Analysis of Oil/Fat based
Products
Fats and Oils
Organic substances
Relatively Non-polar
Slightly soluble in water
Highly soluble in organic solvents
Structure & Types of lipids
Fats & oils – chemically known as
Triacylglycerols (TAG / TG)
Tri-esters of glycerol & fatty acids
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 O
H2 C OH O
HO C R H2 C O C R
Chemical structure of TG:
Glycerol + 3 FA TG
Glycerol:
Simple organic compound
Sometimes called as glycerin
3-carbon molecule containing 3 alcohol groups
FA:
Organic molecules
Contain chains of carbon bound to hydrogen, acid
group (COOH) at one end & a methyl group (CH3) at
other end
Ester bonds hold FA to glycerol with loss of water
Saturated & unsaturated fats
Saturated: FA chain doesn’t contain any carbon-to-
carbon double bonds
Oils typically contain high proportion of UFA
• Unsaturated: FA chain
contains 1/more double bonds
• UFA: MUFA (1 = bond) or
PUFA (2/more = bonds)
Examples of FA, characterized according to
saturation:
FA Name FA Type Notation
Butyric Saturated C4:0
Capric ,, C10:0
Palmitic ,, C16:0
Stearic ,, C18:0
Oleic MU C18:1
Linoleic PU C18:2
linolenic PU C18:3
EPA PU C20:5
Fatty Acid Naming Systems
Chemical reactivity of UFA determine by the
position & no: of double bonds.
High degree of unsaturation: high reactivity
2 types of double bonds:
Methylene interrupted (eg:w-FA):C=C-C-C=C
Conjugated (alternative single & = bonds):
C=C-C=C-C
Cis & Trans Fats
UFA have 2 configurations, defined by the structure at
the double bond;
Cis: H atoms bonding to the C=C are located on same
side of the double bond.
Eg: oleic acid
Trans: H atoms attached to the C=C are opposite to
each other.
Eg: elaidic acid
Mostly UFA double bonds exist in foods in cis form
than trans form.
Melting Point
To at which a solid is converted into liquid.
FA molecules exist in crystalline forms.
Strength of bonding forces between FA determine the
MP of a fat crystal.
FA exhibit unique MP.
Pure substance (single type of FA): sharp MP
corresponding to a defined To.
Determine whether a fat is a liquid/solid/plastic/brittle…
at room To.
Melting point of fats
FA Name FA Type Notation MP (oC)
Stearic saturated C18:0 70
Oleic Cis-unsatu C18:1 14
Elaidic Trans-unsatu C18:2 43
Why???
• MP of Saturated fats are higher than Unsaturated fats.
• Trans fats have higher MP than Cis
• Trans fats exhibit similar characteristics to saturated fats.
Cis- and Trans-Fatty Acids Compared
Chemical Reactions of
Lipids
1. Hydrolysis
Breakdown of lipid molecule / TG into FA &
Glycerol.
Reaction require heat & additional water
molecules.
Glycerol can further break down to “acrolein” &
can produce odorous, irritating fumes from
overheated fats.
Reaction:
Hydrolytic rancidity:
Stored fats become rancid by hydrolysis reaction
with water due to liberation of FFA
Can result due to: heat, naturally present food
enzymes (lipase)
Size of the liberated FA – important in
determining rancidity.
SCFA – objectionable flavors & odors
LCFA – no off-flavors & odors
Eg. Butter – when left out at room To for too
long
Bad smell & taste
(Reason: Many SCFA in butter fat, especially C4-
butyric & C6-caproic, hydrolyze from glycerol.
In digestive tract of human & animals
Fat hydrolyzed by lipase enzyme
FFA released – easily absorbed by GIT
2. Hydrogenation
Forced addition of H atoms to the unsaturated
bonds in an unsaturated fat.
Reaction:
Use H2 (g) at elevated To & under Po.
High Po under suitable catalyst – add H atoms to
the = bond.
Process raises the MP of fats.
Use in food industry to “harden” liquid oils to
semi-solid fats.
Eg. vegetable oils margarine
Side effect – change of the % of Cis-UFA to
Trans-UFA.
Reaction:
Hydrogenation reduce the tendency of a fat to
oxidize due to less unsaturation.
In commercial frying operations:
Use frying oils (corn, peanut) slightly hydrogenated
Provides a longer usable “frying life” for oil by
making it less unsaturated.
Less reactivity with O2 (more stable)
Stages of Hydrogenation:
1. Light hydrogenation – lipid present as liquid
2. Partial hydrogenation – present as fluid / plastic
shortening
3. Complete hydrogenation – solids / flakes
Can stop the process at any stage.
Hence, result 4 types of products:
1. liquid
2. fluid shortenings
3. plastic shortenings
4. solids / flakes
Factors affecting the rate of hydrogenation:
Nature of the substance to be hydrogenated: more
double bonds – high reaction rate
Nature & concentration of the catalyst: high
concentration – high reaction rate. Normally use Cu
& Ni
Hydrogen concentration: high [H2] – high RR
Reaction To: High To – high RR, high trans FA
formation
Pressure: high Po – high RR, low trans FA formation
Agitation: high agitation – high RR, low trans FA
3. Oxidation
Oxygen reacts with double bonds of UFA.
