328 FOOD AND NUTRITIONAL ANALYSIS / Oils and Fats

Inductively Coupled Plasma. Carbohydrates: Sugars

Spectrophotometric Methods; Starch. Food and Nutritional Analysis: Overview; Antioxidants and Preservatives; Oils
and Fats. Ion-Selective Electrodes: Food Applications.
Liquid Chromatography: Amino Acids. Proteins: Foods.
Quality Assurance: Quality Control. Sampling: Theory.
Vitamins: Fat-Soluble; Water-Soluble. Water Determination.

Further Reading
Association of Ofcial Analytical Chemists (AOAC)
(1990) Ofcial Methods of Analysis, 15th edn. Arlington, VA: AOAC.
Codex Alimentarius Commission (1981) Codex Standards
for Processed Fruits and Vegetables and Edible Fungi.

Rome: Food and Agriculture Organization/World Health

Organization.
Eskin NAM (1989) Quality and Preservation of Vegetables. Boca Raton, FL: CRC Press.
Kirk RS and Sawyer R (1991) Pearsons Composition and
Analysis of Foods, 9th edn. London: Longman Scientific
and Technical.
Paquot C and Hautfenne A (1987) Standard Methods for
the Analysis of Oils, Fats and Derivatives, 7th edn. Oxford: International Union of Pure and Applied Chemistry
(IUPAC)/Blackwell Scientific Publications.
Pomeranz Y and Meloan CE (1987) Food Analysis, Theory
and Practice, 2nd edn. New York: AVI.
Robinson
DS
(1987)
Food
Biochemistry
and
Nutritional Value. London: Longman Scientific and
Technical.

Oils and Fats

E W Hammond, Greenisle-Consulting, Northants, UK
& 2005, Elsevier Ltd. All Rights Reserved.

Major Analytes
Lipids that are solid at ambient temperatures are referred to as fats while those that are liquid are
called oils. Two main groups, having signicantly
different natural functions and structures, can be
classied as the neutral lipids (acylglycerides, fatty
acids, alcohols, hydrocarbons, waxes) and polar
lipids (glycolipids, phospholipids). Neutral lipids
generally form part of an energy store, while polar
lipids are functional molecules in the structure of
membranes.
Fatty acids (FAs) form homologous series of
straight chain carboxylic acids, with generally even
numbers of carbon atoms. These vary typically from
C4 to C30, with the majority lying between C10 and
C22. Regio-specific desaturase enzymes yield an important series of FAs that contains one or more double bonds, of the cis isomeric form. For multiple
double bonds, each pair is normally separated by a
single methylene unit (CH2). The series of unsaturated FAs synthesized in plants is different from that
in animals. It is important analytically to recognize
this fact, particularly where nutritional profiling
might be being done, as the difference creates the
requirement by animals for the essential dietary fatty
acids produced by plants. By combining desaturation and chain elongation, animals create two main

series of polyunsaturated fatty acids, the n
6 and

n
3 (measuring from the methyl end) with chain
lengths usually up to C22.
Exceptions to the generality of straight chains are
FAs substituted with hydroxy, keto (oxo), and epoxy
(oxirane) groups and FAs with unusual unsaturation.
Methyl branched FAs occur mainly in bacteria, but
also naturally in the fat of ruminant animals together
with a series of odd-chain fatty acids.
Triacylglycerides (TAGs) are fatty acid triesters of
glycerol. The hydrolysis products of TAGs are FAs
together with diacyl- and monoacyl-glycerides. Due
to acyl transfer diacylglycerides exist in the 1:2(2:3)
and 1:3 forms. Monoacylglycerides can exist in the
1(a) and the 2(b) forms, but are predominantly in the
a form. Despite the potential structural complexity,
most of the TAGs that the food analyst encounters
are composed of a normal range of FAs. There is
considerable conformity of structure in natural
TAGs. This is a bonus for the analyst making differentiation between species of, for instance, seed oils
easier, even after allowing for varietal (cultivar) differences. Table 1 shows typical data for some edible
fats.
Polar lipids are conveniently divided into phospholipids and glycolipids. This is not entirely adequate, but it will serve here to describe the structural
types found in food. Phospholipids fall into two main
groups. The phosphoglycerides carry an sn-3 phosphate with a nitrogenous base such as choline or
ethanolamine (other groups linked to this phosphate
may be inositol or glycerol). In the sphingosyl
phosphatides sphingosine is esteried with, for

