This document provides a review of methods for isolating water soluble and water insoluble polysaccharides. It discusses that water soluble polysaccharides can be isolated from plant materials using hot water extraction and ethanol precipitation. Water insoluble polysaccharides like starch can be isolated using aqueous extraction or chemical treatment, while cellulose and hemicellulose require solvent extraction using reagents like NaOH or ethanol. The document outlines specific extraction methods researchers have used to isolate polysaccharides from various plant sources and characterize their structures.

1. A PROJECT REPORT ON
REVIEW ON ISOLATION OF WATER SOLUBLE AND WATER INSOLUBLE POLYSACCHARIDES
BACHELOR OF PHARMACY
SUBMITTED BY
MD SALIK
(BPH/1026/2011)
UNDER THE GUIDANCE OF
Dr. K. JAYARAM KUMAR
(ASSOCIATE PROFESSOR)
DEPARTMENT OF PHARMACEUTICAL SCIENCES AND TECHNOLOGY
BIRLA INSTITUTE OF TECHNOLOGY
MESRA, RANCHI-835215
2015

2. CONTENTS
 CHAPTER 1: INTRODUCTION
 CHAPTER 2: ISOLATION OF WATER SOLUBLE NON-STARCH
POLYSACCHARIDES
 CHAPTER 3: ISOLATION OF WATER INSOLUBLE NON-STARCH
POLYSACCHARIDES
 CHAPTER 4: CONCLUSION
 REFERENCES

3. INTRODUCTION
 Polysaccharides are polymeric carbohydrates molecules composed of long chains of
monosaccharide units. They are isolated from terrestrial and marine plants or are
principally the exogenous metabolites of some bacteria.
 Polysaccharides are composed of many monosaccharide residues that are joined one
to the other by O-glycosidic linkages.
 When all the monosaccharide in a polysaccharide are the same type the
polysaccharide is called a homopolysaccharide or homoglycan, but when more than
one type of monosaccharide is present they are called heteropolysaccharide or
heteroglycan.

4. Classification of polysaccharides
Polysaccharides by source (Reddy, mohan et al .2011)
 Seaweed extracts: Agars, aliginates, carrageenans
 Higher plant cell wall insoluble: cellulose
 Higher plant cell wall soluble : pectin
 Higher plant seeds: cereal starch, guar gum, locust bean gum
 Higher plant tuber & roots: potato starch, tapioca starch

5. Polysaccharides by structure
 Linear: amylose, cellulose,pectin,alginates
 Short branched: guar gum, locust beangum, Xanthan gum
 Branched-on-branch: amylopectin, gum arabic, arabinoxylan
Polysaccharides by monomers
 Homoglycans: starch, cellulose
 Diheteroglycans: agars, alginate,carrageenans
 Triheteroglycans: Xanthan, gellan, arabinoglycan

6. Polysaccharides by charge
 Neutral: amylose,amylopectin, cellulose, guar gum
 Anionic: Alginates, carrageenans, gellan.
 Cationic: Chitosan
Types of Polysaccharides
Various examples of water soluble polysaccharides are Guar Gum, Dextran(Sluka Peter et
al,2004), Gum Karaya, Heparin etc

7. Gum Karaya
 It is the dried exudation of the Sterculia Urens Tree and other species of Sterculia
 It yields galactose, rhamnose, and galcturonic acid oh hydrolysis
 The major use of Gum Karaya is as a bulk laxative in view of its ability to form a
mucilaginous gel on contact with water
Guar Gum
 Guar gum is a galactomannan, which occurs as a storage polysaccharides
consisting of ( 1→4)- diequatorically linked β-D- mannose monomers,some of
which are linked to single sugar side chains of α-D-galactose attached (Doyle et
al ,2008).
 Guar gum is particularly useful for colon delivery because it can be degraded by
specific enzymes in this region of the gastrointestinal.

8. Structure of Guar gum
Dextran
 Dextran is a complex, branched glucan composed of chains of varying length
 It is used medicinally as an antiplatelet to reduce blood viscosity
 The straight chains consists of α-1,6 glycosidic linkages between glucose molecules,while
branching begin from α-1,3 linkages
 Dextan was first discovered by Louis Pasteur as a microbial product in wine

9. Structure of Dextran.
Water insoluble polysaccharides
Starch
 Starch is a storage carbohydrate consisting of glucose monomers in plants such as cereals, root
vegetables and legumes
 It is comprised of two polymers, namely amylose and amylopectin.
 Most common cereal starches contain 15-30% amylose

10. Pectin
 Pectin is a structural polysaccharide heteropolysaccharide contained in the primary cell walls of
terrestrial plants.
 It was first isolated and described in 1825 by Henri Bracannot
 It is used in food as a gelling agent,particularly in jams and jellies
Cellulose
 Cellulose is a polysaccharide consisting of a linear chain of several hundred to many
thousands of β(1→4) linked D-glucoseunits.
 Cellulose is the most abundant organic polymer on Earth
 Cellulose is mainly used to produce paperboard and paper.
 Cellulose for industrial use is mainly obtained from wood pulp and cotton.

