International Dairy Journal 10 (2000) 119}128

Composition of goat's milk fat triglycerides analysed

by silver ion adsorption-TLC and GC}MS
J. Fontecha *, J.J. RmH os
, L. Lozada , M.J. Fraga, M. JuaH rez
Departamento de Productos La& cteos, Instituto del Frn& o (CSIC), Ciudad Universitaria s/n, 28040 Madrid, Spain

Instituto de la Grasa (CSIC), 41012 Sevilla, Spain
Departamento de Produccio& n Animal, Universidad Polite& cnica, 28040 Madrid, Spain
Received 27 May 1999; accepted 23 March 2000

Abstract
The triglyceride (TG) composition of goat's milk fat (from the milk of "ve herds collected monthly, from November to May) was
studied using AgNO -TLC and GC}MS. Two of the four fractions obtained by TLC contained trisaturated TGs (SSS) and

represented 55% of the total TGs; they were separated by the chain length of short-chain fatty acid (FA). The TGs of the remaining
fractions were identi"ed as mono- (SSM) and polyunsaturated, representing 29 and 16% of the total TGs, respectively. The
distribution of two SSS fractions by carbon number (CN) was unimodal, with maxima at CN40 and CN36 respectively. The
proportions of SSM and polyunsaturated TG were high between CN38}CN44 and CN46}CN54, respectively (mean values, 35 and
52%). One hundred and twenty-four peaks (some containing more that one molecular species of TGs) were detected in the
chromatogram of total fat; the sum of the 30 peaks representing'1 mol% was 70% of the total TGs. One hundred and thirty-seven
molecular species were identi"ed in the total goat's milk fat: 50% SSS, 30% SSM and 20% polyunsaturated. The most important in
quantitative terms were medium-chain TGs containing C8, C10 or C12, and C18 : 1 as unsaturated FA.  2000 Elsevier Science Ltd.
All rights reserved.
Keywords: Goat's milk fat; Triglycerides; Molecular classes; Molecular species; Silver ion adsorption-TLC; GC}MS

1. Introduction
Gas chromatography (GC) with packed or short capillary columns has been used to separate milk fat triglyceride (TG) classes according to their carbon number (CN).
However, to explain the physical properties (melting
point, crystallization behaviour, etc.), nutritive characteristics (action of lipolytic enzymes) or biosynthesis of milk
fat in the mammary gland, the composition must be
known in terms of molecular species of TGs. Given the
high number of fatty acids in milk fat, combined or
hyphenated chromatographic techniques have been indispensable for identifying and quantifying molecular
species of milk fat TGs: thin layer chromatography
(TLC) or high-performance liquid chromatography
(HPLC) without or with Ag> as a pre-separation step,
followed by supercritical #uid chromatography (Man-

* Corresponding author.
E-mail address: jfontecha@if.csic.es (J. Fontecha).

ninen, Laakso & Kallio, 1995), GC with long capillary

column (Steele & Banks, 1994; Myher, Kuksis, Marai
& Sandra, 1988; Gresti, Bugaut, Maniongui & Bezard,
1993; Fraga, Fontecha, Lozada & JuaH rez, 1998), and
HPLC with reverse phase and mass spectrometry (MS)
as detector (Laakso & Kallio, 1993; Ruiz-Sala, Hierro,
MartmH nez-Castro & Santa-MarmH a, 1996).
Quite detailed information on the composition of the
main TGs in cow's milk can be derived from these studies. However, there is very little information on goat's
milk. The use of GC with a short column (Marai,
Breckenridge & Kuksis, 1969; Parodi, 1973; Cerbulis,
Parks & Farrell, 1982; Luf, Stock & Brandl, 1987; Fontecha, DmH az, Fraga & JuaH rez, 1998) showed that the
occurrence of TGs in goat's milk reaches a maximum at
CN38}CN42; beyond this point, the percentage of TGs
decreases but not uniformly, because the values for CN48
and CN50 are close together. With regard to the fatty
acid composition of TGs, in a study using TLC and GC,
Dimick and Patton (1965) demonstrated that the TGs of
goat's milk fat contained no more than one mole of

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PII: S 0 9 5 8 - 6 9 4 6 ( 0 0 ) 0 0 0 2 6 - 1

120

J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

butyrate per mole, a high C18 : 0 content in the high

molecular weight TGs, and a remarkable degree of uniformity in the C16 : 0 distribution. Glass, Jenness and
Lohse (1969) used TLC to show that goat's milk TGs
separate readily into two distinct fractions that have
nearly the same C14 : 0 and C16 : 0 content, but one of
them contains almost all the butyrate, and the other is
much richer in C18 acids.
More recently, Barron, Hierro and Santa-MarmH a (1990)
and Ruiz-Sala et al. (1996) used HPLC}GC to identify
respectively 116 and 181 molecular species of TGs in
goat's milk fat. In both cases it was not possible to
quantify the individual TGs because most of the HPLC
peaks contained more than one molecular species.
The purpose of this work was to study the composition
of the TGs in goat's milk fat from "ve di!erent herds
using a combination of AgNO -TLC and GC}MS.

