Aust. J . Chem.

, 1980,33, 1041-7

Mechanisms of Alkaline Degradation

of Sucrose. Relative Rates of Alkaline
Degradation of Some Sucrose Derivatives

Merilyn Manley-Harris, Wayne Moody and GeofSvey N. Richards

Department of Chemistry and Biochemistry,
James Cook University of North Queensland,
Townsville, Qld. 481 1.

Abstract
An earlier study of the relative rates of degradation of several sucrose derivatives has been refined
and extended. The results support a novel hypothesis to explain the unusual alkali-lability of
sucrose (1). In this hypothesis the first and rate-determining step in the degradation is an SNlcB
displacement effected at the C 1 anomeric centre of the glucose moiety by an oxyanion derived
from the C l ' hydroxyl (lo), to produce 1-(a-D-glucopyranosyl)fructose (11). The latter is then
very rapidly degraded, mainly to lactic acid. The possibility of analogous competing displacements
involving the 3' and 6' oxyanions remains for further investigation.

Introduction
In an earlier paper1 the relative rates of degradation of sucrose and of several
derivative model compounds were determined in 1 N sodium hydroxide at 100°C.
Under these conditions, melezitose and all of the 0-methyl ethers of sucrose that
were studied showed rates of degradation which were much lower than that of
sucrose and these rates were too small to be differentiated. The model compounds
methyl a-D-glucopyranoside and methyl P-D-fructofuranoside also showed negligible
rates of degradation, although raffinose showed a rate of the same order as
sucrose. From these results it was concluded that the mechanism of alkaline
degradation of' sucrose 'probably involves free hydroxyl groups (presumably via
the derived anions) on both glucose and fructose components of sucrose'.
The possibility of the involvement of the C 6 hydroxyl was eliminated on the
basis of the lability of raffinose, It was also concluded that other hydroxyls such
as 1' and 3' were 'unlikely to be involved in a simple intramolecular displacement
at the glycosidic centre of sucrose because of the behaviour of the methyl
fructofuranosides in alkali'. These earlier studies have now been repeated and
extended by using higher concentrations of alkali, pseudo-first-order conditions
(excess hydroxyl ion), a more precise method of analysis for sucrose disappearance,
and some additional sucrose derivatives.

Results and Discussion

For analysis of sucrose in the alkaline degradation solution several methods
were previously used,' always preceded by ion exchange deionization because of the
large ratio of inorganics to sucrose. After deionization the sucrose was analysed by
' O'Donnell, G. W., and Richards, G. N., Aust. J. Chem., 1973, 26, 2041.
 M. Manley-Harris, W. Moody and G. N. Richards

quantitative paper chromatography or by gas chromatography as the trimethylsilyl

ether. We have now devised a method of gas chromatographic analysis which does
not require preliminary deionization and hence avoids inaccuracies associated
with this step. The gas chromatographic method is regarded as accurate in the sense
that it measures actual sucrose, but is rather low in precision and very tedious in
operation. The measurement of sucrose by optical rotation is more precise, provided
that no other optically active compounds are present and to use this method to
measure the degradation of sucrose in alkaline solution we needed to make the
assumption that the products of degradation were not optically active. It is difficult
to validate this assumption by measurement of optical rotation up to completion
of the degradation because of development of colour, but we have compared the
rate values for alkaline degradation of sucrose given by gas chromatography and by
optical rotation. These were k,,, (567 k 12) x and (503C! 6) x lo-' s-' respec-
tively for 4 % sucrose solutions in 3.76 N sodium hydroxide at 100°C. These
values show that the rates given by optical rotation are slightly but significantly
lower than those given by gas chromatography; this suggests that the solution
after completion of the alkaline degradation may have a small positive rotation.
The optical rotation method therefore may introduce a small absolute error in
determination of these rate constants, but has nevertheless been used in all of our
subsequent rate measurements shown in Table 1 and Fig. 1 because of its convenience
and relative precision. In reaching mechanistic conclusions based on these results
we have relied only on major differences in rates. Both of the methods gave a
linear plot of the logarithm of sucrose remaining against time and hence confirm
the pseudo-first-order nature of the degradation in excess alkali.
Table 1 . Reaction rates o f sucrose and derivatives at 100°C
k,,,, observed pseudo-first-order rate constant

Compound Conditions l0'kObs (s-

Sucrose 1 % soln in 5.68 N NaOH

Raffinose
Melezitose
6,6'-Di-0-methylsucrose
4,1',6'-Tri-0-methylsucrose
2,3,4,3',4'-Penta-0-methylsucrose
4,6-Isopropylidenesucrose
2,1f:4,6-Di-0-isopropylidenesucrose 2 % soln in 1.90 N NaOH
Methyl a-D-glucopyranoside
Methyl B-D-glucopyranoside
6 % soln in 5.68 N NaOH
2 % s o h in 1.90 N NaOH
6 % soln in 5.68 N NaOH
6 % soln in 5.68 N NaOH
2 % soln in 5.68 N NaOH
2 % s o h in 5.68 N NaOH
2 % soln in 5.68 N NaOH
2 % s o h in 1.90 N NaOH
6 % s o h in 5.68 N NaOH
6 O/d s o h in 5.68 N NaOH

