EXPERIMENT 1: TRANSPORT ACROSS THE MEMBRANE

Procedure:
A. Diffusion B. Osmosis C. Dialysis
*see lab manual*
TEST FOR NAME OF TEST COLOR REACTION PROCEDURE
“Test for proteins” 1mL dialysate + 1mL 25%
White precipitate or turbid
Proteins trichloroacetic solution.
a. 1mL Fehlings A + 1mL
Presence of reducing Yellow or red precipitate Fehlings B
“Test for sugar” b. heat boiling water bath
sugar
c. + 1mL dialysate. Boil for
1-2 mins.
EXPERIMENT 2: THE BIOMOLECULES IN SOME CELL ORGANELLES
Procedure:
A. Centrifuge at 600xg for 10 minutes – contains nuclei and unbroken cells
B. Centrifuge at 10,000xg for 20 minutes – contains mitochondria of the cell
C. Centrifuge at 15,000xg for 30 minutes – contains lysosome
**
TEST FOR NAME OF TEST COLOR REACTION PROCEDURE
a. 5 drops suspension +
Proteins Biuret test Violet-pink color 10%NaOH.
b. + 5 drops 0.5% cupric
sulfate.
a. 10 drops suspension + 4
drops Molisch reagent
Carbohydrates Molisch Test Purple color b. + 10 drops concentrated
sulfuric acid. DO NOT
SHAKE
a. 8mL suspension in
Sulfuric acid layer: yellow evaporating dish.
with green fluorescence b. Heat to dryness and then
cool + 2mL of ether.
Chloroform layer: bluish red c. Do it three times.
Salkowski Test
Lipids gradually turning to violet- d. Combine all the ether
red extract. Evaporate. +
2mL chloroform.
e. + 10 drops chloroform +
5 drops conc. sulfuric
acid
a. 5 drops of cell
suspension + 5 drops
Nucleic Acids: DNA Disch Test Blue color diphenylamine.
b. Observe for a blue color
result.
c. Add small piece of
 broken porcelain if a
blue color does not
appear to the tube.
d. Warm the test tube in a
water bath for 5 mins.
a. 5 drops Orcinol reagent +
Orcinol Test Green Color 5 drops cell suspension
Nucleic Acids: RNA b. heat in boiling water
bath for 3 minutes.

EXPERIMENT 3: pH AND BUFFER SOLUTIONS

**
Procedure:
A. Measurement of pH B. Buffer System

EXPERIMENT 4: ISOLATION OF GLYCOGEN

**
Procedure:
A. Isolation of glycogen from clamsB. Isolation of glycogen from chicken liver

EXPERIMENT 5: CHEMISTRY OF CARBOHYDRATES

**
Procedure:
A. Hydrolysis of Carbohydrates
B. Reducing or non-reducing sugar - Fehling’s
C. General Tests for Carbohydrates – MAIQ (BBSO)

General Tests for Carbohydrates

TEST FOR NAME OF TEST COLOR REACTION NOTES
Furfural and other
decomposition products
react with alpha-napthol.
General test for
Molisch Test Purple color Not specific test for
Carbohydrates in free or
carbohydrates. However,
combined form
negative result is a good
evidence of absence of
carbs.
sulfuric acid hydrolyses
Another general test for Anthrone Test Dark blue-green color bound sugars and react
carbohydrates with sugars

Test for Polysaccharides.

Differentiate helical Iodine Test Blue color Iodine form complexes
polysaccharides from non- with amylose
 helical polysaccharides.

