Journal of Food Composition and Analysis 63 (2017) 121132

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Journal of Food Composition and Analysis

journal homepage: www.elsevier.com/locate/jfca

Original research article

Changes in the chemical and physical characteristics of cows milk butter MARK
during storage: Eects of temperature and addition of salt

Francisco J. Mndez-Cid, Juan A. Centeno, Sidonia Martnez, Javier Carballo
rea de Tecnologa de los Alimentos, Facultad de Ciencias, Universidad de Vigo, 32004 Ourense, Spain

A R T I C L E I N F O A B S T R A C T

Keywords: The eect of storage at 4 C or 12 C on cows milk butter manufactured without or with salt (2.1%) was ex-
Butter colour amined. Storage of the butter for 9 months scarcely aected the fatty acid contents, and only the amounts of
Butter composition C18:3 n3, C20:5 n3 and C22:2 n6 decreased signicantly in the total lipid fraction. The total amount of free
Cows milk butter fatty acids increased signicantly during storage (from 136167 to 360575 mg/100 g of fat), and both the
Fat autooxidation
addition of salt and the higher storage temperature enhanced the release of fatty acids from lipids. The free fatty
Fat stability
acid contents (360575 mg/100 g of fat) and the values of the parameters indicating lipolysis (acidity value:
Fatty acids
Food analysis 1.892.18 mg KOH/g of fat; degree of acidity: 0.951.09% oleic acid) at the end of the storage period indicate
Food composition that butter undergoes very slight lipolysis. The values of parameters indicating fat oxidation (peroxide value and
Salt content TBA value) increased signicantly during storage (from 0.380.39 to 0.903.75 meq. O2/kg of fat, and from
Storage temperature 0.0840.1 to 0.0930.220 mg malondialdehyde/kg, respectively), and both the addition of salt and the increase
in storage temperature had enhancing eects on the oxidative processes. The b* values increased signicantly
during storage, while a* and L* values decreased. In conclusion, cows milk butter was hardly modied during
refrigerated storage, while both the addition of salt and the increase in storage temperature increased lipolytic
and oxidative changes.

1. Introduction aecting butter during storage is rancidication. This process, caused

Milk and dairy products are important sources of nutrients and
energy in human diets. Milk fat is probably the most complex of all
edible fats, because of its physical and chemical characteristics. It is
mainly present in globules as an oil-in-water emulsion and consists of
triglycerides (98%), diacylglycerol (2%), cholesterol (0.5%),
phospholipids (1%) and free fatty acids (0.1%) (Lindmark
Mansson, 2008). Milk fat contains more than 400 dierent fatty acids
(Collomb and Blher, 2001), including saturated fatty acids (66%),
monounsaturated fatty acids (30%) and polyunsaturated fatty acids
(4%) (Aigster et al., 2000).
Butter is the most widely appreciated and manufactured milk fat
product. It contains at least 80% fat and a maximum of 16% water. It is
an emulsion of water in oil in which the water droplets are dispersed in
the continuous, partially crystallized, fat phase. Butter is an important
product in the dairy industry because of its particular sensory attributes
and nutritional value. The characteristics and quality of butter have
been the object of numerous studies (e.g., Ledoux et al., 2005; Mallia
et al., 2008).
Butter is often stored for long periods of time. The main problem
by lipolysis (release of free fatty acids) and oxidation of the fatty acids,
impairs the avour and lowers the nutritional quality of butter, thus
creating serious problems and economic losses in the dairy and food
distribution industries. Lipolytic changes occur in milk fat as a result of
the hydrolysis of triacylglycerols by lipases (indigenous milk enzymes
and enzymes of microbial origin). Lipolysis triggers the accumulation of
free fatty acids (FFA), which can cause o-avours described as rancid,
butyric, bitter, unclean, soapy or astringent (Ray et al., 2013).
Fat oxidation is a problem in the dairy industry and has important
implications in terms of product value, as it aects the avour, colour,
texture and nutritional value of dairy products (Nawar, 1996). Poly-
unsaturated fatty acids, which contain multiple double bonds with
particularly reactive hydrogen atoms, are prone to oxidation. A high
content of unsaturated fatty acids in milk fat increases the risk of oxi-
dation and production of o-avours (Lin et al., 1996; Im and Marshall,
1998). In dairy products with a high content of unsaturated fatty acid,
oxidation causes metallic, oily or stale avours and a paler colour,
especially after storage (Timmons et al., 2001). The quality of milk and
dairy products (including butter) is the result of a delicate balance
between pro- and antioxidant compounds and processes that are

Corresponding author.
E-mail address: carbatec@uvigo.es (J. Carballo).

http://dx.doi.org/10.1016/j.jfca.2017.07.032
Received 18 February 2017; Received in revised form 27 June 2017; Accepted 18 July 2017
Available online 23 July 2017
0889-1575/ 2017 Elsevier Inc. All rights reserved.
F.J. Mndez-Cid et al.
inuenced by factors such as the fatty acids degree of unsaturation, and
contents of transition metal ions and of antioxidant compounds (e.g.,
tocopherols and carotenoids) (Barrefors et al., 1995).
Oxidative reactions are chemical reactions with low activation en-
ergy and they cannot be stopped by lowering the storage temperature
(Andreo et al., 2003). High temperatures, and exposure to light and
oxygen enhance oxidative processes (Yildiz et al., 2002) and decrease
the nutritional value and customer acceptability.
Salt is sometimes added to butter with the aim of preservation, as
well as to provide a particular desired avour. Sodium chloride has
been shown to be a lipid pro-oxidant factor in dierent foods and food
systems, and chloride has been identied as the active component in
this reaction (Osinchak et al., 1992). Commercial salt also contains
metal ion impurities that enhance its pro-oxidant eect; metals act as
catalysts lowering the activation energy of the lipid oxidation reactions,
thus increasing the rate at which these reactions take place (Frankel,
2012).
Many dierent aspects of the composition and attributes of butter
have been studied. However, no information is currently available as to
how the stability of butter is aected by temperature during re-
frigerated storage or by the addition of salt in the manufacture process.
The main aims of this study were therefore to characterise the lipid
fractions of butter, to investigate the physical and chemical changes
that take place during storage of butter and to examine how storage
temperature and the addition of salt aect these changes.
2. Materials and methods
2.1. Samples and experimental design
The butter used in this study was supplied by Mantequera de Tineo,
S.A. (Tineo, Asturias, Spain). Butter was manufactured using the NIZO
butter process in a Contimab Major (Simon Frres, Crebourg-Octeville
Cedex, France) device. Cream containing 40 2% fat was rstly
pasteurised at 100 C for 15 s. Then, it was cooled to 34 C, and aged at
this temperature for 24 h, in order to crystallise the butterfat. Next,
churning was carried out at 10 C and 4001000 rpm. After draining,
butter was washed and worked. During working, an aromatic starter
culture (Lactococcus lactis subsp. cremoris and Leuconostoc mesenteroides
subsp. cremoris) was added. Salt (2.1%) was optionally added during
working.
Butter pieces (250 g) manufactured with NaCl (2.10%) or without
NaCl were used for this study. Batches of 10 pieces of unsalted butter
and of 10 pieces of salted butter were analysed. Five pieces from each
batch were stored at 4 C and ve were stored at 12 C. Samples from
each batch stored at each temperature were removed for analysis after 0
(i.e. at the beginning of the study), 1, 3, 6 and 9 months. Each sample
consisted of an entire piece of butter. The experiment was conducted in
triplicate with batches manufactured on dierent days in the winter
season.
At the end of each storage time, pieces were removed for mea-
surement of the colour parameters and were then minced and stored at
80 C for no more than 3 months until completion of all analyses.
2.2. Determination of the proximate composition
Moisture and fat contents were assessed using the IDF recommended
standards 80-1:2001 (IDF, 2001) and 9C:1987 (Rose-Gottlieb reference
method) (IDF, 1987) respectively. Salt content was assessed following
the IDF 12:2004 standard (IDF, 2004).
2.3. Determination of fatty acid content of the dierent fractions
For the determination of the fatty acid content of the total fat, about
5 mg of butter were weighed in a test tube. Milk fat was then dissolved
by addition of 50 L of hexane. Lipids were esteried by addition of
122
Journal of Food Composition and Analysis 63 (2017) 121132
100 L 2 N sodium methoxide for 20 min at room temperature. Five-
hundred L of sulfuric acid/methanol (1/1; v/v) was added and the
mixture was allowed to react for 20 min at room temperature (20 C).
Finally, fatty acid methyl esters were extracted by addition of 2 mL
hexane.
For the determination of the fatty acid content of the dierent lipid
fractions, 0.2 g of butter were dissolved in 1 mL of trichloromethane,
and the three dierent lipid fractions (neutral lipids, polar lipids and
free fatty acids) were separated for further analysis in NH2-aminopropyl
minicolumns DISCOVERY DSCNH2 (Supelco Inc., Bellefonte, PA), ac-
cording to the method described by Kaluzny et al. (1985). Neutral lipids
were transesteried just as the total lipids after evaporation of the
solvent. Polar lipids and free fatty acids were transesteried following
the method described by Shehata et al. (1970). The fatty acid methyl
esters from the dierent fractions were identied and quantied by gas
chromatography (Trace GC chromatograph; Thermo Finnigan, Austin,
TX) with split/splitless AI 3000 autoinjection and ame ionisation de-
tection. Samples (1 L) were injected in split mode. The dierent fatty
acids were separated on an Innowax column: length 30 m, internal
diameter 25 mm, lm thickness 0.25 m (Agilent Technologies, Santa
Clara, CA). The temperature of the detector was 250 C and that of the
injector 230 C. The gases used were hydrogen (35 mL/min), air
(335 mL/min) and helium (carrier gas) (30 mL/min). The chromato-
graphic conditions used for fatty acid determination were as follows:
50 C for 1 min; increasing at 5 C/min to 248 C; 248 C for 6 min.
For identication and quantication of the fatty acid methyl esters,
a standard (Sigma Chemical Co., St. Louis, MO) containing the methyl
esters of the following fatty acids was used: butyric (C4:0); caproic
(C6:0); caprylic (C8:0); capric (C10:0); undecanoic (C11:0); lauric
(C12:0); tridecanoic (C13:0); myristic (C14:0); myristoleic (C14:1);
pentadecanoic (C15:0); cis-10-pentadecenoic (C15:1); palmitic (C16:0);
palmitoleic (C16:1); margaric (C17:0); cis-10-heptadecenoic (C17:1);
stearic (C18:0); oleic (C18:1 cis); elaidic (C18:1 trans); linoleic (C18:2);
linolelaidic (C18:2 trans); linolenic (C18:3); arachidic (C20:0); cis-11-
eicosenoic (C20:1); cis-11,14-eicosadienoic (C20:2); cis-11,14,17-eico-
satrienoic (C20:3); arachidonic (C20:4); heneicosanoic (C21:0); behenic
(C22:0); erucic (C22:1); cis-13,16-docosadienoic (C22:2); cis-
4,7,10,13,16,19-docosahexaenoic (C22:6); tricosanoic (C23:0); lig-
noceric (C24:0); and nervonic (C24:1). The standard contained between
2 and 4% of each of these fatty acids. The individual fatty acids were
identied according to their retention times relative to standards. For
quantication of the free fatty acids using external standards, calibra-
tion curves were prepared using dierent concentrations of each methyl
ester. Fatty acids in total lipids, glycerides and phospholipids were
expressed as percentages of total identied fatty acid methyl esters,
while free fatty acids were expressed as mg/100 g of fat.
All samples and standards were injected into the gas chromatograph
at least in duplicate. Repeatability tests were carried out by consecutive
injection of a standard and a sample six times in one day.
Reproducibility tests were also performed by injection of standard and
sample twice a day for 3 days, under the same experimental conditions.
The results of these tests did not dier signicantly (p > 0.05).
2.4. Determination of fat indexes and colour parameters
The fat acidity, iodine, saponication and peroxide values were
assessed according to Spanish Ocial Standards UNE 50.011, UNE
55.013, UNE 55.012 and UNE 55.023, respectively (Spanish
Government Presidency, 1977). The refractive index was measured
with a Standard Abbe refractometer ER-2S (Shibuya Optical Co., Ltd.,
Saitama, Japan) thermostated by a circulating water bath at 40 C. The
TBA value was measured following the method of Tarladgis et al.
(1960), with some modications. Briey, 10 g of sample were homo-
genized with 50 mL of distilled water in a Sorvall Omni Mixer 17150
homogenizer (Ivan Sorvall Inc., Newtown, CT) for 2 min. The mixture
was transferred to a round-bottom ask and subsequently 47.5 mL of
F.J. Mndez-Cid et al. Journal of Food Composition and Analysis 63 (2017) 121132

