FOOD CHEMISTRY

Food Chemistry

OTHER FOOD ANALYSIS METHODS

1 Food Chemistry
 OVERVIEW

The chemistry of methods for the total analysis of the following components are detailed:
Food Chemistry

Carbohydrates analysis

Fibre analysis

Lipid characterization

The material contained below is summary of various analysis methods. It is all examinable.

Further reading on these topics is found in Chapters 11, 12, and 14 of the textbook “Food Analysis” 2nd
Edition by S. Suzanne Nielsen (Editor). Material in this textbook is not examinable.

2 Food Chemistry
 LEARNING OBJECTIVES

AIMS
Food Chemistry

To give a strong understanding of how to analyse food components such as fibre and carbohydrates, and
how to characterize lipids in food.

OBJECTIVES
On completion of “Other Food Analysis Methods”, students will be able to demonstrate a sound
knowledge of:

(1) The methods used to analyse food carbohydrates, including the use of standard methods such as
AOAC and AOCS;

(2) The composition of dietary fibre and the methods used to determine the content of fibre in food,
including the use of standard methods such as AOAC;

(3) Lipid characterization procedures used in food analysis to determine lipid quality.

3 Food Chemistry
 CARBOHYDRATE ANALYSIS
LEARNING OBJECTIVES – CARBOHYDRATE
ANALYSIS
Food Chemistry

AIMS
To give an understanding of the chemistry involved in the analysis of carbohydrates in foods, including total
soluble solids, individual sugars, and starch.

OBJECTIVES
On completion of “Carbohydrate Analysis”, students will be able to demonstrate a sound knowledge of:

(1) The chemical method used to determine the total carbohydrate content of food;

(2) The chemical methods used to determine individual sugars in food;

(3) The chemical methods used to determine starch in food.

4 Food Chemistry
 CARBOHYDRATE ANALYSIS - SUMMARY
INTRODUCTION
Carbohydrates (including sugars and starch) are in all cases expressed as monosaccharides and the tables of
carbohydrate content contain values of available carbohydrate - proximate analysis content. The available
Food Chemistry

carbohydrate consists of the sum of the free sugars (glucose, fructose, sucrose, lactose, maltose and higher
maltose homologues), dextrin, starch and glycogen expressed as monosaccharides. These are the
carbohydrates that are digested and absorbed by man, and which are glucogenic in man.

METHOD
For quantitative work, the sugars can be extracted from the homogenized foodstuffs either by treatment with
80% v/v ethanol in a Soxhlet extraction apparatus for three hours or by boiling with 80% v/v ethanol for 20
minutes. The free sugars in the extracts are examined by two reducing sugar methods or by HPLC to give
the glucose and fructose concentration. (We look at these in detail later in this module). Sucrose in the
extract can be measured by inversion. The residue remaining after extraction of the free sugars is examined
for starch and dextrin content.

Methods of Quantitative Determination:

1. Reducing Sugars
Sugars with a structure containing free aldehyde and ketone groups react as weak reducing agents and are
called reducing sugars.

5 Food Chemistry
 Examples:
Reducing Sugars: all monosaccharides eg: glucose, fructose; the disaccharides maltose, lactose and
cellobiose.

These properties are used to estimate sugars by the measurement of the reduction of Cu(II) to Cu(I).
Fehling’s solution consists of alkaline cupric tartrate and is converted to insoluble cuprous oxide when
Food Chemistry

boiled with a solution of a reducing sugar.

This forms the basis of a number of procedures.

Note: Fehling’s 1: copper(II) sulfate solution
Fehling’s 2: alkaline sodium potassium tartrate

These are kept separate in the dark and mixed just prior to use, to give an alkaline cupric tartrate solution, ie.
Fehling’s solution.

2. Non-Reducing Sugars
Disaccharides such as sucrose and raffinose, and higher oligosaccharides.

These sugars are non-reducing because they consist of simple sugars linked through their aldehydic or
ketonic groups. There is no hydroxyl group on an anomeric carbon.