Produce small organic compounds & generate
undesirable “rancid” odor (oxidative rancidity of fats &
oils)
Nutritional value of original FA – lost
Occur in 3 stages:
1. Initiation
2. Propagation
3. Termination
Initiation Propagation
catalyst O2 RH
RH R* ROO* ROOH + R* ROO*
Odors / “secondary” products
aldehydes ketones alcohols carboxylic acids
Initiation:
Fat molecules form fatty FR in the presence of initiators
(heat, heavy metals, UV light)
Propagation:
FA free radical (R*) reacts with O2.
Produce activated peroxide / peroxyl FR (ROO*)
ROO* reacts with a nearly UFA (RH) to remove H
atom
Generate hydroperoxide (ROOH) from peroxyl FR
& a new FA FR (R*)
Newly form R* again reacts with O2
Cycle repeated
At some point hydroperoxide (ROOH)
decompose into smaller Short chain
organic molecules (aldehydes, ketones..)
These contributes to oxidized & rancid
odors & flavors.
Termination:
Occurs when;
All fat molecules have reacted
Two unstable FR species react with each
other
ROO* + R* ROOR
Anti-oxidant reacts with a radical
AH + ROO * ROOH + A *
Factors affecting the reaction:
FA composition: higher = bonds – high
oxidation, geometry of double bonds (cis rapidly
oxidize than trans)
Temperature: higher the To, greater the
oxidation
To prevent/delay the onset of oxidative rancidity:
Fats & oils should store in cool areas away from
light & metallic containers.
Use of A/O in food products
4. Esterification
Recombination of FFA with glycerol to form
TAG, DAG or MAG by esterification.
Some instances:
Alkaline
TAG + glycerol DAG + MAG
Catalysts
Analytical procedures
applied for processed fats
Iodine Value
Iodine value (IV) - a measure of the average
degree of unsaturation of a lipid
Higher the iodine value - greater the number of
C=C double bonds.
By definition: the grams of iodine absorbed per
100g of lipid.
most commonly used method for determining IV
of lipids - "Wijs method".
Lipid is weighed and dissolved in a suitable
organic solvent, and a known excess of ICl is
added.
Some ICl reacts with the double bonds in
unsaturated lipids, while the rest remains:
R-CH=CH-R’ + IClexcess R-CHI-CHCl-R’ + IClremaining
Amount of Icl reacted is determined by
measuring the amount of ICl remaining after
reaction has gone to completion :
(IClreacted = IClexcess - IClremaining)
ICl remaining is determined by adding excess
KI to the solution to liberate iodine, and then
titrating with a Na2S2O3 solution with starch
indicator to determine the concentration of
iodine released:
IClremaining + 2KI KCl + KI + I2
Na2S2O3
Oil, solvent,
ICl, KI, Starch
I2 + starch + 2Na2S2O3 (blue) 2NaI + starch + Na2S4O6 (colorless)
• Iodine itself - reddish brown color, but not intense
enough to use as a good indicator.
• Thus, starch used as the indicator - forms a molecular
complex with iodine (deep blue color).
While there is any I2 remaining in the
solution it stays blue, but once all of the I2
has been converted to I- it turns colorless.
Concentration of C=C in original sample can
calculate by measuring the amount of
sodium thiosulfate needed to complete the
titration.
Higher the degree of unsaturation, more
iodine absorbed, and higher the iodine value.
The iodine value - a measure of the average
degree of unsaturation of oils
Saponification Value
A measure of the average molecular weight of the
triacylglycerols in a sample.
Saponification - process of breaking down a neutral fat
into glycerol and fatty acids by treatment with alkali:
TAG + 3 KOH Glycerol + 3 FA salts of potassium
Defined as: mg of KOH required to saponify 1 g of fat.
Smaller the saponification number - larger the average
molecular weight of the TAG present.
• Lipid is first extracted and dissolved in ethanol solution
which contains a known excess of KOH.
• This solution is then heated so that the reaction goes to
completion.
• Unreacted KOH is then determined by adding an indicator
and titrating with HCl.
• The saponification number is calculated from a
knowledge of the weight of sample and the amount of
KOH reacted.
HCl
Oil, etanol, excess
KOH,
phenolphthalein
Acid value
A measure of the amount of free acids present in
a given amount of fat.
Defined as: mg of KOH necessary to neutralize
the fatty acids present in 1g of lipid.
The acid value may be overestimated if other
acids are present in the system, e.g. aa, or acid
phosphates.
A good measure of the break down of the TAG
into free fatty acids, which has an adverse effect
on the quality of many lipids.
The lipids are extracted from the food sample and
then dissolved in an ethanol solution containing an
indicator. This solution is then titrated with alkali
(KOH) until a pinkish color appears.
KOH
Oil, ethanol,
phenolphthalein
Peroxide value
Peroxides (R-OOH) are primary reaction
products formed in the initial stages of oxidation.
Gives an indication of the progress of lipid
oxidation.
Commonly used method: ability of peroxides to
liberate iodine from potassium iodide.
Lipid is dissolved in a suitable organic solvent
and an excess of KI is added:
ROOH + KIexcess ROH + KOH + I2
Once the reaction is complete, the amount of
ROOH reacted is determined by measuring the
amount of iodine formed.
This is done by titration with sodium thiosulfate
and a starch indicator:
I2 + starch + 2Na2S2O3 (blue) 2NaI + starch
+ Na2S4O6 (colorless)
Amount of sodium thiosulfate required to titrate
the reaction is related to the concentration of
peroxides in the original sample