SEM: Standard error of the mean. Signicance: R: signicance of the eect of the time of storage; T: signicance of the eect of the temperature; S: signicance of the eect of the salt content; R T: interaction of storage time and temperature;
distilled water, and 2.5 mL of 4 N hydrochloric acid, to adjust the pH to

RS

n.s.
a value of approximately 1.5, were added. Glass beads and a few drops

***

***
of an antifoaming agent were also added. The ask was placed in dis-

RT
tillation equipment, in which the mixture was boiled until 50 mL of

***
distillate were collected. Five mL of the distillate were then mixed in a

*
vial with 5 mL of a 0.02 M thiobarbituric acid solution. The vial was

***

***

***
labelled and heated in a water bath at 70 C for 40 min. The vial was

S
then cooled in a water bath and the absorbance was read at 532 nm.

n.s.
The reading was nally interpolated on a calibration curve (absorbance

***

***
T
vs concentration) prepared with malondialdehyde. The TBA values

***

***

***
were expressed as mg malondialdehyde/kg sample. All parameters

R
were determined at least in duplicate in each fat sample.

SEM

0.40
0.35
0.14
Colour parameters were measured using a portable colorimeter
(Chroma Meter Cr-400 Konica Minolta Sesing, Inc., Osaka, Japan). The
CIELAB space (CIE, 1978) is represented by lightness (L*), redness (a*)

12 C

95.7
81.4
2.47
and yellowness (b*). L* values (from 0 to 100 units) represent the
lightness of colour (a lower value indicates a darker colour, black:

Salted
L* = 0 and white: L* = 100), a* values represent the balance between

92.4
83.3
2.32
4 C
red (> 0) and green (< 0), and b* values represent the balance be-
tween yellow (> 0) and blue (< 0). The a*, b* and L* values were used

12 C

95.4
85.2
0.03
to calculate the chroma, hue angle, E and E values. Chroma (C*) de-

Unsalted
scribes the brightness or vividness of colour (colour saturation, starting
at 0 and increasing thereafter); C* = (a*2 + b*2)0.5. Hue angle (H)

92.5
85.5
0.03
4 C
9
describes the hue or colour angle (dened by its location in a 360 axis;

Total solid, fat and NaCl contents during the storage of cows butter: Eect of temperature and salt content. Values are means of three replicates (n = 3).
0 or 360 = red, 90 = yellow, 180 = green and 270 = blue);

12 C

95.4
83.2
2.63
H = arctan b*/a*. E describes the colour dierence (E =
[(L* L*ref)2 + (a* a*ref)2 + (b* b*ref)2]1/2). The E value re-

Salted
presents enhancement of the redness relative to the yellowness and

91.7
84.6
2.55
4 C
lightness; this parameter was calculated by means of an equation re-
written according to Liu et al. (2003) (E = a*/b* + a*/L*). The col-

12 C

95.1
84.4
0.03
orimeter was calibrated with a white ceramic tile before each series of
measurements. The colour of each piece of butter was measured ve
Unsalted

times at ve dierent points.

91.1
86.6
0.03
4 C
6

2.5. Statistical analysis

12 C

93.2
82.4
2.32
Experiments were done in triplicate, and three dierent samples
Salted

from each salt content, temperature storage and time of storage were
89.9
86.3
2.33
4 C

analysed (n = 3). The eects of the variables storage time, storage

temperature, salt concentration, storage time storage temperature,
12 C

91.6
86.2
0.03

and storage time salt concentration on the dierent parameters were

studied by analysis of variance (ANOVA) implemented using the
Unsalted

General Linear Model (GLM) procedure of SPSS version 19.0 for

88.4
87.8
0.03
4 C

Windows (SPSS Inc., Chicago, IL).

3

12 C

3. Results and discussion

91.3
85.1
1.77

R S: interaction of storage time and salt content; n.s.: not signicant.

3.1. Proximate composition

Salted

87.1
86.7
2.11
4 C
Storage time (months)

The total solid, fat and NaCl contents of the cows milk butter made
12 C

with and without salt and stored at two temperatures are shown in
87.2
90.4
0.03

Table 1. Total solid contents of the unsalted butter immediately after

Unsalted

manufacture (84.2%) are consistent with previously reported data

86.3
88.7
0.03
4 C

(Sagdi et al., 2004; Cronin et al., 2007; Mallia et al., 2008) and are
1

slightly higher than those reported in some specic cases (78.7% in

Expressed as g/100 g of total solids.

butter from cows fed with corn and soybean in the concentrate;
Salted

86.5
90.7
1.65

Middaugh et al., 1988). The values were signicantly (p < 0.001)

Fresh butter

higher in the salted (86.5%) than in the unsalted (84.2%) butter, due to
Unsalted

the contribution of salt to the total solids. Total solid contents increased
Expressed as g/100 g.

signicantly during storage (p < 0.001) and the values were higher in
84.2
92.0
0.03

butter stored at 12 C than in that stored at 4 C; higher temperatures

*** (p < 0.001).

seem to favour water losses during storage. Fat contents of fresh butter
* (p < 0.05).
Temperature

Total Solids

expressed as g/100 g of total solids (9092) were slightly lower than

previously reported values (ranging from 96 to 99%) for butter of
Table 1

NaClb
Fata
Salt

several dierent origins and characteristics (Middaugh et al., 1988;

b
a

Sagdi et al., 2004; Cronin et al., 2007; Mallia et al., 2008). The

123
 Table 2
F.J. Mndez-Cid et al.

Fatty acid composition (% of total methyl esters) of the total lipids during the storage of cows butter: Eect of temperature and salt content. Values are means of the results from three replicates (n = 3).