2. Materials and methods
2.1. Samples and standards
We used Murciana-Granadina adult female goats in
"ve herds, belonging to "ve di!erent breeders in the
Murcia region (Spain) and their size ranged from 98 to
125. Batches were taken once daily from storage tanks
containing milk from the whole herd. Seven batches were
collected for each herd during a milking period of seven
months, starting in November. During the "rst 2 months
after parturition (September) all the milk was used for
feeding the kids. The milk fat was extracted following the
procedure described by Fontecha et al. (1998). All the
goat's milk fat samples from each herd were combined in
equal volumes to obtain "ve mixture samples.
Di!erent dilutions of the fat were prepared in hexane
for chromatographic analysis. To identify the TG, a mixture of synthetic TGs (trilinolein, triolein, tristearin, tripalmitin, trimyristin, trilaurin, tricaprin, and tricaprylin)
was "rst analysed to determine both the best chromatographic conditions and the retention time of these components. The other molecular species were identi"ed
following previous studies carried out in similar conditions (Myher et al., 1988; Hinshaw & Seferovic, 1986;
Kalo & Kemppinen, 1993) and con"rmed by MS. A
reference butter oil of known TG composition, which
had served as test fat in EC collaborative trials (Precht,
1991), was used to determine the response factors
(RF) for quantitative studies of TG composition as described in a previous paper (Lozada, de la Fuente, Fontecha & JuaH rez, 1995). To quantify the relative
percentages of TLC fractions, a solution of the TG
trinanoin (C27, Sigma, Chemical CO., St. Louis, MO)
was added as an internal standard to each TLC fraction
scraped o! from the AgNO -TLC plate before the GC

analysis.

For the determination of fatty acid composition a BCR

reference fat (CRM 164) was used.
2.2. Separation of triglycerides by AgNO3 -TLC
TLC glass plates (20;20 cm) with silica gel (0.25 mm)
(Merck Darmstadt, Germany) were incubated with 20%
aqueous solution of AgNO (Panreac) overnight. The

plates were then activated at 1003C for 30 min and
0.5 mL of the fat solution (12 mg mL\ in chloroform,
containing 0.5}0.8% ethanol) was applied to the plate.
Triolein and tristearin were also applied to the edge of
the plate as qualitative standards. The plate was developed twice in a saturated chamber in chloroform with
15 cm migration. After drying, the plates were sprayed
with a 0.15% ethanol solution of 2,7-dichloro#uorescein
and the bands were visualized under UV light. Bands were
scraped o!, and 20 lL of 3.15 mg mL\ solution of C27
was added to each band as internal standard. The TGs
were extracted four times with 20 mL diethyl ether protected with butylhydroxytoluene (Panreac), "ltered, and
the solvent was evaporated under reduced pressure. The
residue was resuspended in 140 lL hexane and the solution
was used for the analysis of TGs and fatty acids. Fractionation was repeated twice, and the corresponding fractions
were injected in duplicate for GC and GC}MS analysis.
2.3. Gas chromatographic analysis of triglycerides
For the analysis of total TGs in goat's milk fat, 20 mg
of fat was dissolved in 0.5 mL hexane. For the analysis of
both goat's milk fat and fractions obtained by AgNO 
TLC, 0.2 lL was injected into the gas chromatograph.
The triglyceride analyses were performed on an Autosystem Gion 4072042 gas chromatograph (Perkin-Elmer,
Beacons"eld, UK) equipped with an automatic injector
(split/splitless) and programmed temperature. A capillary
column (30 m;0.22 mm i.d.), supplied by Restek
(Bellefonte, PA), Rtx-65 TG (35% dimethyl, 65%
diphenyl polysiloxane) (d "0.10 lm) was used. Experi
mental chromatographic conditions were as follows: the
initial temperature (2203C) was raised to 3203C at a rate
of 153C min\ and then to 3553C at a rate of 73C min\
and then held at this temperature for 20 min. The injector
and detector temperatures were 355 and 3703C, respectively. The pressure at the top of the column was 25 psig,
the split ratio was 1 : 4 and the carrier gas was helium.
The calculated #ow rate was 0.8 mL min\.
2.4. Gas chromatographic analysis of fatty acids
For the preparation of methyl esters of goat's milk fat,
0.1 g of fat was dissolved in 1 mL hexane and 0.05 mL of
2 M potassium hydroxide in methanol was added as
described by Christopherson and Glass (1969). For the
analysis of the fractions obtained by AgNO -TLC the


