LIPID

Soluble in non-polar solvents and insoluble in polar solvents. Lipid is not polymers. Lipids: 1. 2. 3. Fatty acids Neutral fats and oils Waxes

4.
5. 6.

Phospholipid
Sterols Fat soluble vitamins

Fatty Acids
O R C OH Acid Group
Polar End - Hydrophilic End

#1 Carbon
O R C

OH

Non-polar End - Hydrophobic End (F at-soluble tail)

Saturated Fatty Acids

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

Octanoic Acid

Unsaturated Fatty Acids

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

3 - Octenoic Acid

8 5 3 7 4 6 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 (D3) 8:2 (D3,6)

Cis And Trans Fatty Acids

H CH3 (CH2 )7 H (CH 2 )7 O C OH

C C 10 9

Cis 9 - Octadecenoic Acid (oleic)

H CH3 (CH 2 )7 C C H (CH 2 )7

O C OH

Trans 9 - Octadecenoic Acid (elaidic acid)

Polyunsaturated Fatty Acids

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

Linolenic acid:
Arachidonic acid:

Cis, cis, cis 9, 12, 15 - Octadecatrienoic acid

Cis, cis, cis, cis 5, 8, 11, 14 - Eicosatetraenoic acid

Linoleic Acid Linolenic Acid Arachidonic Acid

Naturally-occurring fatty acids

O R CH2 CH
7

CH
6 5

CH 2

CH
4

CH
3

CH 2

OH

1. 2. 3.

Cis form Not conjugated --- isolated double bond. Even numbered fatty acids.

CLASSIFICATION OF FATTY ACIDS PRESENT AS GLYCERIDES IN FOOD FATS

Common Name Systematic Name Formula Common source

I. Saturated Fatty Acids

Butyric Caproic Caprylic Capric Lauric Myristic Palmitic Stearic Arachidic Butanoic Hexanoic Octanoic Decanoic Dodecanoic CH3(CH2)2COOH CH3(CH2)4COOH CH3(CH2)6COOH CH3(CH2)8COOH CH3(CH2)10COOH butterfat butterfat, coconut and palm nut oils coconut and palm nut oils, butterfat coconut and palm nut oils, butterfat coconut and palm nut oils, butterfat coconut and Palm nut oil, most animal and plant fats practically all animal and plant fats animal fats and minor component of plant fats peanut oil

Tetradecanoic CH3(CH2)12COOH Hexadecanoic CH3(CH2)14COOH Octadecanoic Eicosanoic CH3(CH2)16COOH CH3(CH2)18COOH

Common Name

Systematic Name

Formula

Common source

II. Unsaturated Fatty Acids

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

B. Diethenoic Acids
Linoleic 9,12-Octadecadienoic C17H31COOH peanut, linseed, and cottonseed oils linseed and other seed oils peanut seed fats

C. Triethenoid Acids
Linolenic 9,12,15-Octadecatrienoic C17H29COOH C17H29COOH

Eleostearic 9,11,13-Octadecatrienoic

D. Tetraethenoid Acids
Moroctic Arachidonic 4,8,12,15Octadecatetraenoic 5,8,11,14C17H27COOH C19H31COOH fish oils traces in animal fats

Common and Systematic Names of Fatty Acids

Common Name Systematic Name Formula Common source

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

B. Diethenoic Acids
Linoleic 9,12-Octadecadienoic C17H31COOH peanut, linseed, and cottonseed oils linseed and other seed oils peanut seed fats

C. Triethenoid Acids
Linolenic 9,12,15-Octadecatrienoic C17H29COOH C17H29COOH

Eleostearic 9,11,13-Octadecatrienoic

D. Tetraethenoid Acids
Moroctic Arachidonic 4,8,12,15Octadecatetraenoic 5,8,11,14Eicosatetraenoic C17H27COOH C19H31COOH fish oils traces in animal fats

