Professional Documents
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Chapter Ix: Analysis of Fats and Oils
Chapter Ix: Analysis of Fats and Oils
Fats and oils are a heterogeneous group of predominantly hydrophobic compounds. The
distinction between fats and oils does not have a chemical basis. Those fats/oils that remain liquid
at normal (ambient) temperature are generally taken as oil and those that remain solid, fats.
Analysis of fats and oils is carried out for various reasons, viz.:
1. Shelf life study (how long the item will remain without deterioration in quality under a
given set of conditions)
2. Functional quality (e.g., suitability for use in biscuits, bakery, hydrogenation, etc.)
3. Sensory quality (e.g., rancidity)
4. Nutritional quality (e.g., melting point, polyunsaturated fatty acids)
5. As an aid in controlling production operation (e.g., control of hydrogenation, recovery of
oil in mills)
6. Conformance to regulatory standards (e.g., with respect to free fatty acids, saponification
value, peroxide value, moisture)
7. Detection of adulteration (e.g., contamination with mineral oil and argemone oil,
adulteration of dairy ghee with vegetable ghee)
8. Advanced research (e.g., determination of fatty acid profile)
Some of the routine tests carried out on fats and oils are as follows:
1.
2.
3.
4.
5.
6.
7.
8.
Some of the special tests used for particular fats and oils are:
1. Crismer test for rapeseed and mustard oil
2. Reichert-Meissl, Polenske and Kirschner values for dairy ghee
3. Polybromide test for linolenic oils such as linseed oil
Some of the important physicochemical characteristics of common fats and oils are as follows:
FOOD ANALYSIS
Fat/oil
Soybean oil
Mustard seed oil
Maize oil
Sunflower seed oil
Refractive
index at 40C
1.466-1.470
1.461-1.469
1.465-1.468
1.467-1.469
Saponification value,
mg KOH/g oil
189-195
170-184
187-195
188-194
Iodine value,
Wijs
120-143
92-125
103-128
110-143
Unsaponifiable
matter, %
1.5
1.5
2.8
1.5
Rancid oils markedly lower the esthetic value of oil. Such oils also bring about health problems.
In connection with the afore-mentioned points, regulating bodies have set mandatory standards
for edible oils, for example, in Nepal:
Mandatory standards of selected fats/oils
Fat/oil
FFA (as %oleic acid)
Vanaspati
0.50
Refined oil
0.25
Mustard/Rapeseed oil
3.00
PRINCIPLE
Free fatty acids are readily soluble in rectified spirit or absolute alcohol. A suitable amount of oil
is therefore mixed with neutralized rectified spirit to extract free fatty acids and the amount of the
105
FOOD ANALYSIS
latter calculated by titrating with standard NaOH or KOH using phenolphthalein indicator. To
facilitate extraction, the mixture may be warmed to about 70C and swirled vigorously.
Calculation for both acid value and FFA can be carried as follows:
REQUIREMENTS
Neutral alcohol (95%, v/v)64
Titration arrangement
Weighing arrangement
0.1N NaOH
Hot plate
PROCEDURE
Weigh out 10 g of fat/oil in a 250-ml conical flask (by difference)
Add 50 ml of neutral alcohol
Add a drop or two of phenolphthalein indicator
Swirl the contents and place flask on the hot plate
Warm the mixture to about 70C. Swirl well
Titrate warm with 0.1N NaOH to persistent pink color
In case of doubt, tilt the mixture to allow separation of alcohol and fat fractions. Observe
the color of the alcohol fraction for persistent pink color
8. Carry out titration in triplicate
1.
2.
3.
4.
5.
6.
7.
CALCULATION
% FFA =
Acid value =
M=
561.0937 (100 P )
12.683
Saponification value
64
Neutralize the acidity in the with 0.1N NaOH using phenolphthalein indicator
106
FOOD ANALYSIS
Soybean oil:
Rapeseed oil:
189 195
168 181
210 230
The test merits considerable attention in that successful testing is more of an art. There are at least
two titrimetric methods for the determination of saponification value. A relatively easy method
utilizes double indicator, viz., phenolphthalein and bromophenol blue.
When fat is boiled with an excess of alcoholic KOH, the glycerides irreversibly hydrolyze, giving
rise to glycerol and fatty soap (Fig. IX-1). The alkali consumed for this is a measure of
saponification value, and is defined as the number of milligram of KOH needed to saponify one
gram of oil or fat.
