Ahmed Oil and Ghee Industries (pvt) LTD QP-001

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Table of Contents
Sr. Method Page No:
No:
1 FREE FATTY ACID IN FATS AND OILS 3
2 PEROXIDE VALUE 4
3 NEUTRAL OIL IN SOAP STOCK 5
4 HELPHEN TEST (Canola) 6
5 IODINE VALUE 7
6 QUALITATIVE ESTIMATION OF MINERAL OIL 10
7 SOAP CONTNTENTS OF REFINED OIL 11
8 SAPONIFICATION VALUE 12
9 MOISTURE AND VOLATILE MATTER (HOT PLATE METHOD) 13
10 MOISTURE AND VOLATILE MATTER (AIR OVEN METHOD) 14
11 COLOUR DETERMINATION 15
12 CLOUD TEST 17
13 COLD TEST 18
14 VITAMIN TEST 19
15 ACIDITY OF BLEACHING EARTH 20
16 BLEACHABILITY OF BLEACHING EARTH 21
17 QUALITATIVE ESTIMATION OF VITAMIN E 22
18 TITRE TEST 23
19 SPECIFIC GRAVITY OF OIL 25
20 DETERIORATION OF BLEACHING INDEX OF CPO 27
21 UNSAPONIFIABLE MATTERS 28
22 REFRACTIVE INDEX 30
23 QUANTITATIVE ESTIMATION OF VITAMIN A 31
24 INSOLUBLE IMPURTIES 33
25 QUALITATIVE ESTIMATION OF NICKEL 34
26 DETERMINATION PHOSPHOROUS 35
27 DETERMINATION OF IRON IN FATS 37
28 TURBIDITY TEST 39
29 DETERMINATION OF OIL & GREASE 40
30 DETERMINATION OF SOLID FATS CONTENT BY PULSE NMR 41
31 DILATATION OF FAT 43
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32 GASS ANALYSIS 45
33 IDENTIFICATION OF KETONE IN GROUP 46
34 IDENTIFICATION OF NITROGEN GAS 47
35 DETERMINATION OF TRANCE METALS IN FATS (IRON,COPPER,NICKLE) 48
36 HARDNESS OF WATER 50
37 DETERMINATION OF CHEMICAL OXYGEN DEMAND(COD) 52
38 DETERMINATION OF TOTAL SUSPENDED SOLID (TSS) 54
39 QUALITATIVE & QUANTATIVE ESTIMATION OF PHOSPHORIC ACID 55
40 QUALITATIVE & QUANTATIVE ESTIMATION OF CITRIC ACID 56
41 QUALITATIVE & QUANTATIVE ESTIMATION OF BUTYRIC ACID 58
42 QUALITATIVE & QUANTATIVE ESTIMATION OF SULPHURIC ACID 59
43 QUALITATIVE & QUANTATIVE ESTIMATION OF HYDROCHLORIC ACID 60

44 QUALITATIVE & QUANTATIVE ESTIMATION OF MONO ETHANOL AMINE 61

45 QUALITATIVE & QUANTATIVE ESTIMATION OF SODIUM HYDROXIDE 63
46 QUALITATIVE ESTIMATION OF ACETIC ACID 65
47 IDENTIFICATION OF TERT. BUTYLE HYDRO QUIONE (TBHQ) 66
48 DETERMINATION OF FREE CHLORINE 67
49 IDENTIFICATION OF SULPHITE ION 69
50 TESTING AND MEASUREMENTS ( TIN-CONTAINERS ) 70
51 TESTING AND MEASUREMENTS (CARTON) 71
52 TESTING AND MEASUREMENTS ( PLASTIC BOTTLE ) 72
53 INSPECTION & TESTING METHOD OF FILM ROLLS 73
54 TESTING PLASTIC HANDLE 74
35 DETERMINATION OF TRANCE METALS IN FATS (IRON,COPPER,NICKLE) 48
36 HARDNESS OF WATER 50
37 DETERMINATION OF CHEMICAL OXYGEN DEMAND(COD) 52
38 DETERMINATION OF TOTAL SUSPENDED SOLID (TSS) 54
39 QUALITATIVE & QUANTATIVE ESTIMATION OF PHOSPHORIC ACID 55
40 QUALITATIVE & QUANTATIVE ESTIMATION OF CITRIC ACID 56
41 QUALITATIVE & QUANTATIVE ESTIMATION OF BUTYRIC ACID 58
42 QUALITATIVE & QUANTATIVE ESTIMATION OF SULPHURIC ACID 59
43 QUALITATIVE & QUANTATIVE ESTIMATION OF HYDROCHLORIC ACID 60
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44 QUALITATIVE & QUANTATIVE ESTIMATION OF MONO ETHANOL AMINE 61

45 QUALITATIVE & QUANTATIVE ESTIMATION OF SODIUM HYDROXIDE 63
46 QUALITATIVE ESTIMATION OF ACETIC ACID 65
47 IDENTIFICATION OF TERT. BUTYLE HYDRO QUIONE (TBHQ) 66
48 DETERMINATION OF FREE CHLORINE 67
49 IDENTIFICATION OF SULPHITE ION 69
50 TESTING AND MEASUREMENTS ( TIN-CONTAINERS ) 70
51 TESTING AND MEASUREMENTS (CARTON) 71
52 TESTING AND MEASUREMENTS ( PLASTIC BOTTLE ) 72
53 INSPECTION & TESTING METHOD OF FILM ROLLS 73
54 TESTING PLASTIC HANDLE 74

FREE FATTY ACID IN FATS AND OILS

1. DEFINITION:
1.1 The %FFA is the amount of NaOH necessary to neutralize the acid, calculated by wt. of fatty acid of specific
molecular weight.

2. SCOPE:
2.1 Applicable to crude, refined vegetable and marine oils & animal fats.

3. APPARATUS:
3.1 Oil sample bottles, 115 or 230 ml. (4 or 8 oz. ) or 250 ml, conical flasks.
3.2 Burette, 50 ml.

4. REAGENTS:
4.1 Ethyl alcohol 95% (U . S . S. D . Formulas 30 and CA are permitted ). The alcohol must give a definite,
distinct and sharp and point with phenolphthalein and must be neutralized with alkali to a faint but
permanent pink color just before using.
4.2 Phenolphthalein Indicator solution, 1% in 95% alcohol.
4.3 2N Sodium Hydroxide stock solution, 20 gm/ 250 m distilled water.
4.4 0.05N NaOH (25 ml of 2N NaOH/1000 distilled water)

5. PROCEDURE:
5.1 Sample must be well mixed and entirely liquid before weighing.
5.2 Weight 10 grams of sample in conical flask.
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5.3 Add 50 ml of hot neutralized alcohol and 0.5 ml of 'indicator.

5.4 Titrate with alkali shaking vigorously to the appearing of first permanent color of the
same intensity as that of the neutralized alcohol before addition of the sample. The color must persist for
30 second.

6. CALCULATION:
6.1 The percentage of free fatty acid in most types of fats and oil is calculate as Oleic acid, although in Coconut
and palm Kernel oils it is frequently expressed as lauric acid and in palm oil in terms of Palmitic acid.
𝐕 × 𝐌𝐖 × 𝐍 × 𝟏𝟎𝟎
% 𝐅𝐅𝐀 =
𝐖𝐭: × 𝟏𝟎𝟎𝟎

V = Volume of NaOH used

N = Normality of NaOH
Wt: = Weight of sample
MW = Molecular weight of fatly acid
Oleic Acid = 282 gms Palmitic
Acid = 256 gms
Lauric Acid = 200gms

7. REFERENCE:
7.1. American Oil Chemists Society/ AOCS . Ca 5a - 40
PEROXIDE VALUE

1. DEFINITION
1.1 This method determines all substances, in terms of milli equivalents of peroxide per 1000 grams of
sample, which oxidize potassium iodide under the conditions of the test. These are generally assumed to
be peroxides or other similar products of fat oxidation.

2. SCOPE:
2.1 Applicable to all normal fats and oils including margarine. This method is highly empirical and any
variation in procedure may result in variation in results.

3. APPARATUS:
3.1 Pipette, Mohr, measuring solution, 1-ml capacity
3.2 Conical flasks, 250 ml.

4. REAGENTS:
4.1 Acetic acid chloroform solution. Mix 3 parts by volume of glacial acetic acid, reagent grade, with 2 parts by
volume of chloroform.
4.2 Potassium iodide solution, saturated solution of KI, in recently boiled distilled water. Make sure the
solution remains saturated as indicated by the presence of undissolved crystals. Store in the dark. Test
daily by adding 2 drops of starch solution to 0.5 ml of the potassium iodide solution in 30 ml of acetic acid
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chloroform solution. If a blue color is formed which requires more than I drop of 0. I N sodium thiosulfate
solution to discharge, discard the iodide solution and prepare a fresh solution.
4.3 Sodium thiosulfate solution, 0.1 N, accurately standardized.
4.4 Sodium thiosulfate solution, 0.005 N, accurately standardized. This solution into a 250 ml volumetric flask
and diluting to volume with recently boiled distilled water.
4.5 Sodium thiosulfate solution, 0.01 N, accurately standardized. This solution may be prepared by accurately
pipette out 25 ml of the 0.I N solution into a 250 ml volumetric flask and diluting to volume with recently
boiled distilled water. Starch indicator solution. 1.0% of soluble starch in distilled water.

5. PROCEDURE:
5.1 Weigh 5.0 gm of sample into a 250 m conical flask and then add 30 ml of Acetic acid chloroform solution.
Swirl the flask until the sample is dissolved in the solution. Add 0 5 ml of saturated potassium iodide
preferably using Mohr Type measuring pipette.
5.2 Allow the solution to stand in dark for exactly one minute and then add 30 ml of distilled water.
5.3 Titrate with 0.01 N sodium thiosulfate adding it gradually and with constant and vigorous shaking counties
the titration until the yellow color has almost disappeared. Add Ca. 0.5ml of starch indicator solution.
Continue the titration, Shaking the flask vigorously near the end point to color has just disappeared.

6. CALCULATION:
Peroxide value ( milli-equivalents peroxide / 1000 g sample ) = (𝐒−𝐁)×𝐍×𝟏𝟎𝟎0/ 𝐖𝐭:𝐨𝐟 𝐒𝐚𝐦𝐩𝐥𝐞

Where
B = Volume of titration ml of blank
S = Volume of titration ml of sample
N = Normality of sodium thiosulfate solution.

7. REFERENCE: 7.1. American Oil Chemists Society/ AOCS Official Method

NEUTRAL OIL IN SOAP STOCK

1. DEFINITION:
1.1 This method determines the total neutral oil, i.e. unsaponified oil. Any unsaponifiable fatty matter in the
sample will be oil included

2. SCOPE:
2.1 Applicable to all type of soap stock.

3. APPRATUS:
3.1 Separating funnel 500 ml
3.2 Flat bottom flask 250 ml.

4. REAGENTS:
4.1 14% W/V KOH Solution in distilled water.
4.2 Ethyl alcohol (Ethanol) 50% ( Mix 50 ml of 95% alcohol with 50 ml Distilled water).
4.3 Petroleum ether.
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5. PROCEDURD:
5.1 Weigh 8 to 10 gms of the well mixed sample 4-5 gms. Acidulated soap stock ). Into separating Funnel.
5.2 Add 100 ml of 50% alcohol and 50ml of petroleum ether, agitate gently until thoroughly mixed.
5.3 Cool to 20 to 25 C and add 10 m! of the aqueous KOH. Agitate gently until thoroughly mixed but avoid
vigorous shaking.
5.4 Add 25 ml of 50% ethyl alcohol and shake gently until thoroughly, mixed.
5.5 Allow to until the two layers are completely separated. Do not allow the ether oil layer to remain in
contact with the alcohol alkali layers longer than 30 minutes.
5.6 Take ether oil into another 500 ml separating funnel. Make at least 4 more similar extraction, using 50 m!
of pet. Spirit for each extraction. Combine all of the extracts the separatory funnel.
5.7 Wash the ether extract in the separatory funnel with 25 ml portion of 10% alcohol, shaking vigorously
each time until the washings are neutral to phenolpthaline. Tree washing are usually sufficient.
5.8 Filter through filter paper free from fatty matter with pet ether. Wash the paper free from fatty matter
with petroleum ether.
5.9 Draw the ether extract into the weighed flat bottom flask which has been previously dried and cooled in
a desiccator.
5.10 Evaporate the ether on a water bath under a gentle steam of clean dry air. Dry in an air oven at 105°C +2
for 30 minutes.
5.11 Cool in a desiccator to R.T. weight repeat until constant weight is obtained.
5.12 Constant weight is attained when the loss (or –again) in weight does not exceed 0.1% in successive 30
min drying periods.

6. Calculation:
𝐌𝐚𝐬𝐬 𝐨𝐟 𝐎𝐢𝐥 𝐠𝐦𝐬 ×𝟏𝟎𝟎
Neutral Oil % =
𝐌𝐚𝐬𝐬 𝐨𝐟 𝐬𝐚𝐦𝐩𝐥𝐞 𝐠𝐦𝐬

7. REFERENCE:
7.1 American Oil Chemists Society/ AOCS Official Method

HELPHEN TEST

1. DEFINITION:
1.1 This method detects, qualitatively, the presence of cotton seed oil in vegetable, animal fats or oils.

2. SCOPE:
2.1 Kapok oil reacts with an intensity equal to or greater than that of cotton seed oil and fats from animals
which have been fed cotton seed meal many give a positive result.

3. APPARATUS:
3.1 Test tubes, 200-250 × 25 mm
3.2 Oil or saturated brine bath capable of being regulated at 110 to 115 °C.
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4. REAGENTS:
4.1 Prepare a solution of 1-5 % sulfur in carbon disulphide and then add an equal volume of amyl alcohol.

5. PROCEDURE:
5.1 Mix 10 ml liquid in a test tube with an equal volume of the reagents.
5.2 Shake and heat gently in hot water (70-80)°C for a few minutes with occasional shaking until the carbon
disulphide is boiled off and the sample stops foaming.
Caution: Carbon disulphide vapors may be ignited with a hot bath or steam pipe.
5.3 Place the tube in the 110-115 °C bath and hold for 1-2 hrs. A red colour at the end of this period indicates
the presence of cotton seed oil.
5.4 If appreciable quantities of cotton seed oil are present, a positive reaction will be obtained in one hour or
less in a water bath at or near the boiling temperature. For small amounts a 2 hrs time interval may be
necessary at the temperature of 110-115°C .

6. REFERENCE:
6.1 American Oil Chemists Society/ AOCS Official Method Official Method CB 1 - 25

IODINE VALUE

1. DEFINITION:
1.1 This iodine value is a measure of unsaturation of the fatly acids and it expressed in terms of the number
of centigrams of iodine absorbed per grams of samples

2. SCOPE
2.1 Applicable to all oils, fats soap and soap products except those made of fats containing conjugated
system.

3. APPRATUS
3.1 Glass stoppered volumetric flask, conforming to burette of (25 ml)
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3.2 Pipette (Bulb) 25 ml A grade

3.3 Graduated pipette (5 ml)
3.4 Measuring cylinder (25 ml)

4. REAGENTS
4.1 Glacial acetic acid. The permanganate should be applied to be sure that this Specification to be met.
Test: Dilute 2 ml of the acid with 10 ml of distilled water and add 0.1ml of 0.1N KMnO4.The pink color must not
be entirely discharged within 2 hrs.
4.2 Potassium iodide
4.3 Carbon tetrachloride 4.4 Hydrochloric acid
4.5 Soluble starch.

TEST OF SENSITIVITY:
4.3.1 Make a paste with I gm of starch and small amount of cold distilled water
4.3.2 Add, while stirring, 200 ml of boiled water.
4.3.3 Place 5 ml of this solution in 100 ml of water and add 0.05 ml of 0.1 N iodine.
4.3.4 The deep blue color produced must be discharged by
0.05ml of 1N sodium thio-sulfate.