FOOD AND NUTRITIONAL ANALYSIS / Oils and Fats 329

Table 1 Fatty acid and triacylglyceride composition of typical edible fats
% Fatty acid (chain length and number of double bonds)
Edible fat

8:0

10:0

12:0

14:0

16:0

16:1

18:0

18:1

18:2

18:3

20:0

20:1

22:0

22:1

24:0

Palm kernel oil

Palm oil
Soya bean oil
Rapeseed oil

4.9

4.0

47.5

15.8
1.0

tr

7.7
43.8
10.0
4.7

0.5
0.2
0.3

2.4
5.0
3.5
1.7

14.3
38.5
21.0
59.3

3.1
10.5
55.6
21.4

0.3
9.2
9.9

0.4
0.5
0.6

0.3
1.4

0.4

0.3

0.2

% Triacylglyceride (carbon number)

Edible fat

C30

C32

C34

C36

C38

C40

C42

C44

C46

C48

C50

C52

C54

C56

C58

C60

C62

C64

Palm kernel oil

Palm oil
Soya bean oil
Rapeseed oil

1.7

6.9

9.2

21.6

16.5

9.9

8.8

6.5

5.3
0.4
0.2

5.7
9.2
0.7
0.3

2.4
42.8
5.1
1.1

2.5
38.9
27.1
15.1

2.8
8.4
61.3
74.2

0.2
0.3
4.0
6.8

1.2
1.3

0.3
0.6

tr
0.4

0.2

Note: Carbon number relates to the sum of the carbon atoms in the three fatty acid chains of a triglyceride. Thus, C54 might be made
up of 18 18 18, 16 18 20, etc. No distinction is made, in this analysis, between saturated and unsaturated fatty acids of the
same chain length. Data can vary between batches of fat or oil. Large differences can be found between different cultivars and varieties
of the same plant.

instance, phosphorylcholine. Glycolipids feature

widely in nature. The structure is a 1:2 diacylglyceride with the sn-3 hydroxyl glycosidically linked to a
mono-, di-, or trisaccharide. The mono- and di-galactosyldiglycerides are very abundant, particularly in
cereal lipids.
Other common food lipids are the sterols and tocopherols. Sterols are generally present in natural oils
as both the free sterol and sterol fatty esters. Animal
fats exhibit solely cholesterol while plant lipids
contain a range of phytosterols. Tocopherols are
found in vegetable oils as a mixture of four isomers:a-tocopherol (true vitamin E) and b-, g-, and
d-tocopherol. Four unsaturated analogs, tocotrienols,
exist with the same numbering terminology and are
particularly abundant in palm oil.

Sampling, Preparation, Extraction of

Lipids
Obtaining the lipid in suitable form for analysis may
be the most difcult stage of sample analysis. This
step can be responsible for the degradation of lipids
and the formation of artifacts. There are a number of
important considerations to be made if the analytical
results are to give a true representation of fat
composition:
*
*
*
*
*
*

the sampling procedure,

the extraction procedure,
processing temperatures,
the possible exclusion of air,
the lighting conditions,
the surface area of lipid lms during concentration,

*
*

the possible addition of antioxidants, and

the storage temperature.

Representative sampling of a bulk of material is

not easy. Only gram quantities are required for analysis, but this size of sample may have to represent
many tons. The analyst may have to make assumptions about the contents and a conclusion about
homogeneity. Bulk fat storage tanks ideally will be
heated to maintain a temperature of 5151C above
the clear point of the fat (the point at which no
crystalline fat exists). The contents are unlikely to
have been mixed, other than by agitation during
transport and pumping from one tank to another.
Bulk oils can exhibit layering due to density gradients
and if storage temperature drops to, or slightly below, the clear point, the oil will begin to crystallize
causing sedimentation.
Sampling at several different levels and in different
quadrants of a tank is normal. The International
Organization for Standardization developed the International StandardISO-5555 Animal and Vegetable
Fats and Oils Sampling. The British Standard BS 627
(1982) forms the basis of the International Standard.
The Standard includes: sampling technique; methods
and apparatus for sampling; packaging and labeling
of samples; dispatch of samples; preparation of a sampling report. Any sampling from a large bulk of fat
must follow the procedures laid out in the Standard
as the sample and the report are considered to be
legal evidence in disputes.
Smaller quantities of fat (0.525 kg) should be
melted and mixed very well before sampling. For
measurement of the oxidation level, a core sample
should be taken, before melting, with a trier tube