R2-COOCH
R1-COOK
CH2OH
CH2OCO-R1
+ 3KOH
CH2OCO-R3
Mixed triglyceride
HOCH
R2-COOK
CH2OH
Glycerol
R3-COOK
Potassium soap
Sterols
Hydrocarbon
Pigments
KOH
No reaction
(Unsaponifiable matter)
FOOD ANALYSIS
Titration arrangement
PROCEDURE
1. Melt the sample (if not already liquid) and filter warm. Ensure that the sample is free from
moisture and impurities
2. Weigh accurately by difference about 2 g of sample in a 250-ml conical flask
3. Add about 500 mg KOH pellet, a small amount of rectified spirit ( 10 ml), and emulsify
by swirling briefly
4. Add about 30 ml rectified spirit and gently reflux the whole until the oil becomes
transparent (this usually takes 25 min)
5. Add some rectified spirit (if the volume decreases) and continue refluxing till completely
saponified. The oil-alcohol mixture appears transparent at this stage
6. Slightly cool the flask and add a drop or two of phenolphthalein indicator. Intense red color
indicates the presence of excess KOH. If the color does not change, repeat the whole
process using more KOH (e.g. 600-800 mg)
7. Add a drop or two of distilled water. If a milky color develops, the sample contains
significant amounts of unsaponifiable matter or is contaminated with mineral oil
8. add more water (about 50ml) and mix well
9. Neutralize the excess KOH with 0.5N HCl. The pink color should just disappear
10. Add a drop of bromophenol blue indicator and swirl. It should give a blue color
11. Note the reading on the burette (containing the standard HCl) and titrate till a permanent
greenish-yellow color appears. If, during titration, fat-like globules suddenly appear, warm
the flask a little and continue titration to the end point
12. Note the volume of 0.5N HCl consumed (the second reading, that is) and calculate the
saponification value
CALCULATION
Saponification value =
FOOD ANALYSIS
Soybean oil:
Rapeseed oil:
Butter oil:
120-143
94-120
26-38
The test is of tremendous value in vanaspati (hydrogenated oil) plants. It is routinely used for
monitoring the degree of hydrogenation. Iodine value is also used to calculate the amount of
hydrogen used or wasted in vanaspati plants. In general, a drop in I unit of iodine value means to
the vanaspati manufacturer that 0.075 kg of hydrogen has been added to every 1000 kg oil. There
are several methods for measuring the iodine value of fats and oils. Some of the variations and /or
equivalent methods are: Hanus method, Bromine Value method, Rosenmund-Kuhnhenn method,
etc. There are some difference vis--vis reagent preparation in Wijs method also.
PRINCIPLE
Halogens add across the double bonds of unsaturated fatty acids to form addition compounds.
Iodine monochloride (ICl) is allowed to react with the fat in the dark. The amount of iodine
consumed is then determined by titrating the iodine released (after adding KI) with standard
thiosulfate and comparing with blank in which the fat is omitted. The reaction occurring in the
test can be shown in Fig. IX-3.
CH CH
Unsaturated
portion of fat
ICl
Iodine
monochloride
ICl
+
KI
Residual Added after
titration
Na2S2O3 +
Na-thiosulfate
I2
CH CH
I Cl
Addition
compound
KCl
+
I2
Molecular
iodine
2NaI
+ 2Na2S4O6
Na-tetrathionate
65
Dissolve 1g of reagent grade starch in hot water. Transfer the clear fraction into another container. Use only
fresh solution, as it is subject to microbial degradation
66
Dissolve 25g AR grade Na2S2O3.7H2O in distilled water to make 1000ml. Mix the solution thoroughly, allow to
stand for a few days, and then siphon off the clear liquid. Standardize the solution with AR grade potassium
dichromate (K2Cr2O7). Weigh 0.20 to 0.23g of K2Cr2O7 (dried for 2 hrs at 105C). Transfer to a 250-ml beaker
using 150ml of water. Add 2g of KI and mix. Add 20ml of 1N HCl, swirl, and allow to stand for 10 min. start
109
FOOD ANALYSIS
Wijs solution
Measuring cylinder, 25 ml
67
PROCEDURE
1. Weigh accurately by difference suitable quantity of oil using the formula: (20.3 expected
iodine value) grams, in to a clean, dry 250-ml IV flask (see Fig. IX-4)
