Practical 01

Date: 14th June 2014

Determination of salt content of brine

1.1 Introduction

Solutions of sugar and brine are used in preserving canned food. The shelf life of these foods
depends on the concentrations of the solutions used. Therefore, concentration of these salt and
sugar solutions is one of the important factors used in thee evaluation of their quality.

1.2 Literature review

“http://staff.buffalostate.edu/nazareay/che301/lab5.pdf” has indicated an introduction on titration

Mohr method of salt determination;

The AgNO3 solution (~0.02 M) needs to be standardized using NaCl as a primary standard. You
will perform standardization using Fajans method with adsorption indicator and using Mohr
method with chromate indicator. Both titrations are to be done in triplicate.

1.3 Experiments

1.3.1 Principle

58.5 × 𝑂. 1 × 𝑇
% m/v 𝑠𝑎𝑙𝑡 𝑖𝑛 𝑏𝑟𝑖𝑛𝑒 =
5

T = Mean titre of 0.1M AgNO3

1.3.2 Materials

Canned food samples

50 ml burette

1
10 ml and 25ml pipettes 250ml volumetric flask

250 ml titration flask

100 ml volumetric flask

Watch glass

Funnel

Chemicals

(AgNO3,Potassium chromate indicator)

1.3.3 Procedure

0.1M AgNO3 was prepared and stored in a dark place. Then 5ml of brine sample was pipetted
out and diluted with 250ml of distilled water by using volumetric flask.250ml of diluted brine
sample was taken and transferred into titration flask.1ml of potassium chromate was added as an
indicator in to the titration flask and titrated with 0.1 M AgNO3 until a distinct reddish brown
colour appears and persist on brisk shaking. Three replicates were taken to get the mean value.

1.4 Results and calculations

Serial No Burette Burette Result

Initial reading Final reading

1 7ml 8.7ml 1.7ml

2 10ml 11.7ml 1.7ml

3 16ml 17.6ml 1.6ml

2
% (m/v) salt in brine = 58.5 × 0.1 × T
5

T = Mean titer of 0.1 M AgNO3 = 1.7ml

% (m/v) salt in brine = 58.5 × 0.1 × T

5
= 58.5 × 0.1 × 1.7
5
= 1.99%

1.5 Discussion

As the indicator Potassium chromate was used. At the end point of this titration brownish colour
was appeared due to formation of AgCl by reacting with salt. But the colour was persisted due to
AgNO3 remaining at the end point because of no more salt to react.

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Practical 02

Date: 19th June 2014

Food colours

2.1 Introduction
A colour additive is any dye, pigment or substance that can impart colour when added or
applied to a food. Colour additives may be used in foods, drugs and cosmetics. Colour
additives ate used in foods for many reasons, including offsetting colour loss due to storage
or processing of foods and to accommodate variations on natural food colour. Colour such as
turmeric and saffron have been used in food from ancient times to attract consumers.
However, some additives are not suitable for consumption. Therefore, regulations are needed
impose to prevent the use of such additives as metallic coloured salts and non- permitted
coal-tar dyes. Only permitted colours which satisfy purity criteria ate now allowed in
specified foods. Raw or unprocessed meat, poultry, fish fruits and vegetable, tea, coffee,
bread, milk, dried milk, cheese and butter are not permitted to be incorporated with colour
additives. Colours are also prohibited to add baby foods. All food colours are acid soluble.

2.3 Experiments

2.3.1 Principle

Identification of colours was done by a paper chromatography technique after extracting them
using a suitable method.

2.3.2 Materials

100% pure wool thread

300 ml beakers

Water bath petroleum ether

Sample of soft drinks

4
2M acetic acid

2M ammonia solution

2% ammonia in 70% alcohol

Dye samples

Funnel and filter papers

2.3.3 Procedure

Dye sample was acidified using small amount of 2M acetic acid. Observation was recorded.
Beaker was cleaned and small amount of same dye was taken and 2M ammonia solution was
added. Observation was recorded. This procedure was repeated for the each dye sample given for
the experiment.

