LIPID

Soluble in non-polar solvents and insoluble in polar

solvents. Lipid is not polymers.

Lipids:
1. Fatty acids
2. Neutral fats and oils
3. Waxes
4. Phospholipid
5. Sterols
6. Fat soluble vitamins
Fatty Acids

O
R C OH

#1 Carbon
Acid Group
O
R C OH Polar End - Hydrophilic End

Non-polar End - Hydrophobic End

(Fat-soluble tail)
 Saturated Fatty Acids

O
8 7 6 5 4 3 2 1
CH 3 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 C OH

Octanoic Acid
 Unsaturated Fatty Acids
O
8 7 6 5 4 3 2 1
CH 3 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 C OH

3 - Octenoic Acid
O
8 7 6 5 4 3 2 1
CH 3 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 C OH
3, 6 - Octadienoic Acid

Short hand: 8:1 (∆3)

8:2 (∆3,6)
 Cis And Trans Fatty Acids

HH O
CH 3 (CH 2 )7 C C (CH 2 )7 C OH
10 9
Cis 9 - Octadecenoic Acid (oleic)
H O
CH 3 (CH 2 )7 C C (CH 2 )7 C OH
H
Trans 9 - Octadecenoic Acid (elaidic acid)
 Polyunsaturated Fatty Acids

Linoleic acid: Cis, cis, 9, 12 - Octadecadienoic acid

Linolenic acid: Cis, cis, cis 9, 12, 15 - Octadecatrienoic acid
Arachidonic acid: Cis, cis, cis, cis 5, 8, 11, 14 - Eicosatetraenoic
acid

Linoleic Acid

Linolenic Acid

Arachidonic Acid
 Naturally-occurring fatty acids

O
R CH 2 CH CH CH 2 CH CH CH 2 C OH
7 6 5 4 3

1. Cis form
2. Not conjugated --- isolated double bond.
3. Even numbered fatty acids.
CLASSIFICATION OF FATTY ACIDS PRESENT
AS GLYCERIDES IN FOOD FATS
Common Systematic Formula Common source
Name Name

I. Saturated Fatty Acids

Butyric Butanoic CH3(CH2)2COOH butterfat
Caproic Hexanoic CH3(CH2)4COOH butterfat, coconut
and palm nut oils
Caprylic Octanoic CH3(CH2)6COOH coconut and palm
nut oils, butterfat
Capric Decanoic CH3(CH2)8COOH coconut and palm
nut oils, butterfat
Lauric Dodecanoic CH3(CH2)10COOH coconut and palm
nut oils, butterfat
Myristic Tetradecanoic CH3(CH2)12COOH coconut and Palm nut oil, most
animal and plant fats
Palmitic Hexadecanoic CH3(CH2)14COOH practically all animal and
plant fats
Stearic Octadecanoic CH3(CH2)16COOH animal fats and minor
component of plant fats
Arachidic Eicosanoic CH3(CH2)18COOH peanut oil
Common Systematic Formula Common source
Name Name

II. Unsaturated Fatty Acids

A. Monoethenoic Acids
Oleic Cis 9-octadecenoic C17H33COOH plant and animal fats
Elaidic Trans 9-Octadecenoic C17H33COOH animal fats

B. Diethenoic Acids
Linoleic 9,12-Octadecadienoic C17H31COOH peanut, linseed, and
cottonseed oils
C. Triethenoid Acids
Linolenic 9,12,15-Octadecatrienoic C17H29COOH linseed and other seed
oils
Eleostearic 9,11,13-Octadecatrienoic C17H29COOH peanut seed fats

D. Tetraethenoid Acids
4,8,12,15-
Moroctic C17H27COOH fish oils
Octadecatetraenoic
Arachidonic 5,8,11,14- C19H31COOH traces in animal fats
 Common and Systematic Names of Fatty Acids
Common Systematic Formula Common source
Name Name
A. Monoethenoic Acids

Oleic Cis 9-octadecenoic C17H33COOH plant and animal fats

Elaidic Trans 9-Octadecenoic C17H33COOH animal fats

B. Diethenoic Acids
Linoleic 9,12-Octadecadienoic C17H31COOH peanut, linseed, and
cottonseed oils
C. Triethenoid Acids
Linolenic 9,12,15-Octadecatrienoic C17H29COOH linseed and other seed
oils
Eleostearic 9,11,13-Octadecatrienoic C17H29COOH peanut seed fats