The rates of degradation of raffinose (2) (6-0-(a-D-galactopyranosyl)sucrose)

and 4,6-0-isopropylidenesucrose (7), are both similar to those of sucrose under
similar conditions (Table 1). This confirms that the ionization of neither the C 6
hydroxyl (as was earlier indicated') nor the C 4 hydroxyl, is necessary for alkaline
degradation. It is assumed, in connection with the mechanism detailed below, that
these reductions in rates, compared with sucrose, are associated with some slight
steric hindrances introduced by the substituents at the C 4 and/or the C 6 positions.
Alkaline Degradation of Sucrose

Interestingly, the 2,11:4,6-di-0-isopropylidenesucrose(8) was not significantly

degraded after several days in 6 N sodium hydroxide at 100°C, the 2,l'-0-isopropyl-
idene bridge having, therefore, stabilized the sucrose to alkali."

Fig. 1. Degradation of sucrose derivatives

in 5.68 N sodium hydroxide at 100°C.
I, Sucrose;
2, raffinose;
3, 6,6-di-0-methylsucrose;
4, melezitose;
m
5, 2,3,4,3',4'-pentamethylsucrose;
M 6, 4,1',6-tri-0-methylsucrose;
7, methyl a-D-glucopyranoside.

1
50 100 150

Time ( h )

6,6'-Di-0-methylsucrose (4) degraded at about one-quarter the rate of sucrose.

Examination of models indicates that the 0-methyl group at C6' introduces very
little steric hindrance for the transition state required in the mechanism postulated

* This observation incidentally makes possible an improvement in workup for preparation of the
di-0-isopropylidenesucrose. In the procedure for synthesis of the latter compoundZ both the mono-
and di-0-isopropylidene derivatives are obtained and have to be separated by column chromato-
graphy. Our results now show that the mono-derivative could be completely removed by treatment
with 6 N sodium hydroxide at 100°C overnight. The di-0-isopropylidene sucrose would survive
this treatment and would then be recovered in relatively pure form after deionization.
Khan, R., and Mufti, K. S., Cavbohydr. Res., 1975, 43, 247.
 M. Manley-Harris, W. Moody and G. N. Richards

below (Scheme 1). It is possible therefore that the increased stability of the
6,6'-di-0-methylsucrose may indicate some relatively small involvement of the
C6' hydroxyl in alkaline degradation of sucrose by a mechanism analogous to and
competitive with that shown in Scheme 1 (see below). However, these effects are
much smaller than those discussed below relating to the hypothesis shown in
Scheme 1.

t
/
HO-C-H
1
H-C-OH
- LHOH
I
etc.

CH3
I
H-C-OH (12)
CH20H

OH OH (11)
Scheme 1

4,1',6'-Tri-0-methylsucrose (5) was the most alkali-stable of the partially

methylated sucroses studied, its rate of degradation being two orders of magnitude
smaller than sucrose. There are two possible explanations for this stability, viz.
steric hindrance by the methyl groups or the prevention of ionization of one or
more of the three hydroxyl groups which have been methylated. The presence of
the 4 and 6' methoxyl groups introduces little steric hindrance (see above) and we
have already eliminated the 4, 6 and 6' hydroxyls from any other type of major
role in the alkaline degradation. The conclusion then must be that the alkaline
degradation of sucrose requires a free hydroxyl at C l', in its rate determining step.
On this basis it is now possible for the first time to postulate a hypothesis which
will at least partly explain the unusual lability of sucrose to alkali. This mechanism is
shown in Scheme 1 and is classified as SNlcB, the transition state implied in the
first step involving formation of a new and relatively unstrained five-membered ring.
Similar SNlcB displacements have been postulated to explain the alkaline degra-
dation of some other disa~charides,~ but always involving a displacement which
generates monosaccharide derivatives as initial products. The present hypothesis
for sucrose however, appears to be the first instance where an SNlcB reaction in
a disaccharide yields another disaccharide. The 34-fold difference in rate between
6,6'-di-0-methyl- and 4,lf,6'-tri-0-methyl-sucroses is the most convincing evidence
for this hypothesis and this difference must be almost entirely associated with the
methylation of the C1' hydroxyl group. The product (11) is 1-0-(P-D-gluco-
pyranosy1)-D-fructose. This is an analogue of I-0-methyl-D-fructose whose alkaline
degradation has previously been studied in some detail4 and the degradation of (1 1)
would be expected to proceed very rapidly mainly to lactic acid as shown in Scheme 2.
Brandon, R. E., Schroeder, L. R., and Johnson, D. C . , 'Cellulose Technol. Res.' Am. Chem. Soc.
Symp. Ser., 1975, 10, 125.
Kenner, J., and Richards, G. N., J. Chem. Soc., 1954, 1784.
Alkaline Degradation of Sucrose

