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Fatty acid profile of milk from Saanen goats fed a

diet enriched with three vegetable oils
M. Matsushita a, , N.M. Tazinafo a , R.G. Padre a , C.C. Oliveira a ,
N.E. Souza a , J.V. Visentainer a , F.A.F. Macedo b , N.P. Ribas c
a

Department of Chemistry, State University of Maringa, Av. Colombo, 5790, CEP 87020-900 Maringa, Parana State, Brazil
Department of Animal Science, State University of Maringa, Av. Colombo, 5790, CEP 87020-900 Maringa, Parana State, Brazil
c Dairy Livestock Program, Laboratory of the Paranaense Association of the Holstein Breed Cattle Breeders,
Av. Pres. Carlos Cavalcanti, 623, CEP 80510-040 Curitiba, Parana State, Brazil
Received 18 October 2005; received in revised form 21 August 2006; accepted 13 September 2006

Abstract
Characterization of fatty acid profiles and physico-chemical parameters of milk samples from Saanen goats fed diets enriched
with 3% of three different vegetable oils (soybean, canola and sunflower) were carried out. Animals were arranged in a double 3 3
Latin square design and each pair of goats received diets containing one of the oils for 21 days14 days for animal adaptation plus
7 days for milk sampling. Samples were collected twice a day. Milk from animals that received sunflower oil presented the highest
conjugated linoleic acid (CLA) concentrations, whereas animals receiving canola oil had the lowest levels. Animals treated with
soybean oil had the highest monounsaturated and polyunsatured fatty acids concentrations and the lowest concentration of saturated
fatty acids. The n 6/n 3 ratios were 3.90, 5.77 and 4.24 for milk of animals treated with soybean, canola and sunflower oils,
respectively. Significant differences (P < 0.05) between soybean and canola oil and between canola and sunflower oil treatments
were observed. It is concluded that nutritional milk quality can be improved by adding vegetable oils to animal feed.
2006 Elsevier B.V. All rights reserved.
Keywords: Goat milk composition; Polyunsaturated fatty acids; Conjugated linoleic acid; Monounsaturated fatty acids

1. Introduction
Conjugated linoleic acids (CLA), a group of positional and geometric isomers of omega-6, is a essential
fatty acid and has recently been shown to have positive health effects in experimental models related to
suppression of carcinogenesis (Ip, 2001; Parodi, 2002;
Shingfield et al., 2003), antiobesity (Pariza et al., 2001;
Evans et al., 2002; Ritzenthaler et al., 2004), modulation

Corresponding author. Tel.: +55 6 44 3261 3655;

fax: +55 44 32614125.
E-mail address: mmakoto@uem.br (M. Matsushita).

of the immune system (Cook et al., 1993), atherogenesis (Nicolosi et al., 1997) and diabetes (Sebedio et al.,
1999). Although cis-9, trans-11 CLA is considered the
key to this anticarcinogenic effect (Pariza et al., 2001;
Ip et al., 1991), the specific CLA isomer(s) responsible for these benefical biological effects is unidentified
as yet.
Goat milk is an alternative for infants and adults sensitive or allergic to cow milk (Mir et al., 1999) but, in
contrast to cow milk, there is little information on ovine
or caprine milk, trans fatty acid content and compositional variations, although Bickerstaffe et al. (1972) and
Ledoux et al. (2001) have reported the existence of cisand trans-18:1 acids in goat fat.

0921-4488/$ see front matter 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.smallrumres.2006.09.003

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As characteristics of milk fat are influenced by dietary

supplements fed to animals and that diet supplementation with polyunsaturated oils, containing either linoleic
acid or linolenic oil, can increase the CLA content in
milk (Dhiman et al., 1999; Bauman et al., 2000; Jones
et al., 2000), the aim of this study was to determine the
composition and fatty acid profile, including CLA, in
natural milk from goats fed diets enriched with 3% soybean, canola or sunflower oils.
2. Material and methods
2.1. Animals, treatments and sampling
The study was conducted at the Experimental Farm of the
State University of Maringa. First lactating Saanen goats were
distributed in a double 3 3 Latin square, where each pair
received an isoprotein, isoenergetic, isocalcic and isophosphoric ration containing different vegetables oils (Table 1) for 21
days14 days for adaptation period plus 7 days more for
sampling.
Samples were collected twice a day, in the morning
(07:00 h) and afternoon (16:00 h), on day 15, 18 and 21 and
stored in sterile polyethylene flasks. Samples were divided
into two aliquots, one for immediate determination of physicochemical parameters (pH and density) determined immediately
and the samples frozen at 18 C. In the second sample,
aliquots were conditioned in sterile flasks containing bronopol
(2-bromo-2-nitro-propane-1,3-diol) for later total lipids, crude
protein, lactose and total solids analysis.

