Strusovskaya O.G., Artemyeva A.P.
The Northern State Medical University, Arkhangelsk, Russia
Determination of the polysaccharides
in some wild medical plants growing on the islands of the Solovetsky
archipelago
Vegetable polysaccharides (VPS) have a broad spectrum of
pharmacological activity. It is known that VPS have immunomodulatory and anti-atherosclerotic
action [1], inhibit the activity of the cough [2] have a positive effect on the
deterioration of memory, slow down the aging process [3].
The aim of this work was to determine the
total content of polysaccharides in certain types of wild medicinal plants
(WMP) of the islands of the Solovetsky archipelago. Air-dried raw
material of Linnea borealis
(LinnaeaceaeL.), Empetrum hermaphroditum (Empetraceae), Cochlearia officinalis
(Crucifereae L.) and Laminaria
saccharina (Laminariaceae L.), harvested on the islands of Solovetsky
archipelago in the summer of 2011, have been used as objects of research.
Determination
of the total content of polysaccharides in the samples of WMP was performed
using anthrone reagent [4]. For this
purpose, air-dry raw material was crashed and sieved through a laboratory sieve
with an aperture size of 1 mm. Approximately 1.0 g (exact mass) of raw
material (each plant separately) was placed in flat-bottomed conical flask with
a capacity of 100 ml stoppered flask, then 70 ml of 1% solution of sodium
chloride in purified water was added, the flask contents were heated in a
boiling water bath for 1 hour. After cooling, the extract was filtered in
a volumetric flask with a capacity of 200 ml. Extraction was repeated twice
and the combined volume of the filtrate was adjusted to the mark with purified
water (solution A). 2 ml of solution A (for Laminaria
saccharina - 1 ml of solution A) was transferred to a centrifuge tube, 6 ml
of ethyl alcohol 96% (alcohol) was added, stirred and heated in a boiling water
bath for 10 min. After cooling, the contents of the tubes were centrifuged
for 10 min with a speed of 3000 rev/min. The supernatant was decanted and the sediment was slurried with 5 ml of
alcohol and again centrifuged under the same conditions.The supernatant was
decanted, the sediment in the test tube was flushed with hot air to remove
traces of alcohol. 4 ml of anthrone reagent was added to sediment and
heated in a boiling water bath for 10 min. After cooling, the contents of
the tube was transferred with alcohol to a flask with a capacity of 25 ml, the
volume of solution in the flask was adjusted to the mark with the same solvent
(solution B). The optical density of solution B was measured by a
spectrophotometer at a wavelength of 430 nm ± 2 nm. The analysis was carried out in quartz cells with a
layer thickness of 10 mm relative to the reference solution. To prepare
the reference solution 4 ml of anthrone reagent was heated in a flask from
heat-resistant glass in a boiling water bath for 10 min, after cooling the
contents of the flask was transferred with alcohol into a volumetric flask with
a capacity of 25 ml. The volume of solution in the flask was adjusted to
the mark with alcohol. the UV spectra of extracts from the investigated WMP,
obtained in these conditions, shown in Fig. A.
The
content of polysaccharides (X%) in terms of hydrous glucose and absolutely dry
raw material was calculated by the formula:
, where
c - the level of glucose in solution B on the calibration curve, mg/ml;
F - the dilution factor;
0.91 – the coefficient of hydrolysis;
d - a conversion factor of anhydrous glucose in water;
b - a loss in weight during drying WMP,%.
To
prepare anthrone reagent, 100 mg of anthrone (MRTU 6-09-1214-64) previously
recrystallized from benzene, was placed in a flask with a capacity of 200 ml and
adjusted to the mark with concentrated sulfuric acid. Shelf life of the reagent is 5 hours.
To
construct a calibration curve, about 0.1 g of glucose (exact mass) was placed
in a flask with a capacity of 100 ml, was dissolved in purified water and was
brought to the mark with the same solvent (solution A).
0.2, 0.3, 0.4, 0.5, 0.6 ml of solution A was brought in test tubes and was
adjusted to the mark of 0.6 ml with purified water. 4 ml of anthrone reagent was brought in each tube and
placed in a boiling water bath for 10 min. After cooling, the contents of
the tube was quantitatively transferred with alcohol to volumetric flasks with
a capacity of 25 ml, the volume of solution in the flask was adjusted to the
mark with the same solvent (solution B).
The optical
density of the obtained solutions were determined at a wavelength of 430 nm ± 2
nm in quartz cells with a layer thickness of 10 mm. A mixture of 4 ml of anthrone reagent and 0.6 ml of
purified water was used as a reference solution, designed in the same
conditions as the investigated solutions. According to reports received
during the experiment was built a calibration graph plot optical density
against concentration of glucose (Fig. 2).
The
results of determination of the amount of polysaccharides in the samples of WMP
and their metrological characteristics are presented in Table. A.
Thus, the research determined the total content of polysaccharides in the
samples of WMP growing on the islands of Solovetsky archipelago.
It was
found that total content of polysaccharides in air-dried raw Laminaria saccharina is more than
12%. Least of all polysaccharides was defined in the Linnea borealis samples.
References:
1.
Gunter, EA. Cell
cultures of non-traditional plants as producers of polysaccharides / EA Gunter
/ / Chem. and Comput. Simul. Butlerov Com. - 2001.-N5. -
P.11-12
2.
Sutovska M. The antitussive activity of polysaccharides from Althea
officinalis L., var Robusta, Arctium lappa L., var. Herkules, and Prunus
persica L., Batsch / M.Sutovska, G. Nasalova, S. Franova,
A. Kardosova / / Bratisl Lek Listy. - 2007.-N108.-Vol.2. - P.93-99.
3.
Yang Li. Pharmacological
action of Adenophora polysaccharides / L. Yang, L. Chun-hong,
T. Fu-tian, L. Xin-fang / / Chin. J. of Integr. Med. -
Vol. 10. - N. A. - P. 78-80.
4.
Olennikov. DN The method of quantitative determination of the
total content of polysaccharides in the seeds of flax (Linum usitatissimum L.)
/ DN Olennikov, LMTankhaeva / / J. Chemistry of plant material. -
2007.-¹4. -C. 85-90.
Table 1
The results of the quantitative determination of total content of
polysaccharides in the samples of WMP
|
Samples of WMP |
Total content of the polysaccharides, %, ( |
|
|
ε,% |
|
Linnea borealis |
0,84 |
0,022 |
0,056 |
±6,6 |
|
Empetrum hermaphroditum |
1,03 |
0,027 |
0,070 |
±6,84 |
|
Cochlearia officinalis |
2,09 |
0,032 |
0,083 |
±4,03 |
|
Laminaria saccharina |
12,23 |
0,417 |
0,497 |
±5,31 |
)
, %
Fig.1
UV spectra of exrtacts from
WMP
1- Linnea borealis; 2- Empetrum hermaphroditum;
3- Cochlearia officinalis; 4-Laminaria saccharina
Fig. 2
Calibration curve