The hypothesis shown in Scheme I as a major route for alkaline degradation of

sucrose, suggests that any derivative in which C 1' hydroxyl is unsubstituted and
which is free to adopt the transition state should degrade in alkali, but the results in
Table 1 show that 2,3,4,3',4'-penta-0-methylsucrose(6) is only slightly more labile
than the 4,l1,6'-tri-0-methyl ether. The model of the former derivative however shows
extensive hindrance to the adoption of the necessary transition state, especially between
the 0-methyl groups at positions 2 and 3'. It is concluded that this hindrance
slows the reaction shown in Scheme 1, probably by about an order of magnitude.
CH20G CH2 -0G
CH,OG I I
I C=O C -0-
C=O I ll
I CH20H CH-OH
HO-C-H
I
H-C-OH
+
CHO

+ GO-
COOH I
I

Scheme 2
The further verification of the hypothesis will require study of the relative rates
of alkaline degradation of mono-0-methyl ethers of sucrose and this work is now
under way. Deoxy derivatives of sucrose would in many ways be better models for
this work, but in general their synthesis is more difficult than the 0-methyl ethers.
Meanwhile it should be noted that there are two other possible reactions which are
entirely analogous to that shown in Scheme 1 and which could be competitive with
the mechanism shown. These involve the displacement at the glucose anomeric
carbon by the anion of either the C3' or the C6' hydroxyl. Of these, the C3'
mechanism seems particularly feasible. This hydroxyl should be one of the three
most acidic hydroxyls in sucrose (with C 2 and C 1') because of the inductive effects
associated with adjacent acetal carbons. Furthermore, the C 3' oxyanion displacement
would involve the same type of five-membered ring transition state as the C 1'.
The rate of degradation of melezitose [3'-0-(a-D-glucopyranosy1)sucrose] (3) is in
fact of the same order as the relatively stable tri- and penta-0-methylsucroses.
To some extent this may represent inhibition of reaction through the C 3' oxyanion,
but in view of the very low reactivity of melezitose, there must also be inhibition
of the C1' displacement reaction by steric hindrance associated with the bulky
glucopyranosyl group.
The displacement at C 1 by the C 6' oxyanion would involve the formation of a
new six-membered ring in the transition state and this would probably be less
favoured than the five-membered rings associated with the C 1' and C 3' oxyanion
reactions. Accordingly, the conclusion has been reached above (from the
6,6'-di-0-methylsucrose rate) that the C 6' mechanism is of relatively small importance.
 M. Manley-Harris, W. Moody and G. N. Richards

In all of these cases, viz. the 1' displacement (which is strongly supported by our
present evidence) and thz 3' asd 6' displaezments, the product is a glucosyl fructose
derivative which is su3ject to very rapid further degradation, mainly to lactic acid.
The question of the 1att;s two possible displacement mechanisms should be resolved
by our current investigation of the relative alkali-stabilities of the mono-0-methyl-
sucroses.
Experimental
Materials
6,6'-Di-0-methylsucrose was prepared from 6,6'-di-0-tritylsucrose5 by benzylation with benzyl
chlorid- and sodium hydroxide and detritylation. The resultant 2,3,4,11,3',4'-hexa-0-benzylsucrose
was purified by dry column chromatography on silica ge! eluted with dichloromethane/ethyl acetate
mixtures. The amorphous product, which gave a single t.1.c. spot, was methylated by the method
of Hskamori6 to yield 6,6'-di-0-methyl-2,3,4,1',3',4'-hexa-O-benzylsucrose ['H n.m.r. 8 (CD,COCDJ
3.26, s, 3 8 , OCH,; 3.32, s, 3H, OCH,; 5.74, d, lH, J,,, 4 Hz, H 1; 7.1-7.5, m, 30H, arom.].
Debenzylation by Raney nickel, in 95 % ethanol, yielded 6,6'-di-0-methylsucrose as a colourless
syrup.'
4,1f,6'-Tri-0-methylsucrosewas prepared from 6,1',6'-tris-O-trity1-2,3,4,3',4'-penta-O-acetyl-
s u ~ r o s e . ~Boiling in aqueous acetic acid resulted in detritylation accompanied by 0 4 + 0 6
migration of an acetyl g r ~ u p to * ~yield
~ 2,3,6,3',4'-penta-0-acetylsucrose,10~"which was methylated
with diazomethane/boron trifluoride etherate to give 4,1',6'-tri-0-methyl-2,3,6,3',4'-penta-0-acetyl-
sucrose.'," Deacetylation by sodium in methanol yieldcd 4,1f,6'-tri-0-methylsucrose which after
purification by dry column chromatography had the same properties and 'H n.m.r. spectrum
previously reported.' The n.m.r. spectra of its perdeuteromethylated derivative were also
determined.'
2,3,4,3',4'-Penta-0-methylsucrose was prepared as previously described,13 as were 4,6-0-iso-
propylidene-2,3,1',3',4',6'-hexa-0-acetylsucrose and 2,1':4,6-di-0-isopropylidene-3,3',4',6'-tetra-0-
acetyls~crose.~
Sucrose, raffinose pentahydrate, melezitose d~hydrateand methyl a-D-glucopyranoside were
obtained commercially and used as received.