6 Food Chemistry
 FOOD ANALYSIS EXAMPLES
Food products containing a mixture of reducing sugars

The Cu2+ reducing power is referred to as ‘Dextrose Equivalent’ or ‘D.E.’ or ‘K value’.

Food Chemistry

eg: glucose syrups prepared by the hydrolysis of starch.

42% reducing sugars expressed as dextrose on the dry basis : DE 42

OTHER METHODS FOR THE QUANTITATIVE DETERMINATION OF

SUGARS IN FOOD:

These are based on the traditional polarimetry, refractometry, hydrometry and the more modern HPLC, GC,
ion exchange and enzymatic methods.

Polarimetric Methods - optical rotation

Refractometric Methods - refractive index tables

Hydrometric Methods - specific gravity tables

7 Food Chemistry
 Ion exchange chromatography: an anion-exchange column as part of an autoanalyser is now used to
separate and analyse mixtures of sugars.

HPLC and Gas Chromatography

HPLC is a quick, sensitive and powerful tool for both the qualitative and quantitative analysis of sugars in
food products.
Food Chemistry

Enzymic Methods

METHODS OF QUALITATIVE ANALYSIS

1. General Carbohydrate Test

Molisch Reaction:
All carbohydrates, when treated with strong acids, yield furfural derivatives which combined with naphthol.
Red-violet coloured products result.

2. Tests for Reducing Sugars

As mentioned earlier, carbohydrates containing a free aldehyde (-CHO) or ketone (-C=O) group act as
reducing agents. Most are based on the reduction of Cu2+ (cupric ions) to Cu+ (cuprous ions) in alkaline
solution to give a red precipitate of cuprous oxide, Cu2O. A number of tests have evolved using different
substances for preventing the formation of black cupric oxide when CuSO4 is heated with alkali.

8 Food Chemistry
 (a) Fehling’s Solution Test - (simplest qualitative test):
Solutions of cupric sulphate (Fehling’s 1) and alkaline sodium potassium tartrate (Fehling’s 2) are
mixed together and then with the reducing sugar solution, and finally heated. A positive reaction
shows the red precipitate of cuprous oxide or with more dilute solutions of reducing sugar, a green or
red, or a red-yellow colour.
Food Chemistry

(b) Benedict’s Test:

Same as above using Benedict’s reagent (CuSO4, sodium citrate and Na2CO3).

(c) Barfoed’s Test:

In alkaline solution, all reducing sugars precipitate Cu2O; however, in a slightly acidic solution only
monosaccharides do so. The cupric acetate reagent is used.

STARCH DETERMINATION
The residue remaining after the extraction of the food samples with ethanol (extraction of free sugars) is
further examined for starch and dextrin content. Both are hydrolysed with the selective enzyme
glucoamylase to glucose. After filtration of the digest, the liberated glucose is treated with glucose oxidase
and estimated colorimetrically at 37 °C in an autoanalyser.

9 Food Chemistry
 CARBOHYDRATE ANALYSIS – FURTHER READING

INTRODUCTION
Food Chemistry

This study examines chemical and physical methods for analysing various types of carbohydrates in foods.

Further reading on this topic is found in Chapters 11 Carbohydrate Analysis of the supplied textbook “Food
Analysis” 2nd Edition by S. Suzanne Nielsen (Editor). The material contained in the textbook will not be
examined; only that material listed above and below is examinable.

Chapter 11 is divided into a number of sections covering various aspects of the analysis of carbohydrates in
food; however, there are five main themes:

Sample Preparation
Chemical Determination of Total Carbohydrates
Chemical Analysis of Individual Sugars in Food
Chemical Analysis of Starch in Food
Physical Methods for Analysing Soluble Solids in Foods

10 Food Chemistry
 Section 11.1 on Page 169 of the textbook discusses the different types of carbohydrates, including a full list
in Table 11-1 on Page 170 of the textbook. Additionally, the analysis of carbohydrates is introduced, and
the terms total carbohydrate and other carbohydrate discussed. Total carbohydrate is normally determined
by difference with the mass of moisture, total fat, ash, and crude protein subtracted from the mass of the
food. The total carbohydrate content of some foods is summarized in Table 11-2 on Page 171 of the
textbook. Other (complex) carbohydrate, including starch is determined by subtracting the sum of the mass
Food Chemistry

of dietary fibre, sugars and sugar alcohols from the total carbohydrate.