10g of Necto and Fanta sample were taken sufficient amount of petroleum ether was added into
it. It was mixed well and yellow colored solution was discarded. This was repeated until get a
colorless solution. 70ml of 2% ammonia in 70% alcohol was added to it and mixed well. The
mixture was warmed and starch was allowed to settle down. The mixture was filtered and
alcohol was removed from the filtrate using water bath. Small amount of sample was taken and
acidified using 2M acetic acid. The pure wool thread was dipped in to it. Observation was
recorded. Thread was washed with tap water. 2M NH4OH was added to release the dye from the
thread. Dye was concentrated by heating. Finally a paper chromatography was run with the
standard colors. As the solvent a mixture of Butanol, Acetic acid and Water was used with a ratio
of 20:5:12 to run the chromatography for Fanta and Necto sample.

5
2.4 Results and calculations

Result for food colours

Observation

Dyes Distance(cm) in Trial 01 Distance(cm) in Trial 02 Mean

Sunset yellow 1cm 1cm 1cm
Tartrazine 0.4cm 0.3cm 0.35cm
Fanta(Sample) 1.1cm 1.2cm 1.15cm
Green FCF 1.4cm 1.2cm 1.3cm
Carmoisine 1.8cm 2cm 1.9cm
Necto(sample) 1.5cm 2cm
0.3cm 0.3cm 2.05cm
Erythrosine 9cm 9cm 9cm
Green shade 3cm 3.3cm 3.15cm

Calculation

Rf = Distance of the colour substance

Distance moved by solvent colour

Rf(sunset yellow) = 1/9 =0.111

Rf(Tartrazine) = 0.35/9 = 0.038

Rf(Fanta sample) = 1.15/9 = 0.127

Rf(Green FCF) = 1.3/9 = 0.144

Rf(Carmoisine) = 1.9/9 = 0.211

Rf(Necto sample) = 2.05/9 = 0.227

Rf( Erythrosine) = 9/9 = 1

Rf(Green shade) = 3.15/9 = 0.35

6
2.5 Discussion

To extract the colours in soft drinks given, there was a standard procedure according to this
experiment. Before extracting the colours a purification steps were followed. To precipitate
starch and sugars ammonia and alcohol solutions were taken. Then colour compounds were
absorbed to a thread by acidifying the medium. The absorbed colours were related to a separate
container by making medium basic.

However, this method of extraction colours is useful in identification them with a

chromatographic technique. Here paper chromatographic technique was used. RF values were
not match exactly with any given standard colour compound.

In this chromatography as solvent front we use n-butanol, Acetic acid, water mixture in 4:1:5
ratio.

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Practical 03

Date: 12th July 2014

Determination of Sulphur Dioxide content of a food

3.1 Introduction

SO2 is used as food preservative world widely. It may be used in the form of a gas, in solution as
sulphours acid, or as the sulphites of sodium, potassium or calcium; but for the purpose of
regulations the mount present is calculated as SO2, sulphours acid inhibit the growth of moulds,
yeasts and aerobic bacteria and also prevents browning of fruits and vegetables due to enzymatic
reactions. It assists in conserving vit. C but inactivates vit. B by cleavage of the molecule.

3.2 Experiments

3.2.1 Materials

Round bottom flask (250ml)

Condenser
Receiving adaptor
Beaker (500ml)
Burette with stand
Pipette (10ml)
Heating mental Conc. HCl
0.05N Iodine solution
Starch solution
Pumic stones

3.2.2 Procedure

About 25ml of given sample was introduced into a double neck distillation flask. Some
antibumping granules were added into the flask and water was added to volume up to 200ml. the

8
still head of the condenser and a receiver adapter was connected to the distillation flask and
dipped the receiving adapter into a beaker containing about 200ml distilled water and few drops
of starch indicator. Then 20ml of conc HCl was added to the distillation flask using the other
neck and close it quickly. The flask was heat in direct heating. 0.05 N iodine solution from the
burette was added into the distillate in the beaker in sufficient amounts to oxidize the sulphur
dioxide as it distills over maintaining a pale blue colour throughout the distillation. The
distillation and titration was continued until the pale blue colour persists for at least due to
0.10ml of iodine solution. Finally the burette readings were taken.