J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

"nal hexane solution was methylated according to the

same procedure. In both cases, 0.2 lL was injected into
a gas chromatograph. A Perkin-Elmer Model 8420 gas
chromatograph (Beacons"eld, UK) was used, equipped
with programmed temperature vaporizer inlet, #ow splitter and hydrogen #ame ionization detector. The carrier
gas was helium, the split ratio was 1 : 20 and the pressure
at the top of the column was 25 psig. Calculated #ow rate
was 0.7 mL min\. The column was a WCOT silica capillary (50 m;0.22 mm i.d.) containing a Silar 5CP
(50% phenyl, 50% cyanopropyl) stationary phase
(d "0.22 lm) (Chrompack, Middelburg, The Nether
lands). Experimental chromatographic conditions were
as follows: the initial temperature of 603C was maintained for 3 min, then raised to 1903C at a rate of
153C min\. The "nal temperature was maintained for
30 min. The injector and detector temperature was 2003C.

121

Fig. 1. Fractionation of triglycerides of goat's milk fat by AgNO 

TLC. See Table 1 for bands identi"cation.

2.5. Gas chromatography}mass spectrometry

The mass spectrometry of goat's milk fat fractions obtained by AgNO -TLC, was performed with

a Finnigan MAT95 s (Finnigan, Bremen, Germany) highresolution mass spectrometer interfaced with a HewlettPackard 5890 Series II gas chromatograph. The column
and the experimental chromatographic conditions were
the same as in the GC analysis of TGs. Electron impact
(EI) spectra were recorded at 70 eV. Full spectra
(50}1000 amu) were recorded at a scan speed of 2 s decade\ over the entire elution pro"le. Data were analysed
using an ICIS II Data System from Finnigan MAT.

Table 1
Total wt% of AgNO -TLC fractions (mean values and standard devi
ations of the "ve samples obtained from the corresponding herds) of
goat's milk fat
AgNO -TLC

fractions

Wt%

Characterization of triglycerides

38.5$2.62

16.5$1.76

C
D

28.9$4.02
16.0$1.93

Saturated (C6 and longer chain

fatty acids)
Saturated (C4 and longer chain
fatty acids)
Monounsaturated
Polyunsaturated

3. Results
3.1. Triglyceride separation by AgNO3 -TLC: fatty acid
and triglyceride composition of the diwerent fractions
Using AgNO -TLC, the TGs from the goat's milk fat

were separated into four main fractions (Fig. 1); although
fractions A and D included two bands, they were too
close to be individualized. Table 1 gives the relative
amounts (wt%) of material recovered from the di!erent
AgNO -TLC fractions (mean values and standard devi
ations of the "ve goat's milk fat samples obtained from
the corresponding herds) and its characterization.
Table 2 shows the fatty acid composition (mol%) of
TGs of the goat's milk fat and the four fractions obtained
by AgNO -TLC. A large proportion of the fatty acids in

fractions A and B was saturated (95.3 and 94.3%, respectively) and hence the TGs contained in both fractions
were identi"ed as trisaturates (SSS). Fraction B contained 94% of the butyric acid detected in SSS fractions,
while fraction A contained larger amounts of C8, C10
and C12 fatty acids (73, 80 and 74% of the total SSS
fractions, respectively).

The TGs in fraction C contained nearly one-third of

the monounsaturated fatty acids (29%) and hence were
identi"ed as monounsaturated (SSM). The di!erence between this percentage and the theoretical value (33.3%)
could have been due to overlapping with the saturated
fractions, which also contained some monounsaturated
acids (0.9 and 1.5% in fractions A and B, respectively)
and also due to the presence of C18 : 2 and C18 : 2U

in the SSM band (1.2 and 0.8%, respectively, Table 2).

Fraction D contained 35% of monounsaturated fatty
acids, mainly C18 : 1 (78% of the total), 14.6% of C18 : 2
(including 0.8% of C18 : 2U
) and 1.4% of C18 : 3.
Fig. 2 shows the molar distribution of the TGs in
goat's milk fat by CN, degree of unsaturation and, in the
saturated TGs, length of the short fatty acid chain. The
values were calculated by multiplying the molar percentage of each TG class of total goat's milk fat by the relative
proportions of the four fractions obtained by AgNO 
TLC in the corresponding class of TGs calculated from
the GC data. Data are shown from CN28, because the
molar percentages of TG classes CN22, CN24 and CN26
were low (0.09, 0.19 and 0.45% respectively).