Fresh butter Storage time (months)

1 3 6 9

Salt Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted

Temperature 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C
Fatty acids SEM R T S R T RS

*** **
C4:0 2.37 2.36 2.37 2.38 2.36 2.34 2.38 2.38 2.33 2.43 2.36 2.34 2.31 2.48 2.36 2.37 2.27 2.50 0.008 n.s. n.s. n.s.
* ** *
C6:0 1.85 1.86 1.87 1.87 1.87 1.84 1.86 1.86 1.88 1.89 1.87 1.86 1.86 1.93 1.87 1.86 1.83 1.93 0.004 n.s. n.s.
C8:0 1.22 1.22 1.20 1.23 1.22 1.21 1.20 1.21 1.24 1.19 1.21 1.23 1.23 1.18 1.23 1.22 1.22 1.20 0.003 n.s. n.s. n.s. n.s. n.s.
** ** **
C10:0 2.84 2.82 2.74 2.86 2.80 2.81 2.74 2.79 2.91 2.72 2.78 2.86 2.87 2.63 2.85 2.87 2.85 2.69 0.011 n.s. n.s.
** ***
C12:0 3.48 3.50 3.42 3.53 3.48 3.49 3.42 3.48 3.59 3.39 3.45 3.55 3.54 3.30 3.52 3.61 3.50 3.36 0.012 n.s. n.s. n.s.
* * ***
C14:0 11.5 11.7 11.6 11.7 11.6 11.7 11.6 11.6 11.9 11.4 11.6 11.6 11.8 11.2 11.8 12.0 11.6 11.3 0.027 n.s. n.s.
*
C14:1 1.10 1.13 1.13 1.11 1.12 1.11 1.14 1.14 1.12 1.13 1.16 1.14 1.12 1.13 1.12 1.13 1.10 1.10 0.003 n.s. n.s. n.s. n.s.
* *
C15:0 1.09 1.10 1.09 1.09 1.09 1.10 1.09 1.10 1.10 1.08 1.09 1.09 1.09 1.08 1.10 1.10 1.09 1.08 0.001 n.s. n.s. n.s.
**
C15:1 0.27 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.24 0.25 0.25 0.25 0.25 0.25 0.001 n.s. n.s. n.s. n.s.
* * **
C16:0 33.0 33.3 33.1 32.9 33.0 33.3 33.1 33.0 33.2 33.0 32.8 32.8 33.2 33.2 33.4 33.1 33.2 32.9 0.029 n.s. n.s.
* ** *
C16:1 n7 1.82 1.82 1.82 2.08 1.89 1.85 1.83 1.89 1.87 1.80 1.92 1.88 1.88 1.80 1.86 1.83 1.92 1.87 0.010 n.s. n.s.
* ** *** **
C17:0 0.52 0.52 0.52 0.50 0.52 0.51 0.53 0.51 0.50 0.53 0.52 0.51 0.51 0.56 0.52 0.51 0.51 0.55 0.002 n.s.
* ** ***
C17:1 0.29 0.29 0.29 0.28 0.29 0.29 0.29 0.29 0.28 0.29 0.29 0.29 0.28 0.29 0.29 0.27 0.29 0.29 0.001 n.s. n.s.
** *
C18:0 9.84 9.67 9.71 9.67 9.71 9.67 9.66 9.61 9.52 9.82 9.59 9.59 9.57 10.0 9.66 9.72 9.76 9.88 0.020 n.s. n.s. n.s.

124
**
C18:1 n9 24.9 24.5 24.6 24.5 24.8 24.5 24.8 24.9 24.5 25.0 25.0 24.9 24.6 24.9 24.3 24.2 24.8 25.0 0.046 n.s. n.s. n.s. n.s.
C18:2 n6 2.25 2.22 2.31 2.25 2.25 2.28 2.30 2.27 2.26 2.28 2.31 2.29 2.26 2.24 2.28 2.23 2.23 2.24 0.006 n.s. n.s. n.s. n.s. n.s.
*** ** **
C18:3 n6 0.10 0.10 0.09 0.09 0.10 0.09 0.09 0.10 0.09 0.10 0.09 0.10 0.09 0.10 0.10 0.09 0.10 0.10 0.0004 n.s. n.s.
*** * *** ** ***
C18:3 n3 0.32 0.31 0.32 0.31 0.31 0.31 0.32 0.32 0.30 0.31 0.32 0.32 0.29 0.30 0.31 0.29 0.30 0.30 0.001
* ** **
CLA 0.65 0.72 0.83 0.76 0.72 0.77 0.75 0.76 0.66 0.79 0.80 0.74 0.66 0.73 0.70 0.73 0.65 0.80 0.008 n.s. n.s.
* *
C20:0 0.11 0.11 0.16 0.11 0.11 0.11 0.14 0.10 0.11 0.13 0.14 0.11 0.11 0.14 0.12 0.11 0.11 0.11 0.003 n.s. n.s. n.s.
*** **
C20:1 n9 0.14 0.14 0.15 0.14 0.14 0.14 0.14 0.14 0.13 0.15 0.15 0.14 0.13 0.15 0.14 0.14 0.14 0.15 0.001 n.s. n.s. n.s.
* ** * ***
C20:2 n6 0.016 0.015 0.015 0.015 0.015 0.015 0.015 0.016 0.015 0.015 0.016 0.016 0.014 0.015 0.015 0.014 0.015 0.015 0.0001 n.s.
*** *** *
C20:3 n6 0.10 0.10 0.09 0.09 0.09 0.08 0.09 0.10 0.08 0.09 0.09 0.09 0.08 0.10 0.09 0.09 0.08 0.10 0.001 n.s. n.s.
C20:4 n6 0.12 0.11 0.12 0.12 0.12 0.11 0.12 0.12 0.11 0.11 0.12 0.12 0.11 0.11 0.11 0.12 0.11 0.11 0.001 n.s. n.s. n.s. n.s. n.s.
*** *
C20:3 n3 0.011 0.010 0.011 0.003 0.014 0.004 0.012 0.009 0.004 0.009 0.011 0.010 0.004 0.015 0.008 0.015 0.004 0.012 0.001 n.s. n.s. n.s.
** *** *** * *
C20:5 n3 0.012 0.010 0.010 0.013 0.010 0.010 0.010 0.010 0.010 0.010 0.010 0.010 0.009 0.010 0.009 0.010 0.009 0.010 0.0002
** *
C22:2 n6 0.028 0.026 0.027 0.030 0.027 0.026 0.024 0.025 0.026 0.025 0.025 0.026 0.025 0.023 0.027 0.025 0.027 0.024 0.0003 n.s. n.s. n.s.
C24:0 0.070 0.065 0.068 0.068 0.057 0.066 0.068 0.066 0.065 0.068 0.068 0.067 0.064 0.068 0.068 0.067 0.063 0.069 0.001 n.s. n.s. n.s. n.s. n.s.
**
SFA 67.9 68.2 67.9 67.9 67.9 68.1 67.8 67.7 68.3 67.7 67.5 67.7 68.2 67.9 68.4 68.5 68.0 67.6 0.052 n.s. n.s. n.s. n.s.
**
UFA 32.1 31.8 32.1 32.0 32.1 31.9 32.2 32.3 31.7 32.3 32.5 32.3 31.8 32.1 31.6 31.5 32.0 32.4 0.052 n.s. n.s. n.s. n.s.
**
MUFA 28.5 28.1 28.3 28.4 28.5 28.2 28.5 28.6 28.1 28.6 28.7 28.6 28.3 28.5 27.9 27.9 28.5 28.7 0.048 n.s. n.s. n.s. n.s.
* *
PUFA 3.60 3.63 3.82 3.67 3.65 3.70 3.73 3.72 3.55 3.74 3.80 3.73 3.54 3.63 3.64 3.62 3.52 3.71 0.013 n.s. n.s. n.s.