4.6 Potassium dichromate. The K2Cr2O7, is7 is finely ground and dried to constant weight Ca 110 °C .
4.7 Sodium thio-sulfate (Na2S2O3)
4.8 Iodine trichloride
4.9 Mercuric acetate

5. SOLUTION:
5.1 Potassium iodide solution( 15% in distil water)
5.2 Starch sol ( 1% in boiling distill water)
Salicylic acid (1.25 gms/ltr) may be added to preserve the indicator or be kept in a refrigerator.
5.3 Standard potassium dichromate solution. Dry and weight 4.9035gms of K2Cr2O7 in 1000 ml of
volumetric flask and make to volume at 25 °C.
5.4 Sodium thio-sulfate sol. Dissolve 24.8gms/1000 ml distilled water.

STANDARDIZATION:
5.4.1 Pipette out 10ml standard dichromate solution in to a flask.
5.4.2 Add 5 ml of HCI (dil) and 10ml of potassium iodide sol and rotate to mix.
5.4.3 Allow to stand for 5 minutes in dark and add 100 ml of distilled water.
5.4.4 Titrate with sodium thiosulfate the yellow color has almost disappeared.
5.4.5 Add 0.5ml of indicator and continue titration adding thiosulfate soln. Slowly, until the blue
color has just appeared.
5.4.6 The strength of the Sodium thio-sulfate soln Is expressed in terms of its normality.

𝐍𝟐×𝐕𝟐
N1 =
𝐕𝟏
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N1 = Normality of Na2S2O3
N2 = Normality of K2Cr2O7 = 0.1N
V1 = Volume of K2Cr2O7 = 10 ml
V2 = Volume of Na2S2O3

5.5 Wij's SOLUTION.

5.5.1 Dissolve 7.8 gms iodine trị chloride in 1000 ml of glacial acetic acid.
5.5.2 Add 8.5 gm iodine crystals.
5.5.3 KI solution (15 % KI in distilled water).
5.5.4 Starch Solution 1%
5.5.5 Sodium thiosulphate 24.8 gm in 1000 ml distilled water (0.1N).

6. PROCEDURE:
6.1 Oil and fat to be tested is remove any moisture and then is filtered. The temperature should not exceed
15 °C above the titer point of fatty acids.
6.2 Weight the sample accurately in to 250 ml flask.

The weight of sample must be such that there will be an excess or wij, s sol of 50-60%of The amount added, i.e. 100 to
150 % of the amount obsoded.
Table-1 is a convenient guide to the size of sample to weight.

6.3 Add 20 ml of CCI4 to dissolve the sample. Pipette out 25 ml of the Wij's soln into flask and swirl to insure
and intimate mixtures. Stopper the flask and store in a dark place for 30 minutes.
Holman & Green Modification

(If 10 ml of Hofman and Green reagent) is added to the flask then the reaction time is reduced 3 minutes).

6.4 Remove the flask from dark place and add 20 ml of KI (10%) solution and 50 ml dist. water. Titrate with 0.
IN Na2S2O3. Solution adding it gradually and with constant and vigorous shaking continue the titration
until the yellow color has disappeared. Add 0.5 ml of starch solution and continue the titration until the
blue color has just disappeared.

7. CALCULATION

(B−S)×N×EW×100
The iodine value =
1000×W

B = Titration of blank
C = Titration of sample
N = Normality of Na2S2O3 solution
W = Wt. of sample
EW = Eq Wt. of sample = 126.9
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8. NOTE:
8.1 Store samples with an iodine value above 150 for 1hr in a dark place at 25±5°C.
8.2 Mechanical stirring is very satisfactory for agitating during the addition of thiosulfate. Table-1
S.No Iodine Value Sample Weight (Gms) Accuracy (Gms)
1 3 10.58-8.46 +5
2 10 3.17-2.54 0.2
3 20 1.59-1.27 0.2
4 40 0.79-0.63 0.2
5 80 0.40-0.32 0.2
6 120 0.26-0.21 0.2
7 160 0.20-0.16 0.2
8 200 0.16-0.13 0.2

9. REFERENCE:
9.1 American Oil Chemists Society/ AOCS Da 15 – 48 CD 1 – 25
9.2 Laboratory Handbook for Oil & Fat Analysis by L.V Cock on C. Van Rede.
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QUALITATIVE ESTIMATION OF MINERAL OIL

1. DEFINITION:
1.1 The test determines qualitative the presence of mineral oils in oil samples.

2. SCOPE:
2.1 Applicable to oils and Fats

3. APPRATUS:
3.1 Conical flask (250 ml)

4. CHEMICALS:
4.1 KOH Solid
4.2 Ethanol 95%

5. PROCEDURE:
5.1 Take 1 gram samples in a flask.
5.2 2 Add 1 gram KOH solid & 50 ml ethanol.
5.3 Boil on hot plate shaking occasionally until saponification is complete in about 5 minutes.
5.4 Cool and add 25 ml of distilled water.
5.5 In presence of > 0.5% mineral oil , distinct turbidity.

6. REFERENCE:
6.1 American Oil Chemists Society/ AOCS Official Method 28, 285 (1945).
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SOAP CONTNTENTS OF REFINED OIL

1. DEFINITION:
1.1 This method determines sodium content and other basic materials reacting with HCL (Such as Mg Soap,
basic phosphatides) as % Na soap or as % NaOH.

2. SCOPE:
2.1 Applicable to washed oil and under washed oil

3. APPRATUS
3.1 Micro burette 0.02 or O. I ml
3.2 Volumetric flask 2 liters.
3.2 Conical flask 250 ml

4. REAGENTS:
4.1 Acetone (pure)
4.2 0.01 N HCI in acetone prepared by diluting in aqueous HCI should be standardized weakly, when
normality falls below 0.007 discard it.
4.3 Indicator solution
4.3.1 Take 2 liters flask
4.3.2 Add 20 ml water to 1 liter acetone
4.3.3 Add 0.40g Bromophenol Blue
4.3.4 Titrate with 0.1N NaOH to green and then with 0.01N HCL till green yellow.

5. PROCEDURE
5.1 Take perfectly clean flask, check by adding some indicator solution.
5.2 for an under washed oil take 5-10 gms of sample and 25 ml of indicator solution.
5.3 For washed oil weight 40 gms of sample and take 50 ml of indicator solution.
5.4 Titrate, while shaking vigorously with 0.01N HCI till a constant yellow change in Acetone phase is obtained.

6. CALCULATION:
6.1 Soap content as % Na Soap V×304400
Soap Content (PPM) =
Wt:

Where:
V = Volume used (ml of HCL)
Wt: = Weight of sample

7. REFERENCE:
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7.1. Analysis and characterization of Oils, Fats and Products.

SAPONIFICATION VALUE

1. DEFINITION:
1.1 This saponification Value is the amount of alkali necessary to saponify a definite quantity of the sample. It
is expressed as number of milligrams of Potassium hydroxide required to saponify I gm of sample.

2. SCOPE:
2.1 Applicable to all normal fats and oils

3. APPRATUS:
3.1 Flat bottom flask, Air condenser, minimum 650 mm long
3.2 Hot plate with variable heating control

4. REAGENTS:
4.1 Hydrochloric acid 0.5 N accurately standardized for preparation and standardization
4.2 Alcoholic potassium hydroxide
4.3 Take 5 to 10 gms of KOH in a 1 liter flask
4.4 Add 500-750 ml 95% ethanol alcohol
4.5 Boil on a heater under a reflux condenser 0- 60 minutes
4.6 Distilled and collect the alcohol.
4.7 Dissolve 10 gms potassium hydroxide low in carbonate, in 250 ml of the distilled alcohol keeping
temperature below 15.5 °C (60 F ) while the alkali is being dissolved. 4.8 This solution remains clear.
Phenolphthalein indicator solution (0.1% in 95% alcohol)

5. PROCEDURE
5.1 Melt the sample if it is not already liquid, and filter through filter paper to remove any Impurities and last
trace of moisture. The sample must be completely dry.
5.2 Weigh a sample of such size that the back titration is 45% to 55% of the blank. This is usually 4-5 gms. Add
50 ml of alcoholic KOH with a pipette and allow the pipette to drain for a definite period of time.
5.3 Prepare and conduct black determination simultaneously with the same similar in all respects.
5.4 Connect the air condenser and boil gently but steadily until the sample is completely saponified. This
usually requires ca 1 hr. be careful that the vaporing in the condense does not rise to the top of the
condenser or there may be some loss.
5.5 After the flask and the condenser have cooled somewhat but not sufficiently to jell, wash down the inside
of condenser with little distilled water. Disconnect the condenser, add Ca 1ml of indicator and titrate with
0.5N HCI until pink color has just disappear.
5.6 Calculate the saponification value by the given formula
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(V2−V1)×N×EW×1000
S.V =
1000×W S.V
= Saponification value in mg KOH/gm sample.
V2 = Volume of HCL used for blank
V1 = Volume of HCL used for sample
W = Weight of sample taken
N = Normality of HCl = 0.5 NEW = Eq. wt. of KOH = 56.1 gms

6. REFERENCE:
6.1. American Oil Chemists Society/ AOCS Official Method Ca 3C – 91

MOISTURE AND VOLATILE MATTER (HOT PLATE METHOD)

1. DEFINITION:
1.1 The test determines the moisture and volatile mater present in oils & fats.

2. SCOPE:
2.1 Applicable to oils& fats.

3. APPRATUS:
3.1 Clean dry beaker 100-150ml or other similar convenient dish.
3.2 Electric hot plate.
3.3 Desiccator containing on efficient desiccant.

4. PREPARATION OF SAMPLE:
4.1 Because water tends to settle in samples that have softened or melted, care must be taken to mix the
samples thoroughly so as to distribute the water uniformly. Soften with gentle heat (Do not melt) and mix
thoroughly with an efficient mixer.

5. PROCEDURE:
5.1 Weight a clean dry beaker.
5.2 Take 5 to 20 gms sample in a beaker.
5.3 Heat on the hot plate taking care that the temp of the sample does not at any time go about 130°C.
5.4 During the heating rotate the vessel gently by hand to avoid spattering by the too-rapid evolution of
moisture.
5.5 The approach of the end point may be judged by the absence of rising bubbles of steam a by the absence
of foam at the last.
5.6 The heating, however, should momentarily be carried on to incipient smoking but not beyond this point.
Cool and weight.
5.7 Cool to room temperature in a desiccator and weight.

6. CALCULATION:
6.1 Moisture and volatile matters
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Loss in weight ×100

Most, & V.M% =
Weight of sample

7. REFERENCE:
7.1 American Oil Chemists Society/ AOCS Official Method Ca 2B - 38

MOISTURE AND VOLATILE MATTER (AIR OVEN METHOD)

1. DEFINITION:
1.1 This method determines the moisture and volatile matters in Oil, Soap, Cartons and Powder

2. SCOPE:
2.1 This method is applicable to cartons, soap products containing synthetic detergents and powders. It is not
applicable to products containing appreciable amount of sodium silicate, over 1 % Glycerol, and soaps
product from oil such as linseed oil, that tend to oxidize readily. Thus Distillation method must be used for
these product.

3. APPRATUS:
3.1 Air Oven maintained at 105±2°C.
3.2 Porcelain or glass moisture dishes, 6 to 8 cm diameter and 2 to 4 cm depth.
3.3 Desiccator containing an efficient desiccay.

4. PROCEDURE:
4.1 Weight 5 to 30 gms sample into a weighted moisture dish which has been previously dried for 1 hr. at
105±2°C and cooled to room temperature in a desiccator.
4.2 Dry the sample to constant weight in an air oven at 105±2°C. cool to room temperature in a desiccator
and weight.
4.3 Constant weight is attained when successive 1hr.heating periods show a loss (or gain ) of not more than
0.1 %.

5. CALCULATION
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Loss in mass x 100

Moisture and volatile matter % =
Wt:of sample

6. REFERENCE:
6.1 American Oil Chemists Society/ AOCS Official Method Ca, 2C – 25

COLOUR DETERMINATION

1. DEFINITION:
1.1 This method combines color by comparison the color in Lovibond Units as measured on a Lovibond
Tintometer.

2. PRINCIPLE:
2.1 The method consists of matching the color of the light transmitted through a specific depth of sample to
the color of the light, originating from the same source, transmitted through stand color slides.

3. SCOPE:
3.1 Application to all normal oils and fats, providing no turbidity is present in the sample.

4. APPARATUS:
4.1 Lovibond Tintometer
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4.1.1 The instrument consists of a light proof cabinet with dull interior illuminated by dual
tungsten filament lamps positioned at 45 to the sample and reference field. A central integrating
chamber ensures similar illuminating conditions for both field.
4.1.2 The cell holder for examination of liquid samples is capable of holding cells with path lengths
from 0.25 mm to 6 inches.
4.1.3 A rack system of Lovibond color glass filters in red, yellow, blue and neutral tints is located in
the front part of the cabinet.
4.1.4 By creating different combinations of the three primary, colors, red, yellow and blue, the
operator can match any color,
For each primary colors, there is a scale which progresses from the very pale to the very dark. For
example, 0.1 units of red is the palest red color, while 70.0 units is the darkest.
Red Yellow Blue Neutral
0.1-0.9 0.1-0.9 0.1-0.9 0.1-0.9
1-9 1-9 1-9 1.0-2.0-3.0
10-70 10-70 10-70

The neutral tints are used to darken very bright samples in order to bring them within the range of the intensity of the
glass color standard.
4.1.5 The front panel contains 'ON/OFF push buttons together with the two red lampoon
reflectors and time lapse indicator to ascertain lamp life.
4.1.6 The optical system consists of a viewing tube which divides the field of view in two: the left
hand field is for observing the sample, whilst the right hand field presents a white reference standard
in front of which the glass color filters are interposed.
4.1.7 The viewing tube also incorporates a blue correction filter to convert the tungsten
illumination sources to standard Lovibond viewing conditions.
4.1.8 A block of magnesia is placed in the instrument at the proper angle lo reflect the light from
the electric bulb vertically upward through the viewing tube and color glasses. The block must be free
from oil and dirt and positioned to give equal illumination to both sides of the viewing field.
Glass cells, with lengths of light path 1 0 inch (25 4 mm) and 5.25 inch (133.4 mm)

5. OPERATORS:
5.1 All operators shall satisfy the requirements of a color vision test and shall be retested at five-yearly
intervals.
5.2 Operators who normally wear spectacles or contact lenses may continue to wear them.

6. PROCEDURE:
6.1 If the prepared sample is not liquid at room temperature heat it to a temperature of 60+5 degree C and
pour into the cell
6.2 For crude and bleached oils chose a 1 inch (254 mm) cell For refined oils, a 5.25 inch (133.4 mm) cell is
suitable.
6.3 Using the lovibond tintometer, immediately determine the color of the sample by achieving the best
possible match with the standard color slide.
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6.4 Since the onset of eye fatigue is rapid (i.e. after about 10 sec) it is preferable, if a color match cannot be
quickly obtained to repeat the procedure after a few minutes. It is also desirable that the best be carried
out by trained operator
6.5 If the red, yellow and neutral unit do not form a possible match of the color of the oil, use a minimum of
blue units to obtain the suitable match
Express the results in terms of the number of red, yellow, neutral or blue units needed to obtain the match and the cell
used and the Lovibond Model Number.

7.REPRODUCIBILITY OF RESULT
7.1 The difference between the results obtained by two trained operations carrying out procedure described using the
same apparatus, in the same laboratory simultaneously or in rapid succession shall not exceed the value given in table
1.
Reading Acceptable Difference
Up to 0.9 Red 0.2 Red
From 1.0-2.9 Red 0.4 Red
From 3.0-4.0 Red 0.5 Red
From 4.1-12.0 Red 1.0 Red

8. REFERENCE:
8.1. American Oil Chemists Society/ AOCS Official Method Cc, 13 B – 45

CLOUD TEST

1. DEFINITION:
1.1 The cold point is that temperature at which, under the conditions of this test a cloud is induced in the
sample caused by the first stage of crystallization.

2. SCOPE:
2.1 Applicable to all normal animal and vegetable fats.
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3. Apparatus:
3.1 Oil sample bottle, 115 ml (4 Oz)
3.2 Thermometer
3.3 Water bath made up of water, chipped ice and water, or chipped ice, salt and water, depending upon the
temperate required The temperature of the cloud bath shell be not less than 2°C, nor more than 5°C
below the cloud point. A beaker or sauce pan is a convenient container for this purpose.