D. Tetraethenoid Acids
4,8,12,15-
Moroctic C17H27COOH fish oils
Octadecatetraenoic
Arachidonic 5,8,11,14- C19H31COOH traces in animal fats
Eicosatetraenoic
Melting Points and Solubility in Water of Fatty Acids

Melting Point

Solubility in H O
2

Chain Length
CHARACTERISTICS OF FATTY ACIDS

Fatty Acids M.P.(0C) mg/100 ml Soluble in H2O

C4 -8 -
C6 -4 970
C8 16 75
C10 31 6
C12 44 0.55
C14 54 0.18
C16 63 0.08
C18 70 0.04
Effects of Double Bonds on the Melting Points

F. A. M. P. (0C)
16:0 60
16:1 1
18:0 63
18:1 16
18:2 -5
18:3 -11
20:0 75
20:4 -50

M.P.

# Double bonds
 FAT AND OILS

Mostly Triglycerides:

O
O
H2 C O C R
H2C OH HO C R O
O
HC OH + HC O C R + 3 H2O
HO C R
H2C OH O
O
HO C R H2 C O C R

Glycerol 3 Fatty Acids

 GLYCERIDES

H2 C OH O
H2 C O C (CH 2 )16 CH 3
HC OH O
HC OH O
H2 C O C (CH 2 )16 CH 3
H2 C O C (CH 2 )16 CH 3
Monoglyceride (α - monostearin) Diglyceride (α, α' - distearin)
O
H2 C O C (CH 2 )16 CH 3 ( C18 )
O
HC O C (CH 2 )14 CH 3 (C16 )
O
H2 C O C (CH 2 )16 CH 3 (C18 )

Triglyceride (β - palmityl distearin)

Oleic

Palmitic OPP
Palmitic
α - oleodipalmitin
1 - oleodipalmitin
Linoleic
Oleic
LOO
Oleic
α - Linoleyldiolein
1 - Linoleyldiolein
FATS AND OILS ARE PRIMARILY TRIGLYCERIDES (97-99%)

Vegetable oil - world supply - 68%

Cocoa butter - solid fat
Oil seeds - liquid oil

Animal fat - 28% (from Hogs and Cattle)

Marine oil - 4%
Whale oil
cod liver oil
 Fatty Acids (%) of Fats and Oils

Fatty Acids Butter Coconut Cottonseed Soybean

4 3
6 3
8 2 6
10 3 6
12 3 44
14 10 18 1
16 26 11 4 12
16:1 7 1
18:0 15 6 3 2
18:1 29 7 18 24
18:2 2 2 53 54
18:3 2 8
MELTING POINTS OF TRIGLYCERIDES

Triglyceride Melting Point (°C)

C6 -15

C12 15

C14 33

C16 45

C18 55

C18:1 (cis) -32

C18:1 (trans) 15
 WAXES
Fatty acids + Long chain alcohol
Important in fruits:
3. Natural protective layer in fruits, vegetables, etc.
4. Added in some cases for appearance and protection.
Beeswax (myricyl palmitate), Spermaceti (cetyl palmitate)

O O
C30 H61 O C C 15 H31 C16 H33 O C C 15 H31
 PHOSPHOLIPID
Lecithin (phosphatidyl choline)

O
H2 C O C R
O
R C O CH
O CH 3
+
H2 C O P O CH 2 CH 2 N CH 3
O_ CH 3
Phosphatidic Acid Choline
 STEROLS

Male & female sex hormones

Bile acids
Vitamin D
Adrenal corticosteroids
Cholesterol
21
22
H 3C CH 3
18
H 3C 20
CH 3
19 12 17 16
H3C 11 13 14
10 15
1 9
2 8
3 6 7
4
HO 5
 FAT SOLUBLE VITAMINS
Vitamin A:

H3 C CH 3 CH3 CH3
5
8 CH2OH
9 7 3 1
6 4 2

CH3
 CH 3
H 3C CH 3
H 3C
CH 3

Vitamin D2:
H
H
CH 2

HO

Vitamin E:

R1 CH 3
O CH 3
R2 (CH 2 CH 2 CH 2 CH 2 )2 CH 2 CH 2CH 2 CH(CH 3 )2
HO
R3
ANALYTICAL METHODS TO MEASURE THE
CONSTANTS OF FATS AND OILS

1. Acid Value
2. Saponification Value
3. Iodine Value
4. Gas Chromatographic Analysis for Fatty Acids
5. Liquid Chromatography
6. Cholesterol Determination
 1. Acid Value

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
 2. Saponification Value

Saponification - hydrolysis of ester under alkaline

condition.