Melting Points and Solubility in Water of Fatty Acids

Melting Point

Solubility in H O Chain Length

CHARACTERISTICS OF FATTY ACIDS

Fatty Acids C4 C6 M.P.(0C) -8 -4 mg/100 ml Soluble in H2O 970

C8
C10 C12 C14 C16 C18

16
31 44 54 63 70

75
6 0.55 0.18 0.08 0.04

Effects of Double Bonds on the Melting Points

F. A. 16:0 16:1 18:0 18:1 18:2 18:3 20:0 20:4
M.P.

M. P. (0C) 60 1 63 16 -5 -11 75 -50

# Double bonds

FAT AND OILS

Mostly Triglycerides:

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

O O C R O O C R O O C R + 3 H2O

Glycerol

3 Fatty Acids

GLYCERIDES
H2 C HC H2 C OH OH O O C (CH 2 )16 CH3
H2 C HC H2 C O O C OH O O C (CH 2 )16 CH 3 (CH 2 )16 CH 3

Monoglyceride (a - monostearin)
H2 C HC H2 C O O C O O C O O C

Diglyceride (a, a' - distearin)

( C18 ) (C16 ) (C18 )

(CH 2 )16 CH3 (CH 2 )14 CH3 (CH 2 )16 CH3

Triglyceride (b - palmityl distearin)

Oleic Palmitic Palmitic OPP

a - oleodipalmitin 1 - oleodipalmitin
Linoleic Oleic Oleic LOO

a - 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 6 8 10 12 14 16 16:1 18:0 18:1 18:2 18:3

3 3 2 3 3 10 26 7 15 29 2 2

6 6 44 18 11
6 7 2

1 4 1 3 18 53

12 2 24 54 8

MELTING POINTS OF TRIGLYCERIDES

Triglyceride C6 C12 Melting Point (C) -15 15

C14
C16 C18 C18:1 (cis) C18:1 (trans)

33
45 55 -32 15

WAXES
Fatty acids + Long chain alcohol Important in fruits: 1. Natural protective layer in fruits, vegetables, etc. 2. Added in some cases for appearance and protection.

Beeswax (myricyl palmitate), Spermaceti (cetyl palmitate)

O C30 H61 O C C 15 H31
C16 H33 O

O C C 15 H31

PHOSPHOLIPID
Lecithin (phosphatidyl choline)
O O R C H2 C O O C R

CH O O P O_ CH3 O CH CH2 2 N CH3 CH3 Choline

+

H2 C

Phos phatidic Acid

STEROLS
Male & female sex hormones

Bile acids
Vitamin D Adrenal corticosteroids Cholesterol
21 H3C 18 22 20 16 15

CH3 CH3

H3C

HO

12 19 17 H3C 11 13 14 10 1 9 2 8 3 6 7 4 5

FAT SOLUBLE VITAMINS

Vitamin A:

H3 C

CH3
8 9

CH3
5 7 6 4

CH3
3

CH2 OH
2 1

CH3

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

Vitamin D2:
H H CH 2

HO

Vitamin E:
R1 R2 HO R3 CH3 CH 3 O (CH2 CH2 CH2 CH2 )2 CH2 CH2CH2 CH(CH 3 )2

ANALYTICAL METHODS TO MEASURE THE CONSTANTS OF FATS AND OILS

1. 2. Acid Value Saponification Value

3.
4. 5. 6.

Iodine Value
Gas Chromatographic Analysis for Fatty Acids Liquid Chromatography Cholesterol Determination

1. Acid Value
Number of mgs of KOH required to neutralize the Free Fatty Acids in 1 g of fat.

AV =

ml of KOH x N x 56 Weight of Sample

= mg of KOH

2. Saponification Value
Saponification - hydrolysis of ester under alkaline condition.

H2 C HC H2 C

O O C O O C O O C

R R R + 3 KOH

H2 C HC H2 C

O H OH O H + O 3R C OK

Saponification Value of Fats and Oils

Fat Milk Fat Coconut Oil Cotton Seed Oil Soybean Oil Lard

Saponification # 210-233 250-264 189-198 189-195 190-202

2. Saponification Value Determination

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

1.
2. 3. 4. 5.