4. PROCEDURE:
4.1 The sample must be completely dry before making the test. Heat 60 to 75 gms of sample to 130"C
immediately before making the test. Pour Ca 45 ml of the heated fat into an oil sample bottle.
Begin to cool the bottle and contents in the water bath, stirring enough to keep the temperature uniform. When the
sample has rechecked a temperature Ca 10 °C about the cloud point, begin Stirring steadily and rapidly in a circular
motion so as to prevent super cooling and solidification of fats crystals on the sides or bottom of the bottle.
4.2 Form this point on do not remove the thermometer from the sample, since to do so may introduce air
bubbles which will interfere with the test. The bottle is maintained in such a position that the upper levels
of the sample in the bottle and the water in the bath are about the same.
Remove the bottle from the bath ad inspect regularly. The cloud point is that temperature at which that portion of the
thermometer immersed in the oil is no longer visible when viewed horizontally through the bottle and sample.

5. NOTE:
5.1 It is essential that the sample be heated to 130 degree centigrade to destroy any crystal matter.

6. REFERENCE:
6.1 American Oil Chemists Society/ AOCS Official Method Cc, 6 – 25

COLD TEST

1. DEFINITION:
1.1 This method measures the resistance of the sample to crystallization and is commonly used as an index of
the winterization or similar stearin removing process

2. SCOPE:
2.1 Applicable to all normal, refined and dry, animal and vegetable oils

3. APPRATUS:
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3.1 Oil sample test tube I50 mm x 15 mm. These must be completely clean and dry.
3.2 Chipped ice and water bath at 0 °C. Prepare by filling a container (pail or bucket 4 to 5 Ib. capacity).

4. PROCEDURE:
4.1 Filter a sufficient quantity of sample (2000 to 300 ml) through a filter paper and then heat the filtered
portion.Stair the sample while heating and remove from the heat source immediately when the
temperature reaches 130°C.
4.2 Take 10 ml of sample in the test tube and close its mouth with a cotton plug.
4.3 Put the tube containing the sample in the ice on water bath.
4.4 Replenish the ice as often as is necessary to keep the bath solidly packed, otherwise the bath temperature
will not remain at 0 degree centigrade as specified. This temperature is essential.
4.5 At the end of 5.25 hours, remove the tube from the bath and examine closely for fat crystals or
cloudiness. Do not mistake small and finely dispersed air bubbles for fat crystals. To pass the test sample
must be clear, limpid, and brilliant.

5. NOTE
5.1 The purpose of the preliminary heat treatment is to remove traces of moisture and to destroy any crystal
nuclei which may be present. Either will interfere with the test, causing cloudiness or premature
crystallization.
5.2 If it is desired, the test may be continued by reexamination of the sample at hourly intervals after the first
5.25 hour examination. However, in doing this, the sample should be returned to the bath as promptly as
possible after each inspection so that the temperature will not increase by any significant amount.

6. REFERENCE:
6.1 American Oil Chemists Society/ AOCS Official Method Cc, 11 – 53

VITAMIN TEST

1. DEFINITION
1.1 This method determines qualitatively the presence of Vitamin-A in the sample.

2. SCOPE:
2.1 Applicable to all oils and fats

3. APPARATUS:
3.1 Separating funnel 100 ml
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3.2 Measuring cylinder 250 ml

3.3 China dish (500)
3.4 Flat bottom flask (250 ml)
3.5 Reflex condenser

4. REAGENTS:
4.1 Ethyl alcohol 95%
4.2 Sodium sulphate (Anhydrous)
4.3 Take 33 gms Antimony trichloride and dissolve it in 100 ml chloroform by refluxing, transfer the war
solution to a bottle containing anhydrous sodium sulphate.

5. PROCEDURE
5.1 Take 10 ml of sample in a separating funnel.
5.2 Add approx. 15 ml ethanol and shake vigorously.
5.3 Allow to stand for few minutes into two layers.
5.4 Take upper layer of ethanolic portion in 50 ml china dish.
5.5 Keep on water bath and add 5 ml chloroform and mix with glass rod.
5.6 Add Antimony trichloride solution.
5.7 If blue color is appears, it indicates the presence or Vitman A.

6. REFERENCE:
6.1 Pakistan Standard Institution

ACIDITY OF BLEACHING EARTH

1. DEFINITION
1.1 The acidity of bleaching earth is the amount of NaOH necessary to neutralized tha acid, calculated by wt
of HCL.

2. SCOPE:
2.1 Applicable to all type of blenching earth.

3. APPRATUS:
3.1 Beaker 100 ml, 25O ml
3.2 Conical flask 250 ml
3.3 Burette 50 ml
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3.4 Pipette 25 ml
3.5 Volumetric flask 250 ml, 1000ml

4. Reagents:
4.1 Phenolphthalein indicator sol ( 0 1% in spirit)
4.2 2N NaOH (20gm/250ml water)
4.3 0.005N NaOH sol. (25 ml of 2N NaOH/1000 ml spirit

5. Procedure:
5.1 Weight 3-4 gms fullaer’s earth in a 250ml beaker.
5.2 Add 5-20 ml of dis Water, and heat on hot plates.
5.3 Now allow settle for 10 min and then filter the upper liquid portion in conical a flask.
5.4 Now again repeat steps ( 5.20 (5.3) so that 4 washings have been applied to the sample.
5.5 Now wash the filter with 10 ml of distilled water.
5.6The collected filtrate is taken titrate with NaoH (0.005N).

6. CALCULATION:
6.1 Calculate the acidity of bleaching earth as HCI.
𝐕×𝐍×𝐄𝐖×𝟏𝟎𝟎
Acidity=
𝟏𝟎𝟎𝟎×𝐖

V= Volume of NaOH used

N= Normality of NaOH
Wt= Weight of sample EW =
Eq. Wt. of HCI= 365 g7.

7. REFERENCE:
7.1 Soyabean Oil Processing
American Soyabean Association Technical Bulletin

BLEACHABILITY OF BLEACHING EARTH

1. DEFINITION
1.1 This method determine the color of the sample after treatment with a specified bleaching earth under the
condition of this test.
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2. SCOPE:
2.1 Applicable to all type of Bleaching Earth

3. APPARATUS:
3.1 Beaker 300ml
3.2 Thermometer 0-150 °C

4. PROCEDURE:
4.1 Weight 99 gms of refined oil in a 300 ml beaker.
4.2 Heat to about 80 °C
4.3 Now add 1 gm of standard earth and heat to about 110 °C with constant stirring on a magnetic stirrer for
15.
4.4 Don't let the temp go about 110 °C.
4.5 Filter through watsmann no 1 filter paper.
4.6 Note the color of filtered oil in 5.25" cell on Tintometer.
4.7 Repeat the same procedure with the test earth. The test earths bleachability is compared with standard
earth.

5. CALCULATION5:
𝐁𝐂−𝐀𝐂
Bleachability %= ×100
𝐁𝐂

BC = R×10+Y (Colour Before Bleach)

AC = R×10+Y (Colour After Bleach)

6. REFERENCE:
6.1 Soyabean Oil Processing
American Soyabean Association Technical Bulletin
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QUALITATIVE ESTIMATION OF VITAMIN E

1. DEFINITION:
1.1 This method determines presence of vitamin E.

2. SCOPE:
2.1 Applicable to vitamin E containing compounds.

3. APPRATUS:
3.1 Beaker 100 ml.
3.2 Flat bottom flask 250 ml.
3.3 Reflux condenser
3.4 Volumetric flask 250 ml

4. REGENTS:
4.1 Sulphuric acid
4.2 Dehydrated alcohol
4.3 Nitric acid

5. PROCEDURE:
5.1 Test Solution for alpha tocopherol acetate.
Transfer about 275 mg of d-alpha tocopheral acetate, accurately weighed, to a flat bottom flask and dissolve in 25
ml of dehydrated alcohol add 20 ml of dilute sulphuric acid on alcohol (1 in 7) and reflux it for 3 hours, protected
from sunlight cool, transfer to a 250 ml volumetric flask, add dilute sulphuric acid in alcohol (1 in 72) to volume
and mix.
5.2 Identification Test: Take 10 ml of test solution for alpha tocopherol acetate. Add with swirling 2ml of nitric
acid and heat about 75 C for 15 mints a bright red or orange colour.

6. REFERENCE:
6.1 Official Monograehs/Vitamin
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TITRE TEST

1. DEFINITION:
1.1 This method is used to determines the solidification point of the fatty acid

2. SCOPE
2.1 Applicable to fatty acid of all soap & soap products.

3. APPRATUS
3.1 Beaker 100 ml or 250 ml.
3.2 Bottle 175 cm length & 75 cm Dia.
3.3 Test tube 200 cm length & 25 cm Dia.
3.4 Stirrer with one end bent in the form of a loop 18 cm o.d. Glass, Nichrome, Stainless steel or Copper wire
may be used.
3.5 Thermometer, Laboratory thermometers range 0-110 °C or 115 °C.
3.6 Filter paper No. 1.
3.7 Sepratory funnel 500 ml.
3.8 Graduated cylinders 50/100 ml.
3.9 Glass beats, Beaker 500 ml/1000ml.

4. REAGENTS:
4.1 Sulphuric acid (conc) 1:1 V/V with distilled water.
4.2 Distilled water.

5. PREPARATION OF FATTY ACID:

5.1 Dissolve 50 gms of original sample in 500 ml hot water contained in a 1000 ml beaker. (If the sample
contains alcohol, this sample must be completely evaporated from the soap solution before proceeding).
5.2 Add 10 ml of 1:1 Sulphuric acid (conc) & a few glass beads to minimized bumping & heating gently
transfer the content in a sepratory funnel allow to settle fatty matter collect in a clear layer on the top.
5.3 Remove the aqueous acid (lower) layer add 30 ml of hot water , boil gently for a few minutes and again
remove aqueous lower layer.
5.4 Wash the acid three times in this matter Complete the acidification and washing in as short a period as
possible, keeping the funnel covered to prevent oxidation of the acid.
5.5 After washing, allow to settle for a few minutes and then decent the fatty acid carefully so as not to
include any water

6. PROCEDURE:
6.I Take 15 to 20 g of test sample in a 100 ml of beaker & heat gently to remove moisture content.
6.2 Filter the moisture free fatty acid using filter paper No-1 and for this purpose keep the beaker in hot air
oven at 100 +1-3°C.
6.3 Transfer filtered sample to test tube & fit the tube with thermometer & stirrer & ring.
6.4 Fill the water to the designated level (1-2cm above than sample level) & adjust the temperature to 15-20
°C for all samples.
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6.5 Place the tube containing Fatty acid in the assembly (Titer bottle), insert the Titer thermometer in such
manner that it will be equidistant from the sides of the tube.
6.6 Stir with stirring rod in a vertical manner This agitation is started while the temperature is at least 10 °C
above the Titer point.
6.7 Stir until the temperature remain constant for 30 second or begins to rise in less than 30 seconds interval.
Discontinue stirring immediately remove the stirrer or rise it out of the samples and observed the increase
in temperature. The titer point is the highest temperature indicated by the thermometer during the rise.

7. Note:
7.1 Prepartion of fatty acid is not applicable for titer test of palm fatty.

8. REFERENCE:
8.1 American Oil Chemist Society/AOCS Da, 13 – 48
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SPECIFIC GRAVITY OF OIL

1. DEFINITION:
1.1 This method determines the ratio of the weight of a until volume of the sample at 25 °C to the weight of a
unit volume of water at 25°C.

2. SCOPE:
2.1 Applicable to all oils and liquid fats.

3. APPARATUS:
3.1 Specific gravity bottle with well-fitting ground glass joints, Ca 50 ml. Calibrate these bottles as follows:
Clean and dry thoroughly and then will with recently boiled and cooled distilled water at 20 to 23°C . Fill
the bottle to overflowing by holding the bottle on its side in such a manner as to prevent the entrapment
of air bubble Insert the stopper and immerse in the water bath at 25°C±0.2°C. Keep the entire bulb
completely covered with water and hold at specified temperature for 30 minutes. Carefully remove any
water which has exuded the side opening. Remove the bottle from the bath and wipe completely dry.
Weight the bottle and contents. Calculate the weight of water in the bottle by subtracting the weight of
empty bottle from the weight of bottle plus water.
3.2 Water bath maintained 0.2 °C fluctuation.
3.3 Thermometer, any convenient thermometer of suitable range with 0.1 or 0.2 subdivisions. standardize
carefully, preferably by comparison with a thermometer calibrated by the US. Bureau of standards.

4. PROCEDURE:
a) Specific Gravity at 25 / 25°.
4.1 Melt the sample and filer through filter paper to remove any impurities and the traces of moisture. must
be completely dry.
4.2 Cool The sample to 20 to 23 °C and filled the bottle to over flowing, holding the bottle on its side in such a
manner as to prevent the entrapment of air bubble.
4.3 Insert the stopper, immerse, and hold in the water bath at 25 °C+ 02°C for 30 minutes.
4.4 Carefully wipe off any oil which has come through capillary opening and remove from bath. Clean and dry
thoroughly.
4.5 Weight the bottle and contents and calculate the specific gravity as directed in 5.1.

b) Specific Gravity at 60/ 25"C

4.1 The procedure is the same as described in (a except that the method fat is poured in the specific
gravity bottle at 56 to 58 °C.
4.2 The bottle containing the sample is followed to remain in abath at 60±2°C for 30 min. Cool to room
temperature and weight the bottle and contents. Calculate the specific gravity as directed in 5.2.

5. CALCULATION
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W2−W1
5.1 Specific gravity at 25/25 C =
WW[1+(0.000025 ×35)]

W1 = weigh of empty specific gravity bottle

W2 = weight of bottle in oil
W3 = Weight of water

6. Note:
6.1 Fats not liquid at 25°C may be tested at 60/25°C.
6.2 Unless the specific gravity bottles are protected with caps, care must be taken so that no oil of the room is
above 25°C this is very apt to happen. Even the warmth of the hand surrounding the bottle is sufficient to
cause expansion of the contents of the flask.
6.3 Unless the specific gravity bottles are made of glass with a very low coefficient of expansion a correction
must be made for the expansion of the glass. The coefficient of expansion of glass is ca 0.000025.

7. REFERENCE:
7.1 American Oil Chemist Society/AOCS Ea 7 – 95
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DETERIORATION OF BLEACHING INDEX OF CPO

1. DEFINITION:
1.1: The deterioration of bleach ability index (DOBI) is the numerical ratio of the spectrophotometric absorbance at 446
nm to absorbance at 269 nm.

2. SCOPE:
2.1: Applicable to crude palm oil.

3. PRINCIPLE:
3.1: Spectrophotometric measurement of a solution of oil sample in iso-octane or n-hexane (0.5-1.0 % Concentration)
against the solvent. The absorbance values are not corrected in any other way.

4. REAGENTS:
4.1: 2,2,4-Trimethyl Pentane (Iso-Octane) or n-hexane of spectroscopic or equivalent grade.

5. APPARATUS:
5.1: Spectrophotometer, suitable for measurements at 269 nm and 446 nm.
5.2: Quartz cuvettes, 10 nm
5.3: Volumetric flask, 25 ml
5.4: Volumetric flask, 10 ml
5.5: Pipettes, 2 ml

6. PROCEDURE:
6.1: Weigh to the nearest 0.1 mg, 0.1 gm of a completely melted and homogenized oil sample into a 25 ml volumetric
flask. Dissolve with the solvent (4.1) and make up to volume. Fill a cuvette with the oil solution and measure its
absorbance at 269 nm and 446 nm against pure solvent (4.1).
6.2: If the solution is too concentrated dilute it by pipetting two milliliters of it into 10 ml volumetric flask and making
up to volume with the solvent (4.1). Measure the absorbance of this solution at 269 nm and 446 nm against pure
solvent (4.1). Further dilution may be necessary for very high DOBI values.

7. CALCULATION:
𝐀𝐛𝐬𝐨𝐫𝐛𝐚𝐧𝐜𝐞 𝐚𝐭 𝟒𝟒𝟔 𝐧𝐦
DOBI =
𝐀𝐛𝐬𝐨𝐫𝐛𝐚𝐧𝐜𝐞 𝐚𝐭 𝟐𝟔𝟗 𝐧𝐦

Express the results into two decimal places.

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UNSAPONIFIABLE MATTERS

1. DEFINITION:
1.1 Unsaponifiable matters includes those substance frequently found dissolved in fats and oils which cannot
be saponified by the caustic alkali but are soluble in the ordinary fats solvents included are the higher
aliphatic alcohols, sterols, pigments and hydro carbonates.

