Lipids

Components of Lipids
Fatty Acids
Fats, and Oils
Chemical Properties of Triglycerides

Compiled by: Dwika Riandari, M.Si

Teaching material for Proximate Analysis

1
 Introduction

• Definition: water insoluble compounds

• Most lipids are fatty acids or ester of fatty acid
• They are soluble in non-polar solvents such as
petroleum ether, benzene, chloroform
• Functions
• Energy storage
• Structure of cell membranes
• Thermal blanket and cushion
• Precursors of hormones (steroids and
prostaglandins)
2
 Components of Lipids
Waxes
Phospholipids
Fatty Acids
Fats and oils (trigycerides)
Fat soluble vitamins
Terpene
Sterol

3
 Waxes
Fatty acid + 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 O
C30 H61 O C C15 H31 C16 H33 O C C15 H 31

4
Spermaceti source

Carnauba wax source

Bee’s wax
5
“TRIACONTANOILPALMITAT”

LILIN PADA LEBAH MADU

(BEE)
 Phospholipid
Two hydroxyls of the glycerol residue are esterfied with fatty acids
The third hydroxyl is bound to phosphoric acid which in turn is
ester-linked with X-OH, usually an amine alcohol

Lecithin (phosphatidyl choline)

O
H2 C O C R
O
R C O CH
O CH 3
+
H2 C O P O CH2 CH 2 N CH 3
O_ CH 3
Phosphatidic Acid Choline

7
The phosphoric acid end of the molecule is strongly polar
hydrophylic
The fatty acid “tails” are non-polar
This dual structure (amphiphathic) makes the phospholipid valuable
surface active agents and emulsion stabilizers
Phospholipid are important cell wall constituent

8
 Sterols

Male & female sex hormones

Bile acids
Vitamin D
Adrenal corticosteroids
Cholesterol
21
22
H 318
C CH 3
H3C 20

12
CH 3
19 17 16
H3 C 11 13
14 15
10
1 9
2 8
3 6 7
4
HO
HO 5
9
STEROID NUMBERING SYSTEM

18
12 17
11
19 C 13 D 16
1
2 9 14 15
10
A B 8
3 5 7
4 6
 Fat Soluble Vitamins

Vitamin A

H3 C CH 3 CH3 CH3
5
8 CH2 OH
9 7 3 1
6 4 2

CH3

11
 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 CH2 CH2CH 2 CH(CH 3 )2
HO
R3
12
 Fatty Acids

• Long-chain carboxylic acids

• Insoluble in water
• Typically 12-18 carbon atoms (even number)
• Some contain double bonds

corn oil contains 86%

unsaturated fatty acids and
14% saturated fatty acids

13
Saturated and Unsaturated Fatty Acids

Saturated = C–C bonds

Unsaturated = one or more C=C bonds

COOH

Palmitic acid, a saturated acid

COOH

Palmitoleic acid, a unsaturated fatty acid

14
 Structures
Saturated fatty acids
• Fit closely in regular pattern

COOH
COOH
COOH
Unsaturated fatty acids
• Cis double bonds H H
C C

cis double bond COOH

15
 Properties of Saturated
Fatty Acids

• Contain only single C–C bonds

• Closely packed
• Strong attractions between chains
• High melting points
• Solids at room temperature

16
 Saturated Fatty Acids
 
Common Systematic Formula Common source
Name Name

Butyric Butanoic CH3(CH2)2COOH butterfat

Caproic Hexanoic CH3(CH2)4COOH butterfat, coconut, palm nut oils

Caprylic Octanoic CH3(CH2)6COOH Coconut, palm, nut oils, butterfat

Capric Decanoic CH3(CH2)8COOH Coconut, palm, nut oils, butterfat

Lauric Dodecanoic CH3(CH2)10COOH Coconut, palm, nut oils, butterfat

Myristic Tetradecanoic CH3(CH2)12COOH Coconut, palm, nut oil, animal fats

Palmitic Hexadecanoic CH3(CH2)14COOH practically all animal, plant fats

Stearic Octadecanoic CH3(CH2)16COOH animal fat, plant fats

Arachidic Eicosanoic CH3(CH2)18COOH peanut oil

17
 Properties of Unsaturated
Fatty Acids

• Contain one or more double C=C bonds

• Nonlinear chains do not allow molecules to pack
closely
• Few interactions between chains
• Low melting points
• Liquids at room temperature

