EXERCISE 6: ISOLATION OF POLYSACCHARIDE

Myrabelle Lopez
Pro-teens
Date Performed: Date Submitted:

INTRODUCTION

Aside from fat and protein, carbohydrates is one of the macronutrients in our diet which represent the
most important source of energy for the body, and are vital for a varied and balanced diet. It is
significant in its role in the structure and function of cells, tissues and organs, as well in the formation of
carbohydrate structures on the surface of cells ( EUFIC,2019) likewise in the transport of energy and as
a structural material for vast molecules (Stubbings,2008). Generally, carbohydrates are carbon
compounds containing large quantities of hydroxyl groups with the basic general formula Cn(H2O)n .
They are classified into several groups, namely monosaccharides, disaccharides, and polysaccharides,
depending on the number of their monosaccharide units.

The monosaccharides are white, crystalline solids that contain a single aldehyde or ketone functional
group. They are subdivided into two classes -- aldoses and ketoses --on the basis of whether they are
aldehydes or ketones (ChemED, n.d.)They are also classified as tetrose, pentose or hexose. The common
monosaccharides are glucose, fructose, and galactose (Stubbings,2008).

Disaccharides, meaning "two sugars", are commonly found in nature as sucrose, lactose and maltose.
They are formed by a condensation reaction where one molecule of water condenses or is released
during the joining of two monosaccharides. The type of bond that is formed between the two sugars is
called a glycosidic bond and examples are sucrose, lactose and maltose (Barnes and Noble, n.d.).

A long chain of monosaccharides linked by glycosidic bonds is known as a polysaccharide which may be
branched or unbranched, and it may contain different types of monosaccharides. The molecular weight
may be 100,000 daltons or more depending on the number of monomers joined. Starch, glycogen,
cellulose, and chitin are primary examples of polysaccharides (Learning, n.d.).

The major glucose storage polymer in animals is glycogen which is highly branched that permits rapid
release of glucose from glycogen stores. It is a very compact structure that results from the coiling of the
polymer chains causing large amounts of carbon energy to be stored in a small volume making it an
important fuel reserve for several reasons (Blamire,2014). The controlled breakdown of glycogen and
release of glucose increase the amount of glucose that is available between meals. Hence, glycogen
serves as a buffer to maintain blood-glucose levels. Glycogen's role in maintaining blood-glucose levels is
especially important because glucose is virtually the only fuel used by the brain, except during
prolonged starvation. Moreover, the glucose from glycogen is readily mobilized and is therefore a good
source of energy for sudden, strenuous activity. Unlike fatty acids, the released glucose can provide
energy in the absence of oxygen and can thus supply energy for anaerobic activity. It is commonly stored
in muscles and livers of animals (Berg,1970) .
 For this exercise the students were tasked to extract polysaccharide from clams and chicken
liver by precipitation.

RESULTS

A. Isolation of Polysaccharide from Clams

Table 1. Observations on the Isolation of Polysaccharide from Clams

SAMPLE STEPS OBSERVATIONS

20 pcs clams original appearance slimy, yellowish with brown and
green pigments

after grinding same appearance but smaller in

size

after boiling small coagulation, formation of

clumps and greyish appearance
of solution

addition of 10% HOAC (15 drops) most particles settled at the

bottom and coagulated with
turbid appearance

filtering turbid-yellowish appearance

B. Isolation of Polysaccharide from Chicken Liver

SAMPLE STEPS OBSERVATIONS

2.23 g chicken liver original appearance reddish-brown and solid

after grinding reddish-brown but became gel-

like

after boiling formation of light brown clumps

of liver

addition of 10% HOAC (15 drops) coagulation and became turbid

filtering dirty-white solution

C. Precipitation of Polysaccharide

SAMPLE STEPS OBSERVATIONS

Filtrate from tulya Original appearance turbid solution
Addition of 95% alcohol remained the same
After centrifugation dirty white precipitates

Filtrate from chicken liver Original appearance turbid solution

addition of 95% alcohol remained the same
After centrifugation No precipitates formed

DISCUSSION

Both liver and muscles are rich storage of glycogen where it is commonly more concentrated in
liver with around 6-8% while for muscles with approximately 1%.

As shown in both Table 1 and 2, the samples namely the chicken liver and clams were cut in
smaller portions and grinded since this will help in hastening the reaction then it was boiled together
with 40 mL water as a form of extracting method for isolating polysaccharide. According to Nie et al
(2013), polysaccharides are polar and hydrophilic macromolecules and can be extracted with a
strong polar solvent, and water is an ideal solvent which is popularly used in practice. High
temperature can accelerate the dissolution of polysaccharide from the cell wall and make it
easier to dissolve in water. Thus, hot water extraction has long been the traditional and classic
method for polysaccharide extraction however, there is slow extraction rate in this method.

In order to prevent the association of other unnecessary compounds such as proteins,

the boiled solution of tulya and liver was slightly acidified with 10% acetic acid that yielded a
turbid solution with minimal coagulation brought about by the denaturation of the proteins (Jin et
al ,2012). The addition of acetic acid was carefully monitored since an excess acid might cause disruption
on the bonds of the glycogen making it soluble.

Glycosidic bonds in glycogen are resistant to hydrolytic activity of OH at elevated temperature.

In contrast, peptide bonds in proteins, ester bonds in lipids and phosphodiester bonds in ribonucleic
acids undergo hydrolysis at high temperature and in alkaline pH (i.e. in KOH solution). Under these
conditions the glycogen solution, only slightly contaminated with other polysaccharides, fragments of
denatured DNA and low molecular weight compounds, can be obtained. Addition of ethanol at lower
temperature since the polysaccharides have lower solubility in high concentration of alcohol or ether, it
results in glycogen precipitation and allows to obtain relatively purified glycogen. The last step was the
centrifugation where it contained only the pure precipitates of glycogen where only the tulya showed
the presence of precipitates since more glycogen is stored in skeletal muscle overall because of its much
greater mass although glycogen is higher in the liver than in muscle (10% versus 2% by weight)
(Berg,1970).

CONCLUSION

Cumulated

mollusk

REFERENCES

Barnes and Noble. (n.d.). Retrieved from https://www.sparknotes.com/health/carbohydrates/section1/.

Berg. (1970). Glycogen Metabolism. Retrieved October 9, 2019, from

https://www.ncbi.nlm.nih.gov/books/NBK21190/.

Blamire. (2014). Polysaccharide. Retrieved from

http://www.brooklyn.cuny.edu/bc/ahp/LAD/C4c/C4c_polysaccharides.html.

ChemED. (n.d.). Carbohydrates. Retrieved from

https://chemed.chem.purdue.edu/genchem/topicreview/bp/1biochem/carbo5.html.

EUFIC. (2019). The Functions of Carbohydrates in the Body. Retrieved from

https://www.eufic.org/en/whats-in-food/article/the-basics-carbohydrates.

Jin et al (2012). Isolation, structure and bioactivities of the polysaccharides from Angelica sinensis (Oliv.)
Diels: A review. Carbohydrate Polymers, 89(3), 713–722. doi: 10.1016/j.carbpol.2012.04.049

Learning. (n.d.). Biology for Majors I. Retrieved from https://courses.lumenlearning.com/wm-

biology1/chapter/reading-types-of-carbohydrates/.

Nie et al. (2013). Current development of polysaccharides from Ganoderma: Isolation, structure and
bioactivities. Bioactive Carbohydrates and Dietary Fibre, 1(1), 10–20. doi: 10.1016/j.bcdf.2013.01.001

Stubbings (2008). Retrieved from http://www.ausetute.com.au/redsugar.html.