The pH of fresh milk was determined in a pH-meter, with

combined glass electrodes, previously calibrated at 20 C using
Table 1
Composition of the diets
Treatments
Sb
Calcium (%)
Soybean meal (%)
Oats hay (%)
Dicalcium phosphate (%)
Corn (%)
Mineral supplements (%)
Soybean oil (%)
Canola oil (%)
Sunflower oil (%)
Total (%)

2.3. Fatty acid methyl esters analyses

Total lipids were extracted from milk samples after defrosting and centrifugation (Murphy et al., 1995). Triacylglycerols
were submitted by transmethylation to methyl esters using the
5509 ISO method (1978).
Fatty acid methyl esters (FAMEs) were analyzed using a
Shimadzu 14A (Tokyo, Japan) gas chromatograph equipped
with a flame ionization detector and Varian (Varian, Inc. Scientific Instruments, Palo Alto, CA, USA) fused-silica capillary column (100 m 0.25 mm and 0.20 m of biscyanopropil
polysiloxane, CP-Sil 88). Column temperature was programmed from 80 to 220 C at 4 C min1 . The injection port
and detector were maintained at 210 and 230 C, respectively.
Hydrogen at 1.2 ml min1 was used as carrier gas and nitrogen
at 30 min min1 was used as the make up gas in 1:100 split
mode.
Identification of fatty acids was carried out by comparing
relative FAME peak retention times of samples to standards
obtained for Sigma (St. Louis, MO, USA). Peak areas were
determined by a CG-300 computing integrator program (CG
Instruments, Brazil). Data were calculated as normalized area
percentage of fatty acids.
2.4. Quantication of conjugated linoleic acids

2.2. Chemical analysis

Composition

standard buffer solutions at pH 4.0 and 7.0. Prior to analysis, the

samples were shaken for 3 min, according to Cunniff (1998).
Milk density (g/ml) was determined using a Qu`evenne thermolactodensymeter. Crude protein, total lipids, lactose and % total
solids were analyzed using a Milk Infra Red (MIR), Bentley
model 2000 (Bentley Instruments Inc., Chaska, MN, USA).

Ca

Sf

0.55
32.00
30.00
0.45
33.50
0.50
3.00
0.00
0.00

0.55
32.00
30.00
0.45
33.50
0.50
0.00
3.00
0.00

0.55
32.00
30.00
0.45
33.50
0.50
0.00
0.00
3.00

100.00

100.00

100.00

Sb: 3% of soybean oil diet; Ca: 3.00% of canola oil diet; Sf: 3.00% of
sunflower oil diet. Source: Laboratory of Animal Science Department,
State University of Maringa, according to AFRC (1998) and NRC
(1996).

Quantification of conjugated linoleic acids (CLA) was

carried out using a Varian CP 3380 gas chromatograph
equipped with a flame ionization detector and Varian fusedsilica capillary column (100 m 0.25 mm and 0.39 m of
100% bonded cyano-propyl, CP-7420). Column temperature
was kept to 65 C for 8 min and programmed from 65 to 170 C
at 50 C min1 and maintaining it at this value for 40 min,
and from 170 to 240 C at 50 C min1 and maintaining for
28.5 min. The injection port and detector were maintained at
220 and 245 C, respectively. Hydrogen at 1.4 ml min1 was
used as carrier gas and nitrogen at 30 ml min1 was used as the
make up gas in 1:80 split mode.
Identification of conjugated linoleic acids was carried out
by comparing relative CLA peak retention times of samples
to standards obtained for Sigma (O-5632). Peak areas were
determined by Star Software from Varian and CLA content in
mg g1 was calculated as follows:
CLA (mg g1 LT) =

(Ax )(WIS )(CFx )

100
(AIS )(Wx )(1.04)

where Ax is the CLA peak area, AIS the internal standard (tricosanoic acid 23:0) peak area, WIS the the weight (mg) of the
internal standard, Wx the sample weight (mg), CFx the theo-

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retical correction factor calculated from the internal standard