SAMPLE PREPARATION
Section 11.2 on Pages 169-171 of the textbook discusses sample preparation required prior to carbohydrate
analysis, particularly drying or removal of fat from the food.

CHEMICAL DETERMINATION OF TOTAL CARBOHYDRATES IN FOOD

Section 11.3.2 on Pages 172-173 of the textbook details the phenol-sulphuric acid method for determining
the total carbohydrate content of food. The method involves adding an aqueous solution of phenol to an
aqueous solution of the carbohydrate, followed by concentrated sulphuric acid (heat generated). A yellow
colour results which is then measure spectrophotometrically at 490 nm. Standard sugar solution are then
used for quantification. Polysaccharides are broken down to sugars by the acid and heat.

11 Food Chemistry
 CHEMICAL ANALYSIS OF INDIVIDUAL SUGARS IN FOOD
Total Reducing Sugars

Section 11.3.3 on Pages 173-174 of the textbook details wet chemical methods for determining the reducing
sugar content of foods. Methods discussed are:
Food Chemistry

Somogyi-Nelson method (Section 11.3.3.1)

Munson-Walker method (Section 11.3.3.2)

Lane-Eynon method (Section 11.3.3.2)

Standard AOAC methods for each of these methods are listed. These methods are based on the reduction of
copper (II) ions in alkaline solution to copper (I) ions, which then precipitate as brick-red Cu2O. For the
Somogyi-Nelson method, the Cu+ ions reduce an arsenomolybdate complex to produce an intense blue
colour that is measured spectrophotometrically. For the Munson-Walker method, the Cu2O can be
determined:

(1) gravimetrically;
(2) by titration with sodium thiosulphate;
(3) by titration with potassium permanganate; and
(4) electrolytically, using AOAC methods.

12 Food Chemistry
 Munson - Walker Gravimetric Method
Fehling’s solution is boiled with a solution of a reducing sugar for six minutes and then filtered hot. The
precipitated cuprous oxide (Cu20) is filtered off and weighed. The sugar content is calculated from the mass
of cuprous oxide by referring to AOAC tables.
Food Chemistry

Lane-Eynon Method
This method involves the determination of reducing sugars by titration with Fehling’s solution using the
redox indicator, methylene blue. This is an empirical method and standardized experimental conditions and
methods must be rigidly adhered to for satisfactory results.

The classical method uses tables indicating the amounts of invert sugar (mixture of fructose and glucose),
dextrose (glucose), fructose, maltose or lactose equivalent to volumes of reduced Fehling’s solution.

The now accepted and official method (AOAC Method 923.09, 920.183b; 1995) is a constant volume
technique where the conditions for reduction are standardized by the addition of water to give a constant
final volume of 75 ml in the titration flask, and the volume of Fehling’s solution used is calibrated against a
standard reducing sugar solution. This standard solution must be specific for a particular reducing sugar.

The Lane and Eynon method involves the titration of a boiling solution of the Fehling’s solution with a
dilute solution of the sugar sample.

13 Food Chemistry
 Luff-Schoorl Method
This method uses an alkaline reagent containing cupric citrate (Cu2+ ion). After boiling this reagent with a
solution containing reducing sugars, potassium iodide (KI) and acid (sulphuric acid) are added after cooling.
Iodine is liberated from the redox reaction:

2I- + Cu2+ → I2 + Cu+

Food Chemistry

The liberated iodine is thus equivalent to the unreduced copper (Cu2+) ie:
1 mole I2 from 1 mole Cu2+

The liberated iodine (brown-black colour) is then titrated (to colourless) with the reducing agent, sodium
thiosulphate. Thiosulphate equivalent tables can then be used:
I2 → I-
coloured colourless

The Luff-Schoorl reagent is less alkaline than Fehling’s solution. Consequently, it is a weaker oxidizing
agent requiring longer boiling with sample solutions than the Lane and Eynon technique.