SFA: sum of saturated fatty acids; UFA: sum of unsaturated fatty acids; MUFA: sum of monounsaturated fatty acids; PUFA: sum of polyunsaturated fatty acids. SEM: Standard error of the mean. Signicance: R: signicance of the eect of the time of
storage; T: signicance of the eect of the temperature; S: signicance of the eect of the salt content; R T: interaction of storage time and temperature; R S: interaction of storage time and salt content; n.s.: not signicant.
* (p < 0.05).
** (p < 0.01).
*** (p < 0.001).
Journal of Food Composition and Analysis 63 (2017) 121132
F.J. Mndez-Cid et al.
contribution of fat to the total solids content was signicantly lower in
the salted than in the unsalted butter (p < 0.001). Dierences in fat
content relative to previously reported data can be attributed to dif-
ferences in the manufacturing process used in each case.
3.2. Fatty acid contents
The fatty acid contents of the total lipids of the cows milk butter at
time 0, and the changes that occurred during storage for the two salt
contents and storage temperatures are shown in Table 2. The fatty acid
composition of milk fat depends on the animal species (Sagdi et al.,
2004) and feeding (Middaugh et al., 1988; Baer et al., 2001;
Ramaswamy et al., 2001; Mallia et al., 2008), and seasonal and geo-
graphical variations may also have some eects (Collomb et al., 2002;
Ledoux et al., 2005).
In fresh butter immediately after manufacture, the most abundant
fatty acid was palmitic acid (C16:0) (33%), followed by oleic acid
(C18:1 n9) (24.8%), myristic acid (C14:0) (11.5%), stearic acid
(C18:0) (9.8%), lauric acid (C12:0) (3.5%), capric acid (C10:0) (2.8%),
butyric acid (C4:0) (2.4%) and linoleic acid (C18:2 n6) (2.2%). The
presence of salt did not inuence either the fatty acid prole or the
quantity of each fatty acid observed. Despite the wide variation factors
already commented on, the fatty acid prole is consistent with those
reported for cows milk butter and cows yoghurt butter in previous
studies (Middaugh et al., 1988; Sagdi et al., 2004; Ledoux et al., 2005;
Mallia et al., 2008). Other authors, however, reported slightly dierent
proles. Kaya (2000) reported higher contents for C18:1 than for C16:0,
while Samet-Bali et al. (2009) reported higher values for C18:0 than for
C14:0. Percentages of saturated (68%), monounsaturated (28.5%) and
polyunsaturated (3.6%) fatty acids were similar to those previously
reported for cows milk butter (Sagdi et al., 2004; Mallia et al., 2008).
Storage for 9 months scarcely aected the dierent fatty acids
contents. Only the linolenic acid (C18:3 n3) (from 0.32% to 0.29%),
together with eicosapentaenoic acid (C20:5 n3) (from 0.012% to
0.009%) and docosadienoic acid (C22:2 n6) (from 0.028% to 0.024),
which are present in very small proportions, decreased signicantly
after 9 months of storage. There is no available information on the
changes in the fatty acid contents of butter stored for long times under
refrigeration. Our results are consistent with those reported by Mallia
et al. (2008), who only observed very small losses of trans-C18:1 n9
(contents decreasing from 0.32% to 0.24%) and trans-C18:1 n12 (from
0.22% to 0.20%) fatty acids after 8 weeks (a short period of time) of
storage at 6 C. Previous studies of butter fat stability have been per-
formed using the accelerated shelf-life testing method in trials with heat
treatments at 60, 70 and 80 C for periods ranging from 4 to 40 days
depending on the heating temperature (Kaya, 2000; zkanli and Kaya,
2007; Samet-Bali et al., 2009). Signicant losses of some fatty acids
were reported in these accelerated trials. Kaya (2000) reported sig-
nicant losses of linoleic acid (C18:2) (from 22.8 to 27.6%) and lino-
lenic acid (C18:3) (from 69.1 to 77.8%) in samples of cows milk butter
depending on temperature (60, 70 or 80 C) or time of storage (from 5
to 41 days). This author did not observe any loss of oleic acid (C18:1),
thus conrming the higher stability to oxidation of this fatty acid re-
lative to that of linoleic or linolenic fatty acids. Similarly, zkanli and
Kaya (2007) reported losses of C18:2 and C18:3 fatty acids in samples
of butter made from unpasteurised and pasteurised sheeps milk and
heated at 60, 70 and 80 C for 20, 15 and 5 days, respectively. However,
Samet-Bali et al. (2009) did not report any loss of fatty acids in cows
milk butter heated for 360 or 720 h at 60 C.
Glycerides are the main fraction in the total lipids in milk fat, and
the fatty acid prole of the neutral lipids and levels of individual fatty
acids (Table 3) are almost the same as those of the total lipids. As ob-
served for total lipids, storage scarcely aected the levels of the dif-
ferent fatty acids; some signicant losses of CLA (the contents decreased
from 0.69% to 0.54%) and C20:1 n9 (from 0.28% to 0.12%) fatty acids
were observed, and these losses were higher in butter containing salt
125
Journal of Food Composition and Analysis 63 (2017) 121132
(p < 0.001).
The composition of the fatty acids in the polar fraction (Table 4) was
slightly dierent from that of the total and neutral lipids. In the polar
fraction, the short-chain fatty acids C4:0, C6:0 and C8:0 were not de-
tected, C14:0 was more abundant than C18:0, and C18:2 was more
abundant than C14:0 and C12:0. As occurred with the other lipid
fractions, storage produced scarce modications in the amounts of the
dierent fatty acids. In this lipid fraction, C17:1 seems to be the fatty
acid most aected (p < 0.01). Its content decreased from 0.27% to
0.19%, and losses were signicantly inuenced by temperature
(p < 0.001). The CLA content was also aected (p < 0.05) dropping
its content from 0.74% to 0.66%, and the losses were signicantly
higher (p < 0.001) in salted than in unsalted butter, and they were
also signicantly higher (p < 0.001) at the higher storage tempera-
ture. The amount of arachidonic acid (C20:4 n6) also decreased
(p < 0.05) during storage and was aected by the storage temperature
(p < 0.001).
The changes in the free fatty acids in butter during storage are
shown in Table 5. Total free fatty acids increased signicantly
(p < 0.001) from 136167 mg/100 g of fat in the fresh butter to
360575 mg/100 g of fat after storage for 9 months. Temperature sig-
nicantly aected (p < 0.001) the free fatty acid release, and for each
storage time and salt content the free fatty acid content was always
higher in the samples stored at 12 C than in those stored at 4 C. This
temperature-related eect increased with storage time (R T,
p < 0.01). The addition of salt to the butter also enhances lipolysis (S,
p < 0.05).
The contents of each individual free fatty acid increased sig-
nicantly (p < 0.001) during storage, and the dierent free fatty acid
contents increased by a factor that in most cases ranged from 2 to 4. The
eects of temperature and salt on the release of the fatty acids con-
sidered were also observed in most cases. Regarding the abundance of
the dierent free fatty acids, C16:0 was the most abundant, followed by
C18:1, C14:0, C4:0, C18:0, C6:0, C12:0 and C10:0. Data on free fatty
acid contents of butter during the storage are scarce. The abundance of
the dierent free fatty acids in the present study was fairly consistent
with previous observations on cows yoghurt butter both fresh (1 day)
and after 30 days of storage (Senel et al., 2011). In the present study,
however, the abundance of C4:0 and C6:0 free fatty acids in the butter
samples was notable. Senel et al. (2011) reported signicant increases
(p < 0.01) in amounts of capric, lauric, myristic, palmitic, oleic and
linoleic acids in cows yoghurt butter stored for one month. In the
present study, the concentrations of C8:0, C10:0, C12:0, C14:0 and
C18:0 fatty acids in the butter after storage for one month were similar
to those reported by Senel et al. (2011). However, the amounts of C6:0,
C16:0 and C18:1 fatty acids were higher and those of C18:2 were lower
than those reported by Senel et al. (2011). In any case, the release of
short-chain fatty acids during storage could be relevant for the sensory
characteristics, because they could provoke rancid avours in butter.
3.3. Fat characteristics
The changes in the values of the parameters indicating changes in
fat structure and oxidative stability during storage of the butter samples
are shown in Table 6. In accordance with the increase in the free fatty
acid content (Table 5), the parameters indicating lipolysis (acidity value
and acidity degree) increased signicantly (p < 0.001) during storage
time, reaching values between 2.3 and 2.8 times higher at the end of the
storage period than in fresh butter. As occurred with the release of the
fatty acids, for each storage time, higher storage temperatures led to
higher values of these two parameters (p < 0.01); the presence of salt
also produced a slight increase in the values (p < 0.05).
The initial values of the degree of acidity (0.350.42% oleic acid)
are within the range of values (0.150.56% oleic acid) observed for
fresh butter made from both milk and yoghurt from dierent animal
species (Kaya, 2000; zkanli and Kaya, 2007; Samet-Bali et al., 2009).
 Table 3
F.J. Mndez-Cid et al.

Fatty acid composition (% of total methyl esters) of the neutral lipids during the storage of cows butter: Eect of temperature and salt content. Values are means of the results from three replicates (n = 3).

Fresh butter Storage time (months)

1 3 6 9

Salt Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted

Temperature 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C
Fatty acids SEM R T S RT RS

* * *
C4:0 2.31 2.60 2.33 2.67 2.27 2.61 2.31 2.42 2.21 2.51 2.21 2.55 2.24 2.32 2.39 2.66 2.39 2.23 0.024 n.s. n.s.
** * **
C6:0 1.85 1.96 1.90 2.10 1.90 2.01 1.88 1.85 1.86 1.93 1.87 1.94 1.83 1.87 1.95 2.01 1.95 1.83 0.011 n.s. n.s.
** **
C8:0 1.22 1.26 1.27 1.35 1.26 1.27 1.25 1.21 1.24 1.22 1.25 1.24 1.21 1.22 1.28 1.25 1.28 1.22 0.006 n.s. n.s. n.s.
** * *
C10:0 2.87 2.91 3.00 3.09 2.94 2.93 2.94 2.90 2.94 2.85 2.95 2.88 2.86 2.88 3.01 2.85 2.97 2.88 0.011 n.s. n.s.
* *
C12:0 3.63 3.55 3.75 3.70 3.64 3.63 3.67 3.67 3.65 3.59 3.81 3.67 3.59 3.59 3.92 3.55 3.65 3.60 0.020 n.s. n.s. n.s.
C14:0 11.8 11.6 11.9 11.8 12.1 11.9 12.2 12.0 12.1 11.9 12.3 12.0 12.0 12.0 12.3 11.9 12.0 12.5 0.037 n.s. n.s. n.s. n.s. n.s.
*
C14:1 1.15 1.15 1.28 1.20 1.25 1.18 1.25 1.20 1.22 1.21 1.31 1.28 1.30 1.37 1.28 1.15 1.20 1.41 0.013 n.s. n.s. n.s. n.s.
* *
C15:0 1.09 1.09 1.10 1.09 1.15 1.06 1.15 1.11 1.14 1.13 1.15 1.06 1.17 0.91 1.09 1.11 1.13 1.20 0.012 n.s. n.s. n.s.
**
C15:1 0.27 0.22 0.23 0.25 0.22 0.25 0.22 0.27 0.23 0.28 0.23 0.26 0.22 0.24 0.24 0.24 0.23 0.27 0.003 n.s. n.s. n.s. n.s.
*
C16:0 32.8 32.7 32.1 32.4 32.1 32.2 32.4 31.6 32.3 32.9 31.7 31.6 31.7 32.1 31.9 32.8 32.4 31.8 0.074 n.s. n.s. n.s. n.s.
* * * **
C16:1 n7 2.17 2.19 1.90 2.44 1.86 2.38 1.80 1.97 1.90 2.08 1.93 2.28 1.66 1.81 1.78 2.01 1.91 1.94 0.032 n.s.
C17:0 0.53 0.53 0.55 0.52 0.55 0.53 0.54 0.55 0.55 0.55 0.54 0.56 0.55 0.55 0.53 0.56 0.54 0.56 0.003 n.s. n.s. n.s. n.s. n.s.
** *
C17:1 0.29 0.24 0.25 0.30 0.30 0.24 0.25 0.32 0.33 0.20 0.27 0.33 0.32 0.24 0.28 0.25 0.31 0.23 0.006 n.s. n.s. n.s.
* *
C18:0 9.66 9.74 11.9 9.22 12.5 10.66 11.7 11.1 12.2 10.8 12.2 10.7 12.2 11.0 11.3 10.8 12.0 11.81 0.140 n.s. n.s. n.s.