2. SCOPE:
2.1 Applicable to normal animal and vegetable fats and oils. It is not suitable for fats and oil such as the
marine oil containing excessive amounts of unsaponifiable matter.

3. APPARTATUS:
3.1 Erlenmeyer or soxhlet flasks, 100-200 ml.
3.2 Sepratory funnels, 500 ml.
3.3 Flat bottom flask, 250 ml.
3.4 Beaker, 250/500 ml.

4. REAGENTS:
4.1 Ethyl alcohol, 95%
4.2 Aqueous potassium hydroxide solution, 50% KOH by weight.
4.3 Petroleum Ether
4.4 Sodium hydroxide Solution, 0.02N accurately standardized.
(Standardized with any primary standard such as oxalic acid or potassium hydrogen phthalate.)
4.5 Phenolphthalein indicator solution, 1% in 95% alcohol.

5. PROCEDURE:
5.1 Weight accurately Ca 5 gm of well mixed sample into Erlenmeyer or soxhlet flask.
Add 30 ml of alcohol and 5 gm of aqueous KOH. Boil gently but steadily under a reflux condenser for 1 hour or until
completely saponified. Complete saponification is essential.
5.2 Transfer to the sepratory funnel and wash to the 40 ml mark with alcohol. Complete the transfer with
warm and then cold distilled water until the total volume is 80 ml. Wash out the flask with a little pet
ether and add to the cylinder. Cool the cylinder and contents to room temperature ( 20-25°C) and then
add 50 ml of pet ether.
5.3 insert the stopper and shake vigorously for at least one minute and then allow to settle until both layers
are clear.
Collect the lower portion in beaker.
5.4 The pet ether fractions are drawn into and accumulate in a 500 ml separatory funnel.
5.5 Using separatory funnel repeat the extraction using 50 ml portions of pet ether such time at ;east 6 more
times, shaking vigorously with each extraction.
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Caution: there are some cases in which 7 extractions may not be sufficient . this is best judged by making another
extraction and evaporating this separately. There should be no un saponifiable matter in this extract.
5.6 Wash the combined extracts in the separatory funnel 3 times with 25 ml portions of 10% alcohol in
distilled water, shaking vigorously and drawing off the alcohol layer after each wash. Be careful not to
remove any of the ether layer.
5.7 Transfer the ether extract to a weighed flat bottom flask 250 ml and evaporate to dryness on a water bath
under a gentle stream of clean, dry air. Complete the drying to constant weight, preferably in a vacuum
oven at 75-80°C and an internal pressure of not more than 200 mm of mercury. Cool in a desiccator and
weigh.
5.8 After weighing, take up the residue in 50 ml of warm (Ca 50°C) 95 % alcohol contain indicator and
previously neutralized to b faint pink colour. Titrate with 0.02N NaOH to the same colour.

6. Calculations:
6.1.
V×N×MW
W.F =
1000
WF = Weight of fatty acid in the extract
V = Volume of 0.02 N NaOH used
N = Normality of NaOH = 0.02N
MW = Molecular weight of oleic acid = 282

6.2.
(WR−WF)×100
USM % =
W
USM = Unsaponifable matter
WR = Weight of residue
W = Weight of sample

7. REFERENCE:
7.1 American Oil Chemist Society/AOCS Ca 6a – 40
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REFRACTIVE INDEX

1. DEFINITION:
1.1 The refractive index of a substance is the ratio of the speed of' light in a vacuum to the speed in the
substance. For practical measurements including this method, the scales of standard of stand instruments
indicate refractive indices with respect to air rather than vacuum. The index of refraction of oils is
characteristic within certain limits for each kind of oil. It is related to the degree of saturation and heat
treatment.

2. SCOPE:
2.1 Application to all normal oils and liquid fats

3. APPRATUS:
3.1 Refractometor any standard refract meter equipped with the Abbe, Butyro or any other standard scale. A
dipping type refractometer is satisfactory. The temperature of the refractometer must be controlled
within 0.1°C and for this purpose water is preferably provided with a thermostatically controlled water
bath and motor driven pump to circulate water through the instrument. The instrument is standardized,
with a liquid of known purity and refractive index or with a glass prism of known refractive index. Distilled
water, which has a refractive index of 1.3330 at 200 °C is satisfactory in some cases.
3.2 Light source if the refractometer is equipped with a compensator, a tungsten lamp or day light bulb is
permissible. Otherwise a monocratic light such as an electric sodium vapor lamp is require.

4. REAGENTS:
4.1 Toluene or some other fats solvent satisfactory for cleaning the prisms. Lens tissue or cotton is
recommended for cleaning the prisms in order to avoid injury to them.

5. PROCEDURE:
5.1 Melt the sample, if it is not already liquid, and filter through filter paper to remove any impurities and the
last traces of moisture. The sample must be completely dry.
5.2 The temperature of refractrometer is adjusted to 40.0 °C for ordinary fats and oils. For higher melting
point samples. Use a temperature of 60 °C.
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5.3 Be sure the prisms are clean and completely dry and then place several drops of the sample on the lower
prisms close the prisms and tight firmly with the screw had allow to stand for 2 minutes or until with the
sample comes to the temperature of the instrument.
5.4 Adjust the instrument and light to the obtain the most distinct reading possible and then determine
refractive index. Take several readings and calculate the average of all

6. NOTE
Approximate temperature corrections may be made by the following calculations.
Abb'e rcfractometer
R = R’ + K(T’-T)
R = The reading reduced to temperature T
R’ = the reading at T’ °C
T = the standard temperature
T’ = the temperature at which the reading R' is made
K = 0.000365 for fats and 0.000385 for oil

7. REFERENCE: 7.1. American Oil Chemist Society/AOCS Cc 7 – 25

QUANTITATIVE ESTIMATION OF VITAMIN A

1. DEFINIATION
1.1 This method is determines vitamin A in the sample quantitatively.

2. SCOPE:
2.1 Applicable to all samples of oil & fats having vitamin A.

3. APPRATUS:
3.1 Beaker 250 ml
3.2 Separating funnel 250 ml
3.3 Volumetric flask 250 ml
34 Flat bottom flask 250 ml
3.5 Circulating water bath
3.6 Chiller
3.7 Reflex condenser
3.8 Spectrophotometer (UV) with 1 cm quartz cell

4. REAGENTS:
4.1 Hydroquinone
4.2 Ethanol (Analytical grade)
4.3 Potassium Hydroxide
4.4 Ether Anesthetic (Analytical grade)
4.5 Sodium Sulphate Anhydrous
4.6 Iso Propyl Alcohol
4.7 Phenolphthalein
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5. SOLUTION:
5.1 Prepare 50% solution of sodium hydroxide

6. PROCEDURE:
6.1 Weight accurately 10 gms of sample in a 250 ml flat bottom flask then add 80 mg Hydroquinone 30 ml of
Ethanol and 8 ml Potassium Hydroxide solution.
6.2 Boil gently under reflex for 30 minutes using water bath keep temperature of water bath around 70-80°C.
Add 30 ml distilled water while flask is hot and swirl gently to mix the content.
6.3 Transfer the contents of the flask in to separating funnel. Rinse the flask with 50 ml Ether an anesthetic
and add to the separating funnel. Shake well and allow the layer to separate.
6.4 Transfer aqueous layer to another separating funnel and extract with three quantities each of 50ml
anesthetic ether.
6.5 Combine ether extract and wash with 50 ml of cold distilled water till washing become free from alkali,
When checked by Phenolphthalein.
6.6 Filter the Ethanol extract in a flask through filter paper previously wash with Anesthetic Ether with a layer
of 2 gm of anhydrous Sodium sulphate with few ml of Anesthetic Ether.
6.7 Evaporate the ether layer extract to nearly dryness on water bath at 40-50 C. Dissolve the residue in iso
propyl Alcohol.

7. MEASUREMENT OF ABSORBANCE:
7.1 Set blank with Iso propyl alcohol to 0(Zero) at 325 nm.
7.2 Measure the absorbance of the solution in 1cm quartz cell at 325 nm.

8. CALCULATION:
8.1 Vitamin A = A x Factor
While A = Absorbance of sample at 325 nm
Factor = 18.3 (Factor to get Vitamin A in I.U/gram)
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INSOLUBLE IMPURTIES

1. DEFINITION:
1.1 This method determines dirt, meal and foreign substances in soluble in petroleum ether.

2. SCOPE:
2.1 Application to all normal oils and liquid fats.

3. APPRATUS:
3.1 Gooch crucible prepared with a glass-fiber filter without organic filter (What man GF/C) wash the
filter with water, alcohol and ether. Dry to constant weight at 101±°C. Cool in desiccator at room
temperature and weight.
3.2 Filter flask of convenient size and Gooch crucible adaptor.

4. REAGENTS:
4.1 Petroleum ether

5. PREPRATION OF SAMPLE:
5.1 Sample must be thoroughly mixed. If necessary soften with gentle heat (don't melt) and mix with an
efficient mixer.

6. PROCEDURE:
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6.1 Weight accurately 5 20 gms of well mixed sample into a beaker which has been previously dried by
cooled in a desiccator.
6.2 Heat the sample on the hot plate to remove moisture, rotating the beaker gently by hand to avoid
spattering which may result from 100 rapid adulteration of moisture or use the residue from the
moisture and volatile matter determination.
6.3 Add 50 ml of petroleum ether to the sample and heat in a water bath to dissolve the fat. Filter
through the prepared Gooch crucible with the aid of vacuum. Wash the five to seven 10ml portion
of hot petroleum ether, allowing each portion to drain before adding the mixt.
6.5 Dry the crucible and contents to constant weight at 101±°C, cool to room temperature in a desiccator and weight.

7. CALCULATION:
Grain in mass of crucible ×100
7.1 Insoluble impurities % = mass of sample taken

8. NOTE
8.1 Petroleum ether is flammable solvent. A fuming hood should be used at all times when working
with this solvent.
8.2 Samples with higher than usual percentage of insoluble impurities may be difficult to filter, for such
samples, a 2gm sub sample rather than 5gm sub sample can be used.

9. REFERENCE:
9.1 American Oil Chemist Society/AOCS Ca 3a – 46

QUALITATIVE ESTIMATION OF NICKEL

1. DEFINITION:
1.1 This method detects qualitatively, the presence of Nickel in Banaspati or hydrogenated oil.

2. SCOPE:
2.1 Applicable to hydrogenated oil and banaspati.

3. APPRATUS:
3.1 Beaker 100 ml
3.2 Hot Plate
3.3 Separating funnel

4. REAGENTS:
4.1 Dimethyl Glyoximes solution 1% in 95% alcohol.
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4.2 Ammonia hydroxide solution 1:1 ( in distilled water )

4.3 Hydrochloric Acid conc:

5. PROCEDURE:
5.1 Take clean beaker, weight about 10 gm of sample and then add 10ml of HCl (Conc).
5.2 Heat for 2-5 minutes on water bath or Hot plate.
5.3 Cool in a desiccator and then transfer the content in separating funnel, collect lower layer carefully.
5.4 Add 2-3 ml of Ammonium hydroxide and then add 2-3 ml dimethyl glyoxime.
5.5 If pink colour appears, it indicates presence of Nickel.

6. REFERENCE:
6.1 Vogel’s Test book of Quantitative chemical Analysis

DETERMINATION PHOSPHOROUS

1. DEFINITION:
1.1 This method determines phosphorus or the equivalent phosphatides content by ashing the sample in the
presence of Zinc Oxide followed by colorimetric measurement of phosphorus as molybdenum blue.

2. SCOPE:
2.1 Applicable to crude, and refined vegetable oil.

3. APPARATUS:
3.1 Porcelain Crucible 50 cc.
3.2 Watch glass- 75 mm diameter
3.3 Electric hot plate
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3.4 Funnel, glass, short steam, 50 mm diameter

3.5 Filter paper Whatman # 42, 90 m
3.6 Volumetric flask, glass stoppered 50 ml, 100 ml, 250 m, 500, ml
3.7 Pipette 2, 5, 10 and 25 ml
3.8 Pipette, Mohr type, 10 ml. Graduated in 0.1 ml subdivision.
3.9 Spectrometer
3.10 Cuvettes

4. REAGENT:
4.1 Hydrochloric acid, sp. gr. 1.19 4.2 Sodium hydroxide, pallets
4.3 Zinc Oxide.
4.4 Sulphuric acid, sp. gr. 1.84
4.5 Sodium molybdate
4.6 Hydrazinium sulphate
4.7 Potassium di hydrogen phosphate, Dry at 101 °C. for 2 hours before use.

5. SOLUTIONS:
5.1 SODIUM MOLYBDATE M:
Carefully add 140 ml of sulpharic acid (conc) to 300 ml distilled water cool to room temperature and add 12.5 gms
Sodium Molybdate. Dilute to 500 ml with distilled water.
5.2 HYDRAZINE SULPHATE (0.05% W/V):
Dissolve 0.15 gm Hydrazine Sulphate in 1 liter distilled water.
5.3 Potassium Hydroxide:
Dissolve 50 gms Potassium hydroxide in S0 ml distilled water.
5.4 STANDARD PHOSPHATE SOLUTION:
5.4.1 Standard Stock Solution (1000 ppm):
Dissolve 1.0967gm of dry Potassium hydrogen Phosphate in distilled water. Dilute to 250 ml in a volumetric
flask and mix thoroughly. This solution content 0.01 mg of phosphorous per ml.
5.4.2 Standard Working Solution (10 ppm)
Pipette out 5 ml of standard solution into 500 ml volumetric flask and makeup up to the mark with distilled water.
This solution contains 0.01 mg of phosphorus per ml.

6. PROCEDURE:
6.1 Weight the appropriate sample as per table given into a crucible. Add 0.5 gm Zinc Oxide and mix.
6.2 Heat the mixture slowly such that it must not come out of the crucible. When the mask give no smoke
and completely charred transfer the crucible into the muffle furnace at 600°C. Take out the grayish mass
after 2 hrs and cool.
6.3 Add 5ml dist. Water and 5ml conc. HCL through cylinder in the ash and boil gently for 5 mints.
6.4 Filter the solution through Whatman filter paper No. 40 (9cm) into the specified volumetric flask
As per table column B) wash the filter paper by 5x 5ml hot dist. Water into the flask and cool the solution.
6.5 Neutralize to a faint turbidity by adding 50% w/v- KOH drop wise.
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6.6 Add 50% v/v HCL drop wise till Zinc oxide ppt dissolved, add 2ml additional drops. Cool and dilute to the
flask mark by dist. water. Mix the solution thoroughly.
6.7 Similarly prepare a blank (without sample).
6.8 Pipette out the specified aliquot (as per table column C) from the solution prepared in step 6 into a clean
50ml volumetric flask.
6.9 Add 8.0 ml hydrazine sulphate and then 2.0 ml sodium molybdate solution in the order and shake well.
6.10 Loosen the stopper and heat the solution in a vigorously boiling water bath for 10-15mins.
6.11 Cool and dilute to the flask mark with distilled water.
6.12 Now turn on the spectrometer and let it warm up for 1hr. Calibrate the instrument on the clean distilled
water.
6.13 Now note at least 3 absorbance readings of the same solution sample at 650 nm.
6.14 In separate 50ml volumetric flask pipette out 2ml std. P solution and proceed from step 9.

Sample of Wt. of sample To Volume to make Up Aliquot Taken (C) Eqn. To be used
use o (A) to (1B)

Crude under- 3 gms i. 100 ml 10 ml (Abs − 0.002) × 413.22 × 40

gummed SBO ii. Dilute 10 10 ml Wt. of Sample
times as 10 ml to
100 ml
Crude degummed 3 gms 100 ml 10 ml (Abs − 0.002) × 413.22 × 4
SBO
Wt. of Sample
Neutralized SBO 6 gms 50 ml 20 ml (Abs − 0.002) × 413.22

Wt. of Sample
Bleached SBO/PO 6 gms 50 ml 20 ml (Abs − 0.002) × 413.22

Wt. of Sample
RBD Oils 6 gms 50 ml 20 ml (Abs − 0.002) × 413.22

Wt. of Sample

7. REFERENCE:
7.1 American Oil Chemist Society/AOCS Ca 12 – 55

DETERMINATION OF IRON IN FATS

1. DEFINITION:
1.1 This method determines iron in oil and fats by ashing the sample. Iron
11 reacts with 1, 10 phenolphthalein to form an orange red complex.