[Reducing agents reduce another species but are oxidized in the process. Oxidizing agents oxidize another
species but are reduced in the process.]

14 Food Chemistry
 Nonreducing Sugars

Sucrose Determination
Remember, sucrose is a non-reducing sugar. Sucrose can be determined in the absence of reducing sugars
by inverting a portion of the test solution with acid followed by neutralization with alkali and titration by the
Lane and Eynon method, using standard invert sugar solution for calibration.
Food Chemistry

% sucrose = % invert sugar x 0.95

In the presence of reducing sugars, titrations are carried out on portions of the test solution before and after
acid inversion.

Analysis of Specific Mono-, Di- and Oligosaccharides

Section 11.3.4 on Pages 174-178 of the textbook details HPLC, GC and enzymic methods for analysing
individual mono-, di- and oligosaccharides. For HPLC analysis, various stationary phases used to analyses
sugars are discussed in detail. For GC, sugars need to be converted to derivatives that are volatile and able
to be analysed by GC.

Extraction of Free Sugars

Free sugars must be removed from the food matrix prior to HPLC analysis so that possible interfering
components of food are separated from free sugars. This is done by extraction of the dried, lipid-free food
with 80% ethanol (AOAC method 922.02, 925.05; 1995). Purification of this extract prior to HPLC
analysis is often required. Methods to do this are outlined in Section 11.3.1 on Pages 171-172 of the
textbook. After purification of the sugar extract, the aqueous alcohol is removed under reduced pressure
using a rotary evaporator at temperatures of 45-50 °C. The dried sample is then dissolved in water and
analysed by HPLC.

15 Food Chemistry
 CHEMICAL ANALYSIS OF STARCH IN FOOD
Section 11.4.1.1 on Pages 179-180 of the textbook discusses the determination of total starch content. The
most reliable method involves the total conversion of the starch to D-glucose by starch specific enzymes
such as α-amylase and glucoamylase, and measurement of the D-glucose released by spectrophotometric or
HPLC methods. The method involving spectrophotometric analysis (AOAC Method 969.39; 1995) is
Food Chemistry

detailed on Page 180 of the textbook.

16 Food Chemistry
 FIBRE ANALYSIS

LEARNING OBJECTIVES – FIBRE ANALYSIS

Food Chemistry

AIMS
To give an understanding of food fibre, particularly dietary fibre, and the methods used to analyse food fibre.

OBJECTIVES
On completion of “Fibre Analysis”, students will be able to demonstrate a sound knowledge of:

(1) The composition and importance of dietary fibre in food;

(2) Gravimetric methods for determining the fibre content of foods;

(3) Chemical methods for determining the fibre content of foods;

(4) The differences between the methods used to determine dietary fibre.

17 Food Chemistry
 CONTENT – FIBRE ANALYSIS

INTRODUCTION
Food Chemistry

This study examines food fibre and methods used to analyse the different types of fibre, including crude
fibre and dietary fibre. The special nature of food fibre analysis has resulted in it being considered
separately to the carbohydrate analysis detailed later in this module.

Further reading on this topic is found in Chapters 12 Fibre Analysis of the supplied textbook “Food
Analysis” 2nd Edition by S. Suzanne Nielsen (Editor). The material contained in the textbook will not be
examined; only the material summarised below is examinable.