5 g in 250 ml Erlenmeyer.
50 ml KOH in Erlenmeyer. Boil for saponification. Titrate with HCl using phenolphthalein. Conduct blank determination.
SP# = 56.1(B -S) x N of HCl Gram of Sample

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 Iodine chloride

CH Cl

CH I

Excess unreacted ICl

ICl I2 + + KI KCl + I2 + 2 NaI

2 Na2 S2 O3

Na2 S4 O6

Iodine Numbers of Triglycerides

Fatty Acids Palmitoleic Acid # of Double-bonds 1 Iodine # 95

Oleic Acid
Linoleic Acid Linolenic Acid Arachidonic Acid

1
2 3 4

86
173 261 320

Compositions (%) of Fatty Acids of Fats

Fat 1 2 C4 5 C6 C10 C16 C18 C18:1 C18:2 C18:3 C20:4 5 20 20 40 35 40 5 30

3
4 5 6

10
20 10

50
40 20 40 20

40

10

20

20 100

4. GC Analysis for Fatty Acids

1.
2. 3.

Extract fat.
Saponify (hydrolysis under basic condition). Prepare methyl ester (CH3ONa).

4.
5.

Chromatography methyl ester.

Determine peak areas of fatty acids. Fatty acids are identified by retention time.

6.

Compare with response curve of standard.

Fatty Acids Methyl Esters:

Res pons e 18:1

14

16

18:2 18

18:3 20 22

21:1

24

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)

RESPONSE

RETENTION TIME

Oleate-containing triglycerides in olive oil

Fatty Acid Composition Total Acyl Carbons: Unsaturation Equivalent Carbon Number

OL2
O2L OPL O3 OSL

54:5
54:4 52:3 54:3 54:3

44
46 46 48 48

O2P
O2S OPS

52:2
54:2 52:1

48
50 50

OS2

54:1

52

6. CHOLESTEROL DETERMINATION
Enzymatic Determination: Cholesterol Oxidase

Choles terol Oxidase etc. + HO O H2 O2

CH3O

OCH3 NH2

CH3O

OCH3 NH

H 2 O2 + H2N

Peroxidase

HN

+ H2 O

0-Dianisidine (Colorless)

Oxidized 0-Dianisidine (Brown color)At 440 nm

Abs orption at 440 nm

g/ml Cholesterol

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

Spectromertic Absorption Standard Curve of Cholesterol

Abs orption at 4 40 nm

g/ml Cholester ol

Cholesterol by GLC
1. 2. Prepare cholesterol butyrate. 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:

1. Treat sample with H2SO4 or detergent.

2. Centrifuge to separate fat layer. 3. 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.
2.

Initiation.
Propagation.

3.

Termination.

Pentane Formation from Linolenic Acid

14

CH3

(CH 2 )3

CH2

13

CH

12

CH

11

CH 2

10

CH

CH

CH 2

COOH

Initiation (metal)

- H.

CH3

(CH 2 )3

CH2

.CH
12

12

11

10

CH + O2
11

CH

CH

CH

CH 2 n COOH

Propagation CH3 (CH 2 )3 CH2 CH O Propagation O

12

10

CH

CH + H.
11

CH

CH

CH 2 n COOH

10

CH 3

(CH 2 )3

CH2

CH O O H

CH _

CH

CH

CH -

CH 2 n COOH

Hydroperoxide Decomposition

.OH
11 10 9

12

CH 3

(CH 2 )3

CH2

CH

CH

CH

CH

CH

CH 2 n COOH

.
CH 3 (CH 2 )3

. CH
2

O + H C

12

11

10

CH + H

CH

CH

CH

CH 2 n COOH

Termination CH3

(CH 2 )3 Pentane

CH3

ANALYSIS OF FLAVOR QUALITY & STABILITY OF OIL

1.

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

B.

ROOH + 2 HI

I2 +

H2 O +

ROH

C.

I2 + 2 Na2 S 2 O3

2 NaI +

Na2 S4 O6

Peroxide Value =

ml of Na2S2O3 N 1000 Grams of Oil

(milliequivalent peroxide/kg of sample)

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 N OH + H O C CH2 C H O

HS N

OH

HO CH CH

N N

SH + 2 H2 O

CH OH

OH Colored P igment