2. SCOPE:
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2.1 Applicable to oil & fats.

3. APPRATUS:
3.1 Porcelain crucible 50 ml capacity
3.2 Watch glass 70 mm diameter
3.3 Muffle furnace
3.4 Funnel glass, short steam, 50 mm diameter
3.5 Filter paper whatman no 42, 90 mm 3.6 Volumetric flask, 50 ml, 10 ml,
250 ml.
3.7 Transfer pipette 2, 5, 10 and 25 ml
3.8 Pipette, Mohr type, 10 m graduated in 0.1 ml sub division
3.9 Spectrometer, Curettes

4. REAGENTS:
4.1 Hydrochloric Acid, specific gravity 1.19
4.2 Potassium hydroxide pellets
4.3 1. 10-phenanthroline
4.4 Hydroxyl ammonium chloride
4.5 Acetic Acid
4.6 Sodium acetate

5. SOLUTION:
5.1 1. 10-phenanthroline 0.25% Dissolve 0.25 gms of 1. 10-phenanthroline
in 100 ml volumetric flask make the volume up to the mark with
distilled water.
5.2 Sodium acetate 0.2M and 2M please change the raw 0.02M dissolve
2.7216 g sodium acetate in 100 ml volumetric flask.
5.3 Hydroxyl ammonium chloride, 10% dissolve 10 g ammonium chloride in
100 ml of distilled water.
5.4 Standard solution
5.4.1 Standard solution of ammonium iron (11) sulphate. Dissolve 0.7022 g ammonium iron (11)
sulphate in 100 ml distilled water.
5.4.2 Add 5 ml of 1:5 sulphuric acid, and run in cautiously a dilute solution of potassium
permanganate 2g/liter until slight pink coloration remains after stirring well. Dilute to 1 liter and
mix thoroughly 1 ml of this solution contains 0.1 gm of iron.

6. PROCEDURE:
6.1 Weight 3.0-3.5g of sample in to a porcelain crucible.
6.2 Heat slowly on the hot plate until the sample thickens, increase the
heating slowly until the mass is completely charred.
6.3 Ignite the crucible on Muffle furnace for 4 hrs at 600°C, Remove & coo!
to room temperature.
6.4 Add 5 ml of distilled water and 5 ml of Hydrochloric acid to the ash.
6.5 Cover crucible with watch glass and heat to gentle boiling for 5 minutes.
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6.6 Filter the solution into a 100 ml volumetric ask wash the inside of the
wash glass and the sides of the crucible with Ca 5 ml of hot distilled
water using a wash bottle with a fine jet. wash the crucible and the filter
paper with four additional 5 ml portion of hot distilled water.
6.7 Cool the solution to room temperature and neutralize by drop wise
addition of 50% potassium hydroxide solution. Add hydrochloric acid
drop wise.
6.8 Take an aliquot portion of the unknown slightly acid solution containing
iron and transfer it to a 50ml graduated flask.
6.9 Add 5 ml of the 10%hydroxyl ammonium chloride to reduce the iron
solution, adjust the pH of the slightly acid solution to 3-6 with sodium
acetate.
6.10 Add 4ml of 1.10 phenanthroline solution dilute to mark, mix and
measure absorbance after 5-10 mints at 515 mm.
6.11 Take 1 ml, 2 ml, 3 m, 4 ml. 5 ml & 10 ml of standard solution in 100 ml
volumetric flask.
6.12 Prepare standard solution proceeding as described under 6 9 to 6. 10.
6.13 Plot a graph between concentration and absorbance.

7. CALCULATION:
7.1 A = ∑be
A = Absorbance
E= Molar absorptivity
B = Path length of cell
C = Concentration

Aunk ×CK
7.2 CUNK =
AK
Ak = Absorbance of known
Ck = Concentration of Known
Aunk = Absorbance of unknown
Cunk = Concentration of unknown

8. REFERENCE:
8.1 Quantitative chemical analysis be Vogel’s.
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TURBIDITY TEST

1. DEFINITION:
1.1 This method determines turbidity as Nepheometric Turbidity Units (NTU) in sample.

2. SCOPE:
2.1 Applicable to Refined, Bleached and Deodorized oil.

3. APPARATUS:
3.1 Beaker 100 ml
3.2 Sample cell for use with 2100 P Turbidity meter
3.3 Turbidity meter 2100 P

4. REAGENT:
4.1 Turbidity standard kit model 2100 P

5. CALIBRATION OF TURBIDITRMETER:
5.1 Fill the sample cell to the line and then insert in the cell compartment by aligning the orientation mark on
the cell with the mark on the front of the cell compartment. Close the lid and press 1/0.
5.2 Press "CAL" The CAL and so (standard) icons will be displayed. The instrument will count from 60 to 0 (67
to 0 average is on ) read the blank and use it to calculate a correction factor for the 20 NTU standard
measurement.
5.3 The display will automatically shift to the next standard. Remove the sample cell from the cell
compartment.
5.4 Instrument automatically display SI (standard I with the I flashing ) and 20 NTU or the value of the SI
standard for the previous calibration. If the value is incorrect edit the value by pressing the -unite the
number that needs editing flashes. After editing fill a clean sample clean sample cell to the line with well
mixed 20 NTU standard and insert the cell in to the cell compartment.
5.5 The instrument will count 60 to 0 (67 to 0 if signal average is on), measure the turbidity and store the
value the display will automatically shift the next standard. Calibrate similarly using S2 (standard 2 i.e. 100
NTU) and S3 (standard 3 i.e. ) and S3 i.e. (standard 3 i.e. 800NTU).

6. PROCEDURE:
6.1 Take approx. 70 gm sample in a beaker and heat beaker and heat the sample to remove any moisture.
6.2 After this place the sample in a constant temperature bath for 15 minutes which is set for specified
temperature i.e. 20°C.
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6.3 The sample is then filled in to the sample cell, the sample is then allowed to set for several minutes at
25°C.
6.4 Place the sample cell in to the cell compartment of the Turbidity meter, cover the lid and push read to
determine the NTU. The reading should be stable.

7. REFERENCE:
7.1 HACH Company Manual
Sunflower Oil Processing Manual.

DETERMINATION OF OIL & GREASE

1. DEFINITION:
1.1 Dissolved or emulsified oil and grease is extracted from water by intimate content with trichloro trifluro
ethane or chloroform.

2. SCOPE:
2.1 Applicable to water& effluent water

3. SAMPLING & STORAGE:

3.1 Refer to procedure No. SOP-07 AD “Effluent water - Sampling, Storage & Preservation"

4. APPARATUS:
4.1 Separator funnel with stopper
4.2 Pyrex glass basin
4.3 Water bath
4.4 Filter paper
4.5 Measuring Cylinder 250 ml

5. REAGENTS:
5.1 Hydrochloric Acid (1:1)
5.2 Trichloro trifluro ethane/Chloroform
5.3 Sodium Sulphate

7. PROCEDURE:
7.1 Take 250 ml sample & transfer it in a sepratory funnel.
7.2 Acidify with HCL (1:1) generally 5 ml HCL is sufficient.
7.3 Carefully rinse cylinder with 30 ml chloroform and add solvent washings to separatory funnel.
7.4 Shake sepratory funnel vigorously for 2 minutes.
7.5 Let layers separate. Drain solvent layer through a funnel containing approximate 1 gm sodium sulphate.
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7.6 Extract twice more with 30 ml solvent.

7.7 Combined extract in Pyrex glass basin and wash the filter paper with an additional 10-20 ml solvents.
7.8 Evaporate the solvent on water bath at 70°C.
7.9 Cool in desiccator for 30 min and weight.

8. CALCULATION: (a−b)×1000
8.1 Oil & Ghee (mg/ltr) =
Volume of sample

8.1.1 Where
A = Weight of Glass Basin after drying
B = Weight of Empty Glass Basin

9. REFERENCE:
9.1 Aggregate Organic Constituents (5000)

DETERMINATION OF SOLID FATS CONTENT BY PULSE NMR

1. DEFINITION:
1.1 Solid fat content of hard fat as determined by this method is the ratio hydrogen nuclei of the solid phase
to total hydrogen nuclei of sample.

2. TEST METHOD PRINCIPLE:

2.1 Test portion is subject to melting and chilling at O°C in NMR tubes, then tempered at required
temperature solid fat content then measured.

3. SCOPE:
3.1 Applicable to margarine oils, plasticized shortening. hydrogenated & or non-hydrogenated animal and
vegetable fats, and to non-cocoa butter type confectionery fats.

4. APPARATUS:
4.1 NMR Tubes with stoppers
4.2 Water baths
4.3 Aluminum tempering blocks with holes suitable lo hold NMR tubes (see notes no.3 & 4). 4.4. Oven
maintained at 100 °C

5. CALIBRATION:
5.1 To calibrate the NMR, select CALIBRATE from the main menu and then the INSTRUMENT from the sub-
menu. Then select the option: NMR TEST from the INSTRUMENT SUB-MENU. Then select EXECUTE
SHORT FORM. The message Insert NMR TEST SAMPLE 1 (0.00% Solid) appears. Put the reference sample
of' 0% solid and select confirm and enter. In this way do it for 30% and 70% reference sample. A message
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NMR WITHIN LIMIT (OR NOT WITHIN LIMITS)-RECALIBRETE ANYWAY will be displayed. Press YES so that
the changed values are set is the instrument.

6. PROCEDURE- 1 (For fats other than CBS):

INSTRUMENT SETTING:
6.1 Number of scans = 4
6.2 Recycle delay time = 2secs

7. SAMPLE TREATMENT:
7.1 Melt the sample properly and filter it to remove any traces of moisture and foreign matter.
7.2 Fill the sample tubes and as per measuring temperature up to the volume that it should not exceed 4cm
in height.
7.3 Place the tubes in oven at 100°C for 10min cap them and then place in block maintained at70°C for 30
minutes (see notes no: 5).
7.4 Temper the sample tubes at 0°C for 60±2 minutes.
7.5 Transfer sample tubes to each measuring temperature i.e. 20, 25, 30. 40 °C or as per requirement, and
equilibrate for exactly 45 min. (see notes no.2)
7.6 Measure the NMR reading to each measuring temperature.
7.7 Results will be expressed in % SEC and to be reported as N value for corresponding measuring
temperature.

8. PROCEDURE-2 (For CBS): INSTRUMENT SETTING:

8.1 Number of scans = 1
8.2 Recycle delay time = 6secs

9. SAMPLE TREATMENT:
9.1 Melt the sample at 80°C and filter it to remove any traces of moisture and foreign matter.
9.2 Fill the tubes with 2ml of sample, cap it at 60°C for 5 minutes in water bath (see notes no 5).
9.3 Temper the sample tubes at 0°C for 60 + 2 min.
9.4 Transfer sample tubes to each measuring temperature i.e. 20, 25, 30. 40 °C or as per requirement, and
equilibrate for exactly 30-35 minutes. (see notes no.2)
9.5 Measure the NMR reading to each measuring temperature.
9.6 Rules will be expressed in % SFI and to be reported as N value for corresponding temperature.

10. NOTE:
10.1 The room temperature where the low resolution NMR equipment placed should be stable between 15-
30°C on robust table to prevent influence of vibrations large moving metal objects should be kept away
from the instrument as they may interfere with the magnetic field. The magnet should be thermostatic
usually at 40°C to reduce drift.
10.2 After being removed on at 0°C tubes should be carefully and quickly dried before being placed in the
metal blocks in water baths at measuring temperature. Tubes should not be wiped with tissue when
placed from the water bath at the measuring temperature to sample holder of the instrument, because
this may interfere with the results. The holes in the metal block in water baths must be dried frequently
because moisture may condense in holes.
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10.3 SFC values depend on the thermal history of crystallized fat, it is therefore important to adhere to
prescribed method. Temperatures of tempering and measurements are also extremely important and
thermostat settings together with both dimensions of the tubes and the holes in metal blocks should be
strictly observed, the latter in order to ensure the appropriate thermal contact. The influence of
temperature and time are especially important at temperatures where the confectionery fats have a
steep melting profile.
10.4 Diameter of the holes in tempering blocks must not be greater than 5mm of the mean external diameter
of the glass measuring tubes. The depth of the holes should be such that the level of the test sample is
5mm below the outside top surface of the block. Thickness of the metal under the holes and the distance
between the edge of the peripheral hole and the nearest lateral face should be at least 5mm.
10.5 It can be advantageous to use some closure to the tube to limit ingress of dust, solvent vapors or
moisture during tempering steps.
10.6 It is recommended to recalibrate the instrument after every change in parameters.
10.7 Diameter of the holes in tempering blocks must not be greater than 5mm of the mean external diameter
of the glass measuring tubes. The depth of the holes should be such that the level of the test sample is
5mm below the outside top surface of the block. Thickness of the metal under the holes and the distance
between the edge of the peripheral hole and the nearest lateral face should be at least 5mm.

11. SAFETY:
11.1 Persons with a pacemaker should not come within a distance of 30cm of the NMR equipment. Heavy
metal objects should be avoided in the vicinity of the NMR
PPE: Safety goggles, Gloves required during handling of hot oil.

12. REFERENCE:
12.1 American Oil Chemist Society/AOCS Cd 16b – 93 (IUPAC Parallel Method)
12.2 Bruker Pulse NMR Method for CBS

DILATATION OF FAT

1. DEFINITION:
1.1 The dilatation of fat is the isothermal expansion due to change of state from solid to liquid of fat which
has previously been solidified under carefully prescribed conditions. It can be used for the mixture of oil
and fats.

2. SCOPE:
2.1 Applicable to margarine oils, shortening and other fats.

3. APPARATUS:
3.1 Pyrex dilatometer having a volume of 6.5 to 7.5 mm³.
3.2 Dilatometer stopper filled 40 to 50 g led shorts.
3.3 Clamps for holding the dilatometer in the water bath.
3.4 Constant temperature water bath accurate to 2 mm Hg or less.
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4. REAGENTS:
4.1 Sealing liquid, Congo Red (0.1 % in Water) boiled under vacuum.

5. PROCEDURE:
5.1 Filter the melted fat to be investigated if it is not clear. Pour the fat into a 100 ml vacuum flask.
5.2 Add a few pieces of porous pot, heat to 100°C and evacuate for at least 10 min, shaking vigorously at
intervals, until air bubbles no longer escape. Keep the melted fats under vacuum until it has been put in
the dilatometer.
5.3 Using a burette, put exactly 2 ml sealing liquid.
5.4 Weigh the dilatometer with stopper.
5.5 Fill the dilatometer with the prepared liquid fat at 70 °C.
5.6 Firmly insert the stopper, thus causing the sealing liquid to rice in capillary. Make sure that no air remains
in the dilatometer. Alter cooling and drying weight of the fat.
5.7 Immerse the dilatometer to the zero mark in a water bath at the temperature of 60°C. After 20 minutes
read off the height of the sealing air meniscus in the capillary (approximately 800 mm) at equilibrium
temperature of 60°C.
5.8 Remove the dilatometer from the water bath and place it in a mixture of finely ground ice for 1.5 hour.
5.9 Remove the dilatometer from the ice and return it to the water bath at 20 °C. When constant volume is
reached (within 20 min) record the level of the meniscus.
5. 10 Place the dilatometer in a water bath having the temperature at which the next dilatation is to be measured.
When the volume remains constant again note the volume of meniscus.
5.11 Repeat reading a 60 °C. The duplicate values obtained must agree to within 1mm³. Greater differences may be due
to leak in the dilatometer or to air bubbles in the fat.

