RESULTS AND DISCUSSION which was a negative one because the

desirable result was a brick-red

Isolation and General Tests for
precipitate to indicate the changing of
Polysaccharides
ketoses to aldoses which was exhibited
The isolate formed which was a by all monosaccharides and most
turbid white solution was used for the disaccharides. [Refer to table…]
isolation and general tests for
Hydrolysate Description Benedict’s
polysaccharides that used two (Viscosity) Test
procedures such as the Molisch’s test and Acid Fluid; not Clear, Yellow-
I2 reaction. The Molisch’s test which was Hydrolysate viscous orange
a general test for carbohydrates obtained solution
a purple ring at interface. The result Table 2. Hydrolysis of Polysaccharides: Benedict’s Test
obtained was positive because the purple Qualitative Color Reactions
ring at the interface that was produced
signified that the sulfuric acid The qualitative tests for
dehydrolyzed the carbohydrate and carbohydrates were done using four tests
produced an aldehyde. While on the such as Benedict’s test, Barfoed test,
other hand, the reaction with I2 Seliwanoff’s test, and Bial’s-Orcinol test.
produced a turbid white solution which The samples which undergone through
indicated that the result obtained was these procedures are glucose, fructose,
negative as there was no starch present xylose, lactose, sucrose, starch, and the
in the isolate used because the positive hydrolysate. [Refer to table…]
result would produce blue coloration Benedict’s test was a test done to
when reacted with iodine to indicate the detect a reducing sugar (Chhabra, 2014).
presence of starch. [Refer to table…]
In this test, only the fructose and
Description Molisch’s KI/I2 hydrolysate produced a positive result.
Test The fructose exhibited a red solution,
Isolate Turbid white Presence of Turbid that indicates the changing of ketoses to
solution purple ring at white aldoses which was exhibited by all
the interface solution monosaccharides and most
Table 1. Isolation and General Test for Polysaccharides disaccharides. On the other hand, the
Hydrolysis of Polysaccharides hydrolysate produced an orange solution
which means that it contained an
The result of the sample subjected intermediate amount of
to acid hydrolysis was a fluid solution that monosaccharides.
was not vicious.
The second was Barfoed’s test
Benedict’s test was a test done to that determines the difference between
detect a reducing sugar. From the test reducing monosaccharides and
performed in the experiment, the result disaccharides (Chhabra, 2014). The
obtained was a yellow-orange solution positive result of this test would be the
appearance of red precipitate which Carbohy Visible Results
indicate the reduction drate Bened Barfo Seliwa Bial’s
Solution ict’s ed’s noff’s Test
of cupric ions, that were present in the Test Test Test
reagent used, by monosaccharides. Glucose Light Darke Light Clear
However, none of the samples used blue r yellowi yellow
produced a red precipitate which meant shade sh color
of
that all of the samples were negative for
light
this test. blue
The third was Seliwanoff’s test Fructos Forma Clear, Red- Green
e tion of blue orange with
that differentiates ketoses and aldoses red soluti solutio brown
(Chhabra, 2014). Ketoses produced precip on n precip
cherry-red that because of the formation itate itate
of hydroxymethylfurfural, through Xylose Dark Blue Clear, Gray
dehydration, which condenses with the green soluti light soluti
use of resorcinol. An alternative result for soluti on yellow on
on solutio with
this test was the appearance of yellow to
n brick
pink colors by other sugars. Fructose, red
which produced a red solution, was precip
positive for this test. While others, such itate
as glucose, xylose, and starch produced Lactose Black Rema Black
yellow coloration. Moreover, Sucrose layer ined Remain soluti
forma blue ed on
produced a pink clear solution that
tion soluti colorle
indicated the presence of aldose. on; ss;
Lastly was the Bial’s-Orcinol test, neagti negativ
ve e
which was done to distinguish between
Sucrose Clear, Clear, Pink Clear,
pentose and hexose (Chhabra, 2014). light light clear light
Pentoses produce a blue-green solution blue blue solutio brown
by forming furfural through dehydration soluti soluti n soluti
which was then condensed with orcinol. on (no on on
From the test performed, only the color (no
chang color
hydrolysate produced a blue-green
e) chang
solution, therefore, it was the only e)
sample that produced a positive result for Starch Dark Light Pale Pale
the presence of pentose. blue blue yellow yellow
solutio
n
Clear, Clear, Turbid, Clear,
Hydroly Yellow Blue Peach- Blue-
sate - soluti colored green
orang on precipit soluti
e ate on
soluti
on
Table 3. Qualitative Tests for Carbohydrates
Conclusion Based on the Qualitative
Hydrolysab Monosacchari Reducing or
Tests Sugar le or Non- de, Non-reducing
hydrolysab Oligosaccharid
From the results of the tests that
le e, or
were obtained from the experiment, it Polysaccharide
showed that the non-hydrolysable Glucose Non- Monosaccharid Non-reducing
groups were glucose, fructose, and hydrolysabl e
xylose, while the hydrolyzable groups e
were lactose, sucrose, starch, and the Fructose Non- Monosaccharid Reducing
hydrolysate. hydrolysabl e
e
The carbohydrates were also Xylose Non- Monosaccharid Reducing
classified according to the number of hydrolysabl e
monosaccharides that they have. e
Lactose Hydrolysab Oligosaccharid Non-reducing
Glucose, fructose, and xylose were
le e
classified as a monosaccharide, while Sucrose Hydrolysab Oligosaccharid Non-reducing
lactose, sucrose and the hydrolysate le e
were identified as oligosaccharides, and Starch Hydrolysab Polysaccharide Non-reducing
lastly, starch was the only one to be le
classified as a polysaccharide. Hydrolysa Hydrolysab Oligosaccharid Reducing
te le e
The samples were also identified Table 4. Conclusion Based on the Qualitative Tests
based on their reducing properties.
Glucose, lactose, sucrose, and starch Pentose or Aldose or
Sugar Hexose Ketose
were classified as non-reducing sugars
Glucose Pentose Aldose
while fructose, xylose, and the
Fructose Hexose Ketose
hydrolysate were the reducing ones.
Xylose Hexose Aldose
Moreover, the given sugars were
also identified based on the number of Lactose Hexose Aldose
carbons that they have and the functional Sucrose Hexose Ketose
group where they belong. Glucose and
Starch Hexose Aldose
the hydrolysate were classified as
pentoses, while fructose, xylose, lactose, Hydrolysate Pentose Aldose
sucrose, and starch was known as
Table 4.1 Conclusion Based on the Qualitative Tests
hexoses. Another was that glucose,
xylose, lactose, starch, and the Thin-Layer Chromatography
hydrolysate were determined as aldoses, The table presented showed the
whereas fructose and sucrose belonged results of thin-layer chromatography
to ketoses. from the sample, chicken liver.
 Distance 250 0.327 0.324 0.331 0.327
µg/mL
Dextrin Maltose Glucose
125 0.212 0.218 0.215 0.215
Distance µg/mL
Traveled by 3.5 3.5 3.5 Table 6. Absorbance of Glucose at Certain
Solvent Concentrations
Distance The mean absorbance of the
Traveled by 0 0.5 0.9 enzymatic hydrolysate with a value of
the Solute 2.922 was higher than the absorbance
Rf Value 0 0.14 0.26 obtained from acid hydrolysate which
Identity of only acquired a mean of 1.629.
Components
1 2 3 Mean
Table n.1 Thin-layer Chromatography of Chicken Liver
Acid 0.742 2.473 1.672 1.629
Hydrolysate
Hydrolysate Enzymatic 2.207 0 3.637 2.922
Hydrolysate
Acid Enzymatic
Table 7. Absorbance of Hydrolysates
Distance Traveled
by Solvent 3.5 3.5 In relation to the absorbance, the
Distance Traveled mean concentration of enzymatic
by the Solute 0.7 0.8
hydrolysate which was 2586.455 µg/mL
Rf Value D= 0 D= 0 was higher than of acid hydrolysate
G= 0.2 G= 0.23 which only had a mean value of 1411
Identity of Dextrin and Dextrin and µg/mL.
Components Glucose Glucose
Hydrolysate Concentration
Table n.1 Thin-layer Chromatography of Chicken Liver
Acid Hydrolysate 1411 µg/mL
Quantitative Analysis Enzymatic Hydrolysate 2586.455 µg/mL
Table n presented the absorbance Table 8. Concentration of Glucose in Hydrolysates
of glucose at different conditions that
were obtained from the experiment. The From the results obtained in the
sample with a concentration of quantitative analysis, it can be inferred
2000µg/mL obtained the highest mean that cuprous oxide, which produced blue
with a value of 2.165 and the sample with coloration, had a higher concentration on
a concentration of 125µg/mL got the enzymatic hydrolysis which also indicates
lowest mean which was 0.215. that it gave a higher amount of reducing
sugar that were oxidized.
1 2 3 Mean
2000 2.141 2.165 2.188 2.165
µg/mL
1000 1.162 1.218 1.255 1.212
µg/mL
500 0.574 0.563 0.553 0.563
µg/mL

