removal of sodium ions with Amber-
with alkaline solutions.10 Therefore, i t would be lite IR-l2O(H), 0.4 g. of material which was insoluble iii
predicted that the acid formed by the alkaline 757, ethanol and 6.1 g. of 757, ethanol-soluble, non-vola-
degradation of a 8-(1-4) -linked D-mannuronan tile acids were obtained. The latter were converted to their
would have the same configuration a t C2 as that of calcium salts (3.8 g.) which had a Kgluconatsvalue of 0.81
with irrigant A and [ C Y ] ~-11.6'
~D ( c 1.0, water).
P-D-isosaccharinic acid. In contrast, the action I n another procedure, 1 g. of alginic acid was dissolved in
of alkalies on 4-O-substituted-~-glucose forms 500 ml.of oxygen-free water,and an l O O 7 , excess (0.360 g.)
a-~-isosaccharinates,~ and, therefore, it would be of calcium hydroxide was added. The mixture was auto-
expected that a 3-deoxy-2-C-hydroxymethyl- claved a t about 120' for 65 hr., cooled, decationized with
Amberlite IR-12O(H) cation exchange resin, and concen-
pentaric acid of the D-series with the a-configura- trated under reduced pressure. No material which was
tion of C2 would be formed from 4-O-methyl-~- insoluble in 70y0ethanol was present. The acids were re-
glucuronate.ll In partial support of this view i t is converted t o their calcium salts with calcium carbonate.
These salts (0.6 g.) had a Rglucanntevalue of 0.81 with irrigant
observed that the two brucine salts of the desig- -1and [aIz5n -12.8" (c 1.0, water).
nated 3-deoxy-2-C-hydroxymethyl-pentaric acids Kelcoloid 0'5 (8.1 g.) was dispersed in 500 mi. of oxygen-
of the D-series from alginates and 4-0-methyl-D- free N sodium hydroxide solution and kept a t 37" for 7 days.
glucuronate have different X-ray diffraction pat- After removal of sodium ions with hrnberlite IR-lBO(K),
2.3 g. of material which was insoluble in 707, ethanol and
terns. 4.9 g. of 7070 ethanol-soluble, non-volatile acids were ob-
3 - Deoxy - 2 - C - hydroxymethyl - pentarates are tained. The latter were converted t o their calcium salts,
formed in good yields from either alginate esters or 0.5 g., which had a Reluoonate value of 0.81 x i t h irrigatit rl
salts. Rates of production of alcohol-soluble and [piIz5n-12.9' (c 1.0, water).
Thirty-five grams of sodium alginate was dibliersed in 3.3
fragments indicate that alkaline fragmentation of 1. of oxygen-free water a t 80' with stirring. After 12 h r . ,
alginate esters is more rapid than that of salts. 10 g. of calcium hydroxide was added, and the dispersion of
This increase in depolymerization rate is very insoluble calcium alginate was stirred a t 80" for 10 days.
likely due to a 8-alkoxy carbonyl mechanism with At the end of this period, the solution, which had become
clear, was decationized by passing it through :i cvlutnn of
the formation o an a,@-unsaturated acid.3 I t Amberlite IR-120( H ) and concentrated under reduced
has been proposed that the proton on Cc5 which is pressure t o a thick sirup. . portion of this sirup was con-
I
in the a-position to the ester group is removed. verted to calcium salts and a portion was converted to bru-
The free pair of electrons which is left on C5 then cine salts. T o make the brucine salt, a solution of the free
acid was treated with a slight excess of brucine for 18 hr. at
shifts to form a double bond between C4 and C5. 50'. T h e volume of the solution was suiEcient to dissolve
This process ejects the portion of the molecule all the brucine. Excess brucine was removed by chloroform
attached to C4 as a glycoxy anion and breaks the extraction. T h e aqueous solution was concentrated to dry-
polymer chain. This reaction is competitive with ness, and the salts were crystallized out of ethanol.
Equivalent Weight.-The weighed, dried calcium salts
de-esterification and, although it takes place only were dissolved in water, and cations were removed from the
to a slight extent, the exposure of a few reducing
ends in each polymer chain would greatly increase (12) H. Neukom and H. Deuel, Chemistry tr I ~ z d u s l u y ,683 (1988).
the rate of the normal alkaline degradation re- (13) P. Garrick, i b i d . , 1430 (1958).
(14) W. E. Trevelyan, D. P. Procter and J. S. Harrison, Y a l u r c ,
(10) R. L. Whistler and J. N. Behfiller, unpublished data. 166, 444 (1950).
( 1 1 ) R. I.. Whistler and G. S. Richards, THISJOURNAL, 80, 4858 (15) An SO% propylene glycol ester of alginic acid of the Kelco Cu.,
(1'3.58). New York, N . Y .
Jan. 20, 1960 SYNTHESIS
OF AMINO KETONES 459