(equivalent to 1/RRF, where RRF is the relative response factor = (Ax /AIS )(WIS /WES )), WES is the weight of the sample (mg),
1.04 = conversion factor necessary to express results as mg of
fatty acids g1 of lipids rather than as methyl ester (Mendoza
et al., 2005; Padre et al., 2006).
2.5. Statistical analysis
The samples were analyzed in triplicate. Statistical analyses
of the studied variables were interpreted by variance analysis
and mean values were compared by the Tukey test with 5%
of probability, following the mixed model procedure (StatSoft,
USA, 2005).
The model used in the analyses was:
Yijk = + Ti + Pj + eijk
where Yijk is the individual record, the overall mean, Ti the
oil effect (i = 1, 2, 3) and Pj the effect of the jth period (j = 1, 2,
3) and eijk is the residual.

3. Results
When physico-chemical parameters were evaluated
significant differences were observed among treatments (Table 2). Total lipids showed high variations
(CV = 8.32), whereas density was low (CV = 0.06). In

the present work, pH values between 6.56 and 6.58, and

a density of 1.03 g/ml were observed.
Mean values for milk protein (CP) were 3.09, 3.09
and 3.11% for soybean (Sb), canola (Ca) and sunflower
(Sf) oil treatments, respectively (Table 2). Total lipid
(TL) concentrations in milk samples were 4.27, 3.97
and 4.09% for the Sb, Ca and Sf treatments, respectively
(Table 2).
Regarding conjugated linoleic acid (CLA), differences between Sb, Ca and Sf treatments were observed
(P < 0.05, Table 3). CLA concentrations in milk were
25.70, 21.08 and 29.42 mg/100 g sample for Sb, Ca and
Sf treatments, respectively. Values are in agreement with
total lipids (Table 3) and with linoleic acid (Table 4)
determined in the different treatments (Table 4).
Fatty acid composition of milk from Saanen goats
receiving diets with 3% of three different vegetable
oils are presented in Table 4. For the Sb, Ca and Sf
oil treatments, butyric acid (4:0) concentrations were
0.83, 1.06 and 0.82%, respectively, with a CV of 39.71
(P > 0.05).
The Sb treatment differed (P < 0.05) from the Ca and
Sf treatments in total saturated fatty acids (SFA) and
total monounsaturated fatty acids (MUFA), but the Ca
treatment differed from Sb and Sf treatments in total
polyunsaturated fatty acids (PUFA) (Table 4).

Table 2
Proximate composition of milk samples from Saanen goats fed with diet containing 3% vegetable oilsa
Parameters

Soybean (Sb)
pH
Density (g/ml)
Total lipids (%)
Crude protein (%)
Lactose (%)
Total solid (%)

CVb

Oils added to diet

6.56
1.03
4.27
3.09
4.60
12.75

0.10a
0.00a
0.67a
0.26a
0.25a
1.04a

Canola (Ca)
6.58
1.03
3.97
3.09
4.55
12.41

0.15a
0.00a
0.67a
0.22a
0.24a
0.88a

Sunflower (Sf)
6.56
1.03
4.09
3.11
4.59
12.57

0.16a
0.00a
0.79a
0.21a
0.25a
1.08a

0.58
0.06
8.32
3.20
2.25
3.62

a Each value represents the average and its standard deviation from the analysis in triplicate of collected samples in the morning and afternoon
periods, expressed as the whole percentage. Different letters at the same line indicate differences (P > 0.05) according to Tukey test.
b CV: Variation Coefficient.

Table 3
Isomer cis-9 trans-11 octadecadienoic acid (CLA) of milk samples from Saanen goats fed a diet containing 3% different vegetable oilsa
CVb

Oils added to diet

CLAc

(mg/100 g milk)
CLA (mg/g TL)d
CLA (g/100 g (%) TL)d
a
b
c
d

Soybean (Sb)

Canola (Ca)

Sunflower (Sf)

25.70 1.39a
6.02 0.33a
0.92 0.05a

21.08 0.75b
5.30 0.12a
0.84 0.03a

29.42 1.60c
7.92 0.40b
1.10 0.06b

17.45
1.83
0.02

See footnote Table 2.

CV: coefficient of variation.
CLA: isomer cis-9, trans-11 octadecadienoic acid (conjugated linoleic acid).
TL: total lipids.