126
*
C18:1 n9 24.4 24.1 22.5 23.7 22.1 23.1 22.5 23.7 22.1 22.9 22.2 23.6 23.2 24.1 22.7 22.9 22.2 22.8 0.118 n.s. n.s. n.s. n.s.
**
C18:2 n6 2.24 2.25 2.51 2.27 2.35 2.42 2.47 2.38 2.54 2.36 2.63 2.36 2.40 2.28 2.44 2.23 2.43 2.24 0.023 n.s. n.s. n.s. n.s.
* * **
C18:3 n6 0.053 0.053 0.041 0.055 0.036 0.053 0.043 0.055 0.036 0.051 0.035 0.054 0.033 0.046 0.038 0.054 0.041 0.045 0.001 n.s. n.s.
**
C18:3 n3 0.27 0.29 0.30 0.37 0.34 0.37 0.31 0.40 0.32 0.32 0.31 0.41 0.35 0.33 0.39 0.35 0.27 0.34 0.006 n.s. n.s. n.s. n.s.
*** * *** * ***
CLA 0.69 0.69 0.58 0.77 0.55 0.54 0.56 0.64 0.55 0.56 0.55 0.64 0.57 0.53 0.56 0.64 0.54 0.54 0.008
* * *** ***
C20:0 0.13 0.16 0.11 0.16 0.11 0.12 0.10 0.13 0.11 0.13 0.11 0.12 0.11 0.11 0.11 0.12 0.11 0.11 0.002 n.s.
** * *** **
C20:1 n9 0.21 0.28 0.13 0.16 0.12 0.15 0.13 0.16 0.13 0.17 0.13 0.15 0.13 0.13 0.12 0.15 0.13 0.12 0.004 n.s.
* ** *** ** **
C20:2 n6 0.023 0.025 0.025 0.022 0.026 0.026 0.023 0.015 0.029 0.021 0.021 0.013 0.029 0.026 0.024 0.016 0.029 0.024 0.001
* *
C20:3 n6 0.08 0.08 0.08 0.09 0.08 0.08 0.08 0.08 0.08 0.07 0.08 0.08 0.08 0.07 0.08 0.09 0.08 0.07 0.001 n.s. n.s. n.s.
C20:4 n6 0.14 0.14 0.13 0.14 0.13 0.14 0.13 0.16 0.12 0.12 0.13 0.14 0.14 0.13 0.13 0.17 0.12 0.12 0.003 n.s. n.s. n.s. n.s. n.s.
* * **
C20:3 n3 0.007 0.008 0.008 0.006 0.008 0.009 0.010 0.015 0.008 0.009 0.008 0.011 0.007 0.009 0.008 0.009 0.008 0.008 0.0003 n.s. n.s.
* * ** *** *
C20:5 n3 0.015 0.017 0.016 0.015 0.013 0.013 0.014 0.016 0.013 0.012 0.013 0.016 0.013 0.011 0.013 0.017 0.013 0.017 0.0003
* **
C22:2 n6 0.030 0.028 0.029 0.036 0.032 0.032 0.030 0.034 0.034 0.032 0.028 0.041 0.034 0.030 0.029 0.041 0.042 0.028 0.001 n.s. n.s. n.s.
* * ***
C24:0 0.066 0.060 0.048 0.050 0.056 0.063 0.053 0.064 0.057 0.061 0.050 0.062 0.052 0.052 0.051 0.076 0.061 0.043 0.001 n.s. n.s.
***
SFA 68.0 68.2 70.0 68.1 70.6 69.0 70.2 68.6 70.4 69.6 70.1 68.3 69.5 68.6 69.9 69.6 70.4 69.7 0.131 n.s. n.s. n.s. n.s.
***
UFA 32.0 31.7 30.0 31.9 29.4 31.0 29.8 31.4 29.6 30.4 29.9 31.7 30.5 31.4 30.1 30.4 29.6 30.3 0.131 n.s. n.s. n.s. n.s.
***
MUFA 28.5 28.2 26.2 28.1 25.9 27.3 26.1 27.6 25.9 26.8 26.1 27.9 26.8 27.9 26.4 26.8 26.0 26.8 0.133 n.s. n.s. n.s. n.s.
PUFA 3.55 3.58 3.72 3.77 3.56 3.68 3.67 3.79 3.73 3.56 3.81 3.77 3.66 3.46 3.71 3.62 3.57 3.44 0.024 n.s. n.s. n.s. n.s. n.s.

SFA: sum of saturated fatty acids; UFA: sum of unsaturated fatty acids; MUFA: sum of monounsaturated fatty acids; PUFA: sum of polyunsaturated fatty acids. SEM: Standard error of the mean. Signicance: R: signicance of the eect of the time of
storage; T: signicance of the eect of the temperature; S: signicance of the eect of the salt content; R T: interaction of time of storage and temperature; R S: interaction of time of storage and salt content; n.s.: not signicant.
* (p < 0.05).
** (p < 0.01).
*** (p < 0.001).
Journal of Food Composition and Analysis 63 (2017) 121132
 F.J. Mndez-Cid et al.

Table 4
Fatty acid composition (% of total methyl esters) of the polar lipids during the storage of cows butter: Eect of temperature and salt content. Values are means of the results from three replicates (n = 3).

Fresh butter Storage time (months)

1 3 6 9

Salt Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted

Temperature 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C
Fatty acids SEM R T S RT RS

** *** ***
C10:0 1.03 1.17 0.58 0.53 0.80 0.60 0.63 0.54 0.69 0.72 0.82 0.52 0.74 0.70 0.77 0.56 0.87 0.75 0.023 n.s. n.s.
*** *
C12:0 1.72 1.61 0.94 0.95 1.02 1.29 0.87 1.38 0.79 1.28 1.01 1.33 0.81 1.22 1.19 0.95 1.10 1.39 0.037 n.s. n.s. n.s.
** **
C14:0 6.05 6.36 5.50 6.06 5.49 5.86 5.42 5.98 5.37 6.48 6.14 6.61 5.62 6.59 7.03 6.15 5.89 6.70 0.108 n.s. n.s. n.s.
*
C14:1 1.63 1.84 2.13 1.80 2.21 2.13 2.09 1.84 2.07 1.69 2.00 2.12 1.96 1.72 1.67 1.95 2.20 1.62 0.037 n.s. n.s. n.s. n.s.
* **
C15:0 0.97 0.98 1.00 1.19 0.96 1.01 0.98 0.93 1.01 0.98 1.00 1.13 1.02 1.03 1.07 1.06 1.04 0.99 0.012 n.s. n.s. n.s.
C15:1 0.30 0.34 0.32 0.31 0.32 0.33 0.33 0.31 0.32 0.36 0.32 0.34 0.34 0.29 0.32 0.35 0.32 0.31 0.016 n.s. n.s. n.s. n.s. n.s.
* *
C16:0 30.7 31.2 29.1 30.1 29.4 30.7 30.0 30.1 29.7 31.3 30.4 32.2 30.7 31.6 31.6 32.2 30.5 32.6 0.469 n.s. n.s. n.s.
* *** *** *
C16:1 n7 2.26 2.13 1.89 1.46 2.02 2.07 1.95 1.44 1.81 1.98 1.87 1.41 1.99 2.08 1.91 1.92 2.06 1.91 0.032 n.s.
* * *** *** **
C17:0 0.98 0.98 0.55 0.62 1.08 0.56 0.89 0.61 1.06 0.60 0.96 0.64 1.10 0.59 0.93 0.58 1.15 0.57 0.030
** *** *
C17:1 0.27 0.27 0.20 0.24 0.23 0.17 0.22 0.23 0.21 0.16 0.21 0.24 0.30 0.16 0.24 0.24 0.28 0.19 0.019 n.s. n.s.
** *
C18:0 13.3 12.9 12.5 12.6 14.7 13.1 13.0 13.0 13.2 12.6 12.5 13.4 13.4 13.7 12.2 13.3 13.2 12.9 0.198 n.s. n.s. n.s.
* **
C18:1 n9 30.5 29.6 32.2 32.1 30.1 30.4 31.1 31.8 32.0 31.0 30.7 28.9 29.9 30.0 30.3 29.2 30.0 30.1 0.461 n.s. n.s. n.s.