18
 Unsaturated 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 19
Characteristics of Fatty Acids

Fatty Acids M.P.(C) mg/100 ml 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
* Solubility20
Effects of Double Bonds on the Melting Points
Fatty Acids Melting point (C)
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.

x x x
x

# Double bonds
21
 Important Fatty Acids for Nutrition

• EPA (Eicosapentaenoic acid)

20:5(n-3). Omega-3, all-cis –eicosa-5,8,11,14,17
pentaenoic acid

• DHA (Docosahexaenoic acid)

22:6 (n-3). omega-3, all-cis -docosa-4,7,10,13,16,19-
hexaenoic acid

22
Typical fish oil supplements

23
 Fats and Oils
Formed from glycerol and fatty acids

O
CH 2 OH HO C (CH 2)14CH3
O
CH OH + HO C (CH 2)14CH3
O
CH2 OH HO C (CH 2)14CH 3
glycerol palmitic acid (a fatty acid)
24
 Triglycerides (triacylglycerols)
Esters of glycerol and fatty acids

ester bonds
O
CH 2 O C (CH 2)14CH 3 + H 2O
O
CH O C (CH 2)14 CH 3 + H 2O
O
CH 2 O C (CH 2)14CH 3 + H 2O

25
 Properties of fats and oils
• fats are solids or semi solids
• oils are liquids
• melting points and boiling points are not usually
sharp (most fats/oils are mixtures)
• when shaken with water, oils tend to emulsify
• pure fats and oils are colorless and odorless (color
and odor is always a result of contaminants) – i.e.
butter (bacteria give flavor, carotene gives color)

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 Examples of oils
• Olive oil – from Oleo europa (olive tree)
• Corn oil – from Zea mays
• Peanut oil – from Arachis hypogaea
• Cottonseed oil – from Gossypium
• Sesame oil – from Sesamum indicum
• Linseed oil – from Linum usitatissimum
• Sunflower seed oil – from Helianthus annuus
• Rapeseed oil – from Brassica rapa
• Coconut oil – from Cocos nucifera

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 Non-drying, semi-drying and drying
oils
• based on the ease of autoxidation and
polymerization of oils (important in paints and
varnishes)
• the more unsaturation in the oil, the more likely
the “drying” process
– Non-drying oils:
• Castor, olive, peanut, rapeseed oils
– Semi-drying oils
• Corn, sesame, cottonseed oils
– Drying oils
• Soybean, sunflower, hemp, linseed, tung, oiticica oils
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 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 29
 Properties of Triglycerides

Hydrogenation
• Unsaturated compounds react with H2
• Ni or Pt catalyst
• C=C bonds C–C bonds

Hydrolysis
• Split by water and acid or enzyme catalyst
• Produce glycerol and 3 fatty acids

30
 Hydrogenation

O
CH2 O C (CH2)5CH CH(CH2)7CH3
O
Ni
CH O C (CH2)5CH CH(CH2)7CH3 + 3 H2
O
CH2 O C (CH2)5CH CH(CH2)7CH3

31
 Product of Hydrogenation

O
CH 2 O C (CH 2)14CH3
O
CH O C (CH 2)14CH3
O
Hydrogenation converts double bonds in oils to single bonds. The solid
CH
products are used to make O C and(CH
2 margarine other)hydrogenated
2 14 CH 3
items.

32
 Hydrolysis

Triglycerides split into glycerol and three fatty

acids (H+ or enzyme catalyst)
O
CH2 O C (CH2)14CH3
O +
H
CH O C (CH2)14CH3 +3 H2O
O
CH2 O C (CH2)14CH3
CH2 OH O
CH OH + 3 HO C (CH2)14CH3

CH2 OH
33
 Saponification and Soap
• Hydrolysis with a strong base
• Triglycerides split into glycerol and the salts of fatty
acids
• The salts of fatty acids are “soaps”
• KOH gives softer soaps

34
 Soaps
• Process of formation is known as saponification
– Types of soaps:
• Sodium soap – ordinary hard soap
• Potassium soap – soft soap (shaving soaps are potassium
soaps of coconut and palm oils)
• Castile soap – sodium soap of olive oil
• Green soap – mixture of sodium and potassium linseed oil
• Transparent soap – contains sucrose
• Floating soap – contains air
• Calcium and magnesium soaps are very poorly water soluble
(hard water contains calcium and magnesium salts –these
insolubilize soaps)

