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Zoe Angela M. Espinosa, Ma. Pamela Rica M. Fajardo, Emmanuel Joshua M. Garcia, Liana Celina D.
Geluz, Jillian Pamela D. Go, Jennifer Mae C. Heredia
Group 4 2A Medical Technology CHEM600 Laboratory
ABSTRACT
Carbohydrates are generally hydrates of carbon with the common formula (CH 2O)x, ranging from simple sugars such
as glucose and fructose to complex molecules such as starch and cellulose. In the experiment, the solutions of
carbohydrates that were used were glucose, fructose, lactose, xylose, sucrose, and starch. Different qualitative tests
were performed on the different samples namely Benedicts Test, Barfoeds Test, Seliwanoffs Test, Bials Test, Mucic
Acid Test, and Phenylhydrazone Test. Glucose, fructose, lactose, xylose, and sucrose produced a brick-red pptfor
Benedicts Test, indicating a positive result. For Barfoeds Test, only fructose and xylose showed a brick-red pptbut
glucose was also expected to produce a positive result. Seliwanoffs Test generated a cherry red pptfor fructose and
sucrose while Bials Orcinol Test gave a blue green coloration to glucose, xylose, lactose, and starch even when xylose
is the only one expected to give the positive result. Crystals from galactose were observed from the mucic acid test.
Lastly, phenylhydrazone test gave glucose, fructose, and xylose needle-shaped yellow osazone crystals. Lactose also
formed needle shaped crystals. Sucrose did not form osazone crystals. Starch produced some needle shaped crystals.
INTRODUCTION
Qualitative analysis shows whether a particular
substance is present, but does not give an
indication of how much is present [2]. In this
experiment, several qualitative tests were used
to analyze the distinguishing features of standard
carbohydrate solutions.
Benedict's Test detects reducing sugars through
the use of Benedict's reagent [1]. Benedict's
reagent is a chemical reagent named after an
American chemist, Stanley Rossiter Benedict.
When
Benedicts
solution
and
simple
carbohydrates are heated, the solution changes
to orange red/ brick red. This reaction is caused
by
the
reducing
property
of
simple
carbohydrates. Barfoed's test is a chemical test
used
for
detecting
the
presence
of
monosaccharides. It is based on the reduction of
copper(II) acetate to copper(I) oxide (Cu2O),
which forms a brick-red ppt[7].Seliwanoffs test is
a chemical test which distinguishes between
aldose and ketose sugars. It is named after
Theodor Seliwanoff, the chemist that devised the
test. The test reagent, resorcinol in 6M HCl,
dehydrates
ketohexoses
to
form
5hydroxymethylfurfural. Bial's-Orcinol Test or Bial's
Test is a qualitative test which involves a reagent
containing 5-methylorcinolin concentrated HCl
with a small amount of FeCl3. Pentoses undergo
dehydration to furfural that condenses with
orcinol to create a blue-green solution [6]. Mucic
Acid Test employ the principle that aldohexoses
are converted to their corresponding dicarboxylic
acids in the presence of strong oxidizing agents
like concentrated HNO3. The dicarboxylic acidwhich is mucic acid produced from the oxidation
of galactose is relatively insoluble and separates
out as colorless crystals [6]. Osazone or
Phenylhydrazone Test involves a carbohydrate
specific reaction with phenylhydrazine to form
osazone crystals of specific shape which confirms
with
4
drops
of
the
freshly
prepared
phenylhydrazine reagent. The test tubes were
mixed well and were covered with a cotton
stopper. The test tubes were subject to a boiling
water bath for 30 minutes and the time when
crystals first appear was noted. The test tubes
were cooled and the crystals were observed
under the microscope. The shape and color of
each of the crystals were noted.
Benedicts Test
Brick Red ppt (+)
Brick Red ppt (+)
Brick Red ppt (+)
Brick Red ppt (+)
Brick Red ppt (+)
Blue soln (-)
Barfoeds Test
Blue soln (*)
Brick Red ppt (+)
Brick Red ppt (+)
Blue soln (-)
Blue soln (-)
Blue soln (-)
Visible Results
Seliwanoffs Test
Peach soln (-)
Cherry Red ppt (+)
Peach soln (-)
Peach soln (-)
Cherry Red ppt(+)
Peach soln (-)
Crystalline
Structure
Glucose
Needle
shape
Fructose
Needle
shape
Xylose
Needle
shape
Lactose
Needle
shape
Sucrose
No crystals
Starch
Needle
shape
Microscopic
Observation
glycosidic linkage is alpha- for glucose and betafor fructose. Hence, sucrose neither shows
reducing nor mutarotation characters [5].