6. CALCULATION: (R60−Rt−wt)
6.1 Dt = − VT
W

Dt = Dilatation at temperature t
W = Weight of sample
R 60 = Reading at 60°C
Rt = Reading at t°C
Wt = Water and glass connection at t°C (table # 1)
Vt = Oil correction at t°C (table # 2)

Table-1

Height of Meniscus at 60°C

(Water and glass correction)
t 0 100 200 300 400 500 600 700 800 900
20 28 26 25 23 22 20 19 17 15 14
25 26 24 23 21 20 19 17 16 14 13
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30 23 22 20 19 18 17 15 14 13 12
35 20 19 18 17 16 15 13 12 11 10
40 17 16 15 14 13 12 11 10 09 08
45 13 12 12 11 10 10 09 08 07 07
50 09 09 08 08 07 07 06 06 05 05
55 05 04 04 04 04 03 03 03 03 02
60 00 00 00 00 00 00 00 00 00 00
10 33 30 29 28 27 25 24 22 20 19

DILATATION OF FAT
Table-2
Temperature °C Oil Correction (mm³/25g)
20 852
25 748
30 642
35 537
40 430
45 324
50 216
55 108
60 00
10 1060

7. REFERENCE:
7.1 American Oil Chemist Society/AOCS Cd 10 – 57

GASS ANALYSIS

1. DEFINITION:
1.1. This method determines the purity of Hydrogen gas produced by the Gas plant.

2. APPARATUS:
2.1. Orsat apparatus
2.2. Beaker 300 ml
2.3. Measuring cylinder 100 ml
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3. REAGENT:
3.1. Potassium hydroxide solution ( 50 % in distilled water)
3.2. Pyrogallol solution ( 10 gm of pyrogallol / 125 ml of50 % KOH )
3.3. Copper Chloride Solution
3.3.1 Weigh 10 gms of CuCI in a beaker.
3.3.2 Add 50 ml distilled water.
3.3.3 Add 8.5 gm Na₄Cl.
3.3.4 The beaker is heated slightly with stirring.
3.3.5 Add 75 ml NH₄OH (Conc.)

4. PROCEDURE:
4.1. Hydrogen gas is taken in to cylinder by vertical line bubbled in to the bottle.
4.2. Rinse the graduated cylinder three times with hydrogen gas.
4.3. Take the gas into cylinder up to Zero mark.
4.4. Then absorb the gas into CO₂ bulb containing 50% KOH up down three times and note the reading on
cylinder by percentage CO₂ in Hydrogen gas. Again absorb the gas till a constant reading is obtained. Then
absorb the gas similarly in O₂ & C₂ bulb.

IDENTIFICATION OF KETONE IN GROUP

1. IDENTIFICATION
1.1 This method determine presence of ketone group in sample.

2. SCOPE:
2.1 Applicable to all sample having ketone group.

3. APPARATUS:
3.1 Beaker 100 ml
3.2 Test Tube
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4. REAGENTS:
4.1 Sodium Hydroxide
4.2 Potassium Hydroxide

5. SOLUTION:
5.1 Prepare saturated solution of sodium Nitroprusside in beaker, alkaline it with the help of Potassium
Hydroxide.

6. PROCEDURE:
6.1 Take small quantity of solution i.e. Mono Ethyl Ketone & add approximately 01 ml of alkaline Sodium
Nitroprusside red coloration indicates presence of ketone group .

7. REFERENCE:
7.1 Practical Organic Chemistry by Vogel’s.

IDENTIFICATION OF NITROGEN GAS

1. DEFINITION:
1.1 This method is determines presence of nitrogen gas.

2. SCOPE:
2.1 Applicable to Nitrogen gas.

3. APPRATUS:
3.1 Gas jar with lid.
3.2 Key to open the valve of the cylinder.
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3.3 Match box.

4. PROCEDURE:
4.1 Open valve of cylinder slowly with the help of key.
4.2 Collect the nitrogen gas in 2 cylinder.
4.3 Smell the gas & note down the observation.
4.4 Observe the colour of the gas.
4.5 Take burning match stick and put into gas jar having nitrogen gas, since nitrogen gas does not support the
process of burning, it will immediately extinguish the fire.

5. REFERENCE:
5.1 Text book of Chemistry

DETERMINATION OF TRANCE METALS IN FATS (IRON,COPPER,NICKLE)

1. APPARATUS:
1.1. Spectrophotometer VISIBLE RANGE
1.2. Porcelain Dishes 150 ml 400 ml
1.3. Flask with ground glass joint 500ml
1.4. Condenser with ground glass joint
1.5. Volumetric Flasks 100ml, 50, 25m,1 Liter
1. 6.Separating Funnel 250ml, 125m

2. PREPARATION OF SOLUTION OF TIHE SAMPLE:

2.1. Ignite 35 to 50 gms. Sample in a porcelain dish tor individual metals or 100 to 120 gms. For the three
metals - Cool, moisten with Hydrogen Peroxide, evaporate and again ignite Dissolve the Ash in 25 ml
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Hydrochloric Acid (1 : 1) and make up volume to 100 ml. Filter in a dry 100 ml reagent bottle and marks as MAIN
SOLUTION for the estimation Fe, Cu & Ni.
2.2. Alternatively take 100-120 gms sample in a 500ml flask, Add 50 ml of hydrochloric Acid (1:1) and Reflux
for one hour. Transfer the contents lo a separating funnel while warm, shake well allow to
Settle and collect the acid layer in a beaker. Rise the flask twice with 50 ml portions of hot distilled water and wash
the fat in the separating funnel. Collect the washings in the same beaker concentrate the acid extract and make up
volume to 100 ml. Filter in a 100 ml reagent bottle and make as "MAIN SOLUTION".

DETERMINATION OF IRON

1. REAGENTS:
1.1. Potassium thiocynate solution 10% (Dissolve in water) 1.2. Hydrochloric acid 0.5N Approx.
1.3. Standard Iron solution 1ml = 0.1 mg
1.4. Dissolve 0.7022 gms Ferrous Ammonium Sulphate Fes0₄(NH₄)2SO₄ 6H₂0 in 100 ml water. Add carefully
dilute per magnate Solution (0.2%) from the burette while stirring till slight pink colour persists and dilute
to 1 Liter.

2. PROCEDURE:
2.1. Take 25 ml aliquot solution from the main solution in a 100 ml beaker evaporate to dryness. Add 10 ml
Hydrochloric Acid heat and transfer to a 25 ml volumetric flask rinse the beaker with water an add to the flask. Add
5 ml Potassium thiocynate solution. Mix well and measure the absorbance in 1cm cell at 480 nm wave length on a
Spectrometer, within 10 minutes.
2.2. Prepare a standard curve by treating 0.1 ml, 0.3 ml, 0.6ml 1 5 ml and 1.6 ml of the standard iron solution
in the same way.
3. CALCULATION: C×100×1000 C×4000 Iron as Fe (mg/kg)(ppm) = =
25×W W
Where, C = mg of Cu read for the curve
W = Weight of the sample in gms

DETERMINATION OF NICKEL

1. REAGENTS:
1.1. Dimethyl Glyoxime solution 1% in 95% Alcohol
1.2. Ammonium Hydroxide solution 1:1
1.3. Bromine Water standard solution
1.4. Standard Nickel solution 1ml = 0.01 gm Ni
1.5. Dissolve 0.673 gm of pure Nickel Ammonia Sulphate in water and dilute to I Itr. Further dilute 10 ml to
100 ml.

2. PROCEDURE:
2.1. Take 25 ml aliquot portion from the main solution in a 50 ml volumetric flask and 0.5 ml of Bromine water
and Ammonia hydroxide Solution drop by drop till colour of beomine is destroyed add 3 drops more Add 3
ml Dimethyl Glyoxime solution and make up volume to 50 ml.
2.2. Mix and measure the absorbance of the solution at 445 nm wave length within 10 minutes. Prepare a
standardization curve by treating 1m,2 ml, 4 ml, 6 and 10 ml of standard Nickel solution in the same way.
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3. CALCULATION:
𝐂×𝟏𝟎𝟎×𝟏𝟎𝟎𝟎 𝐂×𝟐𝟎𝟎𝟎
Nickel as Ni mg/ kg (ppm) = =
𝟓𝟎×𝐖 𝐖

Where C = mg of Ni read from the curve.

W = Weight of sample in gms.

DETERMINATION OF COPPER
1. REAGENTS:
1.1. Sodium diethyl, Di thio carbamate solution 0.02% in water. Prepare fresh, keep in refrigerator.
1.2. Ammonium hydroxide solution 1:4
1.3. Citric Acid solution 25% in water
1.4. Phenol red indicator
1.5. Chloroform
1.6. Standard COPPER SOLUTION 1 ml, 0.1 mg CU,
Dissolve 0.3927 gm Copper sulphate CuS04.5120 in 1 Liter water. Further dilute 10ml to 100 ml to get 1ml = 0.01
mg CU.
1.7. EDTA Solution 4% in water.

2. PROCEDURE:
2.1. Take 25 ml aliquot portion from the main solution in a 100 ml beaker, add 5 ml Citric Acid solution and
adjust pH to 8.5 by adding ammonia and phenol red indicator.
2.2. Add 15 ml EDTa Solution transfer to a separating funnel. Add 10 ml Sodium Diethyl Di thiocarbamate
solution and add 20 ml Chloroform. Shake allow to settle and transfer the chloroform layer to dry 50 ml
volumetric flask extract with further 10 ml quantities of chloroform and add to the flask. Make up to 50 ml
with chloroform.
2.3. Measure the absorbance of the Solution at 435 nm wave length.
2.4. Prepare a standard curve by treating 1ml, 2ml, 6ml and 10ml Copper Solution. 1ml = 0.01mg. Cu in the
same way.

3. CALCULATION:

C×100×1000 C×4000
Copper as Cu mg/kg (ppm) =
=
25×W W
Where C = mg of Cu read for the curve
W = Weight of sample in gms

3. REFERENCE:
3.1. American Oil Chemist Society/AOCS , Vogel’s

HARDNESS OF WATER
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1. DEFINITION:
1.1. This method determines the hardness in the water sample in terms of Calcium carbonate.

2. SCOPE:
2.1. Applicable to drinking by surfaces water and domestic by industrial wastes.

3. PRINCIPLE:
3.1. Hardness of water is generally due to dissolved calcium and magnesium salts and may be determined by
complexometric titration. In comlexometric titration calcium or magnesium ions. Present in water from a
red complex with Eriochrome Black -T, upon addition of EDTA a colourless complex with Ca⁺⁺² or Mg⁺⁺²
ions are formed at pH 10. End point is detected by the change of colour of the indicator which regains its
blue colour when all of the Ca & Mg. Ions have formed a complex with EDTA.

4. APPRATUS:
4.1. Measuring cycle 100ml
4.2. Conical flask 250 ml
4.3. Pipette 25 ml
4.4. Burette 50 ml

5. REAGENTS:
5.1. EDTA (Disodium - di hydrogen ethylene di amine tetra acetate).
Dissolve 3.7224g of EDTA (Na₂H₂C₁₀H₁₂O₈N₂. 2H₂O) in 1000 ml of dis. H20.
STADARDIZATION OR EDTA SOLUTION
5.1.1. Prepare 0.01 M CaCO₃ in distilled water in 250 ml Volumetric Flask.
5.1.2. Place funnel in neck and add little at time, HCL (1:1) until all CaCO₃ has dissolved.
5.1.3. Add 50 ml distill water and boil for few minutes to expel CO₂.
5.1.4. Cool, add few drops Methyl indicator, & adjust to intermediate orange with 3.N.NH₄OH or HCL
(1:1), as required.
5.1.5. Now make up to 250ml with distill water.
5.1.6. Pipette 10 ml CaCO₃ solution in a 250 ml conical flask.
5.1.7. Add 1 ml buffer solution and 30-40 mg of Erochrome Black- T indicator.
5.1.8. Titrate against EDT A till red colour change to blue.

Calculate the molarity of EDTA as follows:

M₁×M₂
M₂ =
V₂
M₁ = Molarity of CaCO₃
V₁ = Volume of CaCO₃
M₂ = Molarity of EDTA
V₂ = Volume of EDTA
5.2. ERIOCHROME BLACK -T (SOLOCHROME BLACK -T)
5.2.1. Eriochrome Black -T is sodium 1-( 1-hydroxyl- 2-napthylazo )- 6-nitro- 2-napthano-4-
sulphonate.
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5.3. EQUIMOLAR SOLUTION OF MAGNESIUM SULPHATE & EDTA.

5.3.1. Prepare the magnesium complex of EDTA (Na₂, Mg, EDTA) by mixing equal volumes of 0.2M
solution of EDTA(7.4440 g/100ml distill water) and of magnesium sulphate (2.4 g/100 ml
distilled water).
5.3.2. Neutralize with NaOH solution to a pH between 8 & 9 using phenolphthalein indicator.
5.3.3. Take a portion of solution, add a few drops of buffer solution and a few milligram of indicator a
violet colour should be produced which turns blue on the addition of a drop of 0.01M EDTA
solution and red on the addition of single drop of 0.01M (5ml of 0.2M/100 m) magnesium
sulphate solution, this confirms the equal molarity of magnesium and EDTA.
5.3.4. If the solution does not pass this test, it may be treated with more EDTA or with more.
5.3.5. Magnesium sulphate solution until the required condition of equal molarity is attained this an
approximately
0.1M solution.
5.4. AMMONIA-AMMONIUM CIILORIDE BUFFER SOLUTION (pH = 10)
5.4.1. Dissolve 17.5 gm of Ammonium chloride in 142 ml conc. Ammonia solution (sp: gravity 0.88-
0.90).
5.4.2. Add 25ml of equimolar solution of Mg.SO₄ & EDTA.
5.4.3. Dilute to 250 ml with distill water.

6. PROCEDURE:
6.1. Take 50 ml water sample to be tested in a conical flask.
6.2. Add 1 ml buffer solution.
6.3. Add 30-40 mg of indicator mixture.
6.4. Titrate with standard EDTA solution until the colour changes from red to blue.
6.5. The total hardness is expressed in parts of CaCO₃ per million of water sample.
6.6. If the water contains interfering ions e.g. CO, Ni, Cu, Zn, Hg & Mn then 4ml of buffer should be added
followed by 30 mg of hydroxyl ammonium chloride and then analytical grade KCN (caution) before adding
the indicator.

7. CALCULATION:

VE×M×MW×(10)⁶
TH =
1000×VM
TH = Total hardness
VE = Volume of EDTA used
M = Molarity of EDTA
MW = Mol. Wt of CaCO₃ = 100gm
VW = Volume of water sample taken

8. NOTE:
8.1. Somewhat sharper end points may be obtained if the sample of water is first
acidified with dil. HCI, boiled for about a minute to drive off carbon dioxide,
cooled, neutralize with NaOH solution, buffer and indicator solution and
added then titrated with EDTA as above.
8.2. To avoid large titration volumes, use aliquot containing less than 25mg CaCO3.
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7. REFERENCE:
7.1. Vogel’s Text book of quantitative chemical analysis.

DETERMINATION OF CHEMICAL OXYGEN DEMAND(COD)

1. DEFINITION:
1.1. COD is the Quantity of oxygen consumed by organic matter susceptible to oxidation in defined condition.

2. SCOPE:
2.1. Applicable to water & effluent water.

3. APPARATUS:
3.1. One precision Balance (capable of weighing to 0.1mg)
3.2. Two 250 ml Flat bottom Flask.
3.3. Transfer pipette 5, 10 & 25 ml.
3.4. One 50 ml Burette.
3.5. Electric Hot plate.
3.6. Reflux Condensers.
3.7. Measuring Cylinder 25 & 50 ml.
3.8. Magnetic stirrer.

4. REAGENTS:
4.1. Mercury II Sulphate, Hgso4 in powder Form.
4.2. Silver Sulphate Agso4 in powder form.
4.3. Concentrated Sulphuric Acid.
4.4. Mohrs salt.
4.5. Potassium dichromate.
4.6. O-Phenonthroline.
4.7. Ferrous Sulphate.
4.8. Sulphuric Acid Reagent (22gm Agso4/4 kg concentrated H2so4)
4.9. Ferrion indicator solution (1.485 gm o-Phenonthroline &695 mg FeSo4.7H2o in water and dilute to
100 ml).

5. PROCEDURE:
5.1. Place 50 ml sample or a smaller sample portion diluted to 50 ml in the 250 ml refluxing flask.
5.2. Add 1 gm Hgso4 several boiling chips and 5 ml H2SO4.
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5.3. Add 25 ml 0.25 N K2Cr2O7 solution.