122

J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

3.2. Composition of molecular species of main triglycerides

Fig. 3 shows a chromatogram of the goat's milk fat
(from the herd 1 sample), in which a total of 124 peaks
were detected. Table 3 shows the relative retention times
(RRT) and the molar percentages (mean values and standard deviations of the "ve samples obtained from the
corresponding herds) of each peak. Also, where possible
Table 2
Fatty acid molar composition (%) of TGs of goat's milk fat and of the
fractions obtained by AgNO -TLC (values are means of the "ve sam
ples obtained from the corresponding herds)
Fatty Acid

C4
C6
C8
C10
C10 : 1
C12
C12 : 1
C14
C14 : 1# iC15
aiC15#C15
C15 : 1
iC16#C16
C16 : 1
iC17#aiC17#C17
C17 : 1
C18
C18 : 1
C18 : 2
C18 : 3
C18 : 2U

C20
C20 : 1
Others

Total fat

5.09
4.42
4.15
12.91
0.36
5.62
0.21
9.86
0.39
0.83
0.09
25.38
1.41
1.26
0.34
7.17
15.46
2.83
0.35
0.57
0.11
0.05
1.14

Mean values of two replicates.

AgNO -TLC fraction


A

0.56
3.83
6.70
18.57

19.26
6.87
5.81
10.79

8.72
13.54
0.85
2.18

6.96
0.05
10.01
0.73
1.67

30.54

24.26

0.88

1.05

9.57
0.90

7.17
1.47

4.43
3.94
4.57
11.74
0.60
5.21
0.41
7.95
0.69
1.06
0.37
19.64
2.79
0.66
0.60
6.58
24.24
1.19

0.06
0.18

0.12
0.42

2.92

3.37

3.01
2.50
3.98
7.75
1.84
3.80
0.40
5.15
0.86
1.08
0.53
12.30
3.98
0.98
0.73
4.47
27.35
13.80
1.44
0.76
0.28
0.18
2.83

0.76
0.11
0.03
2.43

the table indicates the tentative identi"cation of TGs

contained in each peak. Data are shown from CN28,
because no TG was identi"ed in TG classes CN22, CN24
and CN26.
As indicated, the molecular species of TG were "rst
tentatively identi"ed following previous studies carried
out in similar conditions (Myher et al., 1988; Hinshaw
& Seferovic, 1986; Kalo & Kemppinen, 1993). The con"rmation of identity of a TG species was made on the
basis of GC and GC}MS analysis of the intact TG
recovered from the four fractions obtained by AgNO 
TLC. As an example, Fig. 4 shows the partial chromatograms corresponding to CN36 class TG of the total milk
fat and of the four AgNO -TLC fractions. A total of eight

peaks (49}56) were recognized in the goat's milk fat, some
of them containing contributions from more than one
AgNO -TLC fraction. Table 4 illustrates the method

followed for the identi"cation of individual TGs in the
CN36 class TG. The table indicates the composition of
TG CN36 in each AgNO -TLC fraction determined by

CG}MS, the situation of each peak in relation to the
RRT of total fat, and the molar percentages of the peaks
for the individual AgNO -TLC fractions. The unidenti
"ed peaks (NI) in each band are indicated, but only where
their molar percentages multiplied by the mass proportion of the corresponding band in the total fat were
'0.1%. The CG}MS analysis also provides estimations
on the relative abundance of the molecular species within
each peak. Thus, in Table 3, the most abundant TG in
each peak is underlined.
More than one species of TG was identi"ed in 37 peaks
in the chromatogram in Fig. 3. Some peaks (i.e. peaks 50,
51, 54 and 55, see also Table 4 and Fig. 4) were located in
more than one of the AgNO -TLC fractions. In addition,

more than one TG was identi"ed in some peaks of
chromatograms from each of the AgNO -TLC fractions

(i.e. those corresponding to peaks 49 and 50 in fraction
A and 50}52 in fraction C, Table 4).

Fig. 2. Distribution (mol%) of triglycerides in goat's milk fat according to carbon number and degree of unsaturation. A: saturated (C6 and longer
chain fatty acids); B: saturated (C4 and longer chain fatty acids); C: monounsaturated; D: polyunsaturated.

J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

123

Fig. 3. Capillary GC pro"le of triglycerides in a goat's milk fat sample collected from herd 1.

Eleven peaks, situated at the beginning of each TG

class from CN36 to CN52 (49, 50, 57, 58, 67, 75, 83, 90, 98,
105, and 112 in the chromatogram in Fig. 3), were largely
located in fraction A. The largest peaks in TG classes
CN28}CN38 were preferentially located in fraction B. Of
these, numbers 24, 33, 42 and 51 were the most abundant
in that fraction, where individual molar percentages
ranged from 7 to 16%. The 10 largest peaks in the
chromatogram of band C corresponded to numbers 58,
60, 68, 69, 76, 77, 84, 91, 99 and 106. However, only the
last six peaks (from CN42 to CN52) were preferentially
located in that band. The largest peaks in fraction D corresponded to numbers 73, 80, 85, 92, 107 and 114 (from
CN40 to CN52 in the chromatogram for total fat).
One hundred and thirty-seven molecular species of
TGs were identi"ed: 69 (50%) saturated, 41 (30%) monoand 27 (20%) polyunsaturated. By length of chain, 59
(43%) of the molecular species identi"ed contained
short-chain fatty acids (C4, C6), 101 (74%) medium-chain
fatty acids (C8}C14), and 112 (82%) long-chain fatty
acids (C16}C18).