solution with Amberlite IR-120( a). The equivalent weight with water, and, after standing for 12 hr., the mixture was
was determined by titration of the acidic sirup obtained upon filtered. The filtrate was concentrated under reduced
removal of water with standard sodium hydroxide solution. pressure a t 35-40' and purified by paper chromatography
The value obtained was 100 compared t o a calculated equiva- with irrigant B. A chromatographically pure sirup was
lent weight of 97 for 3-deoxy-2-C-hydroxymethyl-pentaric obtained (53 mg.) which had R,,IB and R117 values of 2.9 and
acid. 1.2, respectively, in irrigant B and a [ a I z 5-0.3" ~ (C 2.7,
Resolution.-An ethanol solution of brucine salts was water). This sirup had chromatographic flow rates in irri-
concentrated t o dryness, and the resulting powder was gants B, C and D which were identical t o those of the nitric
shaken with a little water a t 0". The mixture was then acid oxidation product of a-D-isosaccharinic acid.
stored for 24 hr. a t 0' and centrifuged a t 0'. This process Oxidation of a-D-Isosaccharinic Acid.-Calcium a-D-iso-
was repeated until all the remaining glass was soluble in saccharinate (3.5 g.) was oxidized with nitric acid by Kili-
water a t 0 " . The crystalline residue which was insoluble ani's method.20 The oxidized product was isolated by
in water a t 0 ' had a block m.p. of 163-188' and a X-ray paper chromatography with irrigant B; Rpaloand values
powder diffraction pattern which differed from that of the of the product in irrigant B were, respectwely, 2.9 and 1.2.
brucine salt of the acid formed by the action of lime water The yield was 0.74 g. of sirup which contained 1.2 X
on a 4-O-methyl-~-glucuronate.~~ acid equivalents per gram and had [ C Y ] ~ ~+1.2" D (C 7.4,
3-Deoxy-Z-C-hydroxyrnethyl-~-euythro(orL-thueo)-pen- water). This acid is 2-C-carboxy-3-deoxy-~-glycero-pen-
taric Acid.-The above crystalline, but slightly soluble, taric acid.
brucine salt was converted t o the free acid with Amberlite 3-Deoxy-Z-C-hydroxymethyl-~-erytltro(or D-three)-pen-
IR-l20(H), and the sirup obtained by concentration of the taric Acid (11).-The cold-water-soluble brucine salt from the
solution under reduced pressure a t 35-40' was further above resolution was converted t o the free acid by stirring
purified by paper Chromatography using the descending its solution with Amberlite IR-lBO(H). When this acidic
method and irrigant B. A component was obtained which solution was concentrated under reduced pressure a t 35-
appeared as a single component with irrigant to B, C and D. 40, a sirup was obtained which was purified by paper
This acid had a R,,16value of 3.1-3.4 and a R1I7valueof 1.2- chromatography. The final sirup (0.325 9.) moved as a
1.3. The purified sirupy, free acid thus obtained (1.46 9.) single component with irrigants B, C and D and had RBa16
had6.4 X 10-3acidequivalentsper gramand [ a ] z 6 ~ + 5 . 6 0 ( cand R117 values of 3.1-3.4 and 1.2-1.3, respectively, wlth
1.0, water). irrigant B. It contained 2.0 X acid equivalents per
When a portion of this sirup was oxidized with periodate gram and had [ a I z 6+3.0" ~ ( c 1.3, water).
according to the method of Sowden18and the formaldehyde A portion of the sirup was converted to its brucine salt
was determined by the method of Reeves,lg 0.5 mole of which, when crystallized from ethanol and placed in a capil-
formaldehyde per equivalent of acid was obtained. lary, began to melt a t 162" and decomposed a t about 250'.
Another portion of the acidic sirup was converted to its Anal. Calcd. for C5&2015NI: C, 63.6; H , 6.4; ?J, 5.7.
brucine salt which was crystallized from ethanol. The Found: C,61.7; H , 6 . 4 ; N,5.28, 5.53.
crystals had a block m.p. of 192-193', a capillary m.p. of
177-186" and [ a ] " ~- 18.0" ( c 3.4, water). Another portion (0.1 g.) was oxidized with nitric acid by
the method of Kiliani20 and purified chromatographically
Anal. Calcd. for C62Hs2015N4: C, 63.6; H, 6.4; N, 5.7. with irrigant B. A sirup was obtained which had a
Found: C,58.2; H , 6 . 5 ; h',5.78, 5.67. value of 2.8 and a R117 value of 1.2 with irrigant B and a
A third portion (0.1 g.) was oxidized with nitric acid ac- [ a I z 5 D 4-1.2" ( c 4.3, water). This acid had chromato-
cording t o the method of Kiliani,20that is, by treatment with graphic flow rates with irrigants B, C and D which were
nitric acid (1.4 d.) for 24 hr. a t 35", for 24 hr. a t 45', and identical t o those of 2-C-carboxy-3-deoxy-~-g/3'cero-pentaric
then for 24 hr. a t 50'. The reaction product was diluted acid produced by nitric acid oxidation of a-D-isosaccharinic
acid.
(16) Rga is the rate of movement relative t o t h a t of D-gluconic acid.
(17) Ri is the rate of movement relative to that of ~-glucono-l,4- Acknowledgment.-The authors gratefully ac-
lactone. knowledge the grant from the Kelco Co. and from
(18) J. C. Sowden, Mary G. Blair and Dorothy J. Ruenne, THIS the Department of Health, Education, and We1
JOURNAL, 79, 6450 (1957).
(19) R. E. Reeves, ibid., 6S, 1476 (1941).
fare which helped to support this work.
(20) H.Kiliani, Bey., 18, 631 (1885). IND.
LAFAYETTE,

[CONTRIBUTIOX FROM AVERPLABORATORY,


UNIVERSITY OF NEBRASKA]

Reactions of Amines. V. Synthesis of a-Amino Ketones1'2

BY HENRYE. BAUMGARTEN
AND JAMES R t . PETERSEN
RECEIVEDJULY 13, 959

The treatment of sec-alkyl amines ( I ) with t-butyl hypochlorite in benzene solution followed by succesiive treatments of
the uriisolated intermediates 11-IV with sodium rnethoxide and with dilute hydrochloric acid gave good yields of the cor-
responding a-amino ketone hydrochlorides 17. The infrared spectra of the a-amino ketone hydrochlorides resembled to
some extent the spectra of a-amiuo acid hydrochlorides.

In the first paper3 of this series a new rearrange- NHz NClz N


ment of N,N-dichloro sec-alkyl amines 11, which I - I --f /\ --+
RCHCHiR RCHCHzR RC-CHR
led to the formation of the corresponding cy-
I I1 111
amino ketone hydrochlorides V, was described.
The present communication reports an improved
procedure for the conversion of sec-alkyl amines
1 to a-amino ketone hydrochlorides via I1 and
offerssome measure of the scope of the reaction.
(1) Paper IV, THISJOURNAL, 81, 2132 (1959). OMe
(2) This work was supported in part by grant G-3689 of the Na- IV v
tional Science Foundation.
(3) H. E. Baumgarten and F. A. Bower, THIS JOURNAL,
76, 4561 In the original work the amine was converted
(1954). into the N,N - dichloroamine I1 using aqueous

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