127
* *
C18:2 n6 7.03 7.08 9.04 8.34 7.95 8.24 8.77 8.09 7.90 7.59 8.37 7.64 8.35 7.07 7.33 7.87 7.92 7.01 0.111 n.s. n.s. n.s.
*
C18:3 n6 0.19 0.17 0.18 0.15 0.19 0.18 0.18 0.19 0.18 0.18 0.18 0.17 0.17 0.18 0.18 0.17 0.18 0.18 0.002 n.s. n.s. n.s. n.s.
***
C18:3 n3 0.62 0.54 0.54 0.59 0.52 0.51 0.52 0.57 0.56 0.45 0.51 0.55 0.57 0.42 0.52 0.62 0.53 0.41 0.009 n.s. n.s. n.s. n.s.
* *** ***
CLA 0.74 0.72 0.82 0.78 0.80 0.74 0.81 0.81 0.81 0.68 0.80 0.76 0.83 0.65 0.78 0.78 0.76 0.66 0.008 n.s. n.s.
** *** * *
C20:0 0.17 0.14 0.19 0.18 0.18 0.22 0.17 0.19 0.18 0.20 0.20 0.19 0.16 0.23 0.17 0.16 0.18 0.20 0.003 n.s.
*** * *
C20:1 n9 0.12 0.13 0.15 0.15 0.15 0.17 0.14 0.15 0.14 0.17 0.15 0.17 0.14 0.17 0.14 0.13 0.15 0.16 0.002 n.s. n.s.
*** ***
C20:2 n6 0.062 0.065 0.050 0.069 0.052 0.067 0.055 0.067 0.067 0.056 0.059 0.073 0.066 0.058 0.058 0.075 0.059 0.057 0.001 n.s. n.s. n.s.
** *
C20:3 n6 0.40 0.57 0.62 0.57 0.54 0.55 0.63 0.56 0.63 0.50 0.57 0.50 0.66 0.47 0.46 0.64 0.53 0.42 0.012 n.s. n.s. n.s.
* ***
C20:4 n6 0.62 0.74 0.90 0.76 0.80 0.67 0.86 0.70 0.83 0.58 0.74 0.60 0.80 0.58 0.60 0.71 0.73 0.52 0.017 n.s. n.s. n.s.
* ** *
C20:3 n3 0.037 0.036 0.060 0.063 0.071 0.058 0.056 0.058 0.071 0.058 0.054 0.055 0.063 0.052 0.059 0.062 0.060 0.050 0.001 n.s. n.s.
**
C20:5 n3 0.16 0.17 0.16 0.18 0.17 0.16 0.18 0.19 0.17 0.15 0.18 0.17 0.13 0.16 0.18 0.15 0.14 0.15 0.003 n.s. n.s. n.s. n.s.
*** ***
C22:2 n6 0.066 0.047 0.088 0.071 0.055 0.083 0.073 0.074 0.065 0.073 0.071 0.073 0.063 0.079 0.067 0.083 0.064 0.078 0.001 n.s. n.s. n.s.
* *
C24:0 0.13 0.13 0.19 0.16 0.14 0.15 0.14 0.15 0.15 0.14 0.14 0.17 0.13 0.15 0.13 0.16 0.14 0.13 0.003 n.s. n.s. n.s.
** **
SFA 55.0 55.5 50.6 52.3 53.8 53.4 52.1 52.9 52.2 54.3 53.2 56.2 53.6 55.9 55.2 55.1 54.0 56.2 0.265 n.s. n.s. n.s.
** **
UFA 45.0 44.5 49.4 47.7 46.2 46.6 47.9 47.1 47.8 45.7 46.8 43.8 46.4 44.1 44.8 44.9 46.0 43.8 0.264 n.s. n.s. n.s.
** ** *
MUFA 35.1 34.3 36.9 36.1 35.0 35.3 35.8 35.8 36.6 35.4 35.2 33.1 34.7 34.4 34.6 33.8 35.0 34.2 0.145 n.s. n.s.
* **
PUFA 9.92 10.1 12.5 11.6 11.1 11.3 12.1 11.3 11.3 10.3 11.5 10.6 11.7 9.7 10.2 11.2 11.0 9.53 0.144 n.s. n.s. n.s.

SFA: sum of saturated fatty acids; UFA: sum of unsaturated fatty acids; MUFA: sum of monounsaturated fatty acids; PUFA: sum of polyunsaturated fatty acids. SEM: Standard error of the mean. Signicance: R: signicance of the eect of the time of
storage; T: signicance of the eect of the temperature; S: signicance of the eect of the salt content; R T: interaction of time of storage and temperature; R S: interaction of time of storage and salt content; n.s.: not signicant.
* (p < 0.05).
** (p < 0.01).
*** (p < 0.001).
Journal of Food Composition and Analysis 63 (2017) 121132
 Table 5
F.J. Mndez-Cid et al.

Free fatty acid contents (mg/100 g of fat) during the storage of cows butter: Eect of temperature and salt content. Values are means of the results from three replicates (n = 3).

Fresh butter Storage time (months)

Months 1 3 6 9

Salt Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted Unsalted Salted

Temperature 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C
Fatty acids SEM R T S RT RS

*** *** *** ***

C4:0 6.02 6.72 7.15 9.60 7.45 10.4 9.67 12.6 9.01 14.0 11.8 13.5 12.9 15.3 13.6 15.6 14.3 16.1 0.50 n.s.
*** *** *** *
C6:0 4.87 5.54 5.95 8.29 6.11 8.37 7.47 10.3 7.61 10.5 8.69 11.1 10.2 12.2 10.4 13.0 12.5 14.4 0.39 n.s.
*** *** *** ***
C8:0 1.34 2.00 1.20 1.97 1.04 1.61 1.34 3.80 1.24 2.50 1.44 4.65 2.35 2.76 2.10 3.71 2.42 4.54 0.16 n.s.
*** *** *** *** ***
C10:0 3.90 3.82 3.91 5.16 4.51 4.85 5.03 5.76 5.14 6.84 4.26 5.92 6.05 10.3 6.19 9.42 8.09 13.3 0.33
*** *** *** *** ***
C12:0 4.48 4.29 4.10 5.99 4.89 6.17 5.53 6.07 5.53 8.03 5.46 6.42 6.73 11.8 6.67 10.1 8.77 14.8 0.36
*** *** **
C14:0 7.65 10.7 11.0 10.7 11.8 17.6 12.3 28.4 13.4 25.0 22.2 35.0 24.2 31.1 25.6 45.3 26.3 40.5 1.35 n.s. n.s.
*** *** *** *** **
C14:1 1.02 1.28 1.13 1.39 1.17 1.49 1.30 2.02 1.58 1.88 1.54 2.50 1.95 3.78 2.18 2.64 1.77 4.96 0.13
*** *** **
C15:0 0.54 0.69 0.70 0.70 0.78 1.16 0.80 1.66 0.88 1.61 1.39 2.14 1.53 2.11 1.67 2.96 1.75 2.60 0.08 n.s. n.s.
*** ***
C15:1 0.03 0.03 0.03 0.03 0.03 0.05 0.04 0.08 0.04 0.07 0.07 0.10 0.07 0.12 0.08 0.13 0.09 0.12 0.004 n.s. n.s. n.s.
*** *** **
C16:0 68.3 82.1 85.1 84.0 84.1 139 93.3 175 94.7 183 151 214 166 245 183 290 192 297 8.91 n.s. n.s.
*** *** ** *** ***
C16:1 n7 3.35 3.48 2.04 4.66 2.35 3.25 2.19 4.12 2.59 4.45 4.02 4.59 4.40 7.27 4.43 6.98 4.95 10.5 0.28
*** *** ** **
C17:0 0.11 0.29 0.37 0.34 0.38 0.60 0.39 0.77 0.40 0.83 0.66 0.92 0.70 1.06 0.77 1.08 0.86 1.24 0.04 n.s.
*** **
C17:1 0.02 0.06 0.07 0.07 0.08 0.13 0.09 0.25 0.10 0.16 0.19 0.27 0.08 0.33 0.18 0.33 0.17 0.32 0.01 *** n.s. n.s.
*** *** *** *** **
C18:0 5.26 5.83 6.19 5.80 5.56 10.1 6.57 10.9 6.19 13.1 9.68 13.1 10.8 17.7 10.9 13.9 13.4 20.8 0.58

128
*** *** * * *
C18:1 n9 26.3 34.6 35.6 34.7 37.7 61.5 38.8 76.3 42.6 79.8 79.9 99.1 75.7 104 84.1 90.1 83.7 132 3.83
*** *** ***
C18:2 n6 1.67 2.15 2.26 2.22 2.43 3.67 2.46 4.70 2.69 4.85 4.87 6.52 4.65 7.55 5.10 9.50 5.01 10.5 0.32 n.s. n.s.
*** *** *** *** ***
C18:3 n6 0.09 0.10 0.06 0.09 0.06 0.10 0.06 0.11 0.06 0.12 0.08 0.14 0.09 0.27 0.09 0.14 0.11 0.27 0.009
*** *** *** *
C18:3 n3 0.15 0.20 0.20 0.20 0.21 0.33 0.22 0.41 0.24 0.41 0.39 0.57 0.39 0.55 0.44 0.86 0.41 0.67 0.02 n.s.
*** *** *** **
C20:0 0.16 0.17 0.19 0.21 0.17 0.39 0.23 0.35 0.22 0.48 0.38 0.49 0.43 0.68 0.51 0.50 0.42 0.92 0.03 n.s.
*** *** *** *** ***
C20:1 n9 0.19 0.22 0.29 0.26 0.29 0.52 0.37 0.45 0.35 0.68 0.66 0.89 0.66 0.93 0.70 0.74 0.68 1.18 0.03
*** *** *** *
C20:2 n6 0.06 0.07 0.03 0.07 0.03 0.05 0.04 0.08 0.04 0.07 0.05 0.11 0.07 0.09 0.07 0.17 0.08 0.10 0.004 n.s.
*** *** * *** *
C20:3 n6 0.18 0.22 0.19 0.24 0.19 0.38 0.29 0.44 0.26 0.46 0.30 0.61 0.38 0.62 0.34 0.58 0.38 0.96 0.03
*** *** *** *** **
C20:4 n6 0.22 0.16 0.24 0.24 0.24 0.41 0.26 0.34 0.32 0.52 0.43 0.57 0.51 0.79 0.49 0.56 0.49 1.18 0.03
*** *** ** *** ***
C20:3 n3 0.07 0.06 0.02 0.07 0.04 0.03 0.03 0.09 0.05 0.04 0.04 0.10 0.07 0.08 0.04 0.12 0.06 0.13 0.004
*** *** **
C20:5 n3 0.06 0.03 0.03 0.05 0.03 0.04 0.04 0.07 0.03 0.05 0.06 0.09 0.06 0.10 0.05 0.11 0.06 0.12 0.003 n.s. n.s.
*** *** ***
C22:2 n6 0.58 0.77 0.68 1.10 0.73 0.60 0.86 1.31 0.90 1.00 1.03 1.90 1.15 1.87 1.10 2.12 1.17 2.43 0.07 n.s. n.s.
*** *** *** *** *
C24:0 0.10 0.28 0.16 0.15 0.28 0.70 0.30 0.19 0.34 0.81 0.31 0.47 0.42 0.99 0.47 1.15 0.48 1.35 0.05
*** *** **
SFA 102 124 122 126 125 191 138 250 142 253 206 308 232 339 262 406 267 410 12.0 n.s. n.s.
*** *** * ** *
UFA 34.0 43.7 42.9 45.3 45.6 72.5 47.0 90.8 51.9 94.6 93.6 118 90.3 128 99.4 115 99.1 166 4.67
*** *** * ** *
MUFA 30.9 39.8 39.2 41.1 41.6 66.9 42.8 83.3 47.3 87.0 86.4 107 82.9 116 91.7 101 91.4 149 4.22
*** *** ***
PUFA 3.09 3.76 3.71 4.27 3.97 5.61 4.26 7.55 4.60 7.52 7.25 10.62 7.35 11.9 7.72 14.2 7.77 16.4 0.47 n.s. n.s.
*** *** * **
TFFA 137 167 165 171 171 264 185 341 194 348 299 426 322 467 361 521 366 576 16.6 n.s.

