Determination of composition of alginates by infrared

spectroscopic method
a
M. P. F I L I P P O V and bR. KOHN

^-Institute of Chemistry, Moldavian Academy of Sciences,

277 028 Kishinev, USSE

^Institute of Chemistry, Slovak Academy of Sciences,

809 33 Bratislava

Received 21 February 1974

An infrared spectroscopic method for the determination of the composi

tion of alginates has been elaborated. This method is based upon the de
termination of the ratio of absorbances of bands at 1320 and 1290 c m - 1
(KBr technique, 1 2 mg of potassium alginate, analysis error 3 % ) ,
and 1125 and 1030 c m - 1 (0.1 0.2 mg of potassium alginate, analysis error
8%).
The properties of alginic acids as well as their selectivity of exchange reactions of
metal cations depend on the ratio of D-mannuronic (M) and L-guluronic (G) acid units
in the molecule of alginate [1]. To estimate the ratio M/G, a sufficiently precise, but quite
laborious method has been elaborated [2]. Infrared spectroscopy has been employed
for a rapid qualitative information on the composition of alginates [3]. The estimation
of the ratio of absorbances at 787 and 808 c m - 1 allowed to determine the M/G fraction
within a 10% error [4].
The aim of our investigation was to determine the M/G ratio on the basis of infrared
spectra applying the KBr technique, because films of alginates cannot be prepared in
all the circumstances, and especially with low molecular fragments.
Examined were the infrared spectra of alginates with a high content of D-mannuronic
acid (94% M, 6% G; Mw > 500 000) and L-guluronic acid ( 9 1 % G, 9% M; Mn = 8300,

600 800 1000 1200 %00 1600 1800 v [cm'1]

Fig. 1. Spectra of films of polymannuronic acid (H) and its salts (Cu, Ca, ) .

Chem. zvesti 28 (6) 817-819 (1974) g j 7

 M. P. FILIPPOV, R. KOHX

1 1 1 i 1 1 1 1 -J

| y G

\^^MG

i
n m
$10 080 9*5 1030 1090 1125 1290 1320
i I I I I I I i
N^ M
i i

600 BOO 1000 1200 1 1600 1800 v [cm-1]

Fig. 2. Spectra of potassium alginates of various composition (KBr technique).

G - 9 1 % G, 9% M; MG - 4 8 % G, 52% M; M - 6% G, 94% M.

DP = 47). We did not succeed in preparing convenient films from the polyguluronie
acid. The effect of hydrogen atom exchange in carboxyl group for a metal cation upon
infrared spectra was therefore investigated with polymannuronic acid only. Spectra
of the same film of polymannuronic acid and its Cu, Ca, and salts were measured
without support [5], enabling thus to record also slight alterations in wavenumbers and
intensities of bands involved (Fig. 1). The spectrum of polymannuronic acid markedly
differs from those of its salts; spectra of its salts differ less from each other. Since tablet-
ting of polyuronic acids with KBr results in the exchange of counter-ions [6], a part of
alginate will be present in form. Alginates should be therefore analyzed as salts.
The spectra of potassium polymannuronate and polyguluronate markedly differ in
the 750 to 1400 c m - 1 region. We selected 9 various wavenumbers in the spectra (Fig. 2)
at which the M/G ratio can be in principle determined from absorbance readings. Mixtures
of various M and G content were prepared from the starting potassium alginates and the

A,/A2 i i I l -
1.5

/ '

1.0 yS
JX ^ ^
0.5

\ ^ ^ ^ Fig. 3. Dependence of the Ai/Ao absor

bance ratio on the composition of
0.0 I . I 1 1 alginates.
0 20 40 60 80 % G 100 1- -4l320/-4l290; 2. ^4ll25/-4l030.
00% M 60 40 20 0
Model mixtures; reference alginates.