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Table 4
Fatty acid profile of milk samples from Saanen goats fed a diet containing 3% different vegetable oilsa
Fatty acids

CVb

Oils added to diet

Soybean (Sb)

Canola (Ca)

Sunflower (Sf)

4:0
6:0
8:0
10:0
12:0
14:0
14:1 n 5
15:0
15:1 n 10
16:0
16:1 n 7
17:0
17:1 n 9
18:0
18:1 n 9
18:1 n 7
18:2 n 6
18:3 n 6
18:3 n 3
20:0
20:2 n 6

0.83
1.36
2.09
8.36
3.88
8.27
0.10
0.34
0.69
24.00
0.88
0.37
0.49
13.14
27.58
2.62
3.43
0.34
1.01
0.23
0.16

0.51a
0.54a
0.51a
1.37b
0.69b
1.05a
0.01b
0.03a
0.12a
1.96a
0.09a
0.05b
0.07a
2.80a
2.80a
0.55a
0.54a
0.17a
0.22a
0.06c
0.04b

1.06
1.56
2.35
9.48
4.56
8.84
0.21
0.35
0.70
23.39
0.67
0.39
0.46
12.90
26.95
1.90
2.96
0.39
0.63
0.32
0.17

0.75a
0.83a
0.80a
2.54a
1.37a
1.90a
0.00b
0.06a
0.14a
1.75a
0.12b
0.10b
0.09ab
1.81a
3.03a
0.63b
0.58b
0.26a
0.17b
0.04a
0.08ab

0.82
1.44
2.22
9.22
4.57
9.03
0.22
0.33
0.66
23.36
0.68
0.44
0.42
13.20
24.95
2.40
3.42
0.25
0.94
0.28
0.19

0.35a
0.53a
0.87a
3.19ab
1.62a
0.87a
0.04a
0.03a
0.06a
2.72a
0.09b
0.11a
0.07b
3.06a
2.12b
0.66ab
0.35a
0.13a
0.22a
0.04b
0.09a

39.71
27.60
16.33
10.65
8.48
9.53
43.82
9.17
16.16
4.04
12.23
12.17
8.56
10.15
6.28
24.57
9.05
57.91
16.75
15.79
51.20

SFAc
MUFAd
PUFAe
n 6f
n 3g
PUFA/SFA
n 6/n 3

62.85
32.28
4.87
3.85
1.01
0.08
3.90

3.30b
2.87a
0.83a
0.66a
0.22a
0.02a
0.59b

65.20
30.69
4.11
3.48
0.63
0.06
5.77

4.44a
3.75b
0.91b
0.78a
0.17b
0.02b
1.15a

64.92
30.28
4.81
3.87
0.94
0.07
4.24

2.51a
2.24b
0.63a
0.44a
0.22a
0.01a
0.58b

2.57
4.79
11.72
11.40
16.75
13.01
14.22

a
b
c
d
e
f
g

See footnote Table 2.

CV: coefficient of variation.
SFA: total of saturated fatty acids.
MUFA: total of monounsaturated fatty acids.
PUFA: total of polyunsaturated fatty acids.
n 6: total of omega-6 fatty acids.
n 3: total of omega-3 fatty acids.

For total SFA, a greater palmitic (16:0) concentration

was observed in all treatments, but was not significant
(P > 0.05) (Table 4). Araquidic acid (C:20:0) presented
the lowest SFA concentration at 0.23, 0.32 and 0.28%
for the treatments Sb, Ca and Sf, respectively (P < 0.05).
In MUFA analysis, oleic acid (C:18:1n 9) levels differed (P < 0.05) between Sb and Sf, and Ca and Sf treatments, but there were no differences (P > 0.05) between
Sb and Ca treatments. For the Sb, Ca and Sf treatments,
C:18:1n 9 MUFA had concentrations of 24.95, 26.95
and 27.58%, respectively.
Linoleic acid (C:18:2n 6) was the main PUFA
found and differed (P < 0.05) between Sb and Sf, and
Ca and Sf treatments, but not (P > 0.05) between Sb and
Ca (Table 4). In the present study, n 6/n 3 ratios were