2. Add 10 ml of CCl4 and allow oil to dissolve
3. Add accurately 20 ml of Wijs solution. Swirl once and close the flask with the stopper.
The stopper may be moistened with minimum of 10% KI solution
4. Stand the flask at 15-20C for 30 min in dark
5. Add 15 ml of 10% KI solution, followed by 100 ml distilled water
6. Titrate with 0.1N Na2S2O3 using starch indicator towards the end of the titration (The
mixture turns straw color near the end point. Add two drops of starch solution. The mixture
immediately turns dark blue. Continue the titration until the blue color just disappears)
7. Carry out a blank test upon the same quantities of reagents, omitting the oil, at the same
time and under the same conditions. The excess of reagent remaining for titration in the test
must be 150% of the reagent absorbed
CALCULATION
Iodine value =
12.69
Stopper
N of sod-thiosulfate =
Add a pinch of Na2CO3 and 1ml of chloroform to preserve it from microbial degradation
67
Dissolve 8g iodine trichloride in 150ml glacial acetic acid and mix with 9g iodine dissolved in 350ml glacial
acetic acid. The strength of Wijs solution, as determined by titrating with Na-thiosulfate, should not be less than
0.2N. Store the reagent in a colored bottle in dark. The solution is stable for about 30 day.
110
FOOD ANALYSIS
Note: Wijs solution can be prepared by other methods also, viz., (i) using iodine monochloride,
and (ii) using chlorine gas and resublimed iodine. The latter method is described here.
Preparation of Wijs solution by chlorination
Before anything else, prepare standard sod-thiosulfate, conc sulfuric acid, 10% KI solution and
starch indicator. Assemble pipettes, burettes and other glassware needed for iodometric titration.
Take 13g resublimed iodine in a 1-liter beaker
Add 200ml glacial acetic acid and dissolve by gentle heating (along with stirring). Iodine
dissolves very slowly and the complete dissolution can be carried out in stages by using
small portions of glacial acetic acid
Transfer the dissolved portion to 1-liter volumetric flask
Add more glacial acetic acid (~ 200ml) to the undissolved iodine in the beaker and heat
gently (as previously done) to affect dissolution
Transfer the dissolved portion to the volumetric flask (to pool the solution) again
Carry out this operation until iodine is completely dissolved. However, do not exceed the
total volume of 1000ml. If some space is available, make up the volume to 1000ml by
glacial acetic acid. Mix the solution well. Take out about 25ml solution and set aside (as a
reserve) in a separate flask (you will need this later)
Transfer the bulk iodine solution in a Woulfe bottle and assemble the parts as in Fig. IX-5
Generate chlorine68 and pass through the iodine solution to form iodine monochloride
Continue passing the chlorine until the characteristic color of free iodine is discharged
(solution suddenly lightens because of free chlorine)
Stop passing chlorine and test the Wijs solution for dismantle the assembly for
chlorination
Add small amounts of iodine solution (reserved earlier) until the free chlorine has been
destroyed (the color again darkens). A slight excess of iodine does little or no harm but
excess chlorine must be avoided. Typically, the iodine/chlorine ratio should be 1.1 0.1
and this can be ascertained by determining iodine content and total halogen content as
follows:
Iodine content:
o Take 150ml of Chlorine-saturated water in a 500-ml conical flask and add some
glass beads
o Add 5ml of Wijs solution
o Mix, and heat to boiling for 10 min
o Cool and add 30ml of 2% H2SO4
o Add 15ml of 15% fresh KI solution
o Titrate with 0.1N sod-thiosulfate to starch end point
o Note the titer (say A)
68
111
FOOD ANALYSIS
o
o
o
o
o
o
2A
, the Wijs solution thus prepared should consume approximately
( 3B 2 A )
Conc. HCl
KMnO4
Iodine solution
Chlorine bubbles
FOOD ANALYSIS
Peroxide value of an oil or fat is the amount of peroxides present and expressed as milliequivalents of peroxide per 1,000g of sample.
PRINCIPLE
When a rancid fat or oil sample is treated with potassium iodide after dissolving in an appropriate
solvent, peroxides present in the fat liberate iodine. The test is a volumetric one where I2, formed
from KI in the presence of peroxide is determined by titrating with sodium thiosulfate and
conducting a blank determination.