O
H 2 C OH
H2 C O C R O
O
+ 3 KOH HC O H + 3 R C OK
HC O C R
O
H 2 C OH
H2 C O C R
Saponification Value of Fats and Oils

Fat Saponification #

Milk Fat 210-233

Coconut Oil 250-264

Cotton Seed Oil 189-198

Soybean Oil 189-195

Lard 190-202
 2. Saponification Value Determination

Saponification # --mgs of KOH required to saponify 1 g of fat.

1. 5 g in 250 ml Erlenmeyer.
2. 50 ml KOH in Erlenmeyer.
3. Boil for saponification.
4. Titrate with HCl using phenolphthalein.
6. Conduct blank determination.

56.1(B -S ) x N of HCl
S P# =
Gram of S ample

B - ml of HCl required by Blank.

S - ml of HCl required by Sample.
 3. Iodine Number

Number of iodine (g) absorbed by 100 g of oil.

Molecular weight and iodine number can calculate the

number of double bonds. 1 g of fat adsorbed 1.5 g of
iodine value = 150.
 Iodine Value Determination

Iodine Value = (ml of Na2S2O3 volume for blank - ml of Na2S2O3

volume for sample) × N of Na2S2O3 × 0.127g/meq × 100
Weight of Sample (g)

CH CH + ICl CH CH
Iodine chloride Cl I

Excess unreacted ICl

ICl + KI KCl + I2

I2 + 2 Na2 S 2 O3 Na2 S 4 O6 + 2 NaI

 Iodine Numbers of Triglycerides

Fatty Acids # of Double-bonds Iodine #

Palmitoleic Acid 1 95

Oleic Acid 1 86

Linoleic Acid 2 173

Linolenic Acid 3 261

Arachidonic Acid 4 320

 Compositions (%) of Fatty Acids of Fats

Fat C4 C6 C10 C16 C18 C18:1 C18:2 C18:3 C20:4

1 5 5 20 40 30

2 20 35 40 5

3 10 50 40

4 20 40 40

5 10 20 20 10 20 20

6 100
 4. GC Analysis for Fatty Acids

1. Extract fat.
2. Saponify (hydrolysis under basic condition).
3. Prepare methyl ester (CH3ONa).
4. Chromatography methyl ester.
5. Determine peak areas of fatty acids.
Fatty acids are identified by retention time.
6. Compare with response curve of standard.
 Fatty Acids Methyl Esters:

Response

18:1

14

18:3 21:1 24
16 18:2 20
18 22

Time

GC condition: 10% DEGS Column (from supelco)

Column temperature 200C.
 5. TRIGLYCERIDE ANALYSIS BY LIQUID
CHROMATOGRAPHY

Soybean Oil
Solvent CH3CN/HF
Column 84346 (Waters Associates)

RES PONS E

RETENTION TIME
Oleate-containing triglycerides in olive oil

Fatty Acid Total Acyl Carbons: Equivalent Carbon

Composition Unsaturation Number

OL2 54:5 44

O2L 54:4 46

OPL 52:3 46

O3 54:3 48

OSL 54:3 48

O2P 52:2 48

O2S 54:2 50

OPS 52:1 50

OS2 54:1 52
 6. CHOLESTEROL DETERMINATION

Enzymatic Determination: Cholesterol Oxidase

Cholesterol Oxidase
etc. + H2 O2
HO O

CH3O OCH3 CH3O OCH3

H2 O2 +H2N NH2 Peroxidase HN NH
+ H2 O

0-Dianisidine Oxidized 0-Dianisidine

(Colorless) (Brown color)At 440 nm
 Absorption
at 440 nm

µg/ml Cholesterol

Cholesterol by GLC
1. Prepare cholesterol butyrate.
2. Analyze by GLC.
time in GC - 15 min.
sensitivity - 10-7 g.
Spectromertic Absorption Standard Curve of Cholesterol

