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Carbohydrates are the most abundant class of organic compounds found in living
organisms. The objective of this experiment are to : (a) compare the products of the isolated
glycogen from chicken liver after acid and enzymatic hydrolyses, (b) illustrate the specificity
of the - amylase on the hydrolysis of the isolated polysaccharide, (c) prepare dialyzing bag
used inseparating the products of enzymatic hydrolysis and explain the principle behind it,
lastly, (d) examine microscopically the mucic acid crystals.The glycogen was first isolated
from the chicken liver. The sample produced was described as light yellow solution. The
isolate produced was then subjected to acid hydrolysis. Five mL of concentrated HCl were
added to the isolate and was placed in a boiling water bath. The acid hydrolysate was
described as clear yellow solution and is not viscous. Furthermore, the isolated glycogen
was also subjected to enzymatic hydrolysis. About 2.3 mL of saliva were added to the isolated
carbohydrate. The solution was then introduced to a dialyzing bag ang was suspended
overnight in a small flask filled with 50 mL distilled water. It was then subjected to an open
flame and was concentrated to a volume of 10 mL. The enzymatic hydrolysate was described
as whitish yellow solution and is not viscous. Both the acid and enzymatic hydrolysate were
used in qualitative tests which are done to determine the identity of the monosaccharides
present in the solution. The hydrolysates tested negative on most of the qualitative tests
except the Seliwanoffs test. The Mucic Acis test, which is a test specific for the
determination of galactose, was also performed. Three drops of galactose and lactose and 3
drops of concentrated HNO 3 were mixed in a glass slide. Then, it was passed over an alcohol
lamp in order to dry. The mixture was then cooled down at room temperature. The crystals
formed on the glass slides were examined under the microscope. The two types of sugars,
glucose and lactose exhibited positive results in this test.
The acid and enzymatic hydrolysate In this test, the nitric acid first
tested positive in Seliwanoffs test. The catalyzed the breakdown of lactose into
Seliwanoffs is a distinguishing test between glucose and galactose. Then, it oxidized the
aldoses and ketoses. The presence of cherry two sugars into tetrahydroxyadipic acids
red precipitate indicates the presence of a which eventually precipitates as insolluble
ketohexose. Consequently, Ketopentoses crystals after cooling down at room
yield blue-green series while aldoses and temperature.
disaccharides give no shading change. The
presence of cherry red precipitate indicates
that the hydrolysates contain a ketohexose,
specifically fructose. Meanwhile, the color
change in the acid hydrolysate suggests the
presence of trace amounts of reducing
sugars. [3] Glycogen is a polysaccharide
consist of glucose monomers linked via -
1,4-glycosidic bonds in a helical structure.
Glucose is an example of a reducing sugar.
Sugars thatcontain aldehyde groups that are
oxidised to carboxylic acids are reducing
sugars.