35
 Saponification

O
CH2 O C (CH2)16CH3
O
CH O C (CH2)16CH 3 + 3 NaOH
O
CH2 O C (CH2)16CH3
CH2 OH O
+-
CH OH + 3 Na O C (CH2)16CH3
salts of fatty acids (soaps)
CH2 OH

36
 Learning Check L1

How would the melting point of stearic

acid compare to the melting points of oleic
acid and linoleic acid? Assign the melting
points of –17°C, 13°C, and 69°C to the
correct fatty acid. Explain.
stearic acid (18 C)
oleic acid (18 C)
linoleic acid (18 C)

37
 Solution L1

Stearic acid is saturated and would have a

higher melting point than the unsaturated fatty
acids. Because linoleic has two double bonds, it
would have a lower mp than oleic acid, which
has one double bond.
stearic acid mp 69°C
oleic acid mp 13°C

linoleic acid mp -17°C 38

 Learning Check L2
What are the fatty acids in the following
triglyceride?

O
CH2 O C (CH2)16CH3
O
CH O C (CH2)7CH CH(CH2)7CH3
O
CH2 O C (CH2)12CH3
39
 Solutions L2
What are the fatty acids in the following
triglyceride?
O
Stearic acid
CH2 O C (CH2)16CH3
O
CH O Oleic acid
C (CH2)7CH CH(CH2)7CH3
O
CH2 O C (CH2)12CH3 Myristic acid
40
 Learning Check L3
What are the products obtained from the complete
hydrogenation of glyceryl trioleate?

(1) Glycerol and 3 oleic acids

(2) Glyceryltristearate
(3) Glycerol and 3 stearic acids

41
 Solution L3
What are the products obtained from the complete
hydrogenation of glyceryl trioleate?

2. Glyceryltristearate

42
 Analytical Methods for The
Determination of Characteristics of
Fats and Oils

43
 Analytical Methods

1. Acid Value
2. Saponification Value
3. Iodine Value
4. Gas Chromatographic Analysis for Fatty Acids
5. Liquid Chromatography
6. Cholesterol Determination
44
 Acid Value
Number of mgs of KOH required to neutralize the Free
Fatty Acids in 1 g of fat.

RCOOH + KOH RCOO K + H2O

- +

ml of KOH x N x 56
AV = = mg of KOH
Weight of Sample

45
 Acid Value
The free fatty acid content in a good soybean oil should be
less than or equal to 0.05 %. The average molecular
weight of free fatty acids of the oil is 280 which is the
molecular weight of linoleic acid.
What is the maximum acid value of the good soybean oil?

What is the content (%) of free fatty acids of a soybean

oil if the acid value is 0.3? The average molecular weight
of free fatty acids of the oil is 280.
46
• The free fatty acid content in soybean oil is
0.05%.
• What is the content of free fatty acid in 1g oil in
mg?
• 1g = 1000mg
• What is the 1% of 1000mg in mg?
• What is the 0.1% of 1000 mg soybean oil in mg?
• What are the 0.5% of 1000mg soybean oil in mg
and ug?
47
 Acid Value

0.05 % in 1 gram is 0.5 mg RCOOH in 1 gram of oil

56 mg of KOH reacts with 280 mg of RCOOH
56 mg of KOH / 280 mg of RCOOH = 1 : 5
The 0.5 mg RCOOH reacts with 0.1 mg KOH
Acid Value is 0.1 mg KOH/g oil

48
• What is the content (%) of free fatty acids of
soybean oil if the acid value is 0.3?
• Acid value 0.3 means that 0.3 mg KOH is required
to react with the free fatty acid in1g (1000mg) of
oil
• The 56mg KOH reacts with 280 mg free fatty acid,
56 : 280 =1 : 5
• 1mg KOH reacts with 5 mg free fatty acid
• The 0.3 mg KOH reacts with 1.5 mg of free fatty
acid in 1 gram oil
• 1.5 mg free fatty acid /1000 mg oil x100(%) =0.15
%

49
 Saponification Value

Saponification - Hydrolysis of ester (triglyceride) under

alkaline condition.

O
H2 C OH
H2 C O C R O
O Heat + 3R - +
+ 3 KOH HC OH C OK
HC O C R
O
H2 C OH
H2 C O C R

Definition : mgs of KOH required to saponify 1 g of fat

50
 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.
5. Conduct blank determination.

