Food analysis lab - 1 – 14FP 2004

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Ex. No. 8 ESTIMATION OF ASCORBIC ACID CONTENT IN FOODS

OBJECTIVE
To estimate the total ascorbic acid or vitamin C content in the given food sample.

EQUIPMENTS
Test tubes, volumetric flask, Pipette, Micro Burette, Conical flask, Filter paper

REAGENTS
1) 3% Meta phosphoric acid (HPO3)

Weigh 15 gm of HPO3 crystals and dissolve in distilled water.

2) Ascorbic acid standard:
Weigh accurately 100 mg of 1- ascorbic acid and make up the volume to 100 ml. with 3%
Metaphosphoric acid.
3) Dye solution:
Dissolve 50 mg of sodium salt of 2, 6 -Dichlorophenol indophenol in about 150 ml of hot
distilled water containing 42 mg of sodium bicarbonate. Cool and dilute with water to 200
ml. and store in a refrigerator. Standardize every day.
4) 40 % Formaldehyde.
5) Conc. HCl

DESCRIPTION
Ascorbic acid reduce the 2, 6 Dichlorophenole indophenol dye to a colorless leuco –base.
The ascorbic acid gets oxidized to dehydroascorbic acid. Though the dye is blue colored
compound, the end point is the appearance of pink color. The dye is pink colored in acid
medium.

PROCEDURE
a) Standardization of Dye:
Take 5 ml. of standard ascorbic acid solution and add 5 ml. of HPOз. Fill a micro
burette with dye. Titrate with the dye solution to a pink color which should persist for 15
seconds. Determine the dye factor i.e., mg of ascorbic acid per ml of the dye, using the
formula,

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Food analysis lab - 1 – 14FP 2004
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Dye factor = 0.5 / titer value

b) Preparation of the sample:

Take 10 g of sample blends with 3% HPOз and make up to 100 ml. with HPOз and
filter.
c) Assay of extract:

Take an aliquot (2-10 ml.) of HPOз extract of the sample and titrate with the standard
dye to a pink end point which should persist for at least 15 seconds. Titrate rapidly and make
a preliminary determination of Titrate. In the next determination, add most of the dye
required and then titrate accurately. The aliquot of sample should be such that titer value
should not exceed 3 to 5 ml.

d) Elimination of interference due to SO2:

SO2 when present in sample reduces the indophenols dye and thus interferes in
ascorbic acid analysis. If the sample contains SO2, eliminate the interference by following the
formaldehyde condensation procedure given below to 10 ml. of the filtrate in the test tube,
add 1 ml. of 40 % formaldehyde and 0.1 ml. of HCl keep for 10 min. and titrate.

CALCULATION
Ascorbic Acid (mg / 100g) = Titer value × Dye factor × Volume made up × 100

Aliquot of Extract × Weight of sample

RESULT
Amount of ascorbic acid present in given sample =----------mg/100g.

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