Qualitative Test for Sugars

Same reaction; diff
Differentiate aldoses and Seliwanoff's Test Cherry red color reaction time= slower for
ketoses
aldoses

Distinguish between Brick red precipitate monosaccharide reacts

reducing monosaccharide
Barfoed’s Test formed within 5 quickly
and reducing disaccharide minutes
Reducing ability of sugar Fehling’s Test Red precipitate

Reducing ability of sugar Benedict’s Test Red precipitate

Detect the presence of a Green to blue color

Bial’s or Orcinol Test
pentose within 5 minutes

EXPERIMENT 6: CHEMISTRY OF PROTEINS

**
Procedure:
A. Isolation of Casein from Milk
B. Preparing of casein solution
C. Test for proteins
D. Denaturation Test
TEST FOR NAME OF TEST COLOR REACTION NOTES
Cu+ and alkali are used
to treat cmpds w/ 2 or
Protein Biuret Test Purple color more peptide bonds
Negative test: Blue color
sol’n (fewer than two
peptide bonds)
a.k.a Glyoxylic Acid Test
Presence of indole Hopkins-Cole Reaction Violet Colored complex
group Indole group (like
typtophan)
Presence of White precipitate which hydroxyphenyl group
Millon’s Test
hydroxyphenyl group turns red upon heating (like in tyrosine)
Negative: Yellow color
(proline and
Detect free amino hydroxyproline)
Ninhydrin Test Deep blue color
group in amino acids.
 Proteins must be heated
first. React with alpha
amino group
Amino acids containing
sulfur like Methionine,
Amino acids containing Cysteine & Cystine
Brown or black color on lead
sulfur Sulfur Test
acetate paper Uses lead acetate
paper. Reaction
between H2S gas and
lead acetate paper
yields to lead (II) sulfide
Based on the ability of
Lemon-yellow solution upon aromatic ring to
heating undergo nitration
Characteristic reaction reaction by adding NO2
Xanthroproteic Test Orange color solution in
of proteins containing group to their structure
aromatic rings addition of alkali (NaOH)
Test positive:
phenylalanine, tyrosine,
and tryptophan
EXPERIMENT 7: IDENTIFICATION OF AMINO ACIDS BY PAPER CHROMATOGRAPHY
**
Procedure:
A. Preparation of Chromatography Chamber
B. Preparation of the Paper chromatogram
C. Experiment Proper
Aspartic Acid, Phenylalanine, Proline, Lysine, Unknown solution

EXPERIMENT 8: CHEMISTRY OF ENZYMES

**
Procedure:
A. Preparation of catalase
B. Preparation of amylase
C. Test for activity of catalase
D. Action of Amylase

TEST FOR ACTIVITY OF CATALASE

TEST TUBE COLOR REACTION
First test tube: 2mL catalase solution + 3mL *Glowing splinter = _______
hydrogen peroxide. Blue to green color change
Second test tube: 2mL catalase solution + 3mL *Glowing splinter = _______
sodium sulfide. After a minute, + 3mL 3%
 *+ 1mL 0.5% Benzidine: ________
hydrogen peroxide and mix quickly.

ACTION OF AMYLASE
TEST TUBE COLOR REACTION
First test tube: 5mL 2% starch solution + drops of *Blue color
iodine solution

Second test tube: 5mL starch sol’n + 10 drops

amylase sol’n. Warm at 50C for 2 mins. Do not * After +iodine: ________
boil.

Third test tube: 5mL 2% starch solution + 10

drops amylase sol’n + 10 drops ethanol. Warm * After +iodine: ________
the mixture for 5 mins.
EXPERIMENT 9: ENZYMATIC BROWNING OF FRUITS AND VEGETABLES
**
Procedure:
A. Preparation of soaking solutions
Ascorbic acid, Syrup, Salt solution, Calamansi, Plain water
B. Preparation of Sample

EXPERIMENT 10: CHEMISTRY OF LIPIDS

SOLUBILITY TESTS OF COCONUT OIL
TEST TUBE SOLUBILITY
Distilled water *Insoluble
Ethyl Alcohol * Insoluble
Ether * Soluble
Chloroform * Soluble
5% HCl * Insoluble
5% NaOH * Insoluble