3.90, 4.24 and 5.77 for milk samples from goats receiving the Sb, Sf and Ca oil treatments, respectively.
4. Discussion
In this experiment, the results for physico-chemical
analysis are in agreement with others studies. Data for
pH and density are similar those observed by Chornobai
et al. (1999), who obtained average values of 6.7 and
1.03 g/ml for pH and density, respectively, in milk samples. Protein content is in agreement with Mir et al.
(1999), who found that addition of 4% canola oil to the
diet of dairy goats lead to a 3.64% CP in milk. Dhiman
et al. (2000) observed 2.93% TL in milk of cows fed 4%
soybean oil. Mir et al. (1999) also found 5.42% TL in

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milk of goats that received a diet containing 4% canola

oil. Chornobai et al. (1999) observed a 3.81% TL for
natural samples of goat milk.
Food products from ruminants are a major dietary
source of CLA for humans and there is consensus
that CLAs are intermediates in the biohydrogenation of
linoleic acid. It is generally accepted that CLA in ruminants originates from the incomplete biohydrogenation
of unsaturated fat by rumen bacteria (Griinari et al.,
2000), although recent studies have demonstrated that
cows can also synthesize CLA from trans-11 octadecenoic acid, another intermediate in the rumen biohydrogenation process (Griinari et al., 1998). All cisand trans-isomeric combinations of CLA have been
identified in food, but cis-9, trans-11, octadecadienoic
acid (c-9, t-11 CLA) are the most common, although
the presence of others CLA isomers and their probably concentration vary according to rumen conditions (Ha et al., 1989; Griinari et al., 1997; Kelly,
2001).
The values to CLA indicate significant difference
between treatments, which are in agreement with the
total lipids content and linoleic acid. Giesy et al. (2002)
found 6.46 mg/g TL for the cis-9 trans-11 CLA isomer and 1.28 mg/g TL for the trans-10 cis-12 isomer
in samples of cow milk, where the animals had been
fed 100 g/day CLA, protected by calcium salt (CLA60; Natural Lipids) with 24% cis-9 trans-11 and 35%
trans-10 cis-12 CLA isomer. Dhiman et al. (1999) found
8.6 mg CLA/g TL in samples of milk where the cows had
received a ration with 12% full-fat extruded soybean oil.
Peterson et al. (2002) found 0.80 g CLA/100 g TL in
samples of Holstein cow milk, where the animals had
received a diet with 13.4% full-fat extruded soybean oil.
Lawless et al. (1999) determined CLA in milk of different breeds of dairy cows, finding 17.0 mg CLA/g TL in
Irish Holstein breed, 14.7 mg CLA/g TL in Dutch breed,
18.3 mg CLA/g TL in Montbeliarde breed and 15.5 mg
CLA/g TL in Normande breed.
The fatty acid, SFA, MUFA and linoleic acid composition are in accordance with literature reports.
Chornobai et al. (1999) found 1.58% butyric acid in
the milk of goats fed diets without vegetable oil enrichment. Mir et al. (1999) observed 359.1 mg C:16:0 per
g TL in milk samples of goats treated with 4% canola
oil. Dhiman et al. (2000) reported that when 4% soybean oil was added to the dairy diet of cows, the C:16:0
concentration was 31.8%. Mir et al. (1999) reported
MUFA concentrations of 27.90% C:18:1n 9 in milk of
goats fed diets with 4% canola oil, while Jenkins (2000)
reported a C:18:2n 6 concentration of 1.58% for milk
of cows fed diets with 3.5% canola oil. Depeters et al.

(2000) found 3.29% C:18:2n 6 in milk of cows on diets

of 1.6% of canola oil.
The UK Health Department (Simopoulos, 2004) recommends 4.0 as the maximum value for the n 6/n 3
ratio. In the present study, n 6/n 3 ratio of 3.90, 4.24
and 5.77 were found for milk samples of goats that
received Sb, Sf and Ca treatments, respectively Ca
treatment giving to best results.
5. Conclusion
Among all treatments, soybean oil is recommended
for addition to goats ration due to its cost/benefit ratio,
ease of consumption and market availability.
Acknowledgments
The authors wish to thank CAPES, CNPq and
Fundaca o Araucaria for financial support and J.A. Horst
and D.R. Veiga from the Laboratory of PARLPR of
APCBRH, Curitiba, Pr, Brasil and C. Volpato from the
Laboratory of Nutrition and Animal Science, UEM, Pr,
Brazil for assistance in laboratory analysis.
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Please cite this article in press as: Matsushita, M. et al., Fatty acid profile of milk from Saanen goats fed a diet enriched with
three vegetable oils, Small Rumin. Res. (2006), doi:10.1016/j.smallrumres.2006.09.003