Figure 1. The Standard Curve for Glucose

References: osazone. The visual positive result of this
test was yellow orange crystals same as
HiPer® Carbohydrates Estimation
what was seen on enzymatic phenyl
Teaching Kit (Qualitative). (n.d.).
when it was viewed under the
Retrieved November 24, 2019, from
microscope. On the other hand, acid
http://himedialabs.com/TD/HTBC002.pd
phenyl failed this test and obtained a
f.
negative result due to the absence of
Chhabra, N. (2014). Qualitative tests for yellow orange crystals.
carbohydrates. Retrieved November 24,
2019, from
https://www.slideshare.net/namarta28/
qualitative-tests-for-carbohydrates-
35884145

Mucic Acid Test

The principle involved in this test
was the addition of HNO3, which was a
strong oxidizing agent, to oxidize formyl
and primary alcohol functional groups
into a carboxylic acid. The expected
positive result for this test was the
appearance of rhombic crystals when
viewed under the microscope. The said
characteristic of the positive result was
evident in enzymatic mucic. However,
the result obtained from the acid mucic
did not show an appearance of rhombic
crystals which made it a negative results.
Osazone Test
In this test, the principle involved
was the addition of a strong oxidizing
agent, which was phenylhydrazine, to
oxidize formyl functional group and
secondary alcohol into a carboxylic acid
and ketone, respectively. The oxidation
will produce the product which was
phenylhydrazone and when combined
with phenylhydrazine will produce