Chem. zvesti 28 (6) 8 1 7 - 8 1 9 (1974)

818
COMPOSITION OF ALGINATES

r e l a t i o n s h i p b e t w e e n a b s o r b a n c e s A1JA2 of t h e following b a n d s a n d t h e c o m p o s i t i o n
of a l g i n a t e s w a s p u t i n t o a g r a p h : 780/^810, ^880/^945, 945/1030, ^4IOPO/^4IO3O, ^.1125/^41030,
a n d ^4i32o/^4i290- Of t h o s e listed, t h e b e s t r e s u l t w a s found w i t h -4i32oA4i29o, h a v i n g t h e
m e a n q u a d r a t i c e r r o r 3 % (Fig. 3). Similar result w a s o b t a i n e d also w i t h t h e ^1125/
r a t i o ; t h e e r r o r of analysis increased in t h i s case t o 8 % d u e t o a less s t e e p l y
descending c a l i b r a t i o n c u r v e . E v e n t h o u g h t h i s p a i r of b a n d s is s u i t a b l e for s p e c t r a l
analysis because t h e h i g h a b s o r p t i v i t i e s of b a n d s e n a b l e t o t a k e s p e c t r a w i t h o n l y 0.1
0.2 m g of a l g i n a t e i n t h e K B r pellet w i t h o u t lowering t h e d e t e r m i n a t i o n a c c u r a c y .
Verification of t h i s m e t h o d b y t w o s a m p l e s of a l g i n a t e s of k n o w n M a n d G c o n t e n t
[8] showed a sufficient a c c u r a c y for d e t e r m i n a t i o n of t h e c o m p o s i t i o n of alginic a c i d s
(Table 1).
Table 1

S p e c t r a l analysis of reference a l g i n a t e s

Composition Determined on the basis of

Alginate
(Source) Chemical analysis ^132/^12 ^1125/^
%G %M %G %M %G %M

Laminaria 72.5 27.5 71 29 69 31

hi/perborea;
stipes
Laminaria 38.5 61.5 41 59 38 62
digitata

Experimental

T h e a q u e o u s s o l u t i o n c o n t a i n i n g p o t a s s i u m a l g i n a t e a n d K B r in a 1 : 200 r a t i o w a s
freeze-dried [7]; pellets 12 m m i n d i a m e t e r were m a d e of 150 m g of t h i s m i x t u r e .
T h e p r e p a r a t i o n of a p o l y m a n n u r o n i c acid film w a s described earlier [5]. T h e s p e c t r a
were t a k e n w i t h a 457 P e r k i n E l m e r s p e c t r o p h o t o m e t e r i n t h e 600 t o 1900 c m - 1
r a n g e . Absorbances of b a n d s were d e t e r m i n e d i n t h e s p e c t r a b y t h e m e t h o d of base-line
d r a w n t h r o u g h t h e a b s o r p t i o n m i n i m a a t 850 a n d 1500 c m - 1 .

Acknowledgements. The authors are indebted to Drs A. Haug and 0. Smidsrod (Instituttfor
marin biokjemi, Trondheim, Norway) for the samples of alginates and their chemical analysis.

References
1. H a u g , A. a n d S m i d s r o d , O., Nature 215, 757 (1967).
2. H a u g , A. a n d L a r s e n , ., Acta Ghem. Scand. 16, 1908 (1962).
3. Szejtli, J . , Nahrung 10, 291 (1966).
4. Mackie, W., Carbohyd. Res. 20, 413 (1971).
5. F i l i p p o v , M. P . , Izv. Akad. Nauk Moldau. SSR, Ser. Biol, i Khim. Nauk 1970,
N o . 4, 59.
6. F i l i p p o v , M. P . a n d B u z h o r , A. V., Izv. Vyssh. Ucheb. Zaved., Pishch. Tekhnol. 1972,
N o . 4, 6 1 .
7. Mehlitz, A . a n d M i n a s , ., Ind. Obst-Gemseverwertung 50, 719 (1965).
8. K o h n , R., F u r d a , I., H a u g , A., a n d S m i d s r o d , O., Acta Ghem. Scand. 22, 3098 (1968).
Translated by Z. Votick

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