SFA: sum of saturated fatty acids; UFA: sum of unsaturated fatty acids; MUFA: sum of monounsaturated fatty acids; PUFA: sum of polyunsaturated fatty acids. SEM: Standard error of the mean. Signicance: R: signicance of the eect of the time of
store; T: signicance of the eect of the temperature; S: signicance of the eect of the salt content; R T: interaction of time of storage and temperature; R S: interaction of time of storage and salt content; n.s.: not signicant.
* (p < 0.05).
** (p < 0.01).
*** (p < 0.001).
Journal of Food Composition and Analysis 63 (2017) 121132
 Table 6
Evolution of the parameters which characterise the fat during the storage of cows butter: Eect of temperature and salt content. Values are means of the results from three replicates (n = 3).

Fresh butter Storage time (months)

1 3 6
F.J. Mndez-Cid et al.

Salt Unsalted Salted Unsalted Salted Unsalted Salted Unsalted

Temperature 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C 4 C 12 C
Indices

Acidity 0.70 0.84 0.77 0.84 0.79 0.88 0.84 0.91 0.92 0.94 0.88 1.05
valuea
Acidity 0.35 0.42 0.38 0.42 0.39 0.44 0.42 0.46 0.46 0.47 0.44 0.53
degreeb
Peroxide 0.39 0.38 0.59 0.64 0.64 0.85 0.62 0.78 0.89 1.19 1.09 0.96
valuec
Saponicatio- 231 231 237 243 292 239 269 259 265 255 251 232
n valued
Iodine valuee 35.2 38.4 33.6 34.8 35.6 38.6 33.0 30.8 31.3 32.5 37.2 33.8
Refractive 1.45645 1.45650 1.45650 1.45650 1.45650 1.45650 1.45655 1.45655 1.45655 1.45655 1.45655 1.45655
index
TBA valuef 0.084 0.100 0.073 0.082 0.084 0.093 0.133 0.186 0.155 0.325 0.126 0.207

Storage time (months)

6 9

Salt Salted Unsalted Salted

129
Temperature 4 C 12 C 4 C 12 C 4 C 12 C
Indices SEM R T S RT RS

*** ** * **
Acidity 0.95 0,98 1.89 1.96 1.94 2.18 0.05 n.s.
valuea
*** ** * **
Acidity 0.48 0.49 0.95 0.98 0.97 1.09 0.02 n.s.
degreeb
*** *** *** *** ***
Peroxide 1.30 2.52 0.90 1.68 1.72 3.75 0.08
valuec
Saponicatio- 237 246 281 258 247 245 2.87 n.s. n.s. n.s. n.s. n.s.
n valued
*
Iodine valuee 37.3 33.5 40.3 37.6 33.1 34.6 0.53 n.s. n.s. n.s. n.s.
Refractive 1.45655 1.45655 1.45655 1.45655 1.45655 1.45655 0.000003 n.s. n.s. n.s. n.s. n.s.
index
*** ** *** *** ***
TBA valuef 0.158 0.217 0.093 0.141 0.176 0.220 0.01

SEM: Standard error of the mean. Signicance: R: signicance of the eect of the time of storage; T: signicance of the eect of the temperatures; S: signicance of the eect of the salt content; R T: interaction of time of storage and temperature;
R S: interaction of time of storage and salt content; n.s.: not signicant;
a
Expressed as mg KOH/g of fat.
b
Expressed as % of oleic acid.
c
Expressed as meq O2/kg of fat.
d
Expressed as mg KOH/g of fat.
e
Expressed as % of I2 absorbed.
f
Expressed as mg malondialdehyde/kg.
* (p < 0.05).
** (p < 0.01).
*** (p < 0.001).
Journal of Food Composition and Analysis 63 (2017) 121132
F.J. Mndez-Cid et al. Journal of Food Composition and Analysis 63 (2017) 121132

a*: redness; b*: yellowness; L*: lightness; C*: chroma; H: Hue angle; E: total colour dierence; E: fraction of redness relative to those of yellowness and lightness. SEM: Standard error of the mean. Signicance: R: signicance of the eect of the
time of storage; T: signicance of the eect of the temperature; S: signicance of the eect of the salt content; R T: interaction of time of storage and temperature; R S: interaction of time of storage and salt content; n.s.: not signicant.
As already mentioned, previous reports on the stability of butter fat

RS

n.s.
involve accelerated trials carried out at high temperatures, and no in-

***

***

***
**

**

**
formation is available regarding the changes produced during re-

RT
frigerated storage. The values obtained in the present study after sto-

n.s.
***

***

***

***

***
**
rage of the butter samples for 9 months (0.951.09% of oleic acid) are
slightly lower than those reported by Samet-Bali et al. (2009) for tra-

***

***

***

***

***
**
S

*
ditional Tunisian cows butter oil after 37 days of storage at 60 C (1.5%

***

***

***

***

***

***
T
oleic acid), but higher than those observed by zkanli and Kaya (2007)

*
*** for butter oils made from unpasteurised and pasteurised sheeps milk
***

***

***

***

***

***
R

(values from 0.230.38% oleic acid after 818 days of storage at tem-

0.003
peratures ranging from 60 to 80 C). In view of the nal values observed
SEM

0.06
0.38
0.33
0.34
0.15
0.33
for these two parameters, and on comparing them with the values of
5.77

these parameters in other dairy fatty products, such as cheese stored/

0.33
12 C

30.5
76.0
23.7

18.1
104

ripened for long periods of time (Collins et al., 2003), it can be con-
cluded that butter undergoes very slight lipolysis both during long-term
5.23
Salted

0.28

storage and during accelerated trials performed at high temperatures.

35.4
79.5
25.2

14.5
4 C

102

This may be because the milk lipoprotein lipase (LPL) that accounts for
most of the lipolytic activity in milk is normally associated with the
5.44

0.25
12 C

23.0
75.3
30.9

17.7

casein micelle (Deeth, 2006) and is almost absent in the lipid fraction.
100

Moreover, LPL is unstable to heat, and the usual pasteurisation treat-

Evolution of the color parameters during the storage of cows butter: Eect of temperature and salt content. Values are means of the results from three replicates (n = 3).

Unsalted

5.85

ments in the dairy industry (100 C for 15 s in the present study) cause
0.24
24.7
82.0
35.9
99.4
13.7
4 C

the almost total inactivation of LPL in milk. LPL therefore scarcely

9

causes lipolysis in pasteurised dairy products. Thermostable lipases

5.55

0.29
12 C

from microbial origin can be present in butter (Adams and Brawley,

40.0
79.8
25.8

13.7
102

1981; Christen et al., 1986). However, no appreciable enzymatic ac-

tivity is expected at the moisture contents observed in butter.
5.01
Salted

0.25
33.8
82.8
27.5

10.5
4 C

101

Peroxide values that indicate primary oxidation increased sig-

nicantly (p < 0.001) during storage of the butter from initial values
5.63

of 0.380.39 meq O2/kg fat to values comprising a range from 0.9 to

0.24
12 C

29.9
79.3
32.9
99.9
14.0

3.75 meq O2/kg fat after 9 months for the dierent storage tempera-
Unsalted

tures and salt contents. The initial values observed in the present study
5.55

0.22

are in the range (0.251.21 meq O2/kg fat) of those observed by other
30.2
83.5
37.7
98.5
13.7
4 C
6

authors in dierent types of butter immediately after manufacture

(Kaya, 2000; zkanli and Kaya, 2007; Samet-Bali et al., 2009; Senel
5.49

101.2

0.27
12 C

32.4
81.8
28.2

10.8

et al., 2011). However, the values observed after 9 months of re-

frigerated storage are much lower than those reported in the mentioned
4.72
Salted

studies that ranged from 8.2 meq O2/kg fat in traditional Tunisian
0.23
37.3
82.2
28.3
99.6
10.5
4 C

butter oil after 37 days of storage at 60 C (Samet-Bali et al., 2009) to

around 14 meq O2/kg fat in butter oils made from unpasteurised and
5.63

0.23
12 C

25.2
82.3
35.9
99.0
10.9

pasteurised sheeps milk after dierent storage times (418 days) at

high temperatures (6080 C) (zkanli and Kaya, 2007). In the present
Unsalted

4.96

study, the increase in storage temperature from 4 to 12 C led to a

0.20
27.0
82.2
35.9
97.9
13.8
4 C

signicant increase (p < 0.001) in fat oxidation, and the addition of

3

salt also increased the peroxide values for each storage time
5.47

0.25

(p < 0.001). Both eects increased with the time of storage (R T,

12 C

35.5
82.6
30.4

9.40
100

p < 0.001; R S, p < 0.001). In this regard, it has been reported that
lipid oxidation is exponentially related to temperature (Labuza and
4.85
Salted

0.22
35.5
85.2
30.6
99.1
7.62
4 C

Bergquist, 1983). Also, sodium chloride is proven to be a lipid pro-

Storage time (months)

oxidant per se and also due to metal impurities, which lower the
5.62

activation energy of the lipid oxidation reactions (Frankel, 2012).