Chapter 12 is divided into a number of sections covering various aspects of the determination of fibre in
food; however, there are 5 main themes:

Importance of Dietary Fibre

Composition of Dietary Fibre
Gravimetric Methods of Fibre Analysis
Chemical Methods of Fibre Analysis
Comparison Dietary Fibre Analysis Methods

18 Food Chemistry
 IMPORTANCE OF DIETARY FIBRE
Section 12.1.1 on Page 191 of the textbook reviews the role dietary fibre plays in human health, and is a
good introduction to dietary fibre.
Food Chemistry

COMPOSITION OF DIETARY FIBRE

Section 12.1.2 on Page 191 of the textbook outlines the two definitions of dietary fibre depending on
whether resistant starches are included. Dietary fibre is composed of cell wall polysaccharides, non-cell-
wall polysaccharides and lignin. Section 12.1.3 on Pages 191-192 of the textbook discusses the composition
of these food components, and should be thoroughly studied.

GRAVIMETRIC METHODS OF FIBRE ANALYSIS

Three gravimetric fibre analysis methods are discussed in Section 12.3.3 on Pages 193-194 of the textbook.
Crude fibre analysis, and acid and neutral detergent fibre analysis methods are discussed and their
limitations highlighted. The most important section to review is the discussion of the AOAC method for
determining the total, insoluble and soluble dietary fibre in food (AOAC Method 991.43; 1995). This is the
standard method for determining the dietary fibre content of food. The experimental procedure is well
outlined making it easier to implement in the laboratory.

19 Food Chemistry
 CHEMICAL METHODS OF FIBRE ANALYSIS
Section 12.3.4 on Pages 195-196 of the textbook outlines the alternative analysis procedure to the
gravimetric methods. Chemical methods for fibre analysis involve determination of all nonstarch
monosaccharides and lignin. Monosaccharides are determined colorimetrically or using GC or HPLC. The
Englyst-Cummings procedure is an alternative to the gravimetric AOAC method for dietary fibre highlighted
Food Chemistry

above, except lignin is not included in the analysis. The levels of lignin in food is often small and thus the
Englyst-Cummings procedure can be used to determine fibre content of such foods. Additionally, resistant
starch is not included in this fibre measurement, but a separate analysis is included to determine resistant
starch alone.

COMPARISON DIETARY FIBRE ANALYSIS METHODS

The merits of the AOAC method and the Englyst-Cummings procedure for dietary fibre determination are
discussed in Section 12.4 on Page 197 of the textbook. The points discussed are useful for making a
judgement about which methods to use particularly since the Englyst-Cummings procedure requires the least
amount of time, technical skill and specialized equipment relative to the AOAC and other methods.
However, the Englyst-Cummings procedure requires access to GC and HPLC equipment, and knowledge of
such chromatographic procedures.

20 Food Chemistry
 LIPID CHARACTERIZATION
LEARNING OBJECTIVES - LIPID
CHARACTERIZATION
Food Chemistry

AIMS
To give an understanding of the physical and chemical methods used to characterize food lipids.

OBJECTIVES
On completion of “Lipid Characterization”, students will be able to demonstrate a sound knowledge of:
(1) The standard methods used to characterize fats and oils;

(2) The methods used to characterize the quality of bulk fats and oils and extracted lipids;

(3) Selected methods used for measuring lipid oxidation in food;

(4) The methods used to analyse lipid fractions from food, focussing on fatty acid composition of
triglycerides, including the use of fatty acid methyl esters (FAME), and cholesterol composition.

21 Food Chemistry
 CONTENT - LIPID CHARACTERIZATION

INTRODUCTION
Food Chemistry

This study examines the different analyses and tests used to characterize bulk fats and oils, lipid oxidation,
and the composition of lipids in foods.

Further reading on this topic is found in Chapters 14 Fat Characterization of the supplied textbook “Food
Analysis” 2nd Edition by S. Suzanne Nielsen (Editor). The material contained in the textbook will not be
examined; only the material summarise below is examinable.

Chapter 14 is divided into a number of sections covering various aspects of the characterization of fat in
food; however, there are 4 main themes:

Introduction to Methods of Analysis

Characterization Methods for Bulk Fats and Oils
Methods for Analysing Lipid Oxidation
Methods for Analysing Lipid Fractions from Foods

Not all the material in Chapter 14 will be studied in this module. The relevant sections will be highlighted
below.