REACTION TO IODINE TO TEST FOR UNSATURATED FATTY ACIDS

TEST TUBE COLOR REACTION
*
First test tube: 2mL CHCL + palmitic acid
3

Second test tube: 2mL CHCL3 + oleic acid *

Third test tube: 2mL CHCL3 + cottonseed oil *

TEST FOR RANCIDITY

TEST TUBE REACTION
 *
First test tube: Fresh coconut oil +
phenolpthalein
Second test tube: Fresh coconut oil + methyl *
orange

Third test tube: Fresh coconut oil + pH paper *

TEST FOR RANCIDITY

TEST TUBE REACTION
*
First test tube: Rancid coconut oil +
phenolpthalein
Second test tube: Rancid coconut oil + methyl *
orange

Third test tube: Rancid coconut oil + pH paper *

EMULSIFICATION OF OIL
TEST TUBE REACTION
First test tube: 2mL H2O & 6 drops olive oil *

Second test tube: 2mL H2O, 5 drops olive oil & 12 *

drops 10% Na2CO3

Third test tube: 2mL H2O, 5 drops olive oil, 5 *

drops oleic acid & 5 drops 10% Na2CO3
Fourth: 2mL H2O, 4 drops olive oil & 12 drops 2% *
egg white
Fifth: 2mL H2O, 5 drops olive oil & 12 drops *
saturated soap solution

Reactions with different Reagents: Lipids

Blue to violet color
Liebermann-Buchard Changes to emerald green upon
Cholesterol plus acetic anhydride addition of acetic anhydride and
Reaction
and concentrated sulfuric acid. concentrated sulfuric acid to a
chloroform solution
Sulfuric acid layer: yellow with green
Lipids Salkowski Test
fluorescence
Chloroform layer: bluish red
gradually turning to violet-red
 Test for Glycerol Acrolein Test Black color followed by evolution of
fumes. Note pungent odor.

EXPERIMENT 11: ANALYSIS OF BILE

**
Tests for bile
Test for Name of test Color reaction

Test for Bile salts Pettenkofer’s Test *Red color

Test for Bile pigment Fouchet’s Test *Green color due to oxidation of
bilirubin to biliverdin
Van der Bergh Test *Red color

Test for pH

EXPERIMENT 12: CHEMISTRY OF RNA

**
Procedure:
A. Preparation of RNA from yeast
B. Solubility of yeast RNA
C. Acid Hydrolysis
SOLUBILITY TESTS OF YEAST RNA
TEST TUBE SOLUBILITY/INTERPRETATION
Cold water *

Hot water *

Alcohol *

5% HCl *

1% NaOH *

ACID HYDROLYSIS OF RNA

Test for Name of test Color reaction

Test for pentose Bial’s Test Green to blue color within 5

 minutes

Test for purine base. *Formation of precipitate

Test for purine base
Hydrolyzate plus ammonia
water. Then add AgNO3

Reducing ability of sugar Benedict’s Test Red precipitate

Test for inorganic phosphate *Ammonium Molybdate Test *Yellow precipitate

EXPERIMENT 13: CARBOHYDRATE DIGESTION

TEST FOR SALIVA CONSTITUENTS

Test for Name of test Color reaction

Protein Biuret Test Violet-pink color

General test for Purple color

Molisch Test
Carbohydrates in free or
combined form

Test for inorganic phosphate 10% Ammonium Molybdate *Yellow precipitate

Test

Silver nitrate

TEST FOR SALIVA DIGESTION

Test for Name of test Color reaction

Test for Polysaccharides.

Differentiate helical Iodine Test Blue color Iodine form complexes
polysaccharides from non- with amylose
helical polysaccharides.

Reducing ability of sugar Benedict’s Test Red precipitate

DIGESTION OF STARCH BY PANCREATIC AMYLASE

Test for Name of test Color reaction
 Test for Polysaccharides.
Differentiate helical Iodine Test Blue color
polysaccharides from non-
helical polysaccharides.

Reducing ability of sugar Benedict’s Test Red precipitate

EXPERIMENT 14: PROTEIN DIGESTION

EXPERIMENT 15: LIPID DIGESTION