330 FOOD AND NUTRITIONAL ANALYSIS / Oils and Fats

(refer to BS 809) or other hollow tube. This sample is

gently melted under nitrogen and mixed before
determination of, e.g., peroxide value (PV).
Emulsied and plasticized fat spreads are already
homogeneous, easily sampled, and have published
standard methods for their processing (IUPAC 1979).
For other food products, it is important that a
homogeneous sample of the food is obtained by
careful blending.
The ester bonds and unsaturation in lipids are
vulnerable to hydrolysis or oxidation, so that any
manipulations or procedures (and subsequent storage) must take account of these facts. Lipid hydrolysis is mainly an issue of residual active lipase
enzymes or microbial contamination. If fresh samples
become bruised or are frozen, tissue damage releases
hydrolytic enzymes. Lipid oxidation is a worse problem, particularly with highly unsaturated marine oils.
Susceptibility to oxidation is enhanced in the presence of even trace (mg kg
1) amounts of copper,
nickel, and iron. Lipid oxidation is a free-radical
process catalyzed by light of wavelengths below
400 nm. Therefore, care should be taken during
process and storage of highly unsaturated samples.
The extraction of the lipids is an important procedure in analysis. The procedure used will depend
upon the structure of the matrix containing the lipid
and the composition and physical state of the lipids.
For instance, the matrix may contain emulsied water as in a low-calorie fat spread. This can generally
be separated into fat and aqueous layers by warming
at 451C. Fresh meat contains a signicant amount of
water and will require homogenization in mixtures
of methanol and chloroform, followed by partition
of the extract against salt solution. The lipids are
found in the lower chloroform phase. However, the
partial solubility of some of the more polar lipids
needs to be considered as does partition at the water/
organic interface. These can result in quantitative
losses if care is not exercised when collecting the
organic extracts.
Fresh green vegetables should rst be homogenized
in 2-propanol to deactivate lipases, which otherwise
might be activated by some organic solvents. The
solids are further extracted with chloroform/2-propanol. The lipids in both extracts are combined.
The fat in coffee whiteners is encapsulated within
a water-soluble carbohydrate matrix, making it
unavailable to extracting solvents. Dry powder extraction will typically achieve only 15% of the fat.
To obtain the total fat:
1. Disperse powder in warm (401C) water and follow by solvent extraction. This will extract 95
100% of the fat in a gentle way. Or

2. Strong acid hydrolysis (WeibullBerntrop procedure) followed by solvent partition. This is aggressive
and can lead to fat oxidation.
The lipids in cereal ours (e.g., wheat) are highly
structural and difcult to extract quantitatively:
1. Total lipid is estimated by gas chromatography
after acid hydrolysis of the our in the presence of
a known amount of heptadecanoic acid internal
standard.
2. Nonstarch lipid is extracted from our with cold,
water-saturated, n-butanol under standard conditions.
3. The residue from extraction (2) can be used to
obtain the starch lipid by resuspending the solids in
methanol, centrifuging, and recovering the solids.
4. The nal solids are extracted with water-saturated
n-butanol at 951001C for 60 min, repeating this ve
times to yield the starch lipids.
Marine organisms present particular problems due
to high levels of lipid unsaturation together with high
water and protein contents. The following procedure
can provide a protected environment under a carbon
dioxide blanket: The tissue is cut into small pieces at
water/ice temperatures and homogenized in a kitchen
food processor, together with enough pelleted solid
carbon dioxide to fully freeze the material to
B
401C. The powdered sample is extracted with
chloroformmethanol mixtures that have been precooled with cardice to
501C. This whole mixture is
left to reach ambient temperature with occasional
mixing and then processed using precautions against
lipid oxidation.

