EXPERIMENT

DETERMINATION OF BETA-CAROTENE

Introduction
In the body, beta-carotene converts into Vitamin A (retinol). We need Vitamin A
for good vision and eye health, for strong immune system and healthy skin.
Beta-carotene is an antioxidant and it protects the body from damaging
molecules called free radicals. Free radicals damage cells through a process
known as oxidation. Over time, this damage can lead to number of chronic
illness. There is good evidence that eating more antioxidants from food boost
our immune system.

Principle
The amount of Beta-carotene the given unknown sample can be determined by
comparing the color of its known standard read at 540nm.

Requirements
Beta-carotene standard

Petroleum ether

Various food items (Carrot, pumpkin, etc.)

100ml Beakers

Burette and Pipette

Funnel

Filter paper

20x150 mm test tubes/test tube rack

Spectrophotometer or UV-VIS
Procedure

Standard concentration:

100mg of Beta-carotene is dissolved in 100ml of petroleum ether.

Preparation of Standard

Prepare the following solutions in five test tubes. Thoroughly mix each with
vortex.

Test Volume of Vol. of Volume of Concentratio Absorbance at

Tube working sample petroleum n (mg) 540 nm
standard extract ether (ml)
(ml) (ml)

Blank - - 3.0 -

S1 0.1 - 2.9 0.1

S2 0.2 - 2.8 0.2

S3 0.3 - 2.7 0.3

S4 0.4 - 2.6 0.4

S5 0.5 - 2.5 0.5

UK - Given Make up ?
volume according to
the given
volume

In series of test tube pipette out aliquots of working standard of Beta-carotene

with known concentration. Make up the tube volumes up to 3.0 with petroleum
ether and then read the content at 540nm in spectrophotometer against blank
reagents.

2
Preparation of food samples

1. Weigh out about 10 g of the solid food samples and make it slice.

2. Transfer the content into mortar and pestle and ground well.

3. Add 20ml of petroleum ether to the ground samples and dissolve.

4. Then filtered out the content using filter paper and use the filtrate as source

5. Pipette out 0.3 ml of filtrate to the unknown test tube and follow rest of the
procedure as per standard prepared.

Data Analysis

1. Prepare a standard curve (Beer’s Law) of the standard concentrations vs.

absorbance.

2. Use the standard curve to determine the Beta-carotene concentration of the

food samples to be tested.

Results

The concentration of Beta-carotene present in given sample is_____ mg/g.