General Method for Measurement of Alkaline Degradation

The alkaline degradations were performed on solutions of the sucrose derivative in 5.68 N
NaOH. The solutions were contained in stainless steel reaction vessels sealed with tight-fitting
Teflon caps, and were suspended in an oil bath maintained at 100f 0 . l 0 C . In experiments where
air was displaced from the solution and tube by nitrogen, no significant effect on the rate of
degradation was detected. At intervals samples were taken, accurately weighed, and neutralized
to p H 6-7 with 60% aqueous acetic acid. They were then diluted to a known volume with
water and their optical rotations recorded at 589 nm on a Perkin-Elmer 141 polarimeter. The data
were then fitted by a linear least squares method to a plot of Inx,,, against time and kobscalculated
from the slope. Optical rotations determined for sucrose solutions containing sodium acetate in
various amounts showed that the effect of sodium acetate on optical rotation was negligible in the
experimental range used.
Otake, T., Bull. Chem. Soc. Jpn, 1972, 45, 2895.
Jansson, P. E., Kenne, L., Liedgren, H., Lindberg, B., and Lonngren, J., Chem. Commun., Unit..
Stockholm, 1976, 8.
Lindley, M. G., Birch, G. G., and Khan, R., Carbohydr.. Res., 1975, 43, 360.
Suami, T., Otake, T., Ogawa, S., Shoji, T., and Kato, N., Bull. Chem. Soc. Jpn, 1970, 43, 1219;
Lemieux, R. U., and Barrette, J. P., J. Am. Chem. Soc., 1958, 80, 2243.
Bredereck, H., Zinner, H., Wagner, A., Faber, G., Greiner, W., and Huber, W., Chem.
Ber., 1958, 91, 2824.
' O Buchanan, J. G., Cummerson, D. A., and Turner, D. M., Carbohydr. Res., 1972, 21, 283.
l' McKeown, G. G., Serenius, R. S . , and Hay\vard, L. D., Can. J. Chem., 1957, 35, 28.
Manley-Harris, M., and Richards, G. N., Carbohydr. Res., in press.
. I 3 O'Donnel!, G. W., and Richards, 6. N., Aust. J. Chem., 1972, 25, 407.
Alkaline Degradation of Sucrose

Analysis by Gas Chvomafography

In order to verify the assumptions made in assaying the sucrose derivatives by polarimetry
during alkaline degradation, a parallel run was done on sucrose by a gas chromatographic method.
A 10-pl aliquot of the sucrose-sodium hydroxide reaction solution was taken by syringe and
accurately weighed into a vial. A 20-p1 sample of a trehalose solution (2.385 % in dimethyl sulfoxide)
was then weighed into the vial and the mixture neutralized with 1 % aqueous acetic acid until the pH
was in the range 6-7. The sample was then dried at 44"C/0,1 mm over phosphoric oxide for
several hours. The vial was sealed with a Teflon-faced septum, 50 p1 of dry redistilled Me2S0
injected and the vial shaken vigorously to disperse the sugar-sodium acetate mixture. Silylating
reagent (200 pl, Trisil; Pierce Chemical Co.) was injected, and the mixture heated for c. 5 min on
a metal block at 100°C with occasional shaking.
Gas chromatography was carried out on a 1500 by 2 mm steel column, packed with 3 % SE30
on Gas Chrom W and operated isothermally at 230°C. Sucrose and trehalose had retention times
of 6 . 1 and 8.1 min respectively and the amount of sucrose present was calculated by reference to the
arealweight ratios obtained from standard sucrose-trehalose mixtures.

Acknowledgments
Experimental assistance was provided by Mr D. Halbert and Mr L. Poncini.
Financial support was provided by the Australian Research Grants Committee and
by the Sugar Research Institute, Mackay.

Manuscript received 17 January 1980