Now, milliequivalent peroxide = milliequivalent thiosulfate at the equivalence point
Again, milliequivalent = (strength volume), when volume is in milliliter
Therefore, PV = milliequivalent thiosulfate / kg sample
REQUIREMENTS
Oil or fat sample
Weighing arrangement
PROCEDURE
1. Weigh accurately (by difference) 5g of fat or oil sample in the Iodine flask
2. Add 25ml of solvent and displace the air with CO2
3. Add 1ml of KI solution, stopper the flask, and allow it to stand for 1min (with gentle
shaking)
4. Add 35ml of distilled water and a few drops of starch indicator. Appearance of blue color
on addition of starch indicates presence of free iodine
5. Titrate the liberated iodine with 0.01N or 0.1N sod-thiosulfate until the blue color just
vanishes
6. Carry out a blank determination simultaneously (omitting oil)
7. Calculate Peroxide value using following equation:
PV (meq/kg) =
N (VS - VB ) 1000
Wt. of sample (g)
Where,
N = normality of sod-thiosulfate, VS = sod-thiosulfate consumed by sample (ml), and VB =
sod-thiosulfate consumed by blank (ml).
69
70
113
FOOD ANALYSIS
Crystal size
5m
20-50m
1-2m
Melting point
Lowest
Highest
Intermediate
Stability
Least stable
Most stable
Intermediate
The stability of fat is related to the polymorphic form and the associated melting point. The
melting points of -, -, and forms of tristearin are 55C, 64C, and 73C, respectively.
Polymorphic transformations occur from to to and are irreversible. When fat is cooled
rapidly the polymorph is produced, which is usually quickly converted to the form. These
polymorphic forms also affect the appearance and texture of fat. form gives a smooth texture
whereas form results in a very coarse granules. It is therefore very important to control the
balance of polymorphic forms in the production of fat and fatty foods like margarine (needs
form), ghee (needs form), etc.
Because of the reasons described above, melting point as such is not very reliable for establishing
identity of the fat and oil. However, it is extensively used in controlling process operation (e.g.,
hydrogenation), quality control, and determining suitability of fat for a particular purpose. The
methods used for the determination of melting point vary considerably. A typical method used in
vanaspati manufacture is the open-tube capillary method. The melting point is therefore defined
by the specific conditions of the method by which it is determined.
PRINCIPLE
The temperature at which the oil or fat softens or becomes sufficiently fluid to slip or run as
determined by the open-tube capillary-slip method.
REQUIREMENTS
Capillary tubes71
71
Thin-walled with uniform bore capillary glass tubes opn at both ends with following dimensions:
Length = 50-60mm
114
FOOD ANALYSIS
PROCEDURE
1. Melt the sample and filter it through a filter paper to remove any impurities and last traces
of moisture
2. Make sure that the sample is absolutely dry
3. Mix the sample thoroughly
4. Introduce a capillary tube into the molten sample, so that a column of the sample, about
10mm long, is sucked into the tube
5. Chill the tube containing the sample immediately by touching the tube, against a piece of
ice until the fat solidifies
6. Place the tube in a small beaker and hold it for one hour either in a refrigerator or in water
maintained at 4-10C
Thermometer
Capillary
Rubber band
Heat source
Fat
Theile tube
Chilled water
115
FOOD ANALYSIS
Reichert-Meissl value: It is the number of milliliters of 0.1N NaOH required to neutralize the
steam-volatile, water-soluble fatty acids distilled from 5g sample of fat under precise conditions
specified in the method. This test measures the quantity of C4 and C6 fatty acids.
Polenske value: It is the number of milliliters of 0.1N NaOH required to neutralize the steamvolatile, water-insoluble fatty acids distilled from 5g sample of fat under precise conditions
specified in the method. This test measures the quantity of C8 to C14 fatty acids.
Kirschner value: It is a measure of steam-volatile, water-soluble fatty acids forming watersoluble silver salts (which is the property of butyric acid). In recent years, this analysis is not
usually done.
PRINCIPLE
Steam-volatile fatty acids can be collected by saponification and steam-distillation of oil. Reichert
Meissl value can be determined by titrating the steam condensate (that contains water-soluble
fatty acids) with0.1N NaOH. Polenske value can be determined by eluting the fatty acids
adhering on the condenser with neutral ethanol and titrating with 0.1N NaOH. Determination of
Kirschner value involves neutralization of the water-soluble fatty acids with barium hydroxide,
preparation of their silver salts, separation of the water-soluble butyric acid salt by filtration,
liberation of butyric acid by acidification, separation by steam distillation, and quantification by
titrating again with barium hydroxide.