0.22
12 C

27.7
85.0
37.4
98.6
10.6

The TBA test measures a secondary product of fat oxidation (mal-

Unsalted

ondialdehyde [MDA]). The TBA values in the present study increased

4.85

0.20

signicantly (p < 0.001) during the rst months of storage, reaching

27.9
87.8
34.1
98.2
8.30
4 C
1

maximum values at 36 months, and then decreasing after 9 months of

storage. The initial values (0.080.1 mg MDA/kg) are slightly lower
2.83
Salted

0.12
28.2
91.7
33.2
94.9
0.00

than those reported for fresh butter in previous studies (Kaya, 2000;
Fresh butter

zkanli and Kaya, 2007). The values in the present study after 9 months
Unsalted

of storage are, however, much lower than those reported by Kaya

2.62

0.12
27.0
93.4
28.3
95.3
0.00

(2000), and zkanli and Kaya (2007), for butter oils after dierent
times of storage at high temperatures (6080 C). In the present study,
Colour parameters

the storage temperature (p < 0.01) and the addition of salt

*** (p < 0.001).
** (p < 0.01).

(p < 0.001) had a signicant eect on the TBA values, and both fac-
* (p < 0.05).
Temperature

tors enhanced the formation of malondialdehyde. The levels of mal-

ondialdehyde decreased from 6 to 9 months of storage; secondary
Table 7

Salt

products of oxidation such as malondialdehyde are unstable and they

E
C*
H
b*
L*
a*

can be further degraded or combine with other substances. The values

130
F.J. Mndez-Cid et al.
of the parameters indicating fat oxidation (peroxide value and TBA
value) at the end of the refrigerated storage (9 months) are much lower
than those reported for these two parameters at the end of the storage
of butter oil in accelerated trials of stability carried out by storing at
higher temperatures (6080 C) for less than 40 days (Kaya, 2000;
zkanli and Kaya, 2007; Samet-Bali et al., 2009). It can therefore be
concluded that accelerated stability trials carried out at high tempera-
tures maximise the oxidation processes that occur during storage of
butter fat. This seems to be due to the exponential eect of temperature
on the rate of lipid oxidation already commented on in the present
work.
The saponication value, which indicates the average molecular
size of the fatty acids, did not vary signicantly (p > 0.05) during
storage under any of the conditions (storage temperatures and salt
level) assayed. This is consistent with the low variation in the fatty acid
contents during storage. The initial values (around 231) are consistent
with those reported by Kaya (2000), and zkanli and Kaya (2007) in
butter made from cows and sheeps milk. No data are available in the
literature on this parameter measured after storage.
The iodine value, which indicates the degree of unsaturation of the
constitutive fatty acids, scarcely varied during storage, which is also
consistent with the scarce losses of unsaturated fatty acids reported
during the storage period. The initial values of this parameter in the
present study (3538% of I2 absorbed) are also reasonably consistent
with those reported by Kaya (2000) for cows milk butter (35.1% of I2
absorbed) and cows yoghurt butter (35.5% of I2 absorbed); this author
did not report any data for this parameter during or at the end of the
storage period.
Finally, the refractive index remained relatively constant during
storage of the butter samples in the present study, which is consistent
with the scarce modication of the degree of unsaturation and chain
length of the fatty acids during storage. The observed values (1.4565)
are also consistent with those reported by Kaya (2000) and zkanli and
Kaya (2007) for butter made from cows and sheeps milk.
3.4. Colour parameters
The changes in colour parameters during storage of the butter
samples are shown in Table 7. The a* values decreased signicantly
(p < 0.001) during storage, from initial values of 2.62 to 2.83 to
nal values of 5.23 to 5.85. The decrease was more marked at the
higher storage temperature (p < 0.001), while the eect of salt was less
marked (p < 0.05). Regarding the b* parameter, the values increased
during the rst 6 months of storage (from 2728 to 3040), but later
decreased between the 6th and 9th months of storage (until nal values
of 2335). The presence of salt had a marked eect on this parameter
(p < 0.001), whereas the eect of temperature was weak (p < 0.05).
The L* values of the butter decreased signicantly (p < 0.001) during
the storage period, from around 9193 to around 7582. Both salt
(p < 0.001) and temperature (p < 0.001) had an enhancing eect on
this decrease throughout the storage period.
Little information about the colour of butter is available in the lit-
erature. The values reported here for the dierent parameters (a*, b*
and L*) are within the range of those reported by Samet-Bali et al.
(2009), with negative values for a*, values of 2540 for b* and of
80100 for the L* parameter. Samet-Bali et al. (2009) reported values of
colour parameters for traditional Tunisian butter made from cows milk
and stored at 60 C for 37 days. When trends in the values of the three
parameters (a*, b* and L*) in the present study are compared with
those reported by Samet-Bali et al. (2009), it can be observed that the
trends for the a* and b* parameters are very similar in the two studies,
although it also seems that changes take place more slowly during
prolonged refrigerated storage than during storage at higher tempera-
tures for shorter times. Samet-Bali et al. (2009) reported an initial de-
crease in the a* values from 0 to 8.5 in the rst 5 days of storage, with
the values remaining constant between days 5 and 23, and then
131
Journal of Food Composition and Analysis 63 (2017) 121132
increasing strongly at the end of the storage. The values of the b*
parameter increased at the beginning of the heating (during the rst
day) and then decreased slowly until the 25th day and then decreased
strongly until day 28. The trend observed in the present study for the L*
parameter is dierent to that reported by Samet-Bali et al. (2009).
These authors reported a large increase in the value from 50 to 83
during the rst day of treatment at 60 C, followed by a slight increase
up to 87 on day 5, and then minimal changes until the end of the heat
treatment.
The decrease in the b* values observed at the end of the storage
period and that occurring earlier in the accelerated trial performed by
Samet-Bali et al. (2009) seems to be related to the loss of yellow pig-
ments, mainly -carotenes, as a consequence of oxidation processes.
Kaya (2000) reported that oxidation of the chromophores that turn the
butter from yellow to light yellow takes place at peroxide values close
to 10 meq O2/kg fat. Much lower peroxide values were reached in the
present study at the end of the storage period (0.903.75 meq O2/kg
fat), which may explain the very slight decrease in the b* value ob-
served at the end of the storage period.
Since the values of C*, H, E and E are calculated from the a*, b*
and L* values, the changes in the values of chroma, hue angle, total
colour dierence, and fraction of redness relative to those of yellowness
and lightness during the storage period reect the changes that oc-
curred in the redness, yellowness and lightness.
The chroma values (28.3 in unsalted and 33.2 in salted fresh butter)
increased in unsalted butter during the rst month of storage and later
stayed relatively unaltered until the end of the storage period. Values of
chroma in salted butter decreased throughout the whole storage period,
the decrease being higher in the samples stored at 12 C. The H values
around 90 in fresh butter denote the yellow tone of butter. The H
values increased during storage and this increase was higher in the
salted samples and in the samples stored at 12 C. The total colour
dierence increased during storage and this increase was generally
higher in the salted samples and in the samples stored at the higher
temperature. The fraction of redness relative to those of yellowness and
lightness decreased during storage; again, changes in this parameter
were more pronounced in salted butter and in butter stored at 12 C.
4. Conclusions
Storage of butter samples for 9 months at low (4 C or 12 C) tem-
perature scarcely aected the dierent fatty acid contents, which is not
consistent with the high losses of C18:2 and C18:3 fatty acids observed
by other authors in accelerated trials performed by heating at high
temperatures for short periods of time. It can therefore be concluded
that temperature is more important than time of storage in relation to
loss of fatty acids, and that losses are almost negligible during storage at
low temperatures.
During storage, butter undergoes very slight lipolysis that may be
due to the enzymatic (both endogenous and of microbial origin) ac-
tivities being hindered by low moisture contents. Fat oxidation is no-
table during butter storage and it is enhanced by higher storage tem-
peratures and the presence of salt. However, fat oxidation during
storage at low temperatures is much less intense than previously ob-
served in accelerated trials. Accelerated trials performed at high tem-
peratures seem maximise the oxidation processes that occur during the
storage of butter fat.
The trends in values of the a* and b* parameters were very similar
for storage at high and low temperatures; however, in samples stored
under refrigeration for long periods, the changes in these values seem to
take place more slowly than during storage at higher temperatures for
shorter periods.
Acknowledgements
The authors are grateful to the Xunta de Galicia (Regional
F.J. Mndez-Cid et al.
Government), Spain for nancial support provided under the
Consolidation and restructuring program of competitive research units:
Strategic Research Partnerships (2009/060). The authors also thank the
University of Vigo for nancial support (Contracts Program with re-
ference Research Groups, Call 2009, Reference 09VIA12).
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