Standard Methods and Quality

Assurance
Analytical precision and accuracy require standardization of methodology. Techniques require proving
with standards and reference materials and should be
tested regularly by blind inclusion of these materials
into normal analytical practice. In this way the statistical variability of results can be built up providing
improved condence. The analyst can draw
upon standard methodology from international authorities such as the International Union of Pure and
Applied Chemistry (IUPAC), British Standards
Institution, American Oil Chemists Society (AOCS),
International Organization for Standardization
(ISO), Deutsche Gesellschaft fur Fettewissenschaft,
and Association of Ofcial Analytical Chemists
(AOAC). The priority is to standardize the chosen

FOOD AND NUTRITIONAL ANALYSIS / Oils and Fats 331

methodology in the laboratory and ensure consistent

practice.
Quality assurance methods commonly used are for
the determination of free fatty acid (FFA), PV, iodine
value (IV), anisidine value (AnV), Rancimat induction time, and trans value (TV).
Crude oils may be up to 15% FFA, while rened
oils will be o0.1%. Measurement of FFA is normally
done by titration of an etherethanol solution of the
fat with standardized aqueous sodium or potassium
hydroxide, in the presence of phenolphthalein indicator (see ISO 660: 1983). This method is very accurate, using the molecular weight of oleic acid (282)
for all calculations. Accuracy is improved by using
the average molecular mass of the fatty acids in the
fat, calculated from the FA composition (e.g., for
palm oil 256 is used, for palm kernel/coconut oil 200
is used). The results are expressed either as acid
value; the number of milligrams of potassium hydroxide required to neutralize 1 g of the fat, or,
FFA%; the percentage concentration of oleic acid
equivalent to the free acids present.
PV is normally assessed using the Wheeler procedure: AOCS (1984) method CD 8-53 (73); and
IUPAC method 2.501. The determination should
ideally be carried out under tungsten lighting as
ultraviolet light liberates iodine from potassium iodide in solution, in the presence of oxygen, creating
high blanks. The liberated iodine is titrated with
standard sodium thiosulfate. Results are expressed in
Standard International (SI) Units. Some laboratories
express the result in different units, which may be
related to the SI units by the factors shown in the
Table 2.
The interpretation of results requires some knowledge of the sample and sample processing history.
For instance, low values do not necessarily indicate
good quality fats; organic lipid peroxides are not
stable at temperatures above 501C so that in deep fat
fryers operating at up to 1801C, one would not expect to nd much peroxide. The half-life of peroxides
can be extremely short under frying conditions and
the sample requires a standard amount of time after
removal from the fryer (typically 30 min) before
analysis providing more meaningful results.

IV measures the level of unsaturated components

in a fat equivalent to a weight of iodine reacting with
100 g of the sample. The sample is reacted under
standard conditions with a known amount of iodine
(iodine trichloride as Wijs solution) in the dark
(BS684 section 2.13, 1981; and ISO 3961, 1979).
Unreacted iodine is liberated chemically and backtitrated with sodium thiosulfate to yield an IV. A
minimum IV can be calculated from the FA composition determined by gas chromatography. The titrated IV may be higher due to the presence of other
iodine-positive components. The technique assays
unreacted reagent therefore, it becomes essential that
the amount of Wijs reagent used is very accurately
measured. A purpose-designed, exact-volume dispenser is better than using a burette. Precision of
technique is very important to the reproducibility of
IV measurement. A common reason for error is incorrect ratio of fat and reagent. For the reaction to be
complete and the back-titration to yield a satisfactory value, there must be a minimum excess level of
Wijs reagent of B50%. The weight of sample fat
used for the estimation depends upon the expected
IV and may be calculated from the equation below.
For a totally unknown sample this amount may be
difcult to judge requiring a test IV to be carried
out:
Weight of sample gm

12:69  M  VR
approx IV

where M is the molarity of sodium thiosulfate and

VR the volume of Wijs reagent used.
Alternatively, where an estimated IV is available,
the weight of sample (25 ml Wijs reagent used) may
be obtained from Table 3.
AnV measures the level of carbonyl compounds,
particularly aldehydes, in a fat (IUPAC method
2.504). The fat, in 2,2,4-trimethylpentane solution,
is treated with r-anisidine reagent prepared with anhydrous acetic acid. The level of reaction products is
determined spectrophotometrically at 350 nm. The
reagent is particularly toxic and skin contact must be