4. Discussion
The SSS TGs represented 55% of the total TGs
(Table 1), a value higher than in cow's milk (47%, Fraga
et al., 1998). All the SSSs of cow's milk fat are located in
a single TLC band in most of the references consulted;
however, as in the present case, Myher et al. (1988) and
Fraga et al. (1998) reported that they can be separated
into two bands di!erentiated mainly by the chain length
of the short-chain fatty acid. The SSSs that contain
butyric acid (fraction B) only represented 30% of the
total SSS, which was much lower than that reported for
cow's milk (46%, Fraga et al., 1998) as a consequence of
the lower butyric acid content of goat's milk fat.
Monounsaturated TGs of cow's milk were separated
into two TLC fractions in most of the references consulted. This separation was mainly controlled by the chain
length of the short-chain fatty acid (Fraga et al., 1998) or
by the geometric con"guration of the unsaturated fatty
acids (Parodi, 1980; Myher et al., 1988; Laakso & Kallio,
1993). However, using similar experimental conditions as

124

J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

Table 3
TG composition (mol%) of goat's milk fat and identi"cation of TGs according to data of AgNO TLC-MS and the relative retention time (RRT) to C27

Peak
no.

CN

RRT

mol% SD

Triglycerides identi"ed

13
14
15
16
17
18
19
20
21

C28

1.066
1.074
1.089
1.096
1.110
1.126
1.144
1.160
1.179

0.13
0.26
0.35
0.23
0.05
0.16
0.07
0.07
0.02

0.011
0.016
0.018
0.066
0.024
0.021
0.006
0.005
0.002

8,8,12/8,10,10
6,10,12
4,10,14
4,8,16

22
23
24
25
26
27
28
29
30

C30

1.205
1.216
1.234
1.242
1.253
1.275
1.286
1.300
1.319

0.36
0.39
1.13
0.13
0.25
0.17
0.13
0.10
0.05

0.028
0.009
0.077
0.026
0.017
0.022
0.014
0.009
0.017

8,10,12/10,10,10
6,10,14
4,10,16/4,12,14

31
32
33
34
35
36
37
38

C32

1.345
1.362
1.377
1.396
1.415
1.431
1.453
1.475

0.86
1.02
1.37
0.60
0.28
0.23
0.16
0.09

0.038
0.088
0.034
0.028
0.023
0.031
0.023
0.008

8,10,14/8,12,12/10,10,12
6,10,16/6,12,14
4,12,16/4,14,14/6,8,18 : 1
4,10,18 : 1

39
40
41
42
43
44
45
46
47
48

C34

1.504
1.515
1.525
1.552
1.569
1.584
1.596
1.614
1.638
1.663

0.83
0.79
1.17
2.46
0.50
0.11
0.38
0.34
0.31
0.11

0.166
0.194
0.237
0.165
0.040
0.010
0.031
0.025
0.023
0.017

8,10,16/8,12,14/10,10,14
6,10,18/6,12,16/6,14,14

49
50

C36

1.697
1.718

2.73
2.32

0.182
0.205

51

1.746

3.29

0.409

52
53
54
55
56

1.764
1.786
1.810
1.834
1.857

1.25
0.55
0.49
0.36
0.25

0.061
0.023
0.023
0.048
0.013

8,12,16/8,14,14/10,12,14
6,12,18/6,14,16/8,10,18 : 1/10,
10,16 : 1
4,14,18/4,16,16/6,12,18 : 1/6,
14,16 : 1
4,14,18 : 1/4,16,16 : 1
4,15,18/4,16,17
4,16,17/4,16 : 1,16 : 1
4,16,17/4,14 : 1,18 : 2

1.892
1.921

3.69
3.43

0.276
0.301

1.946
1.971
1.983
1.994
2.005
2.012
2.038
2.062

2.00
3.14
0.04
0.03
0.37
0.42
0.48
0.21

0.109
0.172
0.012
0.007
0.024
0.037
0.024
0.044

2.097

5.20

0.513

2.128

2.39

0.135

57
58

C38

59
60
61
62
63
64
65
66
67
68

C40

4,10,i15

4,8,18 : 1
6,10,15

Peak
no.

