Measurement of Lipids

Contents
1. Introduction (definition ,characteristics)
2. Function of lipids in foods
3. Determination of fats in food
4. Quality evaluation of edible oils
 Introduction
Lipids
1) The term lipid is used to denote fats and fatlike
substances and is synonymous with the terms lipoids
or lipins.
2) It includes a great varieties. The main component is
triglycerides(>95%), others include phospholipids,
wax, fatty acids, cholesterin etc.
3) Lipids are usually defined as food components that
are insoluble in water but soluble in organic solvents.
4) Essential structural component of living cells (along
with proteins and carbohydrates).
5) exist in the free form(such as fats or oil),or associated
( combined with carbohydrates,or protein in food).
Characteristics of lipids

1. Insoluble in water, but soluble in organic solvents ,

such as ethyl oxide(乙醚),petroleum ether(石油醚),
chloroform(氯仿),acetone (丙酮).
2. Most of lipids contain ester bond, and main
section is fatty acid ester.
3. Be produced by organism, and can be used by
organism(different from mineral oil).
4. Except the phospholipids(磷脂), sphingomyelin(鞘
磷脂), etc.
 Function of lipids in foods
1.Provide caloric.
2. Provide essential fatty acids(linolenic acid亚麻酸,
linoleic acid亚油酸)
3.Carrier of fat-soluble vitamin(VE,VA,VD)
4. Provide glossy appearance, smooth taste.
5. Provide savory in the fry food.

Lipids content is one of indexes of evaluating food

quality , and food nutrition, food processing and
storage.
 Determination of fats in food
 Soxhlet extractor method

 Extraction of crude fats ethyl ether in acid(酸性乙醚提取法）

 Extraction of crude fats by ethyl ether in

Alkali (碱性乙醚提取法）（罗兹－哥特里法）

 Babcock Method and Gerber method(巴布科氏法和盖勃氏法)

 Extraction of fats by Chloroform-Methanol (氯仿－甲醇抽提法）

 索氏提取法

Principle

样品 乙醚或石油醚 蒸去溶剂 粗脂肪

Procedure:
1)Preparation of sample:
Solid sample –grinding
Semi-solid sample or liquid sample– adding sand–
dried—grinding
2) Preparation of filter paper tube
3) Put sample ready into the filter paper tube.
4) Extract the fats with solvents
5) Weighting
6) Calculation
Attention
1) Crude fats include : phospholipids, pigments,wax,
volatile oil, etc.
2) Sample should contain greater amount of free
fats, little associated fats.
3) Samples should be dried(decrease the efficiency,
increase nonfat substance)
4) If more dextrin or saccharide is included in
sample, it should be remove by water
5) Ethyl ether should be purified: no water, no
peroxide, no alcohol.
6) It should be heated in water bath, be careful of
firing.
 Determination of fats in food

Soxhlet extractor method

Principle

Ethyl ether or
sample Distill Crude fat
Petroleum ether
Extraction of crude fats ethyl ether in acid(酸性乙
醚提取法）
Principle:
 sample +HCl +H2O Hydrolysis by heating released
associated fats to exist in the free form.
 Hydrolyzed sample +ethanol +ethyl ether +petroleum
ether Extract Crude fats
 Recycle solvents, and weight
Procedure
 Sample preparation
 Hydrolysis
heating at 70-80℃ in a water bath for 40-50min
 Extract
ethanol , ethyl ether, petroleum ether,
 Weighting
 Calculating
Attention:
 sample without high saccharides and phospholipids
 Sample include: liquid sample, semi-solid sample,
sample difficult to be dried, and sample that can not to
be analyzed by soxhlet extractor.
 It can determine the associated fats in foods.
 ethanol can make protein precipitated, retard
emulsion, promote fats together, and solute
carbonhydrates.
 Addition of petroleum ether can decrease the solubiltiy
of ethanol in ethyl ether, and favorable for phase
separation.
Extraction of crude fats by ethyl ether in alkali(碱
性乙醚提取法）（罗兹－哥特里法）

Principle
Sample + NH3.H2O+ ethanol Hydrolysis Released associated fats in foods

Hydrolyzed sample +ethyl ether +petroleum ether extract fats

Recycle solvents, dried, and weighting

Procedure
 Adding NH3.H2O into sample, mixed, and heated for
5min in a 60℃ water bath.

 Adding ethanol to reaction system, mixed and cooled.

 Extraction with ethyl ether and petroleum ether.

 Recycle solvents ,dried at 100-105℃， and weighting,.

Attention
 The method is used in the determination of fats in the milk
product

 NH3.H2O can destroy the colloidal medium(胶体介质） and the

membrane of fat globule（脂肪球）.

 ethanol can make protein precipitated, and retard emulsion,

and solute the alcohol –soluble substances.

 Addition of petroleum ether can decrease the solubiltiy of

ethanol in ethyl ether, and favorable for phase separation
 Babcock Method and Gerber
method(巴布科氏法和盖勃氏法)
Principle:

Milk product +concentrated H2SO4 solute nonfat

components in milk, destroy the member of fat globule
and emulsion, release fats Heat and centrifugate
adipose layer was seperated record the
volume of fats
Attention
 Control the concentration of H2SO4, if it is too high ,
the milk will be carbonized, if it is too low, the fat
globule can not be destroyed completely.

 Sample without high saccharides and chocolate.