5.4. Attach the flask to the condenser and start the cooling water.
5.5. Add the Acid Reagent ( 30-70 ml ) through the open end of the condenser.
5.6. Reflux the mixture for 2 hours.
5.7. Cool and wash down the condensor with distilled water.
5.8. Dilute the mixture to about twice its volume with distilled water and cool to room temperature.
5.9. Titrate versus Ferrous ammonium sulphate ( 0.1 N ) using ferrion Indicator.
5.10. At the end point the sharp colour change from blue green to reddish brown appeared.
5.11. Reflux in the same manner a blank consisting of distilled water.

7. CALCULATION:

COD (mg/Ltr) = (a−b)×N×100

Volume of sample

Where:
COD = Chemical Oxygen Demand from dichromate
a = Blank Reading b
= Sample Reading
N = Normality of the ferrous Ammonium Sulphate

8. REFERENCE:
8.1. PTA Environmental Management Program Laboratory Manual.
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DETERMINATION OF TOTAL SUSPENDED SOLID (TSS)

1. DEFINITION:
1.1. Total non- filterable residue is the retained material on a standard glass fiber disk after filtration of
well mixed sample of water or waste water. The residue is dried at 103 105°C. The difference between the
total residue and total filterable residue provides an estimate of the total non-filterable residue.
2. SCOPE:
2.1. Applicable to water & effluent water.

3. APPARATUS:
3.1. Measuring cylinder 500 ml.
3.2. Glass Fiber Filter Disks.
3.3. Filtration Apparatus.

4. PROCEDURE:
4.1. Dry the glass fiber filter paper in an oven at 103-105°C for 1 hour store in desiccator or until needed.
4.2. Take 250 ml well mixed sample in a measuring cylinder and filter it under vacuum through glass Filter
paper.
4.3. Carefully remove the filter paper from funnel. 4.4. Dry it for at least 1 hour at 103-105°C
4.5. Cool in a desiccator and weigh.
4.6. Repeat the dry cycle until a constant weight is obtained.

7. CALCULATION:
(𝐚−𝐛)×𝟏𝟎𝟎𝟎
TSS =
𝐕𝐨𝐥𝐮𝐦𝐞 𝐨𝐟
𝐬𝐚𝐦𝐩𝐥𝐞
Where:
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TSS = Total suspended solids a

= Weight of filter + residue b
= Weight of filter paper

8. REFERENCE:
8.1. PTA Environmental Management Program Laboratory Manual.

QUALITATIVE & QUANTATIVE ESTIMATION OF PHOSPHORIC ACID

A. QUALITATIVE ESTIMATION:
Phosphoric acid is identified by the reaction with AgNo3 Add silver Nitrate solution (0.1% in H2O). To phosphoric
acid , yellow ppt of Agpo4 is obtained. This yellow ppt is soluble in diluted HNO₃ and NH₄OH like many salts of silver.

B. QUANTATIVE ESTIMATION:
1. DEFINITION:
1.1. Phosphoric acid is estimated quantitatively by simply determining its percentage acidity. The percent
acidity is the amount of NaOH necessary to neutralize the acid ,calculated by wt. of the acid of specific
molecular weight.

2. SCOPE:
2.1. Applicable to commercial and analytical grade phosphoric.

3. APPARATUS:
3.1. Beaker 100 ml .
3.2. Volumetric flask, 250 ml or 100 ml.
3.3. Pipette 5 ml or 10 ml.
3.4. Conical Flask, 250 ml.

4. REAGENT:
4.1. Phenolphthalein indicator solution (0.1% in spirit).
4.2. 2.0N NaOH Solution (20 g/250 ml distill water).
0.05 N NaOH (25ml of 2N NAOH/1000 ml distilled water.
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5. PROCEDURE:
5.1. Weight 2 to 5 gm of sample and dilute to 250 ml or 100 ml min a volumetric flask.
5.2. Pipette out 2 to 10 ml of the sample solution in a volumetric flask.
5.3. Add few drops of phenolphthalein indicator solution and titrate with 0.05N NaOH.

6. CALCULATION :

V×EW×N×V₁×100
Acidity % =
1000×W×V₂

V = Volume of NaOH used

V₁ = Volume of total solution
V₂ = Volume of solution pipette out
EW = Eq. wt. of phosphoric acid=49
N = Normality of NaOH
W = wt. of sample taken

7. REFERENCE:
7.1. Vogel’s Text book of Quantitative Chemical Analysis.

QUALITATIVE & QUANTATIVE ESTIMATION OF CITRIC ACID

A. QUALITATIVE ESTIMATION:
Citric Acid + CaCl₂ = White ppt

B. QUANTATIVE ESTIMATION OF CITRIC ACID:

1. DEFINITION:
1.1. Citric acid is estimated quantitatively by simply determining its acidity. The % acidity is the amount of
NaOH necessary to neutralized the acid, calculated by wt. of the acid of specific molecular weight.

2. SCOPE:
2.1. Application to commercial and analytical grade Citric acid.

3. APPARATUS:
3.1. Beaker 100 ml.
3.2. Volumetric flask 250 ml or 100 ml.
3.3. Pipette 5 ml or 10 ml.
3.4. Conical flask 250 ml.

4. REAGENTS :
4.1. Phenolphthalein indicator solution ( 0.1% in spirit ).
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4.2. 2.0N NaOH solution (20gm/250ml distilled water).

4.3. 0.05N NaOH (25 ml of 2N NaoH /1000 ml distilled water).
STANDARIZATION OF NaOH 4.3.1.
Weight clean dry conical flask.
4.3.2. Weight about 0.1 gm of potassium hydrogen phthalate which is already dried in oven for 4 hrs
at 105°C & cool in desiccator and dissolve in distilled water.
4.3.3. Add few drops of phenolphthalein indicator.
4.3.4. Titrate with NaOH solution of which strength is to be determined.
4.3.5. Calculate the normality of NaOH by using formula.

W×1000
N=
EW×V
N = Normality of NaOH
W = Wt. of KH- phthalate taken
EW = Eq. wt. of KH-phthalate = 204.22
V = Volume of Naoh cosumed.

5. PROCEDURE:
5.1. Weight 2 to 5 gms of sample and dilute to 250 ml or 100 ml in a volumetric flask.
5.2. Pipette out 2 to 10 ml of the sample solution in to a conical flask.
5.3. Add few drops of phenolphthalein and titrate with 0.05N NaOH solution.

6. CALCULATION: V×EW×N×V₁×100
Acidity % =
1000×W×V₂
V = Volume of NaOH used
V₁ = Volume of total solution
V₂ = Volume of solution pipette out
EW = Eq. wt. of Citric acid = 64 as anhydrate, 70 as monohydrate.
N = Normality of NaOH
W = wt. of sample taken

7. REFERENCE:
7.1. Vogel’s Text book of Quantitative Chemical Analysis.
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QUALITATIVE & QUANTATIVE ESTIMATION OF BUTYRIC ACID

A. QUALITATIVE ESTIMATION:
Butyric acid is identified by its characteristic odor.

B. QUANTITATIVE ESTIMATION:
1. DEFINITION:
1.1. Butyric acid is estimated quantitatively by simply determining its percentage acidity. The percent acidity is
the amount of NaOH necessary to neutralize the acid, calculated by wt. of the acid of specific molecular
weight.

2. SCOPE:
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2.1. Applicable to commercial and analytical grade Butyric acid.

3. APPARATUS:
3.1. Beaker 100 ml.
3.2. Volumetric flask.
3.3. Pipette 5 ml of 100 ml.
3.4. Conical flask 250 ml.

4. REAGENTS:
4.1. Phenolphthalein indicator solution ( 0.1 % in spirit)
4.2. 2.0 N NaOH (20g/ 250 ml distilled water ).
4.3. 0.05 N NaOH ( 25 ml of 2N NaOH / 1000 ml distilled water )

5. PROCEDURE:
5.1. Weight 2-5 gms of sample and dilute to 250 ml or 100ml i8n a Volumetric flask.
5.2. Pipette out 2-10 ml of sample solution in a conical flask.
5.3. Add few drops of phenolphthalein indicator and titrate with 0.05 N NaOH solution.

6. CALCULATION: V×EW×N×V₁×100
Acidity % =
1000×W×V₂
V = Volume of NaOH used
V₁ = Volume of total solution
V₂ = Volume of solution pipette out
EW = Eq. wt. of Butyric acid = 88
N = Normality of NaOH
W = wt. of sample taken

7. REFERENCE:
7.1. Vogel’s Text book of Quantitative Chemical Analysis.

QUALITATIVE & QUANTATIVE ESTIMATION OF SULPHURIC ACID

A. QUALITATIVE ESTIMATION:
1 – 2 ml sulphuric acid + 3ml of 10 % Barium chloride solution = White ppt of BaSO₄ (insoluble mineral acids like
HNO₃ and HCL).

B. QUANTITATIVE ESTIMATION:
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1. DEFINITION:
1.1. Sulphric acid is estimated quantitatively by simple determining its percentage acidity. The percent acidity
is the amount of NaOH necessary to neutralize the acid , calculated by wt. of the acid of specific molecular
weight.

2. SCOPE:
2.1. Applicable to commercial and analytical grade Sulphuric acid.

3. APPARATUS:
3.1. Beaker 100 ml
3.2. Volumetric flask, 250 ml or 100 ml.
3.3. Pipette 5 ml or 10 ml .
3.4. Conical flask, 250 ml.

4. REAGENTS:
4.1. Phenolphthalein indicator solution (0.1% in spirit)
4.2. 2.0N NaOH solution (20 g / 250 ml distilled water)
4.3. 0.05N NaOH (25 ml of 2 N NaOH / 1000 ml distill water)

5. PROCEDURE:
5.1. Weight 2 – 3 gms of sample and dilute to 250 ml or 100 volumetric flask.
5.2. Pipette out 2 – 5 ml of sample solution in a conical flask.
5.3. Add few drops of phenolphthalein indicator and titrate with 0.05 N NaOH solution.

6. CALCULATION:

V×EW×N×V₁×100
Acidity % =
1000×W×V₂
V = Volume of NaOH used
V₁ = Volume of total solution
V₂ = Volume of solution pipette out
EW = Eq. wt. of Sulphuric acid = 49
N = Normality of NaOH
W = wt. of sample taken

7. REFERENCE:
7.1. Vogel’s Text book of Quantitative Chemical Analysis.

QUALITATIVE & QUANTATIVE ESTIMATION OF HYDROCHLORIC ACID

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A. QUALITATIVE ESTIMATION:
A.1. Hydrochloric acid + Silver Nitrate (0.1% in water) = Silver Chloride white precipitate.
A.2. When NaOH is added to this precipitate it becomes soluble due to the formation of di amine silver
chloride.

B. QUANTITATIVE ESTIMATION:
1. DEFINITION:
1.1. Hydrochloric acid is estimated quantitatively by simply determining its percentage acidity. The percent
acidity is the amount of NaOH necessary to neutralize the acid, calculated by wt. of the acid of specific
molecular weight.

2. SCOPE:
2.1. Applicable to commercial and analytical grade hydrochloric acid.

3. APPARATUS:
3.1. Beaker 100 ml
5.2 Volumetric flask, 250 ml or 100 ml
3.3 Pipette 5 l or 10 ml
3.4 Conical flask, 250 mil

4. REAGENTS:
4.1. Phenolphthalein indicator solution (0 1%, in spirit)
4.2. 2.0N NaOH solution (20gm/250ml distilled water)
4.3. 0.05 N NaOH (25 ml of 2N NaOH/1000 ml distilled water)

5. PROCEDURE
5.1. Weight 2-3 gms of sample and dilute to 250 ml or 100 ml volumetric flask
5.2. Pipette out 2-10 ml of sample solution in a conical flask.
5.3. Add few drops of phenolphthalein indicator and titrate with 0.05 N NaOH solution.

6. CALCULATION

V×EW×N×V₁×100
Acidity % =
1000×W×V₂
V = Volume of NaOH used
V₁ = Volume of total solution
V₂ = Volume of solution pipette out
EW = Eq. wt. of Hydrochloric acid = 36.5
N = Normality of NaOH
W = wt. of sample taken

7. REFERENCE:
7.1. Vogel’s Text book of Quantitative Chemical Analysis.
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QUALITATIVE & QUANTATIVE ESTIMATION OF MONO ETHANOL AMINE

A. IDENTFICATION:
Mono ethanol Amine is identified by the reaction of Ferric Chloride. Add Ferric Chloride solution to Mono
ethanol amine diluted Red ppt obtained.

B. QUANTITATIVE ESTIMATION:
1. DEFINITION:
1.1. Mono ethanol amine is estimated quantitatively by simply determining its percentage. The percentage is
calculated by neutralized mono ethanol amine with HCL.

2. SCOPE:
2.1. Application to commercial and analytical grade liquid mono ethanol amine.

3. Apparatus:
3.1. Beaker 100 ml
3.2. Volumetric flask, 250 ml or 100 ml
3.3. Pipette 5 ml or 10 ml
3.4. Conical flask, 250 ml

4. REAGENTS:
4.1. Phenolphthalein ( 0. 1 % in spirit)
4.2. Methyl Orange (0.1% in water)
4.3. HCL 0.IN in distilled water
4.4. Methyl Red (0.1% in water)

4.4.1. Concentrated HCI is 36-37% its normality is first calculated to make 0.1N HCL from it.

N = D×P×1000
100×EW

D = Density of HCL given as Kg/Liter

P = Percentage acidity given on the bottle label
EW = Eq wt. of HCL i.e. 36.5 g
N = Normality in grams equivalent / liter

4.4.2. Now to make 0.1 N solution in 1000 ml of volumetric flask

N₁×V₁
V₂ =
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N₂

V₂ = Volume of HCL conc. Required

V₁ = Volume of 0.I N solution
N₁ = Normality of which solution is to be prepared
N₂ = Normality of HCL concentrated standardize the prepared sol with Na₂S₂0₃ anhydrous.
4.4.3. Standardization of HCL
a. Weight accurately of 0.8g of primary standard anhydrous sodium carbonate that previously has been heated at a
temperature of about 250 °C for 15 hrs. b. Dissolve in 100 ml of distilled water.
c. Add 2 drops of methyl red.
d. Titrate with the acid with constant stirring until the solution becomes faintly pink.
e. Heat the solution to boiling, cool, and continue the titration.
f. Heat again to boiling, and titrate further as necessary until the faint pink colour is no longer affected by continued
boiling
g. Calculate the normality as following:

W×1000
N=
EW×V
N = Normality of HCL calculated
W = Weight of Na₂S₂0₃ taken
EW = Eq. wt. of Na₂S₂0₃ i.e. 52.99g
V = Volume of HCL used

Methyl Orange may also be used for this titration which shows a colour change from pale yellow to pink. No heating is
required when methyl orange is used.

5. PROCEDURE:
5.1. Weight 2 - 4 gms of liquid mono ethanol amine in 250 ml or 100 ml volumetric flask.
5.2. Titrate 2-5 ml portion, diluted with water, with standard 0.1N HCl using methyl orange indicator.
5.3. The colour change from orange to red.
5.4. The volume of HCL used is Vн ml.

6. CALCULATION:

Vн×N×EW×V₂×100
Percent MEA =
1000×W×V₁

Vн = Volume of HCL used for hydroxide

N = Normality of HCI
EW = Eq. wt. of Mono Ethanol Amine i.e. = 61
V = Total volume of Mono Ethanol Amine solution made EW
= Weight of sample of Mono Ethanol Amine taken.

7. REFERENCE:
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7.1. Vogel’s Text book of Quantitative Chemical Analysis.

QUALITATIVE & QUANTATIVE ESTIMATION OF SODIUM HYDROXIDE

A. QUALITATIVE ESTIMATION:
A.1. Sample + KH – Antimonate = Sodium pyro antimonate (Na₂H₂Sb₂O₇) (white crystalline ppt).

B. QUANTITATIVE ESTIMATION:
1. DEFINITION:
1.1. NaOH is estimated quantitatively by simply determining its percentage basicity. The percent basicity, is
the amount of HCL necessary to neutralize the base, calculated by wt. of the base, calculated by wt of
base of specific molecular wt. Sodium carbonate may be present in commercial NaOH whose percentage
is determined to get the actual NaOH percentage.