RRT

mol% SD

Triglycerides identi"ed

2.154
2.181
2.192
2.206
2.221
2.245

2.34
0.81
0.48
0.85
0.53
0.70

0.128
0.150
0.056
0.081
0.030
0.038

4,18,18/6,16,18 : 1/8,16,16 : 1
4,18,18 : 1

2.308
2.333

4.64
2.14

0.510
0.257

2.344
2.364
2.392
2.405
2.432
2.476

1.61
1.01
0.69
0.44
0.96
0.26

0.257
0.072
0.042
0.031
0.085
0.054

10,14,18/10,16,16/12,14,16
10,14,18 : 1/10,16,16 : 1/12,14,
16 : 1
8,16,18 : 1
6,18,18 : 1
8,16 : 1,18 : 1/10,14,18 : 2
6,18 : 1,18 : 1
8,17,18 : 1/8,16 : 1,18 : 2
6,18 : 1,18 : 2

2.515

2.69

0.403

84

2.545

4.13

0.434

85
86
87
88
89

2.587
2.603
2.618
2.638
2.686

1.61
0.17
0.13
0.80
0.37

0.304
0.109
0.069
0.063
0.019

2.722
2.763

1.64
2.49

0.077
0.150

2.806
2.838
2.854
2.874
2.896
2.934

1.58
0.25
0.36
0.36
0.26
0.41

0.068
0.021
0.034
0.062
0.075
0.047

69
70
71
72
73
74
75
76

4,12,18/4,14,16
6,10,18 : 1/8,10,16 : 1
4,12,18 : 1
4,i15,16
4,ai15,16
4,15,16

8,14,16/10,12,16/10,14,14
6,14,18/6,16,16/8,12,18 : 1/
10,10,18 : 1/10,12,16 : 1
4,16,18/6,14,18 : 1/6,16,16 : 1
4,16,18 : 1/4,18,16 : 1
4,17,18/4,16 : 1,18 : 1
4,17,18/4,16,18 : 2
4,17,18
4,16 : 1,18 : 2
8,14,18/8,16,16/10,14,16/
12,14,14
6,16,18/10,12,18 : 1/10,14,16 : 1

Mean values of "ve herds, three replicates per herd.

Carbon number.
Standard deviation.
Underlined TG are the more abundant in the corresponding peak.

C42

77
78
79
80
81
82
83

4,14, ai15
4,14,15

CN

90
91

C44

C46

92
93
94
95
96
97

4,18 : 1,18 : 1
4,18,18 : 2
4,18 : 1,18 : 2

8,18,18/10,16,18/12,14,18/12,
16,16/14,14,16
8,18,18 : 1/10,16,18 : 1/12,14,
18 : 1
8,18 : 1,18 : 1/10,16 : 1,18 : 1
8,18 : 1,18 : 2
12,16,18/14,16,16
10,18,18 : 1/12,16,18 : 1/14,14,
18 : 1
10,18 : 1,18 : 1
10,18 : 1,18 : 2
14,16,17

98
99
100
101
102
103
104

C48

2.977
3.026
3.068
3.084
3.129
3.165
3.239

1.09
2.13
0.55
0.50
0.44
0.55
0.34

0.100
0.079
0.072
0.045
0.033
0.052
0.040

14,16,18/16,16,16
14,16,18 : 1
12,18 : 1,18 : 1

105
106
107
108
109
110
111

C50

3.289
3.350
3.403
3.426
3.480
3.526
3.589

0.79
2.24
0.92
0.54
0.44
0.37
0.36

0.174
0.300
0.080
0.044
0.091
0.077
0.125

16,16,18
14,18,18 : 1/16,16,18 : 1
16,16 : 1,18 : 1
14,18 : 1,18 : 1

112
113
114
115
116
117
118

C52

3.676
3.751
3.826
3.869
3.939
4.024
4.073

0.30
1.15
1.53
0.33
0.44
0.13
0.18

0.123
0.411
0.306
0.062
0.203
0.085
0.124

16,18,18
16,18,18 : 1
16,18 : 1,18 : 1

119
120
121
122
123
124

C54

4.175
4.272
4.368
4.459
4.507
4.606

0.11
0.25
0.43
0.38
0.11
0.08

0.061
0.130
0.224
0.112
0.063
0.028

16,18 : 1,18 : 2
16,18 : 2,18 : 2
18,18,18 : 1
18,18 : 1,18 : 1
18 : 1,18 : 1,18 : 1
18,18 : 1,18 : 2

J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

125

Fig. 4. Partial gas chromatograms of triglycerides (CN36 zone) of goat's milk fat (GMF) and of the AgNO -TLC fractions: A: saturated (C6 and

longer chain fatty acids); B: saturated (C4 and longer chain fatty acids); C: monounsaturated; D: polyunsaturated. Peak numbers are identi"ed in
Tables 3 and 4.