22 Food Chemistry
 INTRODUCTION TO METHODS OF ANALYSIS
Sections 14.1 and 14.2 on Pages 219-221 of the textbook summarizes the classifications for lipids, why
analyses of lipids are important, the standard methods of analyses for lipid characterization, and the lipid
Food Chemistry

content of foods.

Table 14-1 on Page 220 of the textbook summarizes the standard methods to be discussed throughout
Chapter 14 of the textbook as part of Module 7. The three organization with standard methods are:

American Oil Chemist’s Society (AOCS);

AOAC International (AOAC);

International Union of Pure and Applied Chemists (IUPAC).

All the fat characterization analysis methods are controlled by these organizations through their standard
methods of analysis.

23 Food Chemistry
 CHARACTERIZATION METHODS FOR BULK FATS AND OILS
Before any analyses can be performed on bulk fats and oils, some form of sampling and sample preparation
is required. Sampling procedures such as AOCS Method C 1-47 (AOCS 1996) are available. If drying of
the sample is required then the AOAC Method 981.11 is used (AOAC 1995). Any sample storage will need
to exclude, heat, light and air to prevent lipid oxidation.
Food Chemistry

A number of chemical and physical methods used to commonly characterize food lipids are summarized in
the textbook.

Refractive Index
Refractive index is used to control hydrogenation and as a purity measurement, and is summarized in
Section 14.3.2 on Page 222 of the textbook.

Melting Point
Many types of melting points (depending on the analysis method used) are possible, each with its own
application, often depending on the preference in different countries. The disadvantages and methods of
analysis are outlined in Section 14.3.3 on Page 222 of the textbook.

Smoke, Flash and Fire Points

Section 14.3.4 on Page 222 of the textbook details definitions of each, and how they are measured in the
laboratory. These test give an indication of the volatile organic material in fats and oils.

24 Food Chemistry
 Cold Test
This test measure amount of crystallization at 0 °C, and is a measure of the resistance of the oil to
crystallization and, thus is a measure of the winterizing process. Further details are summarized in Section
14.3.5 on Page 222 of the textbook.

Cloud Point
Food Chemistry

The temperature at which a cloud forms due to crystallization in a liquid fat is the cloud point. Section
14.3.6 on Pages 222-223 of the textbook details the procedure.

Colour
The Lovibond (most commonly used method) and spectrophotometric methods for colour measurement are
detailed in Section 14.3.7 on Page 223 of the textbook, and are used by the food industry as quality
measurements for fats and oils.

Solid Fat Index and Solid Fat Content

Two methods for determining the amount of solids in a fat are outlined in Section 14.3.11 on Page 225 of the
textbook. The proportion of solids to liquids in a fat impact on the functional properties of a food such as
mouthfeel, and so is an important measure for a fat.

Consistency
Consistency is a measure of plasticity, hardness, creaminess and spreadability, and is determined using the
penetrometer method. Section 14.3.12 on Page 225 of the textbook summarizes this quality measure.

25 Food Chemistry
 Polar Components in Frying Fats and Oils
The deterioration of frying fats and oils is monitored by determining the polar components. Polar lipids are
separated from the nonpolar compounds using column chromatography, while the composition and quality
of the polar lipids are determined using thin later chromatography. Section 14.3.13 on Page 226 of the
textbook outlines the importance of polar compounds in frying fats and oils.

Iodine Value/Number
Food Chemistry

The iodine value or number measures the degree of unsaturation (number of carbon-carbon double bonds) in
a fat or oil. The procedure involves reacting iodine with carbon-carbon double bonds with a mechanism
similar to that for the hydrogenation of oils. The procedure is detailed in Section 14.3.8 on Page 223 of the
textbook, together with applications. It is particularly important for characterizing oils and for monitoring
the hydrogenation process during the refining of oils. See summary of QC tests later in Unit 3.