 Extraction of fats by Chloroform-Methanol
(氯仿－甲醇抽提法）
Principle

Chloroform-Methanol associated fats filter

Sample
Free fats

Recycle solvents,adding petroleum ether extract

Distill the petroleum ether, weighting

Attention
Suitable for sample with high associated
fats, especially phospholipids. Such as
fish,eggs, meat etc.
 for dried sample, some water should be
added.
 Fractionation of extracted lipids
TLC : one of the main analytical tools in lipid
research and analysis.
Column Chromatography:if silicic acid column was
selected,Neutral lipids, such as hydrocarbons,
glycerides, sterols, and sterol esters, are eluted with
chloroform, the remaining polar lipids are eluted
with methanol.
GC: fatty acid compositions in the oils and fats
HPLC: different components in the mixed lipids
Quality evaluation of oil and fats
1. Transparency(透明度)
2. Color(色泽)
3. Odour(气味)
4. Taste(滋味)
5. Relative density(相对密度)
6. Refractive Index(折射率)
7. Smoke point(烟点)
8. Melting point(熔点)
9. Freezing point(凝固点)
10.Determination of impurity(杂质测定)
11.Determination of acid value(酸价测定)
12. Determination of phospholipids(磷脂测定)
13.Determination of soap (含皂量测定)
14.Determination of saponification value(皂化价的定)
15.Determination of unsaponifiable matter(不皂化物测定)
16. Determination of iodine value(碘价测定)
17.Determination of peroxide value(过氧化值测定)
18.Determination of carbonyl value (羰基价测定)
 Transparency(透明度)

1) Observe the transparency of oil sample under

incandescent light when sample was kept at a
definite temperature for a definite time.
2) determination:
Oil sample in 100ml colorimetric cylinder—kept
20℃ for 24h—observe under incandescent
light—transparent or not
3) if the oil sample is cool, it must be heated to
50℃, and then cooled to 20℃ to observe.
 Color (色泽)

1) Color is a important indicator of edible oil.

2) Determination
Chromometer
 Odour(气味) and Taste(滋味)

1) Different oil have their special odour and taste.

2) Determination:
oil sample in beaker—heat to 50℃--stirring –
smell and degust
 Relative density(相对密度) and
Refractive Index(折射率)

1) Relative density and refractive Index is

physical characteristics of oil.

2) Determination
it can analyzed by densitometer and Abbe
refractometer.
 Smoke point(烟点)
Definition
The temperature when the smoke produce firstly.
Smoke point is a indicator of oil stability in the
air.

• Determination
apparatus for determining smoke point
 Melting point(熔点)

Definition
Temperature when solid fats were transformed to
liquid.

2) Fats sample in a water bath with heat

indicator—heat (0.5℃/min)– transparent liquid
–record the temperature
 Freezing point(凝固点)

 Oil freezing process

 Temperature when liquid oil was transformed
to solid. It is kept at some point for a little wile.
 Determination
Oil sample- saponification-- fatty acids-- dry at
130℃--freezing point determination
 Determination of impurity(杂质测定)

Definition
Residuals that are insoluble in organic solvents, such
as petroleum ether. They are earth, sandstone, etc.

Determination
Dissolve samples in the petroleum ether—filter –
separation of impurity and oil—dry impurity –weighting
 Determination of acid value(酸价测定)
Definition
Amount (mg)of KOH that needed to consume the the
quantity of free fatty acids in 1g of oil.

Determination
sample dissolved in ethanol and ethyl ether—titrate
the free fatty acids with alkali – calculate the alkali
consumed.
 Determination of phospholipids(磷脂测定)

Structure
Characteristic
Effect on oil quality

Determination
• Charateristic
 Amphiphatic molecule
 Polarity

• Effect on oil quality

 Increase the water content because of its hydrophilic nature,
and promote hydrolysis of oil.
 Easily be oxidized.
 Foam was produced when being heated.
 Determination
precipitation
Principle

Phopholipids +H2O centrifugation Washing precipitation with Acetone

Collecting precipitation

Dry and weighting

 Determination of soap (含皂量测定)
Definition
Soap remained in the oil after oil refined. It was produced
by water washing incompletely.
Determination
1) Principles
titrate soap in the oil with strong acid.
2) Procedure
Sample—ethanol + petroleum ether—adding hot
water(80℃),indicator—adding acid drop by drop—
record the amount of acid consumed--calculation
 Determination of saponification value
(皂化价的定)
Definition
Amount of KOH(mg) that needed in the saponifying
1g of oil.

Determination
Oil + KOH—glycerol + soap,
use HCl to titrate the excessive KOH
calculate the consumed KOH
 Determination of unsaponifiable
matter(不皂化物测定)
Definition
 Substances that can not be saponified.
 They includes pigments,vitamins, wax, sterol, etc.
 they are not dissolved in water, but soluble in ether.
Determination

Oil +KOH--saponifying– extract unsaponifiable matter

with ethyl ether—dry -weighting
 Determination of iodine value(碘价测定)

Definition
Amount of iodine (g) that 100g of oil can absorbed.

Determination
 CH3…CH2=CH2-COOH + ICl - …CHI-CHCl…
 Excessive KI was added to act with residual ICl,
KI + ICl =KCl +I2
 I2 was titrated by Na2S2O3
I2 + 2Na2S2O3—Na2S4O6 +2NaI
 Determination of peroxide value
(过氧化值测定)
Stale oil
 Hydrolysis( water, enzyme)—fatty acids
 Oxidization (O2)—acid,aldehyde,ketone, etc.

Determination of peroxide value(mmol/kg)

 Peroxide can oxidize KI to produce free I2
and then I2 was titrated by Na2S2O3,
 It was expressed as mmol O2/kg oil.
 Determination of carbonyl value
(羰基价测定)
Definition
 all amount of Aldehyde and ketone;
 Mmol /kg oil

Determination
Carbonyl substances +DNP-hydrozine(苯肼） –
hydrazone(腙） + OH- quinone(醌 ) --440nm
red color
谢
谢