EXPERIMENT #9: BLOOD

I. Objectives
a. explain the results of different chemical tests for blood; and

b. describe the crystals obtained from a blood sample

II. Material

BEAKER EVAPORATING DISH PIPETTE

WATER BATH MICROSCOPE GLASS SLIDE

III. MSDS
 Chemical Chemical Molecular Melting Boiling point Hazards
name formula weight point
Benzidine  
C12H12N2 184.24 g/mol 120 deg cel 401 deg cel Cause eye
and skin
irritations
Hydrogen Solution N/A 108 degl cel -33 deg cel Corrosive,
Peroxide may cause
fire and
burns
Sodium NaCl 58.44 g/mol 801 deg cel 1,456 deg cel Skin and eye
Chloride irritation

Acetic acid C2H4O2 60.06 g/nol 17 deg cel 118 deg cel Skin
irrritations
Guiac C15H26O 222.37 g/mol 40-50 deg cel 92 deg cel Respiratory
irritations
Diethyl ether (C2H5)2O 74.12 g/mol -116.3 deg 34.6 deg cel Eye unjury
cel and burns

IV. Procedure
I. Chemical test for blood

A. Benzidine Test

1.To 1 mL of Benzidine reagent, add 10 drops

of diluted blood.

2.Add 5 drops of 3% hydrogen peroxide.

3.Observe the result.

B. Hemin Test
1.Place a drop of blood on a glass slide.
2.Add a drop of water, stir, and spread uniformly by using the tip of a cover glass.
3.Add a drop of 0.1% NaCl solution.
4.Dry over a low flame.
5.Place cover glass in position and add underneath a drop of concentrated acetic acid.
6.Warm as before until bubbles are formed.
7.Cool and examine the crystal under the microscope.

C. Guiac Test

1.To 1 mL diluted blood, add new drops of guiac

solution, until the resulting solution becomes turbid.

2.Add 1% H2O2dropwise until a color reaction

appears.Test each test tube with blue litmus paper. In
what test tube do you notice a change in color of the
litmus paper? Into what substance are fats
hydrolyzed before they can be utilized by the body?

D. Confirmatory Test

1.Acidify 2 mL of dilute blood with glacial acetic acid.

2.Add 5 mL ether and shake the solution.

3.Separate the layer by means of clean, dry pipette, and

transfer it to a small evaporating dish and heat the dish over
a hot water bath without flame. Heat the ether extract to
dryness. (CAUTION: Ether is highly flammable; the drying
process should be done without flame.)

4.Add a drop of 3% hydrogen peroxide.