Absorption
at 440 nm

µg/ml Cholesterol

Cholesterol by GLC
1. Prepare cholesterol butyrate.
2. Analyze by GLC.
time in GC - 15 min.
sensitivity - 10-7 g.
 LIPID CONTENT ANALYSES

1. Gravimetric Method
(1) Wet extraction - Roese Gottliegb & Mojonnier.
(2) Dry extraction - Soxhlet Method.

2. Volumetric Methods (Babcock, Gerber Methods)

 1. Gravimetric Method

(1) Wet Extraction - Roese Gottlieb & Mojonnier.

For Milk:
1) 10 g milk + 1.25 ml NH4OH mix. solubilizes
protein and neutralizes.
2) + 10 ml EtOH - shake. Begins extraction, prevents
gelation of proteins.
3) + 25 ml Et2O - shake and mix.
4) + 25 ml petroleum ether, mix and shake.
 (2) Dry Extraction - Soxhlet Method.

Sample in thimble is continuously extracted with ether

using Soxhlet condenser. After extraction, direct
measurement of fat
- evaporate ether and weigh the flask.

Indirect measurement - dry thimble and weigh thimble and

sample.
Soxhlet Method.
 2. Volumetric Method (Babcock, Gerber Methods)

Theory:
2. Treat sample with H2SO4 or detergent.
3. Centrifuge to separate fat layer.
4. Measure the fat content using specially calibrated bottles.

Methods:
1. Known weight sample.
2. H2SO4 - digest protein, liquefy fat.
3. Add H2O so that fat will be in graduated part of bottle.
4. centrifuge to separate fat from other materials completely.
 REACTIONS OF FATS

Hydrolytic Rancidity:

1. Triglyceride -> Fatty acids

Specially C4 butyric acid (or other short chain fatty
acids) are the real problem.

2. By lipase.
 LIPID OXIDATION

Major flavor problems in food during storage are mainly

due to the oxidation of lipid.

Lipid Oxidation - free radical reactions.

1. Initiation.
2. Propagation.
3. Termination.
Pentane Formation from Linolenic Acid
14 13 12 11 10 9
CH 3 (CH 2 )3 CH 2 CH CH CH 2 CH CH CH 2 COOH
n
Initiation (metal)
- H.

12 11 10 9
CH 3 (CH 2 )3 .
CH 2 CH CH CH CH CH CH 2 n COOH
Propagation + O2
12 11 10 9
CH 3 (CH 2 )3 CH 2 CH CH CH CH CH CH 2 nCOOH
O
Propagation
.
O + H.

12 11 10 9
CH 3 (CH 2 )3 CH 2 CH CH CH CH CH - CH 2n COOH
O
Hydroperoxide
Decomposition O _ .OH
H

12 11 10 9
CH 3 (CH 2 )3 CH 2 CH CH CH CH CH CH 2n COOH
.
O

O
CH 3 (CH 2 )3 CH2
. 12
+ H C CH CH CH CH CH
11 10 9
2n COOH

Termination + H .
CH 3 (CH 2 )3 CH 3
Pentane
 ANALYSIS OF FLAVOR QUALITY & STABILITY OF OIL

1. Peroxide Value

O O
A. KI + CH 3 C OH HI + CH 3 C OK

B. ROOH + 2 HI I2 + H2O + ROH

C. I2 + 2 Na2 S 2 O3 2 NaI + Na2 S4 O6

Peroxide Value = ml of Na2S2O3 × N × 1000

(milliequivalent peroxide/kg of sample) Grams of Oil
2. Active Oxygen Method (AOM)

Determined the time required to obtain certain

peroxide value under specific experimental conditions.
The larger the AOM value, the better the flavor
stability of the oil.
3. TBA Test.
To determine the rancidity degree of meat or fish product.

HS N OH O O
+ C CH 2 C
N H H
OH

HS N OH HO N SH
+ 2 H2O
N N
CH CH CH
OH OH
Colored Pigment