REQUIREMENTS
Fat sample
Glycerol
50% NaOH
Titration arrangement
116
FOOD ANALYSIS
Weighing arrangement
RM-Polenske-Kirschner apparatus72
Phenolphthalein indicator
Heating arrangement
PROCEDURE
1. Melt the fat sample if solid but do not heat above 50C
2. Weigh 50.01g of fat sample into a Polenske flask
3. Add 20g of glycerol and 2ml of 50% NaOH solution from a burette which is protected
from CO2 pick up. Wet the tip of the burette before adding alkali to free it of carbonate
deposit and reject the first 0.5ml of NaOH
4. Heat the mixture over a low flame with wire gauze until the liquid becomes clear and the
fat has saponified. Do not overheat at this stage which causes discoloration
5. When all the fat has saponified, cover the flask with a watch glass, and allow to cool
6. Add 93ml of boiling distilled water which is free of CO2 and mix. The solution must be
completely clear at this stage and pale yellow in color. If the solution is not clear which
indicates incomplete saponification, or if it is darker which indicates overheating, repeat
the procedure with a fresh sample. An old sample of oil or fat may behave similarly
7. Add 0.1g of pumice powder and 50ml of dilute H2SO4
8. Connect to the distillation apparatus (Fig. IX-7)
9. Warm the mixture until any insoluble material which may be present melts
10. Increase the heat and distil 110ml of solution in 19 to 21 min. The distillation is considered
to begin when the first drop forms in the shill-head
11. Stop heating soon after 110ml has distilled over, and replace the graduated flask by a
measuring cylinder to collect drippings from the condenser
12. Close the graduated flask with the stopper. Do not mix
13. Place the flask in a water bath at 15C for 10 min and ensure that the 110ml graduation is
below the water level
14. Mix and filter through a 9cm Whatman No. 4 paper. Reject the first 2-3ml of the filtrate
and collect the rest in a dry flask
15. Wash the condenser, still head and the 110ml graduated flask with three lots of 15ml
distilled water passing each washing through the measuring cylinder, 100ml flask and
stopper
16. Filter through the same filter paper ensuring that all insoluble matter is transferred to the
paper. Discard the filtrate. Do not mix with filtrate of the distillate got in the previous step.
The filtrate should be free from water insoluble fatty acids
17. Pipette out 100ml of the filtrate to a dry 250ml conical flask and titrate with 0.1N NaOH
using phenolphthalein as indicator
18. Calculate RM value as follows:
RM value = (Sample titer Blank titer)ml N of NaOH 11
The factor 11 has been obtained as follows:
72
Apparatus consisting of flat-bottom boiling flask, still head (10.7cm wide and 18cm high), condenser (52cm long
with 30cm cooling length and 7cm entry tube) and a receiver (with graduations at 100ml and 110ml)
117
FOOD ANALYSIS
110ml
100ml
FOOD ANALYSIS
Where,
Tk and Tb = sample and blank titer respectively in Kirschner value determination
Tr and Tb = sample and blank titer respectively in the RM value determination
9.6.2. BOUDOUIN TEST
This test is useful in the detection of adulteration of dairy ghee with vanaspati ghee. The test is
based on the color reaction between sesamolin (a compound present in sesame oil) and furfural In
Nepal, use of sesame oil in vanaspati is mandatory. Dairy ghee containing sesamolin gives a
positive Baudouin test, thereby indicating the presence of vanaspati ghee.
PRINCIPLE
The development of pink color with furfural solution in the presence of hydrochloric acid
indicates the presence of sesame oil. The color is produced on account of reaction with sesamolin
present in sesame oil.
REQUIREMENTS
Glass-stoppered test tube/measuring cylinder
Furfural solution73
Oil sample
PROCEDURE
1. Take 5ml of the oil or melted fat in a 25-ml measuring cylinder (or test tube) provided
with a glass stopper
73
119
FOOD ANALYSIS
2.
3.
4.
5.
Chloroform
Ethyl alcohol
Liquid bromine
Diethyl ether
PROCEDURE
1. Pipette 1ml of oil into a boiling tube (wide-mouthed, 100ml cap)
2. All 5ml chloroform and about 1ml of bromine drop-wise till the mixture becomes deep red
in color
3. Cool the test tube in an ice water-bath
4. Add about 1.5ml of rectified spirit drop-wise while shaking the mixture until the
precipitate which was first formed just dissolves
5. Add 10ml diethyl ether
6. Mix the contents and place the tube within the ice water-bath for 20 min
7. Appearance of precipitates indicates the presence of linseed oil
The sensitivity of this test is about 1% if linseed oil in other other oils. The test has some
limitations. It is not suitable for test in mahua oils. Besides, marine oils, which contain
polyunsaturated fatty acids, also give insoluble polybromide precipitate.