Table 3 Weight of sample to be used for IV determination

(25 ml Wijs reagent used)
Table 2 Relationship between SI units and other units used for
peroxide value

Expected IV

Weight sample
(g)

Weighing accuracy
(7g)

Units of PV

Factor

Millimole active oxygen per 2 kg lipid SI units

Milliequivalents of active oxygen per kilogram lipid
Millimole active oxygen per kilogram lipid (Lea value)
Milligram active oxygen per kilogram lipid

 1.0
 1.0
 0.5
 8.0

05
620
2160
6180
81130
4130

4.0
1.0
0.4
0.25
0.15
0.10

0.0005
0.0005
0.0005
0.0001
0.0001
0.0001

332 FOOD AND NUTRITIONAL ANALYSIS / Oils and Fats

avoided absolutely. Good-quality fats should show

values of less than 10.
The Rancimat induction time is the time taken for
a fat under test conditions to reach an approximately
maximal rate of oxidation. The test is a variant of the
Swift active oxygen method using a device called the
Rancimat (Metrohm, Switzerland). A constant rate
of air is bubbled through the fat sample held in a
thermostatically controlled cell normally at 1001C.
The data produced give a measure of potential shelflife for a fat. At 1001C, a typical biscuit shortening
gives a time of 50 h, cocoa butter 4140 h, and sunower oil B12 h. The technique is reliable. All glassware must be very clean and a blank (i.e., without
fat) analyzed. Samples are determined in duplicate
and any showing large deviations repeated.
As part of the nutritional analysis of fat in food, it
may be important to determine the level of trans unsaturation (TV) present. The measurement of total
trans fatty acids is conveniently done by infrared (IR)
spectroscopy but needs careful calibration with
known standard solutions of pure trielaidin and
tristearin (99%). Above 10% trans, the accuracy is
good and the measurement is done on the TAGs.
Below 10% trans, interference is common, reducing
accuracy and TV is measured either by IR of the
methyl esters, or, preferably by gas chromatography.
Calibration in this case must also use methyl esters.
The cis absorption is small in IR, but isolated trans
double bonds show a relatively strong IR band absorbing at B967 cm
1. The AOAC published a
method (AOAC 28.05228.067) for TV. Isomerized
or oxidized fats cause errors because conjugated species show strong absorption close to the isolated
trans bond and interfere with the correct allocation
of baseline. Samples and standards are usually analyzed in solution (200 mg ml
1), traditionally in carbon disulde. A calibration line is drawn for different
amounts of trielaidin mixed with tristearin, a
solution of pure tristearin being in the reference
beam at all times. The line should be straight. For
each standard and sample record, a straight baseline
is drawn joining the minima at 1000 and 925 cm
1.
TV of samples are read from the calibration graph.
For TV analysis by gasliquid chromatography
(GLC) of fat to methyl esters (FAMEs) the ofcial
AOCS method prescribes the cyanopropylpolysiloxane phase SP2340. The column should be 60 m 
0.25 mm fused silica with a 1 m retention gap of silanized fused silica with a lm thickness of 0.2 mm.
Such columns will provide adequate separation of cis
and trans FAME for most situations involving food
fats (except fats containing hydrogenated sh oils).
However, there is peak overlap and for exacting
analyses, where information is required on the iso-

meric nature of the trans FAME, even these columns

are not adequate. For separation that is more
complete, a column of SP2560, 0.2 mm lm on
100 m  0.25 mm fused silica should be used. In
both column cases, the carrier gas can be helium
(linear velocity 20 cm s
1) or hydrogen (40 cm s
1).
Column temperature should be optimized, using
standards, between 1751C and 2001C isothermal.
Otherwise, faster results can be obtained by programming the temperature between 1001C and 2251C
with some sacrice to optimum resolution. The
samples should be injected using the on-column
technique in preference to split injection. Cis/trans
ratios calculated from this type of chromatography
will, in general, be more accurate than those
obtained via IR techniques. However, this is true
only below 15% trans content.

