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Test tube 1 contained arginine as indicated by the presence of a red color complex when reacted with alpha naphthol and bromine water. Test tube 2 did not produce a red color, indicating the absence of arginine. Millon's reagent is used to detect the presence of tyrosine. It is prepared by dissolving mercuric nitrate and nitric acid, then adding water. When drops of Millon's reagent are added to a test solution containing tyrosine and heated, a red or red precipitate forms, indicating a positive result. Cholesterol must be dissolved in chloroform for the Liebermann-Burchard test because cholesterol is a nonpolar compound and lipids are

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45 views

Post Lab Discussion Reviewer

Test tube 1 contained arginine as indicated by the presence of a red color complex when reacted with alpha naphthol and bromine water. Test tube 2 did not produce a red color, indicating the absence of arginine. Millon's reagent is used to detect the presence of tyrosine. It is prepared by dissolving mercuric nitrate and nitric acid, then adding water. When drops of Millon's reagent are added to a test solution containing tyrosine and heated, a red or red precipitate forms, indicating a positive result. Cholesterol must be dissolved in chloroform for the Liebermann-Burchard test because cholesterol is a nonpolar compound and lipids are

Uploaded by

Angelica Camille B. Abao
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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POST LAB DISCUSSION

6.1
Test Tube 1 – red color complex
Test Tube 2 – absence of red color complex

Both students followed the procedure given by the instructor yet they obtained different
results. What do you think is the reason?

Tube 1 – indicates the presence of arginine. The guanidium reacted with alpha naphthol
+ bromine water – red color product.
Tube II – No red color product means absence of arginine.

6.2 What is the composition of Millon’s reagent? Describe their significance in the
test.
Millon’s Reagent
Intended use: Millon’s reagent is an analytical reagent used to detect the presence of
tyrosine, the only amino acid containing a phenol group.
Composition
Ingredients Mercuric Nitrate 160.0g
Mercurous Nitrate 160.0g
Conc. Nitric Acid 400.0ml
Distilled water 600.0ml
*Formula adjusted, standardized to suit performance parameters.

What is the composition of Millon’s reagent? Describe their significant in the test.
➢ Millon’s reagent is prepared by dissolving mercuric nitrate in nitric acid, and then
adding water to dilute it.
➢ The reagent is used in Millon reaction test wherein few drops of the reagent are
added to the test solution, and then heated gently boiling.
➢ When a red-colored solution or red precipitate forms, this means that tyrosine is
present. (+ result).
8.1 Why is there a need to dissolve cholesterol in Chloroform in the
Liebermann-Burchard test?
➢ Mixtures of chloroform and methanol have a had wide use as lipid
extractants. Optimum lipid extraction should result when the tissue is
homogenized with a mixture of chloroform and methanol which, when mixed
with the water in the tissue, would yield a monophasic solution.
➢ The resulting homogenate could be then be diluted with water and/or
chloroform layer of which should contain the lipids purified lipid extract should
be obtained when the chloroform layer is isolated.
➢ Chloroform is first added to dissolve cholesterol and make it easier for the
reaction to proceed.
➢ Cholesterol is dissolved in chloroform because it is a nonpolar compound.
Lipids in hydrophobically associated form are extracted with relatively non-
polar solvents such as ethyl either, chloroform, or benzene.
➢ Direct chloroform extraction is advantageous, as this extract can be used
directly for color development.

8.2 Differentiate Salkowski from Liebermann-Burchard test, include the difference


in the reagents used, chemical reactions and endpoints, and the color
production.
Lierbermann-Burchard Test Salkowski Test
Detects the presence of: Cholesterol Cholesterol
Reagents Acetic Anhydride Sulfuric Acid
Sulfuric Acid Chloroform
Chloroform
Type of Reaction Colorimetric Method Colorimetric Method
Chemical reaction/end point The acetic anhydride is used Concentrated sulfuric acid
as solvent and dehydrating reacts with cholesterol form
agent, and the sulfuric acid is bi-sulphonic acid of bi-
used as dehydrating and the choloestadiene.
oxidizing agent. The -OH
group of cholesterol reacts with
the reagents and the
unsaturation found in the
adjacent fused ring.
Color Reaction Production of red, then blue, Choroform layer – a play of
and finally bluish green color. colors from bluish red to
cherry red and purple is
noted.
Acid layer – a green
flourescence
8.3 What is the purpose of the filter paper moistened with ammoniacal silver
nitrate solution in the Acrolein test?
➢ Fumes of SO2 can arise from the reduction of Potassium Bisulfate and therefore,
cause false positive results since they also have irritating odors. Healing abruptly
and presence of organic compounds that can reduced Potassium Bisulfate must
be avoided.
➢ Heating will cause polymerization of the acrolein, leading to a darkening of the
solution or in this case, the filter paper.
9.0 What is the classical method to check the quality of DNA product? Discuss its
principle.
To assess NA quality, a Nanodrop spectrophotometer can be used
1. Nucleic acids absorb UV at 260 nm (A260)
2. The more light absorbed, the higher the concentration
3. NA purity assessment:
A260 – quantitates RNA, ssDNA, and dsDNA
A260/A280 – for protein contamination; measures purity of nucleic acids
Pure DNA = 1.8
Pure RNA = 2.0
Lower Ratio = presence of contaminants
A260/A230 – for salt contamination; with pure nucleic acids ranging from 2.0 to 2.2

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