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