Additional Analyses
GLC is applied to the analysis of fats and fat components in several ways. Typical data for GLC is
shown in Table 1. This includes the analysis of FA
composition after derivatizing the FAME (or propyl
esters for butter fat) and the direct analysis of TAG
composition. GLC is used for the analysis of sterols,
hydrocarbon contaminants, pesticides, and volatile
products produced during fat processing and refining.
For general FAME analysis it is adequate for most
work to use wide bore (0.53 mm ID) silica (quartz)
columns. A 30 m length with bonded FFAP phase
(lm thickness 1.0 mm) produces a robust column of
excellent long-term stability. Other phases are suitable, such as silar 5C, silar 10C, CPSil88. Better
resolution may be obtained with columns of 0.32
and 0.25 mm ID (lm thickness 0.5 mm) but sample
loading capacity will be lower and they might be
more prone to damage by mishandling (see Figures 1
and 2).
Capillary GLC columns are excellent for the analysis of TAGs. It is essential that a precolumn (retention gap) of uncoated, silanized silica tube (1.0 m 
0.53 mm ID) be attached to the analytical column.
The injection technique of choice for both FAME and
TAGs should be on-column. Split injection almost
always leads to quantitative errors in these analyses.
Both helium and hydrogen are adequate carrier gasses. A 710 m bonded phase (OV1, lm thickness
0.1 mm) 0.53 mm ID column is used with a temperature stability of at least 3501C, but aluminum-clad
columns are avoided. A temperature program
is used, starting at B1001C rising to 3501C
at B101C min
1. While the carrier gas velocity for

FOOD AND NUTRITIONAL ANALYSIS / Oils and Fats 333

16:0

1800
1600

16:1

1400

12:0

600

14:1

10

12

14

16 18 20
Time (min)

22:6

800

24:0
24:1

1000

16:2
18:0
18:2
18:3
20:0
20:1
20:2
20:3
20:4
22:0
20:5
22:1

1200
14:0

mV

resolution and reasonable quantitative analysis. Samples are injected using cold on-column injection. The
column start temperature is 2751C with a 2 min hold,
rising to 3251C at 21C min
1. a-Cholestane is used
as an internal standard in this analysis.
High-performance liquid chromatography (HPLC)
is routinely used for the compositional analysis of
lipid classes, TAGs, and tocopherols. However, there
can be problems for the quantitative detection of
lipids other than tocopherols because most lipids do
not have a suitable chromaphore, and therefore cannot be detected spectrophotometrically. Evaporative
technology such as nebulizing mass detectors and
the quartz/ame ionization transport system has to
be employed.
For lipid class separation a 10 cm  4 mm plain
silica (3 mm particle size) column is adequate,
providing separation across the range from sterol esters and TAGs through to phospholipids and galactolipids in B30 min.
For TAG separation by reversed-phase HPLC, two
10 cm Lichrosphere RP-18, 5 mm columns are linked
together. Ideally, column temperature should be kept
constant at 251C. It is advisable to analyze as many
known materials as possible and construct a graph of
equivalent chain length by plotting total carbon
number against elution time. Peak types overlap because a double bond is almost equivalent to a chain
shortening of two carbon atoms. The rather complex
separation achieved yields much information about
the TAG structure and is a composite of chain length
and unsaturation, being more complex than the hightemperature polar phase GLC procedure. These data
can be used to nger print fats.

18:1

FAME columns should be optimal for maximum

resolution, it is advantageous to have the carrier gas
velocity for TAG analysis at twice optimal to minimize on-column polymerization.
Free and esteried sterols generally occur together
in natural oils but are analyzed after hydrolysis as
part of the nonsaponiable fraction. To reduce chromatographic losses the sterols are derivatized to
the trimethyl silyl ethers prior to GLC. Trimethylsilyimidazole is used for the preparation of
TMS derivatives. A 25 m  0.53 mm (or 0.34 mm)
fused silica column with a 1.0 mm bonded lm of 5%
phenyl-methyl silicone, tted with a 1 m  0.53 mm
silanized retention gap, is adequate for good

22

24

26

C52

Figure 1 FAME gas chromatography: fatty acid methyl esters

of a sh oil; 30 m  0.53 mm FFAP (see text for details). Peak
names are shown as numbers separated by a colon. The number
before the colon is the chain length, that after is the number of
double bonds.