Fraga et al. (1998), the SSMs in goat's milk fat were

located in a single TLC fraction, probably because of the
low molar percentages of butyric acid in the SSM fraction (4.4%, Table 2) in comparison with cow's milk fat. In
any event, separation of the SSMs has not always been
observed in cow's milk (Lund, 1988; Kemppinen & Kalo,
1993).
The percentage of polyunsaturated TGs (16%) was
lower than that was found in cow's milk (20%) by Fraga
et al. (1998), whereas the percentage of monounsaturated
TGs was similar in both the species. These results con"rm that goat's milk contains a lower concentration of
total unsaturated TGs (45%) than cow's milk (52%;
Fraga et al., 1998); a slightly low inter-species di!erence
was reported by Ruiz-Sala et al. (1996) (51 vs. 55%).
The study of the fatty acid composition of the TGs
fractions indicated that the individual saturated fatty
acids of goat's milk fat are distributed into saturated,
mono- and polyunsaturated TGs in a very similar proportion (mean values: 66, 25 and 9%, respectively). This
distribution agrees with that theoretically calculated as-

suming that the saturated fatty acids are included in the

di!erent TLC fractions independent of their chain length.
The saturated TGs have a unimodal molar distribution with maxima at CN40 and CN36 in fractions A and
B, respectively (Fig. 2). In fraction B, the sum of TG from
CN22 to CN38 made up 90% of the total fraction, as
expected in a band that mainly contains butyrates. The
proportion of SSSs in short-chain TGs (CN28}CN38)
found in this study (60%) was higher than in cow's milk
containing 49% of SSSs in short-chain TGs (Fraga et al.,
1998). This di!erence links up with the need to obtain
a TG composition with the appropriate melting point
(Gresti et al., 1993) to allow the fat to be secreted. Thus,
although the percentage of saturated TGs is higher in
goat's milk than the cows milk, they have a shorter chain
length.
The distribution of SSMs was similar to that of total
TGs in goat's milk fat. Most of the SSMs were found in
TGs CN38}CN50, and only 17% were of long chain.
The proportions of polyunsaturated TGs (band D) increased with the CN and represented from 39 to 89% in
the long-chain TGs. This distribution also favours an

126

J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

Table 4
Location of the C36 TGs of the goat's milk fat in the AgNO -TLC fractions according to the relative retention time (RRT) to C27

Peak no.

49

CN

C36

RRT

1.697

50

1.718

51

1.746

52

1.764

53

1.786

54

1.810

55

1.834

56

1.857

AgNO -TLC fractions


A

8,12,16/8,14,14/10,12,14
(6.98)
6,12,18/6,14,16
(4.20)

8,12,16
(0.86)
6,14,16
(1.84)
4,14,18/4,16,16
(16.10)

NI

4,15,18/4,16,17
(1.70)
4,16,17
(Tr)
4,16,17
(0.77)

8,10,18 : 1/10,10,16 : 1
(2.24)
6,12,18 : 1/6,14,16 : 1
(1.36)
4,14,18 : 1/4,16,16 : 1
(2.49)
NI

NI

4,16 : 1,16 : 1
(1.37)
4,14 : 1,18 : 2
(1.95)
NI

Carbon number.
NI, not identi"ed TGs (see text).
In parentheses are the molar percentages in each fraction of TG identi"ed.

appropriate melting point because a higher degree of

unsaturation will o!set the e!ect of a longer chain on the
melting point of TGs.
In qualitative terms, the chromatogram for TGs in
caprine's milk fat shown in Fig. 3 is similar to that for
bovine's milk published by Fraga et al. (1998) using
the same experimental conditions. All the TGs listed in
Table 3 have been identi"ed previously in cow's milk fat
(Myher et al., 1988; Gresti et al., 1993), but a comparison
with the papers cited suggests that there are considerable
quantitative di!erences from one species to another. Similar "ndings were reported by Barron et al. (1990) and
Ruiz-Sala et al. (1996) studying simultaneously cow's and
goat's milk. Using the same procedure (HPLC and GC),
the two groups of workers respectively identi"ed 116 and
181 molecular species of TG in milk from both species.
Sixty three in the "rst case and 84 in the second case
coincided with the ones identi"ed in the present study.
Most of the TGs described by these authors and not
identi"ed here are unsaturated TGs, particularly TGs
containing trans-fatty acids or branched-chain TGs.
As in previous studies it was not possible to quantify
the TGs in terms of molecular species because, as mentioned, some peaks in chromatogram in Fig. 3 contained
more than one molecular species. According to Table 3,
only the three peaks (67, 75 and 84, located in TGs CN40,
CN42 and CN44, respectively) individually exceeded 4%
of the total content. All the TGs identi"ed in these peaks
contained one of the three acids C8, C10 and C12; in all
cases C18 : 1 was the unsaturated acid in the monounsaturated TGs. Mass spectrometry analysis showed that
TGs 10,14,16; 10,16,16 and 10,16,18 : 1 are probably the