11. ISOLATION OF WATER SOLUBLE POLYSACCHARIDES
 Water soluble polysaccharides are isolated from various parts of the plants mainly fruits, roots,
pulp , seeds and leaf using a hot water extraction regime followed by ethanolic precipitation
 Generally for isolation of water soluble polysaccharides raw materials are coarsely grounded
and boiled/washed with hot water for different time periods
 Generally cold centrifugation is done at 4 ᵒC- 8 ᵒC at 8000 rpm for 30 min.
 After centrifugation the supernatant is stored at 4 ᵒC in a freezer for at least 12 hours
 Supernatant is precipitated using alcohol/acetone
 Precipitated polysaccharides are removed by centrifugation.
 For further purification the crude polysaccharide is subjected to dialysis, followed by Gel
Permeation Chromatography/ Size exclusion chromatography using different chromatographic
column.

13. Extraction of water soluble non starch polysaccharides from plants
Name of
author
Name of Plant Plant part used Method of extraction Reagents used
Tao et al
(2012)
Polygala tenifolia Roots The dried P tenifolia was refluxed in a
soxhlet apparatus with toluene-ethanol for
6 hours. Then it was filtered,concentrated
and cooled down. Then 95% ethanol was
added slowly until the concentration is
50% and kept for 1 hour. Then calcium
chloride was added and kept overnight to
precipitate tannin. The supernatant was
obtained by centrifugation and
ultrafiltration was done.
Toluene-ethanol
95% ethanol
CaCl2

14. Name of
author
Name of Plant Plant part
used
Method of extraction Reagents used
Huang et al
(2012)
Cassia obtusifolia Seeds C.obtusifolia was purchased from the
market and grounded. Approx 5 gm of
finely ground powder samples were stirred
in distilled water and followed by boiling
it. After filtration water soluble
polysaccharide was precipitated with 95%
ethanol and collected by centrifugation.
Then the precipitates was dissolved in
water and dialysed to remove the small
molecules. The dialysed solution was
freeze dried to yield polysaccharide
powder
95% ethanol
0.05 N NaOH

15. Name of
author
Name of Plant Plant part
used
Method of extraction Reagents used
Xie et al
(2015)
Ampelopsis
megalophylla
Leaves Dried leaves of A.megalophylla were
chopped into small pieces and mixed with
95% ethanol and then extracted with
distilled water. The aqueous extract was
filtered and polysaccharide were
precipitated by 95% ethanol , then
keeping it overnight and the precipitated
material were collected by
centrifugation. The extracts were then
deproteinised . Finally the precipitate was
washed with absolute ethanol, acetone
and ether and crude polysaccharide of
A.megalophylla was obtained
95% ethanol
Acetone
Ether

16. ISOLATION OF WATER INSOLUBLE POLYSACCHARIDES
 Water insoluble polysaccharides are divided into two groups e.g. Starch and Non-starch
polysaccharides
 Starch is a storage plant polysaccharide. It is isolated using aqueous extraction method or
through chemical treatment
Schematic representation of isolation
of starch

17.  Non-starch water insoluble polysaccharides are cellulose, linin, hemicellulose etc. These
water insoluble polysaccharides are soluble in organic solvents e.g. methanol, ethanol
Schematic representation of isolation of water insoluble non-starch polysaccharides

18. Extraction process of Water Insoluble Polysaccharides
Name of
author
Name of Plant Plant part used Method of extraction Reagents used
Peng et al.
(2004)
Ganoderma
tsugae
Fruits
Mycelia of G .tsugae were obtained through
submerged cultivation and extraction. After
being defatted 100g mycelia were extracted
with 0.2 M phosphate buffer and the residue
was extracted with 2% NaOH. The
supernatants were neutralized with acetic acid
Both the precipitates after neutralization
were re-purified and finally after vacuum
dried for 7 days the white powder coded as
GM5-1 for the water insoluble polysaccharide
were extracted
0.2 M Phosphate
buffer
2 % NaOH