2. SCOPE:
2.1. Applicable to commercial and analytical grade, liquid and solid Sodium hydroxide.

3. APPRATUS:
3.1. Beaker 100 ml.
3.2. Volumetric flask, 250 ml or 100 ml.
3.3. Pipette 5ml , or 10ml.

4. REAGENTS:
4.1. Methyl red (0.1% in water)
4.2. Methyl orange (0.1% in water)
4.3. Phenolphthalein (0.1% in water)
4.4. HCL (0.I N in water)
4.4.1. Concentrated HCL is 36-37% so its normality is first calculated to make 0.1 HCI from it.

D×P×1000
N=
100×EW

D = Density of HCI given as kg/liter

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P = Percent acidity given on the bottle label

N = Normality in grams equivalent/ liter
EW = Eq. wt. = 36.5gm

4.4.2. Now to make 0.1 N solution in 1000 ml of volumetric flask

V = Volume of HCL concentrated Required
V₂ = Volume of 0.1N solution
N₁ = Normality of solution is to be prepared
N₂ = Normality of HCL concentrated
Standardize the prepared solution with Na₂S₂0₃

4.4.3 STANDARDIZITION:
a. Weight accurately about 0.8g of primary standard anhydrous sodium carbonate that previously has been heated at a
temperature of about 250-270°C for 1.5 hrs. b. Dissolve in 100 ml of distilled water.
c. Add 2 drops of methyl red TS.
d. Titrate with the acid with constant stirring until the solution becomes faintly pink.
e. Heat the solution to boiling, cool, and continue the titration.
f. Heat again to boiling, and titrate further as necessary until the faint pink, color is no longer affected by continued
boiling.
g. Calculate the normality as follow.

Wx 1000
N=
EW ×V
N =Normality of HCL calculated
W =Weight of Na₂CO₃ = 52.99gm
EW =Volume of HCL used

Methyl orange may also be used for this titration which shows a colour change from pale yellow to Pink.
No heating is required when methyl orange is used.

5. PROCEDURE:
5.1. Weight 2-4 gms or solid and 3-5 gms of liquid sodium hydroxide in a 250 ml or 100 ml volumetric flask.
5.2. Titrate 2-5 ml portion, dilute with an equal volume of water, standard 0.1N HCI using methyl Orange
indicator.
5.3. The colour change orange to red.
5.4. The volume of HCI used in V ml.
5.5. Titrate another 5 -10 ml portion, diluted with on equal volume of water, when standard 0.1 N HCl using
phenolphthalein indicator.
5.6. The colour change from pink to colour less.
5.7. The volume of HCI used is Vo ml.
5.8. Calculate the volume of acid used for hydroxide by first calculating the volume used for carbonate & then
subtracting from the total.
VC = 2(V-V₀)
Vн = V-2(V-V₀)
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Where VC = Volume used for Carbonate

Vн = Volume used for Hydroxide

6.CALCULATION:
6.1.
Vн×N×EW×V₂×100
Percentage Basicity =
1000×W×V₁

Vн = Volume of HCL used for hydroxide

N = Normality of HCI
EW = Eq. wt. of NaOH = 40gm
V₂ = Total volume of NaOH solution made EW
= Weight of sample of NaOH taken.

6.2. To calculate the % basicity of Na₂CO₃ we use the same formula but instead of VH,VC is used EW is taken 52.99g
instead of 40 g.

7. NOTE:
7.1. To obtain satisfactory result by this method the solution titrate must be cold (as near 0°C as is Practicable)
and loss of carbon dioxide must be prevented as for as possible by keeping the tip of the burette
immersed in the liquid.

8. REFERENCE:
8.1. Vogel’s Text book of Quantitative Chemical Analysis.
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QUALITATIVE ESTIMATION OF ACETIC ACID

A. QUALITATIVE ESTIMATION:
Acetic acid is identified by the reaction with C₂H₅OH & Conc. H₂SO₄. Add Ethyl Alcohol & conc. Sulphuric Acid to
Acetic Acid, gives fruity smell.

B. QUANTITATIVE ESTIMATION:
1. DEFINITION:
1.1. Acetic acid is estimated quantitatively by simply determining its percentage acidity. The percent acidity is
the amount of NaOH necessary to neutralize the acid, calculated by wt. of the acid of specific molecular
weight.

2. SCOPE:
2.1. Applicable to commercial and analytical grade Acetic acid.

3. APPARATUS:
3.1. Beaker 100 ml.
3.2. Volumetric flask, 250 ml or 100 ml.
3.3. Pipette 5ml or 10ml
3.4. Conical flask, 250 ml
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4. REAGENTS:
4.1. Phenolphthalein indicator solution (0.1% in spirit).
4.2. 2.0 N NaOH Solution (20 g/250 ml distilled water).
4.3. 0.05N NaOH (25 ml of 2N NaOH/1000 ml distilled water ) For standardization of NaOH.

5. PROCEDURE:
5.1. Weigh 2-5 gm of sample and dilute to 250 ml or 100 in a volumetric flask.
5.2. Pipette out 2 to 10 ml of the sample solution in a conical flask.
5.3. Add few drops of phenolphthalein indicator solution and titrate with 0.05N NaOH solution.

6. CALCULATION:

V×N×EW×V₂×100
Percentage Acidity =
1000×W×V₁

V₂ = Volume of Solution pipette out.

V₁ = Volume of total solution
N = Normality of NaOH
V = Volume of NaOH used
EW = eq. wt. of butyric acid = 60.
W = wt. of sample taken

7. REFERENCE:
7.1. Vogel’s Text book of Quantitative Chemical Analysis.

IDENTIFICATION OF TERT. BUTYLE HYDRO QUIONE (TBHQ)

1. DEFINITION:
1.1. This method is determines presence of TBHO (Antioxidant) in sample.

2. SCOPE:
2.1. Applicable to all samples of oil & fats in which TBHQ 1 is used as A antioxidant

3. Apparatus:
3.1. Test tube or centrifugal tube
3.2. Beaker
3.3. Volumetric Flask 25 m
3.4. Electrical Balance
3.5. Graduated Pipette

4. REAGENT:
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4.1. Methanol
4.2. Dimethyl Amine.

5. SOLUTION:
5.1. Prepare 25% solution of Dimethyl Amine in water using 25 ml Volumetric flask.

6. PROCEDURE:
6.1. Weight approx. gm of sample in a test tube or centrifugal tube.
6.2. Add 1 ml of Methanol in test tube with the help of pipette.
6.3. Add few drop of 25% SOLN of dimethyl amine in it. Pink to red colour is produced which indicates
presence of TBHQ in sample.

DETERMINATION OF FREE CHLORINE

1.DEFINITION:
1.1. This method is determines quantitatively presence of free chlorine in sample.

2. SCOPE:
2.1. Applicable to all sample contains free chlorine.

3. APPRATUS:
3.1. Beaker 100 ml
3.2. Volumetric Flask 100 ml or 250 ml
3.3. Pipette 5 ml or 10 ml
3.4. Conical Flask
3.5. Iodine Flask with stopper.
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4. REAGENTS:
4.1. Glacial acetic acid solution in distilled water.
4.2. Potassium iodide
4.3. Soluble starch
4.4. Hydrochloric acid
4.5. Sodium thiosulphate

5. SOLUTION:
5.1. Potassium iodide ( 15% in distilled water)
5.2. Starch solution (1% is boiling distilled water)
5.3. 1:1 Solution of glacial acetic acid in distilled water.
5.4. Standard potassium dichromate solution
5.5. Sodium thiosulphate solution. Dissolve 24.8 g/100 ml distilled water.

6. STANDARDIZATION OF SODIUM TIHIOSULPHATE:

6.1. Take standard potassium dichromate solution. (Dry & weight 0.05 to 0.1 g in iodine flask & dissolve it in 10
to 15 ml of distilled water)
6.2. Add 5ml Hydrochloric acid (conc.) & 10ml of 15% potassium iodide solution & shake well to mix.
6.3. Allow to standard for 5 minutes in dark and add 100 ml of distilled water.
6.4. Titrate with sodium thiosulphate till the yellow colour almost disappeared.
6.5. Add 0.5 ml of starch indicator & continue to titration by adding sodium thiosulphate solution slowly, until
the light green colour appeared.
6.6. The strength of the sod. thio sulphate solution is expressed in term of its normality.

Eq.Wt.×W
CALCULATION: N =
V×100
N = Normality of sodium thiosulphate ( Na₂S₂0₃)
Eq. wt. = Equivalent weight of potassium dichromate (K₂Cr₂O₇) = 49
W = Weight of potassium dichromate (K₂C₁₂O₇)
V = Volume of sodium thio sulphate consumed

7. PROCEDURE:
7.1. Take 5 to 10 gm of the sample in 100 or 250 ml volumetric flask and make the volume up to the mark with
distilled water. Shake well & allow to stand for 5 minutes.
7.2. Pipette 5 to 10 ml this diluted sample in conical flask or iodine flask & acidify with few drops of acetic acid
(1:1).
7.3. Add 20 ml of potassium iodide (15%soln ) & titrate with standard solution of 0.1N ( Na₂S₂0₃) until the
yellow colour almost disappeared. Add 0.5 ml of starch indicator& continue the titration until blue colour
has just appeared.
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8. CALCULATION:

V×N×EW×V₁×100
% of free chlorine =
1000×W×V₂

V₂ = Volume of Solution pipette out.

V₁ = Volume of total solution
N = Normality of Sodium thiosulphate
V = Volume of sodium thiosulphate used
EW = eq. wt. of Chlorine = 35
W = wt. of sample taken

9. REFERENCE:
9.1. Vogel’s Text book of Quantitative Chemical Analysis.

IDENTIFICATION OF SULPHITE ION

1. DEFINITION:
1.1. This method is determines presence of sulphite ion in sample.
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2. SCOPE:
2.1. Applicable to all samples having sulphite ion.

3. APPRATUS:
3.1. Beaker 100 &250 ml
3.2. Test tubes
3.3. Glass rod

4. REAGENTS:
4.1. Sodium carbonate
4.2. Barium chloride
4.3. Potassium Dichromate

5. SOLUTION:
5.1. Sodium carbonate extract of sample: Prepare sodium carbonate extract of sample by dissolving approx.
2- 3 gm sample is freshly prepared sodium carbonate solution in distilled water.
5.2. Barium chloride solution: Prepare 5% Barium chloride solution by dissolving 5 gm of Barium chloride in 10
ml of distilled water.
5.3. Potassium dichromate solution: Prepare 1% potassium dichromate solution by dissolving 1gm Potassium
dichromate in 100 ml of distilled water.

6. PROCEDERE:
6.1. Take small quantity of sodium carbonate extract of sample in a test tube and add Barium chloride in it
white ppt of Barium sulphate indicate presence of sulphite in sample.
6.2. Take small quantity of sodium carbonate extract of sample in a test tube and add Potassium dichromate
solution in it green colouration conforms suphite in sample.

7. REFERENCE:
7.1. Qualitative and quantity inorganic analysis by Wasi Askari.
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TESTING AND MEASUREMENTS ( TIN-CONTAINERS )

1. PURPOSE:
1.1. To describe methods for the testing and measurement of empty tin containers.

2. SCOPE:
2.1. Applicable to all empty tin containers.

3. METHOD:
3.1. Sampling

Lot Size Sample Plan

4000 80 Double

2000 80 - -

3.2. Colour and Printing:

3.2.1. Compare tins with reference standard and note any variation. For varnish 5H pencil is
applied on the body if there is no scratch the varnish is ok. Physical appearance and
finish.
3.2.2. Check tins and observe is cleanliness, rust and finish of tin plate.

3.3. Measurement:
3.3.1. For all measurement, Vernier caliper, micrometer, measuring scales and pie glass are
used. All readings are recorded in millimeters.

3.4. Leakage:
3.4.1. Inverted tins are dipped into water if the tin has leakage air bubbles are produced
only continuous bubbles are taken in to account.

3.5. Weigh:
3.5.1. Weight of every tin is taken by keeping it on top load. Electronic balance ( 1 gm
division ) weight are recorded in grams.

3.6. Strength of Cleat and handle:

3.6.1. Strength of cleat and handle is tested by using testing machine in which 25 KG weight
is applied for 30” if cleat and handle remains unbroken it is supposed to have
sufficient strength.
4. REFERENCE:
4.1. Unity Foods Limited Method.
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TESTING AND MEASUREMENTS (CARTON)

1. PURPOSE:
1.1. To describe method of the testing of cartons.

2. SCOPE:
2.1. All types of cartons

3. DEFINITION:
3.1 S.S : Stainless steel
3.2 MM : Millimeter
3.3 GSM : Gram per square meter

4. SAMPLING:
4.1.
Lot Size Sample
700 30
1000 30

4.2. Printing and Colour:

4.2.1. Match with the standard paper to determine the colour of paper and printing.

4.3. Dimension:
4.3.1. Measure the external length, width and height with the help of measuring tape and /or S.S
ruler record the reading in inch or mm.
4.3.2. Weigh the carton on top load balance and record the weight in gram.
4.3.3. Measure the depth of fluting and thickness of cartons with the help of Vernier caliper record
the reading in mm. Count the ply and number of fluting per feet of the box.

4.4. Moisture:
4.4.1. Refer to Method No. CAM-13B

4.5. G.S.M:
4.5.1. Cut a piece of 100 sq. cm. of the box obtain its weight on top load balance ( +/-0.05 gm ):
Multiply the weight with 100 to obtain the G.S.M.

4.6. Pasting:
4.6.1. Pile up the liner of the box if it is clearly removed the pasting is not proper.
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4.7. Strength of Carton:

4.7.1. From a box and put equal sized weight of 50 kg on it observe it after 24 hours if the sides show
no sign of deformation the strength In sufficient.

5. REFERENCE:
5.1. Unity Foods Limited Method.

TESTING AND MEASUREMENTS ( PLASTIC BOTTLE )

1. PURPOSE:
1.1. To describe method of the testing of plastic bottles.

2. SCOPE:
2.1. Applicable to PET & HDP bottles.

3. SAMPLING:

Product Lot Size First Sample for evaluch

HDPE 500-1200 100 bottles *5 bottles from each 20 units

PET 1200-3200 100 bottles 5 bottles from each 25 units
PET 3201-10000 200 bottles 10 bottles from each 50 units

4. DEFINITION:
4.1. HDPE : High density polyethelen
4.2. PET : Poly ethelene Triplate

5. METHOD:
5.1. Physical appearance & colour:
5.1.1. This includes colour of bottles , colour & alignment of labels (if applicable), de flashing and
overall appearance.
5.2. Dimension of bottles & other components:
5.2.1. Measure all dimensions with the help of Vernier calipers, S.S Scale and / or height gauge.
Record all reading in millimeter or in centimeters.
5.3. Leakage:
5.3.1. Maintain the air pressure in the tank not less than 7 lbs per square inch.
5.3.2. Fit the Cork on the mouth of the bottle & open the valve & feel the leakage of air with touch of
hand. Keep the compressor running during testing.

6. REFERENCE:
6.1. Unity Foods Limited Method.
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INSPECTION & TESTING METHOD OF FILM ROLLS

1. DEFINITION:
1.1. This method described the method of testing film rolls.

2. SCOPE:
2.1. Applicable to all type of film rolls.

3. PROCEDURE:
3.1. Sample a no. of the cartons from the lot according to the sampling procedure and CAM-035 and perform
Following analysis on each film using appropriate instruments.

TESTS INSTRUMENTS UNIT

3.2. Colour of film Standard film -
3.3. Outline Measuring scale mm
3.4. Printing standard film - 3.5.
Colour of printing Standard film -
3.6. Treatment Scotch Tape - 3.7.
Thickness Micrometer mm
3.8. When all the characteristics are in accordance with the specification put a green card QCT-01-P on
the lot.
3.9. If the lot shows deviation from specification then reject the lot and put a red card QCT-01-R on the
lot.
3.10. If the lot shows deviation from specification but it could be acceptable after segregation then place
yellow QCT-01-H card which indicates that the lot is under consideration.

4. REFERENCE:
4.1. Unity Foods Limited Method.
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TESTING PLASTIC HANDLE

1. OBJECTIVE:
1.1. To evaluate the strength of handle.

2. APPARATUS:
2.1. Handle testing machine.

3. PROCEDURE:
3.1. Fix the handle in the jaw of machine.
3.2. Switch on the machine.
3.3. Apply the pulse for three successive time & observe the physical condition of the handle.
3.4. The handle should sustain the face of 3rd pulse.

4. REFERENCE:
4.1. Unity Foods Limited Method.