9.5.4. TEST FOR THE PRESENCE OF ANIMAL FAT BY MICROSCOPIC EXAMINATION
Animal body fats such as beef tallow and lard have been shown to contain trisaturated glycerides.
On crystallization these glycerides exhibit a characteristic crystalline appearance when viewed
under microscope. The procedure recommended by Williams Sutton for the microscopy of fat
crystals have been suitably modified and given.
120
FOOD ANALYSIS
REQUIREMENTS
Fat sample
Test tubes
Ice water-bath
Filtration unit
Ethyl alcohol
Glycerin
Microscope
PROCEDURE
1. Take about 2 g of melted fat samples in test tubes
2. Add 10 ml of diethyl ether and mix
3. Plug the tubes with cotton and allow to stand for 30 min in ice water or 24 hrs at 20C
(slow crystallization gives bigger crystals). In certain cases it is preferable to first
crystallize with a stronger solution of fat from a mixture of ether and ethyl alcohol (1:1). In
such cases separate the crystals by filtration and recrystallized in ether
4. Place the crystals on a drop of glycerin previously taken on a microscopic slide
5. Cover the crystals immediately with cover glass
6. Examine the crystals under 160 and finally 400 magnifications. The typical appearance
of beef tallow crystallized into characteristic fan like tufts, the ends of which are more or
less pointed can be seen. Lard crystals are chisel-shaped. Hydrogenated fats deposit
smaller size crystals. The size and shape of the crystals depend upon the strength of
solution, amount of fat taken and the time allowed for crystallization
9.6.5. TEST FOR PRESENCE OF ARGEMONE OIL
Argemone (Argemone maxicana L.), yellow poppy, is a wild herb, which grows in mustard field
and bears capsules full of brown black seeds. Because of its resemblance with black mustard, it is
often used as an adulterant. The oil is reported to cause glaucoma, dropsy and sometimes total
blindness due to the presence of alkaloids namely, sanguinarine and dihydrosanguinarine.
PRINCIPLE
The hydrochloric acid extract of the oil sample containing argemone oil when subjected to TLC
for separation of alkaloid gives fluorescent spot under UV light.
Standard argemone oil extract
Pear-shaped flask
Diethyl ether
Chloroform:Acetic acid:Water =
70:20:10 (v/v)
121
FOOD ANALYSIS
PROCEDURE
1. Take 10 ml sample in a separating funnel and dissolve in 15 ml Diethyl ether
2. Add 5 ml conc HCL and shake vigorously for 2 3 minutes. Allow to separate. Contents
of the separatory funnel may be heated cautiously over the vent of heating water bath for
some time for quick separation
3. Transfer the acid layer to a 25 ml beaker
4. Place the beaker into a boiling water bath and evaporate till dryness
5. Dissolve the residue obtained after evaporation of hydrochloric acid in 1 ml of a mixture
of chloroform and acetic acid (9:1)
6. Spot on TLC plate with the help of spotting capillary. Spot side by side standard
Argemone oil extract (0.1 % in ether)
7. Develop the plate in (a) Butanol:Acetic acid:water; or (b) Hexane:Acetone mixture
8. Allow the solvent front to move up a distance of 10 cm
9. Allow the plate to dry
10. Place the plate under UV light in the visualization chamber
11. Bright yellow or orange yellow fluorescent spots having Rf similar to the standard
argemone oil will confirm presence of argemone oil. The spot gives blue florescence under
UV-light if plate is sprayed with 20% aqueous sodium hydroxide solution
The method is very sensitive and can detect argemone oil upto 50 ppm level.
9.6.6. KRIES TEST FOR RANCIDITY IN FATS/OILS
Kries test is a very rapid test for the assessment of rancidity in fats and oils. It can be carried out
quantitatively as well as quantitatively. The qualitative method involves vigorous mixing of 5ml
of oil with 5ml of 0.1% phloroglucinol solution (in diethyl ether), adding 5ml of conc HCl and
observing for pink color as the evidence of incipient rancidity.
PROCEDURE FOR QUANTITATIVE METHOD
1.
2.
3.
4.
5.
Absorbance values below 0.15 indicate no rancidity. Absorbance values greater than 0.2 denote
incipient rancidity, and absorbance values around 1.0 show that the sample is highly rancid.
122