3.3 Results and calculations

Initial Burette reading (ml) Final Burette reading (ml

20.0 16.2

Amount of iodine consumed to for the reaction – (20.0 – 16.2) = 3.8 ml

1.0 ml of 0.05N iodine solution = 0.0016g of SO2
If the amount of sample is X ml

Titer x 0.0016 x 106

The amount of sample in ppm =
X
3.8 x 0.0016 x 106
The amount of sample in ppm =
25
= 243.2 ppm

3.4 Discussion

The maximum allowable strength of sulphur dioxide is 50 ppm. Cordial is consumed after
diluting with water in a ratio of 1:4. Therefore maximum amount of Sulphur dioxide that can be
present in cordial sample is 250ppm. But according to our experimental results SO2 content is
243.2 ppm. So the SO2 content is higher and may be due to many reasons; bad processing
conditions, combination of other preservatives such as benzoic acid with SO2, experimental
errors such as dissolving of SO2 in water and also inability of collecting all the gas.

9
In this practical for the extraction of SO2 from the food sample we acidify it by adding conc HCl
and heated.

-Na2S2O5 + H2O 2NaHSO3

-2NaHSO3 + H+ 2Na+ + SO2 + H2O

SO2 reacts with iodine and reduce iodine into iodide (I-) while oxidize SO2 to SO4. Therefore the
blue color complex which forms with starch and iodine will disappear when iodine converts to
iodide.

10
Practical 04

Date: 05/07/2014

Determination of Benzoate content of Fruit Juice (Cordial)

4.1 Introduction

Derivatives of benzoic acid are used as a preservative in some foodstuffs as fruit juices, diabetic
jams, beer etc. preservative means a substance which when added to food is capable of
inhibiting, retarding or arresting the process of fermentation, acidification o other decomposition
of food. The preservatives regulations permit any preservative to be present only within the
prescribed limits. Benzoic acid added to a foodstuff can be directly extracted from it using
diethyl ether but it should be collected by gentle evaporation of ether as benzoic acid sublimates,
sublimation is carried out using a specially designed apparatus made of metal, which keeps on a
water bath with a well- fitted small beaker while keeping the sample with benzoic acid in that
beaker.

4.2 Literature review

Food Act No. 26 of 1980, REGULATIONS made by the Minister of Health in terms of section
32 of the Food Act No.26 of 1980 in consultation with the Food Advisory Committee has been
indicated the maximum level of benzoate that can be used with cordial with 25% minimum fruit
juice. According to that it is 600ppm.

4.3 Experiments

4.3.1 Materials

Raw materials Equipment & Instrument/

Glass ware Chemicals/ Reagents Sample of Cordial

11
Beakers (100ml, 250 ml)

Measuring cylinders (25 ml)

Filter funnel, Standard and Filter papers

Water bath

Desiccators

Burette and Burette stand

Metal holder

round bottom flask Acetone

Diethyl ether

Conc. HCl

Anhydrous Sodium sulphate

0.05M NaOH

Phenol red

4.3.2 Procedure

Around 10 g of cordial sample was weighed in to 100ml beaker and it was mixed with 10ml
distilled water. Then 10ml of conc. HCl was added to the above mix. This acidified mixture was
transferred to a 250ml diethyl ether to it and mixed well with a glass rod. Then the diethyl ether
layer was decanted to another beaker. The same sample mixture was extracted with two more
25ml ethyl ether portions and collected. The diethyl ether was filtered through anhydrous sodium
sulphate. The diethyl ether layer was evaporated up to 10ml in a low temperature to prevent
sublimation of benzoic acid. The whole apparatus was kept in a water bath for about 20 minutes.
The round bottom flask was taken out and cooled about 15 minutes in a desiccator and dissolved
it in 2.0ml acetone and 2.0ml distilled water. Finally it was titrated with 0.05M NaOH using
phenol red as the indicator.