most abundant in goat's milk. In cow's milk, using

the same experimental conditions (Fraga et al., 1998), the
peaks for the corresponding RRTs made up only 3.6% of
the total fat. Similarly, Barron et al. (1990) found the
greatest di!erence between goat's and cow's milk in the
TGs with partition numbers from 38 to 42, which contained, among others, the cited TGs. And again, when
GC analyses in TG class of goat's and cow's milk fat were
compared (Fontecha et al., 1998), the largest quantitative
di!erences were found for TGs CN40, CN42 and CN44.
There were other six peaks of over 2.5% located in the
TG classes from CN36 to CN44. Four of these (49, 57, 58,
and 83) contain one or more of the fatty acids C6, C8,
C10 and C12. Particularly abundant are the TG species
containing two identical acyl moieties (10,14,14; 6,16,16
and 14,14,16). The other two peaks (51 and 60) preferentially contain 4,16,16 and 4,16,18 : 1, respectively. In
cow's milk, Gresti et al. (1993) also observed a high
proportion of 6,16,16 which was higher than the corresponding random proportion; however, the high proportion of 10,14,14 was not detected in cow's milk, probably
because it contained less C10. In cow's milk from the six
peaks with equivalent RRT only the last two peaks which
contain butyric acid (molar percentages of total fat 4.4
and 2.2, respectively) are comparable to those of goat in
quantitative terms (Fraga et al., 1998).
Further 21 peaks with molar percentages '1% were
found, three of them (85, 92 and 114) were located preferentially in band D. The majority of these TGs contain
two molecules of oleic acid; TG 16,18 : 1,18 : 1 was the
most abundant polyunsaturated TG, as in cow's milk
(Fraga et al., 1998). This TG is also the most abundant in

J. Fontecha et al. / International Dairy Journal 10 (2000) 119}128

human milk, where it constitutes about 10% of all TGs

(Currie & Kallio, 1993).
The proportions of the peaks where the TGs
16,18 : 1,18 : 1 and 14,18 : 1,18 : 1 were located (114 and
108) in the present study were about 2.5}3 times smaller
than in cow's milk (Fraga et al., 1998). These TGs are
negatively correlated with the butter "rmness (Bornaz,
Fanni & Parmentier, 1993). Other medium-chain TGs
associated with texture (10,12,18 : 1 and 8,16,18 : 1,
located in peaks 68 and 77) exhibited higher proportions
in goat's milk than in cow's milk.
As regards the variability of TG proportions in the
di!erent herds, the standard deviations for each TG class
from CN28 to CN48 were low; the corresponding variation coe$cients (VC) ranged from 10 to 12%. However,
TGs CN50, CN52 and CN54 showed high VC (mean
values: 21, 44 and 45%, respectively) as a consequence of
the high standard deviations in each peak. There has
been little published on the variation in molecular species
of TGs of cow's milk fat, but the data of Bornaz, Novak
and Parmentier (1992) using HPLC show signi"cant regional and seasonal variations, especially for unsaturated
TGs CN38, CN40, CN50 and CN52, which is consistent
with our own results. Also, in a study of seasonal variation
Hinrichs, Heinemann and Kessler (1992) found that most
of the more-variable TGs contained C18 : 1.
5. Conclusions
The distribution of TGs in goat's milk fat by CN and
the degree of unsaturation can be successfully determined
using a combination of the AgNO -TLC technique and

capillary GC. Fifty "ve percent of total goat's milk fat
TGs are trisaturated, and a third of these contain butyric
acid. The bulk of monounsaturated TGs are of medium
chain length; only 17% are of long chain. The percentage
of polyunsaturated TGs (16% of total TGs) increases
with the CN and is high (mean value 52%) in TGs
CN46}CN54. The higher proportion of short-chain TGs
in the trisaturated TGs as compared to more unsaturated
TGs in cow's milk fat relates to the need to obtain a TG
composition with the appropriate melting point to allow
the fat to be secreted.
As in previous studies, it was not possible to quantify
molecular species of TG since some peaks in the
chromatogram contained more than one TG. However,
137 molecular species were identi"ed: 50% trisaturated,
30% mono- and 20% polyunsaturated. The most important in quantitative terms were the medium-chain TGs
containing C8, C10 or C12, and the C18 : 1 as unsaturated fatty acid. Although the qualitative chromatographic pro"le of goat's milk fat TGs is similar to that of
cow's milk, there are noticeable quantitative di!erences
in the TGs of low or medium molecular weight associated with the short- and medium-chain fatty acids that
are most abundant in either animal species.

127

Acknowledgements
This work was supported by a grant from Comunidad
de Madrid (Project 06G/049/96) and from FundacioH n
Alfonso MartmH n Escudero.

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