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FR5A-Isolation and Characterization of Starch in Potato Sample
FR5A-Isolation and Characterization of Starch in Potato Sample
FR5A-Isolation and Characterization of Starch in Potato Sample
ABSTRACT
Gluten, the intact protein, was separated from wheat flour by dissolving the starch that accompanied it in the process
of making wheat flour. The intact protein was tested to describe its structure and functional groups. Such test used
were to predetermine the presence for amides and aromatic side chains
1
In Xanthoproteic Test, the protein solution (Na2CO3) to the diazo reagent. Note the
is slowly treated with 10 drops appearance of the red coloration.
concentrated Nitric Acid (HNO3). Observe
caution for handling acids. Before
proceeding to the next step, take note of
the color of the solution. Next slowly add RESULTS AND DISCUSSION
10 drops of Sodium Hydroxide (NaOH) 1. Isolation of Gluten
and again note the color of the sample. Crude Gluten was obtained by washing the
dough to the water. The crude gluten obtained
d. Millon’s Test was from wheat flour, an ingredient to produce
The sample is treated with 5 drops of bread particularly leavened bread. Wheat flour
Millon’s reagent and be noted for its color has two major composition, starch and crude
reaction. gluten. In order to separate gluten from starch,
certain separation techniques must me used to
e. Hopkins-Cole Test separate the two components. The best way to
Slowly treat the gluten sample with 20 separate the two is by means of solubility.
drops Hopkins-cole reagent and mix well. Starch is a white odorless powdery
Then incline the tube add slowly along the carbohydrate (C6H10O5)x which is a chief storage
side 20 drops of concentrated sulfuric form of carbohydrates in plants [2]. Starch is
acid. (H2SO4). Observe caution. Note the soluble in water while gluten is insoluble. Hence
color of the sample. gluten can be separated by using the solubility
rule.
f. Sakaguchi Test Gluten is an elastic mixture composite of
The gluten sample is treated with 10% proteins glutenin and gliadin. Gluten has
Sodium Hydroxide (NaOH) and 10 drops chemical groups that make up its characteristics.
of 0.02% Naphthol Solution. Let the Gluten undergoes a series of Qualitative analysis
sample stand for 3 minutes and then add to further understand the structure of gluten.
3 drops 2% NaOBr. Again note the color
of the solution. 2. Qualitative Color Reaction of
Gluten
g. Nitroprusside Test In table 1, the results of the color reactions
For the nitroprusside test, add 0.5mL 3M are tabulated and noted. 0.5g of gluten was
of Sodium Hydroxide (NaOH) and 0.25 mL placed in ten test tubes together with 1mL of
2% Nitroprusside Solution. Note the Distilled water.
change in color to a red solution.
TABLE 1. RESULTS OF COLOR REACTION OF
h. Fohl’s Test GLUTEN
Add 5 drops of 30% Sodium Hydroxide COLOR REACTION TEST OBSERVATION/S
(NaOH) and 2 drops of Lead Acetate BIURET TEST Violet
(Pb(CH3COO)2) to the sample then NINHYDRIN TEST Blue Violet
proceed by placing it in a water bath. Note XANTHOPROTEIC TEST Yellow
for the appearance of dark or brown MILLON’S TEST White
sediments HOPKIN’S-COLE TEST Violet (interface)
SAKAGUCHI TEST Light Yellow
i. Test for Amides NITROPRUSSIDE TEST Red
Add 1mL 20% NaOH to 10 drops of the FOHL’S TEST Brown precipitate
gluten sample then proceed by placing it TEST FOR AMIDES R-B (Basic)
in the water bath. Test for the evolution of PAULY TEST Pale Yellow
gas by placing moisten red and blue
litmus paper over th mouth of the tube. a. BIURET TEST (Test for Peptide
Bonds)
j. Pauly Test
First step is by preparing a diazo reagent The biuret test, it is basically the most
by mixing 3-5 drops 1% sulfosalicylic acid fundamental test for proteins, a general
with 3 drops of 5% Sodium Nitrite test for proteins [1]. The test is used for
(NaNO3). Add 5 drops of the sample and the presence of peptide bonds. When a
3-5 drops of 10% Sodium Carbonate protein synthesizes with Copper (II)
Sulfate, test will confirm a protein is
2
present in an unknown forming a protein-
copper complex which has a violet
coloration.
REFERENCES
From Books
[1] Bettelheim, Frederick A., Landersberg
Joseph M. (2001), Laboratory
Experiments for Organic and Biochemistry
Philadelphia: Harccourt College Publishers.
P.201
[2] Merriam-Webster’s Medical Dictionary
(2006). Springfield Massachusetts:
Merriam-Webster Incorporated
[3] Copeland, Robert.(1994). Methods of
Protein Analysis NewYork: Chapman &
Hall,p.46
[4] Gluten, Britannica Concise Encyclopedia
(2002) U.S.A.: Encyclopædia Britannica