Practical 12 - Estimation of sodium Benzoate in Food Sample

Aim: To estimate the sodium benzoate contents in fruit and vegetable products.

Theory and Principle: Sodium benzoate is used as a class II preservative in preservation

of pickles, sauces, squashes etc. Its estimation is important as the quantity of class II
preservative shall not exceed maximum limit prescribed under FPO. In a sample
containing NaCl, the benzoic acid is converted into water soluble sodium benzoate by
addition of NaOH. When the Sodium benzoate solution is acidified with excess of HCl,
water insoluble benzoic acid is formed which is extracted with chloroform. The chloroform
is removed by evaporation and the residue containing benzoic acid is dissolved in alcohol
and then titrated with standard sodium hydroxide.

Reagents, glassware and apparatus required

1. Sodium chloride (standard salt solution)

2. Sodium hydroxide 0.05 N: Dissolve 40.005g NaOH pellets in water
and dilute to one litre in a volumetric flask. Then dissolve 5 ml of the
above solution to 100 ml with water to make 0.05N NaOH.
3. H2SO4 and HCl,
4. Chloroform and Phenolphthalein indicator
5. Distillation flask and condensers, conical flask, volumetric flask,
pipette, burette and separating funnel.

General procedure for sample preparation:

1. Mix the sample (ketchup, sauce and pickle) thoroughly and grind.
Use 50-100g sample for estimation.
2. Transfer to 500ml volumetric flask.
3. Add powdered NaCl (15g) to saturate.
4. Make the solution alkaline to litmus paper by adding 10 % NaOH and
dilute to 500ml with saturated salt solution.
5. Shake thoroughly and allow standing for 2 hrs with frequent shaking.
Filter through Whatman No.4 paper or centrifuge.
6. For sample containing fat, extract with ether before determination.

Procedure for sodium benzoate estimation

1. Take 100 ml of filtrate in 500 ml volumetric flask.

2. Neutralize to litmus paper with dilute HCl (1+3) and add 5 ml of HCl
in excess.
3. Extract carefully by mixing the sample with chloroform in separating
funnel using 70 ml, 50 ml, 40 ml and 30 ml chloroform successively.
4. To avoid the formation of emulsion, shake the funnel continuously.
5. Draw off carefully clear chloroform solution after each extraction and
wash the chloroform layer with water.
6. Transfer the combined chloroform extract from the separating funnel
to a 250 ml conical flask. Rinse the funnel with chloroform.
7. Distill at low temperature to approximately ¼th of the original
volume and evaporate to dryness at room temperature on water bath
till only a few drops remain.
8. Dry the residue overnight in desiccator containing H 2SO4 (until no
acetic acid odour is detected in ketchup sample) and dissolve in 50 ml
alcohol.
9. Add 12-15 ml of water and 1-2 drops of phenolphthalein indicator
and titrate against 0.05 N NaOH.
10. Calculate using following expression and express as ppm sodium
benzoate or benzoic acid.