Saponification Value/Number
Saponification is the hydrolysis of neutral fats or oils to glycerol and fatty acids. The procedure to determine
the saponification value, which is a measure of the amount of saponification, is detailed in Section 14.3.9 on
Page 224 of the textbook. See summary of QC tests in Unit 3.

Free Fatty Acids and Acid Value

Fat acidity is due to free fatty acids and is measured using an acid base titration. Free fatty acids arise from
lipolysis of triglycerides. Section 14.3.10 on Pages 224-225 of the textbook outlines the procedure for
consideration. This method is also used to analyse food lipids extracts as a means of determining the level
of lipolysis and hydrolytic rancidity of a foods with a high oil content. Free fatty acids are monitored in any
process that uses bulk oils for cooking and frying such as in the manufacture of potato chip and crisps. See
summary of QC tests later in Unit 3.

26 Food Chemistry
 METHODS FOR ANALYSING LIPID OXIDATION
Lipolysis, hydrolytic rancidity, lipid oxidation and oxidative rancidity are briefly discussed. A review of
lipid oxidation in Unit 5 is needed prior to studying the analysis methods. A number of measures of lipid
oxidation are detailed in Section 14.4 of the textbook.

Peroxide Value
Food Chemistry

This is one of the most common tests for lipid oxidation and is detailed in Section 14.4.2 on Page 227 of the
textbook. Determining the peroxide value involves a titration of the peroxide or hydroperoxide groups in
compounds present in the lipid component of food, which are the initial products of lipid oxidation.
Additionally, this method is used to analyses food lipid extracts as a means for determining the oxidative
stability of foods that have a high oil content, such as nuts.

Hexanal Determination
Measurement of headspace hexanal using GC is now being used as a measure of lipid oxidation. Section
14.4.4 on Pages 227-228 of the textbook summarizes the procedure. It is being used in meat analysis.
Hexanal together with pentanal may be correlated with sensory analysis of lipid oxidation.

Thiobarbituric Acid (TBA) Test

This test measures malonaldehyde, the secondary product of lipid oxidation and has been shown to correlate
better with the sensory analysis of lipid oxidation than does the peroxide value. TBA reacts with
malonaldehyde to produce coloured compounds that are then measured spectrophotometrically. This
reaction is not specific to malonaldehyde, with other TBA reactive substances (TBARS) existing. The
procedure is discussed in Section 14.4.5 on Page 228 of the textbook. This test is commonly used to
measure lipid oxidation in meat products.

27 Food Chemistry
 METHODS FOR ANALYSING LIPID FRACTIONS FROM FOODS
Fatty Acid Composition and Fatty Acid Methyl Esters
The fatty acid profile is now a routine indicator used to represent the composition of lipids in foods. The
lipid is normally extracted from the food and saponified with alkali. The fatty acids released by this base
hydrolysis are not volatile enough to be analysed by GC, so derivatization to their fatty acid methyl esters
Food Chemistry

(FAME) is done prior to GC analysis. There are various methods for methylating the released fatty acids,
with the standard method involving methanol and boron trifluoride summarized in Section 14.6.1 on Pages
230-231 of the textbook, along with applications of the use of the method in food analysis.

Cholesterol Analysis
Section 14.6.4 on Pages 232-233 of the textbook summarizes the analysis of cholesterol in foods. The
AOAC Method 976.26 (AOAC 1995) is outlined and is similar to other methods used. Common to these
procedures is an extraction of the lipid from the food, followed by saponification of the lipid. The
nonsaponifiable fraction is then derivatized, and the resultant trimethylsilyl (TMS) ethers are quantified
using capillary GC.

TLC Separation of Lipid Fractions

Thin layer chromatography is used to rapidly analyse the various lipid fractions in a food lipid extract. The
procedure is outlined in Section 14.6.5 on Page 233 of the textbook.

Food Chemistry Lecture Notes

28 Food Chemistry