Carbohydrates are polyhydroxyaldehydes

(ketoses) with the general formula of (CH2O)n

where n equals 3 or more. Carbohydrates are
divided into three general classifications
depending on the carbohydrate molecules they
contain; They may be called a monosaccharide,
oligosaccharide or a polysaccharide. [1]
Carbohydrate test reagents are divided into 2
general classes based on the type of reaction
involved. The first class is a 2-step analysis
consisting the use of dehydrating acids followed
by condensation reagents. These principles are
found in the Molischs, Anthrone, Bials, and
Seiwanhoffs tests. The second class uses
solutions containing copper (II) ions The
carbohydrate reduces the copper (II) ions into
copper (I) oxide. Reducing sugars are aldoses
containing either a free aldehyde group or a
cyclic hemiacetal. [1]
The amount of carbohydrates can be measured
by Nelsons method. It is based on the capacity
of free-reducing sugars in a carbohydrate sample
to reduce Copper (II) in an alkaline solution. [1]
The amount of free-reducing sugars in the
sample is directly related to the molybdenum
blue formed via a series of oxidation/reduction
reactions, and is measured colorimetrically. [2]
Thin-layer Chromatography is achromatography
in which compounds are separated on a thin layer
of adsorbent material, typically a coating of silica
gel on a glass plate or plastic sheet. [2] It gives
better resolution and entails faster development
compared to paper chromatography and

identifying unknowns. Partial characterization and

tentative identification of unknown can be carries
out by comparing its Rf value to that of a
standard. [1]
The objectives of this experiment were to isolate
polysaccharides from plant sources and explain
the principle involved, perform the general tests
of polysaccharides and compare products of acid
and enzyme hydrolysis of the isolated
carbohydrate, illustrate the specificity of alphaamylase on the hydrolysis of the isolated
polysaccharide, perform thin-layer
chromatography on the carbohydrate
hydrolysates, correlate data obtained from the
color tests and thin-layer chromatography of the
carbohydrate hydrolysates perform qualitative
tests for carbohydrates based on furfural
formation and oxidation, examine microscopically
the different osazone and mucic acid crystals,
classify unknown carbohydrate based on the
results of the different qualitative tests and
determine the amount of reducing sugars using
Nelsons test and explain the principle involved.
[1] Crisostomo, A., Daya, M., de Guia, R., Farrow,
F., Gabona, M., Liu, I., Pea, G., Pea, L.,
Santiago, L., Santiago, M., Sarile, A., Torres, P.,
Vargas, A., & Ysrael, M. (2010). Laboratory
Manual in General Biochemistry. Quezon City:
C&E Publishing, Inc.
[2] No Author. Thn Layer Chromatography
Procedures. Retrieved May 11. 2016 from
http://orgchem.colorado.edu/Technique/Procedur
es/TLC/TLC.html