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4.4 Results and calculations

Weight of the Sample (g) Titration Reading (ml)

10.12 g 0.18 ml

1 ml of 0.05M NaOH = 0.0061 g of benzoic acid

Amount of Benzoic acid in = Titer x 0.0061 x 106
The X g of sample X

= 0.18 ml x 0.0061 x 106

10.12 g
= 108.5ppm

4.5 Discussion

According to the food act the maximum allowable level of benzoate is 600ppm. So according to
the result given by this practical the product is having a lower level of its maximum allowable
usage.

Although benzoate is used as a preservative, it is carcinogenic. So this method is useful to find

the risk presence of benzoate in foods.

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Practical 05

Date: 28/06/2014

Analysis of vitamin C content in lime juice.

5.1 Introduction

Vitamin C (ascorbic acid) is commonly found in fruits such as citrus fruits, pineapple,
strawberry, black current and vegetables such as tomatoes, pea, cabbage, cauliflower and carrots.
Physiologically ascorbic acid is involved in number of reactions, particularly in the formation of
collagen, the connective tissue protein. Therefore deficiency of vitamin C causes several diseases
including scurvy (failure of normal connective tissue formation) which later leads to internal
bleeding, painful joints and failure of wound healing. Ascorbic acid plays an important role in
the absorption of the iron and its transport in the blood stream. Normal cooking, processing and
storage can destroy ascorbic acid found in vegetables and fruits.

5.2 Literature review

http://www.vitamincfoundation.org/usda.html indicates the vit C content of different food

categories. According to that the vit C content of lemon juice is 46mg/gram.

5.3 Experiments

5.3.1 Materials

Lime

Knife and cutting boars

250 ml conical flasks 2

25 ml measuring cylinder 1

100 ml measuring cylinder 1

14
Burette with stand petridish

Motar and pestle

Filter funnel

Filter papers

Chemicals; indophenol dye solution

3% metaphosphoric acid

5.3.2 Procedure

First of all, 10ml of the standard vitamin C solution was pipetted into a conical flask and added
10ml of metaphosphoric acid solution. Then the dye solution was titrated from the burette until a
faint red colour persists for at least 15 seconds. By using the reading, the milligrams of vitamin C
equivalent to 1ml of the dye solution was calculated.

This factor was used to calculate the vitamin C content of the sample in milligrams per 100
grams or 100ml of sample. (The vitamin C concentration in the standard solution is 0.40mg/ ml)

5.4 Results and calculations

Result for vit c

Observation
Standard solution

Initial burette Final burette reading

Serial No reading Result
1 0.15ml 1.6ml 1.45ml
2 3.2ml 4.6ml 1.4ml
3 4.7ml 6.0ml 1.3ml

15
Mean value = 1.4ml

Sample solution

Initial burette Final burette reading

Serial No reading Result
1 7.0ml 7.85ml 0.85ml
2 8.0ml 8.85ml 0.85ml
3 11.0ml 11.8ml 0.80ml

Mean value = 0.85ml

Calculation
Standardization of dye solution
Vit C concentration of the standard solution= 0.40mg/ml
10 ml of std solution contain = 0.4×10mg of Vit C
=4mg of Vit C
Mean Volume of dye in standardization titration =1.4ml
1ml of dye solution equivalent to =4mg/1.4ml
=2.86 mg of Vit C

Estimation of Vit C content of the sample

Mean volume of titration = 0.85ml of Dye
10 ml of extract contain =0.85×2.86mg of Vit C

100
100ml of extract contain = 0.85 × 2.86 × 𝑚𝑔 of Vit C
10

100 100
100g of sample contain =0.85 × 2.86 × × 10 of Vit C
10

=243.1 mg per 100g of sample

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5.5 Discussion

The results taken from this practical is bit lower than the literature. The possible reasons may be
errors in measuring the sample with measuring cylinder and chemical instability of vit C.
although meta phosphoric acid was added to stabilized the vit C, it is readily lost with vigorous
handling.

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