Saponification number =[ (B – S) x N of HCl x 56] /gram of sample

B - ml of HCl required to titrate KOH in Blank.

S - ml of HCl required to titrate excess KOH by Sample.

51
 Saponification Value

O
CH2 O C (CH2)6 CH3
O
A Tricaprylin (MW= 450)
CH O C (CH2)6 CH3
O
CH2 O C (CH2)6 CH3

O
CH2 O C (CH2)16 CH3
B Tristearin (MW= 890) O
CH O C (CH2)16 CH3
O
1Gram of Oils A and B CH2 O C (CH2)16 CH3
52
 Saponification Value
1 mol TG 3 mol KOH required

MWKOH = 56 g/mol, therefore weight of 1 mol KOH = 56000 mg

1 g TG => 1 g / MWTG (g/mol) mol

1 mol TG 3x 56000 mg KOH required

1 g TG / MWTG X mg KOH required

168000
X  SN 
MWTG 53
 Saponification Value
168000
X  SV 
MWTG

• What is the MWTG ?

O O O

H2C O C-R H2 C O C - R1 H2 C O C - R1
O O O

HC O C- R HC O C - R1 HC O C - R2
O O O
H2 C H2 C O C - R3
H2C O C-R O C - R2
54
 Saponification Value
168000
X  SV 
MWTG
• Which one’s MW
should be taken?
O O
O
H2 C O C - R1 H2 C O C - R1
H2C O C-R O O
O
HC O C - R1 HC O C - R2
HC O C- R O O
O
H2 C H2 C O C - R3
H2C O C-R O C - R2
55
 Saponification Value
168000
X  SV 
AMWTG
• The Answer is the Weighted Average MW

O O O

H2C O C-R H2 C O C - R1 H2 C O C - R1
O O O

HC O C- R HC O C - R1 HC O C - R2
O O O
H2 C H2 C O C - R3
H2C O C-R O C - R2
56
Sample A has large molecular weight triglyceride (e.g. MW.890).
Sample B has small molecular weight triglyceride(e.g. MW.450).
In one gram of sample, number of triglyceride in B is about two times
more than number of triglyceride in A.

Less mg of KOH is needed to saponify sample A than sample B.

Therefore, saponification value of A is about half of that of sample B

Avogadro’s Number (N) = 6.02 x 1023 / mol

57
 Learning Check

A 5.00 grams of exotic tropical oil was saponified

with excess KOH. The unreacted KOH was then
titrated with 1.00 N HCl. The blank required 40 mL
of HCl and the sample required 20 mL.
•Please calculate the saponification value of the oil.
•Please calculate the MWTG
•How many base should be added to saponify 1000 g
of oil?
58
Saponification Values 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
 
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 Iodine Value

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.

60
 Iodine Value
O
CH2 O C (CH2 )7 CH CH (CH2 )7 CH3
Triolein (MW= 884) O
CH O C (CH2 )7 CH CH (CH2 )7 CH3
O
A CH2 O C (CH2 )7 CH CH (CH2 )7 CH3

O
CH2 O C (CH2)7 CH CH CH2 CH CH (CH2)4 CH3
Trilinolein (MW= 878) O
CH O C (CH2)7 CH CH CH2 CH CH (CH2)4 CH3
O
B CH2 O C (CH2)7 CH CH CH2 CH CH (CH2)4 CH3

61
 Iodine Value Determination

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

(ml of Na2S2O3 volume for blank - ml of Na2S2O3

volume for sample)  N of Na2S2O3  0.127g/meq  100
Iodine Value =
Weight of Sample (g) 62
 Determination of Double Bonds per
Molecule

Number of Double Iodine Value x Molecule Weight

=
Bonds per Molecule 2 x 127 x 100

63
 Double Bond Determination

The unknown compound has molecular weight

of 878 and iodine value of 173. Determine the
number of double bonds in the unknown
compound

64
 Learning Check
• Fatty acids A and B have only one double bond per
molecule. The molecular weights of A and B are 150
and 300, respectively. The hypothetical iodine value
of Compound A is 150. What is the Iodine value of
compound B?

• Triglycerides A and B have the very similar molecular

weights of about 878. The compound A has 6 double
bonds per molecule and has iodine value is 174. The
compound B has 3 double bond per molecule. What
is the iodine value of the compound B?

65
 THE END

PS: GOOD LUCK ON YOUR TEST!

66