19. Name of
author
Name of Plant Plant part
used
Method of extraction Reagents used
Wang et al.
(2012)
Panax japonicus Rhizome
P. japonicus rhizomes were refluxed with
85% ethanol in a water bath. After
incubation, mixtures were centrifuged.
The insoluble residue was dried in an
oven and extracted. After elute was
concentrated in vacuum four times the
volume of ethanol was added to the
water and kept overnight in a
refrigerator to precipitate polysaccharide.
The precipitate was separated by
centrifugation and air dried
0.1 M NaOH
85% ethanol

20. Name of
author
Name of Plant Plant part
used
Method of extraction Reagents used
Ding et al.
(2011)
Solanum nigrum L Seeds
Coarsely powdered of dried SNL was pre
extracted twice with 95% ethanol and
then refluxed twice with 80%
ethanol.The degreased powder were
dried.The aqueous extract was filtered and
the residue was extracted with 0.5 M
NaOH solution.The extraction was
condensed and precipitated with 95%
EtOH to obtain the crude polysaccharide
0.5 M NaOH
95 % EtOH

21. CONCLUSIONS
Conclusion of this review can be divided into two parts
 Isolation of water soluble polysaccharides
 Isolation of water insoluble polysaccharides
Isolation of water soluble polysaccharides
In general isolation of water soluble polysaccharides is done through precipitation method. This method has several
steps. Among these steps boiling time, pre-precipitation centrifugation, polysaccharide precipitation and post-
precipitation centrifugation are needed to be optimized for each sample. Petroleum ether, EtOH, n-BuOH are used for
the isolation of water soluble polysaccharides.Among other methods alcohol precipitation method is most effective
because of its recycle flexibility.

22. Isolation of water insoluble polysaccharides
 Isolation of water insoluble polysaccharides is done through solvent extraction method. This
method is needed to needed to be optimized for each sample. Starch is isolated either by simple
aqueous extraction method or chemical treatment method. NaOH and 0.5 M KOH are widely
used for the isolation of water insoluble polysaccharides.

23. REFERENCES
 Xin, T., Zhang, F., Jiang, Q, Chen, C, Huang, D., Li, Y., Shen, W. & Jin, Y., (2012)
Extraction, purification and antitumor activity of a water-soluble polysaccharide
from the roots of Polygala tenuifolia . Carbohydrate Polymers, 90,1127– 1131
 Huang, Y., Chaw, C., Hsiang, T. Y. (2012). Composition, characteristics, and in-
vitro physiological effects of the water-soluble polysaccharides from Cassia seed.
Food Chemistry, 134, 1967-1972.
 Peng, Y., Zhang, L., Zhang, Y., Xu, X., Kennedy, J.F (2004) .Solution properties of
water-insoluble polysaccharides from the mycelium of Ganoderma tsugae
.Carbohydrate Polymers 59 (2005) 351–356
 Wang, R., Chen, P.,Jia, F., Tang, J,. Ma, F., Xu, B (2012) . Characterization and
antioxidant activities of polysaccharides from Panax japonicus C.A. Meyer
.Carbohydrate Polymers 88 (2012) 1402– 1406

24.  Ding,X., Zhu, F,. Gao, S (2011) .Purification, antitumour and immunomodulatory activity of water-extractable and
alkali-extractable polysaccharides from Solanum nigrum L. Food Chemistry 131 (2012) 677–684
 Dong, Q., Yao, J., Fang, Ji (2003) . Structural characterization of the water-extractable polysaccharides from
Sophora subprostrata roots . Carbohydrate Polymers 54 (2003) 13–19
 Xin,T., Zhang, F., Jiang, Q., Chen, C., Huang, D., Li, Y., Shen, W., Jin.,Y (2012) . Extraction, purification and
antitumor activity of a water-soluble polysaccharide from the roots of Polygala tenuifolia . Carbohydrate
Polymers 90 (2012) 1127– 1131
 Palacios ,I., Garcia- Lafuente, A., Guillamon, E., Villares, A (2012) .Novel isolation of water-soluble
polysaccharides from the fruiting bodies of Pleurotus ostreatus mushroom . Carbohydrate Research 358 (2012)
72-77
 Lin, L., Zhuang, M., Zou, L., Lei, F., Yang, B., Zhao, M (2012). Structural characteristics of water-soluble
polysaccharides from Rabdosia serra (MAXIM.) HARA leaf and stem and their antioxidant capacities . Food
Chemistry 135 (2012) 730–737
 Chen, G., Ma, X., Liu,, S., Liao,Y., (2012) .Isolation, purification and antioxidant activities of polysaccharides
from Grifola frondosa . Carbohydrate Polymers 89 (2012) 61– 66
 Liu, C., Jiang, H., Zhou, H., Li, P., Wang, F (2015). Isolation, structural characterization and neurotrophic activity
of a polysaccharide from Phellinus ribis .Carbohydrate Polymers 127 (2015) 145–151