2000
1800

C54

C50

1400

8.0

10.0

12.0

14.0

16.0

18.0

20.0

22.0

C58

600

C56

800

C42
C44
C46
C48

1000

C36
C38
C40

1200

C26
C28
C30
C32
C34

Intensity (mV)

1600

24.0

Time (min)
Figure 2 Carbon number gas chromatography: triglycerides of a mixture of 1:1 butter fat and cocoa butter; 8 m  0.53 mm OV1 (see
text for details). Peak names are shown as carbon numbers which are the sum of the carbons in the three fatty acid chains, excluding
the glycerol carbons.

334 FOOD AND NUTRITIONAL ANALYSIS / Fruits and Fruit Products

For chocolate fats in particular, it is often necessary to know the composition of the TAGs based
simply upon the level and position of unsaturation.
The SOS (1-saturated, 2-monounsaturated, 3-saturated) type TAGs are responsible for the particular
physical properties of cocoa butter and its equivalents. The information can be obtained from the
reversed-phase method mentioned above, but a simpler prole is obtained by using 5% silver nitrateimpregnated silica as a 25 cm column.
Tocopherols are analyzed in their unesteried form
and separations are very easy by HPLC. The use of
uorescence detection provides very accurate results.
The oil is diluted with eluent (3% tetrahydrofuran in
isooctane) to B10 mg ml
1. Pure standards are analyzed (2 mg ml
1) to obtain retention data, while pure
a-tocopherol is used as an external standard for every
two samples. A 15 cm column of 5 mm Chromegasphere is equilibrated with the isocratic solvent at
1.5 ml min
1. Injection volume is 50 ml. The spectrouorimeter excitation is 290 nm and emission is
330 nm. Full separation should be readily achieved,
but the column will require conditioning by pumping
chloroformmethanol (2:1; v/v) for 10 min every 30
samples. After this conditioning, the column activity
must be regenerated by pumping normal eluent for
at least 20 min.

See also: Carbohydrates: Starch. Derivatization of

Analytes. Extraction: Solvent Extraction Principles. Fluorescence: Food Applications. Gas Chromatography:
High-Temperature Techniques. Lipids: Fatty Acids; Polar
Lipids. Liquid Chromatography: Food Applications.
Mass Spectrometry: Food Applications. Quality Assurance: Quality Control; Internal Standards. Sample Handling: Comminution of Samples. Sampling: Theory.

Further Reading
Firestone D (Associate Chapter Editor) (1997) AOAC
Ofcial Methods of Analysis chapter 28, Oils and Fats,
p. 507. Washington: AOAC.
Duchateau GSMJE, van Oosten HJ, and Vasconcellos MA
(1996) Analysis of cis- and trans-fatty acid isomers in
hydrogenated and rened vegetable oils by capillary gas
liquid chromatography. JAOCS 73(3): 275282.
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Fruits and Fruit Products

R J Pither, Campden and Chorleywood Food Research
Association, Chipping Campden, UK
& 2005, Elsevier Ltd. All Rights Reserved.
This article is reproduced from the previous edition, pp. 1525
1531, & 1995, Elsevier Ltd.

well as being good sources of dietary ber, mineral

salts, and vitamins, especially vitamin C. The organic
acids found in fruits essentially balance with the
sugars to give them their characteristic avors and
are therefore also a vital component.

Major Components
Introduction
Fruits have long been valued as a foodstuff owing to
their pleasant avor and aroma, and attractive appearance; however, the use of fruits as a staple in the
diet has only become possible in the past century
through the development of canning and other bulk
preservation techniques, and improvements in transportation and storage systems. The increased awareness of the importance of a nutritionally balanced
diet has also contributed to the increased consumption of fruit and fruit products.
Fruits are an important low-fat energy source containing varying proportions of sugars and starches as

The major component of fresh fruit is water, with

levels ranging from 65% (in avocado) to 93% (e.g.,
in melons, strawberries). Dried fruit products can
contain up to 25% water while fruit juices can vary
between 85% and 93%. The second most important
component in terms of percentage content is carbohydrate, with reducing sugars, primarily a mixture of
glucose and fructose, being the main soluble carbohydrate. Most fruits contain relatively low levels of
sucrose; however, it is the primary sugar in peaches,
apricots, bananas, pineapple, and certain citrus
fruits. Starch, cellulose, hemicellulose, and pectin
are also found in varying proportions in fruit.