Color Reactions of Myoglobin

Objectives:
To demonstrate some of the reactions of a heme pigment.
To understand the effect of ascorbic acid to the protein.
Materials:

25g Beef
100mg Sodium Nitrate
100mg Sodium hydrosulfate or sodium dithionite
100mg Ascorbic Acid

Procedure:
1. Grind fresh meat.
2. Homogenize meat in 100ml of cold water. Keep cold (4C) while
blending. Generally, 25 grams of ground beef in 100ml of cold
water will produce enough supernatant for all the tests required.
3. Centrifuge homogenate at 1000 X g for 10 minutes
4. Decant supernatant and filter through glass wool.
Treatments:
Prepare two 5ml samples of each of the specific chemical
treatments. One sample of each should be placed in a hot water bath
(75C) for 1 hour, the other held at room temperature.
1. Color reactions in fresh meat:
a. 30 mg sodium hydrosulfite or sodium dithionite
b. Supernatant only
c. 15mg NaNO2
d. 50mg NaNO2
2. Color reactions in cured meat:
a. 30mg Ascorbic Acid
b. Add 15mg NaNO2 and mix, then add 30 mg
ascorbic acid
In each case note the initial color after addition of the reagents and
any color changes following mixing and following the start of heating.
Observe against a white light if necessary to see color changes. Record
the water bath temperature.
Guide Questions:
1. What types of reactions are responsible for the reactions?

2. What are the effect of the pigments use?

Determination of the concentration of vitamin C in fruits/vegetables by using the

DCPIP test
Objective:
Materials:
Procedure:
Guide Questions: