OIL AND FAT

ANALYSIS
NOORHIDAYAH ISHAK
Oil and Fat Analysis

Why oil and fat products need to be analyze before it been

distribute to the consumer?
To ensure the quality control in the process
To determine the cause of product failures
To ensure products function and design
 Oil and Fat Analysis

8 types of oil and fat analysis:

Non fatty impurities
Melting, solidification and consistency
Composition analysis
Flavor, randicity and stability
Color and appearance
Refining and bleaching
Performance testing
Nonstandardized methods
 Non fatty impurities

Non fatty
impuritie
s

Soap Moisture Trace

Impuritie material
s analysis analysis analysis
analysis

Insoluble Filterable Turbidimeter

Hot
Vacuum
plate Air
oven
oven Karl
Fisher
Non fatty impurities (Moisture analysis)

Hot plate method

- 10 g sample heated in beaker with gentle
agitation on hot plate until smoking begins
Moisture and volatile matter = The loss in
weight between the beginning and the
ending sample
Non fatty impurities (Moisture analysis)
Air oven method
accurate, more reliable than hot plate
time consuming
approximately5 grams of a representative
sample is weighed into a dried,
tared
moisture dish and dried in the oven for
30 minutes at 101C ( 1C). This
procedure is repeated until a constant weight
is determined
Theloss in weight is calculated as the moisture
and volatile matter
Non fatty impurities (Moisture analysis)

Vacuum oven method

-similar to air oven method except
-vacuum method no to exceed 100 mm of Hg
at temperature 20 25 deg above b.p of
water at the operating pressure until
constant weight is obtained in successive 1h
drying periods.
Non fatty impurities (Moisture analysis)

Karl Fischer method

-Karl Fischer is an analytical technique used
to determine moisture
Non fatty impurities (Moisture analysis)
Karl Fischer method

The titration endpoint is a color change from yellow to brown.

 Non fatty impurities

Non fatty
impuritie
s

Soap Moisture Trace

Impuritie
analysis analysis material
s analysis
analysis

Insoluble Filterable Turbidimeter

Hot
Vacuum
plate Air
oven
oven Karl
Fisher
Non fatty impurities (Impurities analysis)

Foreign material in incoming crude edible oils

must be removed to produce an acceptable
finished product

Methods to remove impurities:

insoluble impurities
filterable impurities
turbidimeter impurities
Non fatty impurities (Impurities analysis)

Insoluble impurities
- Dirt, seed fragments and etc
insoluble in kerosene or
petroleum ether
Utilizes the residue from
the moisture and volatile
matter

-After wash with warm

kerosene, the crucible is dried
and weighed
Non fatty impurities (Impurities analysis)

Filterable impurities evaluation method

-During processing, undissolved impurities might be present
such as salt which formed after the reaction
-Methods: filter a standard quantity (500 g) of heated (70
90 deg.) oil sample with vacuum suction funnel through
Whatman No. 2 filter disc.
Non fatty impurities (Impurities analysis)

Turbidimeter impurities
-Amount of undissolved impurities in oil can
be determined using this method
-method: sample will be poured into
prescribed sample bottle to eliminate air
bubbles. Then the sample will be heated to
70oC and allowed to stand at 5 10 mins in
controlled temp. oven
Non fatty impurities (Impurities analysis)

Turbidimeter impurities cont.

-The turbidimeter readings can be converted to ppm of
impurities using a calibration curve
No suitable for meat fats and products containing emulsifiers
contain impurities that cannot be effectively measured by
this method
 Non fatty impurities

Non fatty
impuritie
s

Soap Moisture Trace

Impuritie
analysis analysis material
s analysis
analysis

Insoluble Filterable Turbidimeter

Hot
Vacuum
plate Air
oven
oven Karl
Fisher
 Trace material analysis
Metals in oils and fats can cause the deterioration of the
product quality

Metals usually present in edible oils and fats through the

processing level

Most prevalent metals found: copper, iron, calcium,

magnesium, sodium, lead, zinc, and nickel

Can be trace by analysis using ICP (Plasma emission

spectroscopy
 Non fatty impurities

Non fatty
impuritie
s

Soap Moisture Trace

Impuritie
analysis analysis material
s analysis
analysis

Insoluble Filterable Turbidimeter

Hot
Vacuum
plate Air
oven
oven Karl
Fisher
 Soap analysis

In caustic refining, NaOH is added into the oil to react

with free fatty acid and produce soaps

Traces of soap in oil after refining and bleaching could

poison hydrogenation catalyst.

2 methods available for determining soaps:

- Conductivity procedure (Cc 15-60)
- Titration procedure (Cc 17-79)
 Oil and Fat Analysis

8 types of oil and fat analysis:

Non fatty impurities
Melting, solidification and consistency
Composition analysis
Flavor, rancidity and stability
Color and appearance
Refining and bleaching
Performance testing
Nonstandardized methods
 Melting, solidification and consistency
Most frequently and routinely analysis performed

Helps with quality control and product development

good indications of the mouth feel, overall general
behaviour at cool, ambient and elevated temperature
Melting point analysis

Natural fats do not have true melting points

Melting range or melting interval is most appropriate
terms
Several melting point procedures have been standardized
by AOCS
Melting point analysis

Melting Point
Analysis

Capillary melting Mettler dripping

Wiley melting point
point point
 Melting point analysis

Capillary melting point

- Similar with procedure used to determine the melting
point of pure organic compound
- Heated until the fat become clear
 Melting Point
Analysis

Capillary melting Mettler dripping

Wiley melting point
point point
 Melting point analysis
Wiley melting point:
-A tube of 3.5 cm internal diameter is half-filled with boiled water then cautiously
topped off with boiled alcohol so that the latter remains above the water which it
forms an interface.
-A cold disc of fat added to the tube and when disc softens to become spherical
globule melting point
Melting point analysis
Wiley melting point cont:
- Disadvantages:
- -> subjective interpretation in order to determine end
point
- -> Constant attention needed to determine the
endpoint
 Melting Point
Analysis

Capillary melting Wiley melting point Mettler dripping

point point
 Melting point analysis
Dripping point
-use METLLER TOLEDO instrument
-Method: 17 drops of melted fats will be put
in the sample cup
-Put in a freezer for 15 mins
-Cold solidified sample warm through the
furnace until it becomes fluid enough to
flow
-When the sample flow, it trip a
photoelectric circuit that records the
temperature on digital read out.
Melting point analysis

Dripping point
Melting point analysis

Advantaged of Dripping point

had become a standard in most fats
and oil laboratory
fast, reliable result
good correlation with Wiley result
ability
to analyse products with low
melting points
 Solidification Analysis

Solidification of mixed fatty acids: point where a balance

is attained between the heat generated and heat lost,
which will vary according to rate of crystallization.
Types of evaluation:

Solidification
Analysis

Titer test Cloud point Cold test

Solidification Analysis

Titer test: measure the solidification

point of fatty acid
- For edible oils and fats, commonly
specified for low iodine values hard fat
refers to as titer stocks
For inedible fats, used for soap making or
as a raw material for fatty acid
manufacture
 Solidification
Analysis

Titer test Cloud point Cold test

Solidification Analysis

Cold test: the ability of an oil to resist

crystallization by determining the time (in
hours) required for the oil to become
cloudy at 0C (32F).
Method: dry filtered oil is placed in a
sealed 4-ounce bottle and submerge into
an ice bath.
If the oil remains clear for 5.5 h, the oil pass
the cold test.
 Solidification
Analysis

Titer test Cloud point Cold test

 Solidification Analysis

Cloud point: is obtained by stirring a

sample of fat while it is being cooled until
the oil has clouded enough to block a light
beam of known intensity.
The cloud or congeal point values are
more closely related to consistency than
melting points
Consistency Analysis
Consistency: impression of resistance
Plasticity: relates to the capacity of the product to be
molded
Fat become firmer as the solid content increases
Butter and margarine depends upon the consistency of fat
portion and its ability to spread on bread
 Consistency Analysis

Consistency evaluation method:

-consistency ratings
-penetrations
 Consistency Analysis

Consistency rating:

-pressing a finger into the product/ squeeze the

product in the hand

-Still applicable and effective

-required an experienced evaluator

 Consistency Analysis
Penetration
- Widely used method to measure the
consistency of plasticized shortening
or margarine
- Using grease penetrometer
- Measured at different temp. to
determine the plastic range of the
sample
- Perfect plastic range: same
penetration at all temp
 Oil and Fat Analysis

8 types of oil and fat analysis:

Non fatty impurities
Melting, solidification and consistency
Composition analysis
Flavor, rancidity and stability
Color and appearance
Refining and bleaching
Performance testing
Nonstandardized methods
 Composition analysis
Usually done by using wet chemical analysis (classical method)

Saponification
value

Iodine value

refractive index

fatty acid
Composition composition

analysis calculated
iodine value

Emulsifier
analysis

Tocopherol
analysis
Antioxidant
analysis
 Composition analysis

Saponification value
- Measure the alkali-reactive groups in fats and oils
- Useful in predicting type of glycerides
- Glyceride with short-chain fatty acid have higher
saponification values than long-chain fatty acid

- Example: vegetable oils and animal fats has same range of

saponification value: 180 - 200
 Saponification Value
Saponification - hydrolysis of ester under
alkaline condition.

O
H2 C OH
H2 C O C R O
O +
+ 3 KOH HC OH 3 R C OK
HC O C R
O
H2 C OH
H2 C O C R
 Composition analysis
Iodine value
- Measure the unsaturation of fatty acid
- But, cannot define specific fatty acid

Refractive index
-measure the change in unsaturation as the fat or oil is
hydrogenated
-A refractometer with temperature control at 25 deg. is used
-A refractive index decrease with no. of double bonds present
in fatty acids
Differential between saturate and
unsaturated fatty acid

Saturate Unsaturated
Fatty acid chain are straight Fatty acid chain are bent in some place

No double bond Contain double bond

Solid in room temperature Liquid in room temperature

Palmitic fatty acid Oleic fatty acid

Butter, palm nut oils, coconut Olive oil, fish oils, peanut, linseed,
and cottonseed oils
 Composition analysis
Fatty acid composition
- Determined by using GC

Calculated iodine value

- Measure the unsaturation in fatty acid
- Calculated from fatty acid composition analysis
 Composition analysis
Triglyceride structure
HPLC procedures used to determine relative amounts of glycerides present
in a fat

O
O
H2 C O C R
H2 C OH HO C R
O
O
HC OH HC O C R + 3 H2O
+ HO C R
H2 C OH O
O
HO C R H2 C O C R

Glycerol 3 Fatty Acids

Composition analysis
Antioxidant analysis
-To measure the amount of synthetic antioxidant used in fat
and oil products
-Ensure the amount of synthetic antioxidant is beyond the
safety limit (e.g: US FDA allow max. limit of 100 ppm for
meat products)
-Use GC or HPLC for analysis
 Composition analysis

Tocopherol analysis
Vegetable oils contain tocopherols, which are natural
antioxidants that retard oxidative rancidity.
The instrumentation and procedures evaluated have
involved colorimeters, paper chromatography (PC), TLC,
column chromatography (CC), GLC, GLC/mass
spectrometry (GLC/MS), and HPLC separation
techniques
 Oil and Fat Analysis

8 types of oil and fat analysis:

Non fatty impurities
Melting, solidification and consistency
Composition analysis
Flavor, rancidity and stability
Color and appearance
Refining and bleaching
Performance testing
Nonstandardized methods
 Flavor, rancidity and
stability
Customer use organoleptic evaluations to judge the
quality of fat and oil

Organoleptic:
evaluation of oil products has long been recognized as
the most sensitive method of assessing quality, but it is
also recognized that these evaluations generally lack
precision and reproducibility
 Flavor, rancidity and stability

Flavor analysis method

- Sensory evaluations
- Volatile flavor analysis
 Flavor analysis method
Sensory evaluations
-most common method of grading finished oil quality

-Experienced taster/ flavorist rate the flavor of the oil

according to established intensity scale.

-time consuming, expensive

 Flavor analysis method

Volatile flavor analysis

- Use GC analysis
- E.g:
Besides lactic acid, acetaldehyde, acetone, and 2-butanone contribute most to
the typical aroma and flavor of yogurt.
Extended storage of yogurt causes off-flavor development, which is mainly
attributed to the production of undesired aldehydes and fatty acids during lipid
oxidation
 Rancidity analysis
The major cause of rancidification of oil and fats
are:
1) Oxidation
2) Hydrolysis

Factors that contribute to the rancifidication and

formation of off-flavors:
1) Temperature
2) Light
3) Moisture
4) Metals
5) Oxygen
Rancidity
The sharp pungent odours mixed
with stale and musty odours (unpleasant smell)
Hydrolytic Rancidity.
Hydrolytic rancidity results in the formation of free fatty
acids and soaps (salts of free fatty acids) and is caused by
either the reaction of lipid and water in the presence of a
catalyst or by the action of lipase enzymes.

Oxidative Rancidity.
Oxidative rancidity results from more complex lipid
oxidation processes which involve free radical reactions.
The processes are generally considered to occur in three
phases: 1) initiation or induction phase, 2) propagation
phase, and 3) termination phase.
 Rancidity
Analysis to measure:
a) Peroxide value
b) Anisidine value
c) Free fatty acid and acid value
 Rancidity analysis

1. Peroxide Value major cause by oxidation of lipid

Hydroperoxides formed by the reaction between oxygen and the

unsaturated fatty acids are the primary products of this reaction.
Hydroperoxides have no flavor or odour but break down rapidly to
form aldehydes, which have a strong, disagreeable flavour and
odour
 Rancidity analysis

2. Anisidine value

The anisidine value measures the amount of and

unsaturated aldehyde present in the oil.
The method is based on present of acetic acid, p-
anisidine reacts with aldehydic compound in an oil,
producing yellowish reaction products.
The color intensity depend on the amount of
aldehydic compound present.
3. Acid Value

The flavors resulting from FFA developed

depend on composition of fat thus, rancidity
analysis can be measure by FFA
Number of mgs of KOH required to neutralize
the Free Fatty Acids in 1 g of fat.

ml of KOH x N x 56
AV = = mg of KOH
Weight of Sample
Stability analysis
Stability of a fat or oil generally accepted as the
storage life of the product until rancidity become
apparent
Most fats and oil products are tested for flavor
stability as a part of quality control program to ensure
customer specification limits are satisfied
3 types of method to evaluated stability
1) Active Oxygen Method
2) Oil stability index
3) Schaal oven test
4) Pastry flavor test