Food Chemistry 127 (2011) 147152

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

The mineral composition (Ca, P, Mg, K, Na) in cheeses (cows, ewes and goats)
with different ripening times using near infrared spectroscopy with a bre-optic
probe
I. Gonzlez-Martn a,, J.M. Hernndez-Hierro a, I. Revilla b, A. Vivar-Quintana b, I. Lobos Ortega a
a
b

Departamento de Qumica Analtica, Nutricin y Bromatologa, Facultad de Ciencias Qumicas, C/Plaza de la Merced s/n, 37008 Salamanca, Espaa, Spain
rea de Tecnologa de los Alimentos de la Universidad de Salamanca en Zamora, Spain

a r t i c l e

i n f o

Article history:
Received 18 June 2009
Received in revised form 25 October 2010
Accepted 29 December 2010
Available online 8 January 2011
Keywords:
Mineral composition
Cheeses
Near infrared spectroscopy
Determination

a b s t r a c t
The results of this work show that it is possible to rapidly quantify calcium, phosphorus, magnesium,
potassium and sodium in unknown cheeses elaborated with percentages (0100%) of milk from different
species (cow, ewe, goat) by direct application of the bre-optic probe on the sample without previous
destruction or treatment of the sample. Of the total number of samples, 170 were used to develop the
calibration models using the Modied Partial Least Squares (MPLS) regression method and 57 samples
were used for external validation. The multiple correlation coefcients (RSQ) and prediction corrected
standard errors (SEP (C)) obtained for calcium (0.74, 0.64), phosphorus (0.69, 0.29), potassium (0.86,
0.13), and sodium (0.92, 0.71) in g/kg respectively and magnesium (0.72, 30.9) in ppm, indicated that
the models developed allow the determination of Ca, P, K, Na and Mg in unknown samples of cheeses
of varying compositions up to 6 months of ripening.
2011 Elsevier Ltd. All rights reserved.

1. Introduction
The consumption of cheese is of great nutritional interest due in
particular to its composition of micronutrients. The mineral content of the milk depends on numerous factors, such as genetic
characteristics, the stage of lactation, environmental conditions,
type of pasture and soil contamination among others (Anguita,
1996; Cichoscki, Valduga, Valduga, Tornadijo, & Fresno, 2002;
Gambelli, Belloni, Ingrao, Pizzoferrato, & Santaroni, 1999; Lucas
et al., 2006a, 2006b; Park, 2000; Patio, Faisal, Cedres, Mendez, &
Guanziroli-Stefani, 2005). The heavy metal in the cheese is fundamentally related to the manufacturing practices and possible contamination from the equipment during the process (Mendil, 2006;
Moreno-Rojas, Pozo-Lora, Zurera-Cosano, & Amaro-Lopez, 1994).
The minerals like sodium, potassium, calcium and magnesium
and the anionic constituents, phosphate, citrate, chloride, carbonate and sulphate are found in the milk or associated to the casein
micelle (Anguita, 1996; Moreno-Rojas et al., 1994). Their participation has been demonstrated in the coagulation, drainage of whey
and the texture of the curds and properties such as the stability
to heat and the capacity to coagulate depend on them (Patio
et al., 2005). Particularly during cheese ripening some of the mineral salts may migrate from the central part towards the external

Corresponding author. Tel./fax: +34 23 294483.

E-mail address: inmaglez@usal.es (I. Gonzlez-Martn).
0308-8146/$ - see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.12.114

layer of the cheese block or vice versa due to the pH gradient effect
causing changes in the concentration of some elements in the nal
product (Moreno-Rojas et al., 1994).
The analysis of the mineral composition in milk and cheeses has
been performed by inductively coupled plasma atomic emission
spectrophotometry (Prieto, Franco, Gonzlez, Bernardo, & Carballo,
2002), ion exchange liquid chromatography, instrumental neutron
activation analysis (Gambelli et al., 1999), atomic absorption
spectrophotometry, atomic emission spectrophotometry (Lucas,
Andueza, Rock, & Martin, 2008; Moreno-Rojas, Amaro-Lopez,
Garcia-Gimeno, & Zurera-Cosano, 1995; Moreno-Rojas, AmaroLopez, & Zurera-Cosano, 1992) and ame and graphite furnace
atomic absorption spectrometry (Mendil, 2006).
In the last decades techniques like near infra-red (NIR) spectroscopy have been developed for the determination of majority parameters in cheese (Rodrguez Otero, Hermida, & Centeno, 1997;
Mazerolles, Duboz, & Hugot, 2000), control of ripening, or sensorial
characteristics (Ortiz, Sarabia, Garcia-Rey, & De Castro, 2006;
Karoui, Pillonel, Schaller, Bosset, & De Baerdemaeker, 2007). The
prediction of the mineral composition in fresh and freeze-dried
cheeses by NIR has been performed by (Lucas et al., 2008), previously grinding the samples of cheese.
Recently, the use of NIRS technology employing a remote reectance bre-optic probe has been studied for the analysis of the percentage of milk (cows, ewes and goats) used in the elaboration of
cheeses with different ripening times and also for the determination of peptides or texture (Gonzalez-Martin et al., 2007; Revilla

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I. Gonzlez-Martn et al. / Food Chemistry 127 (2011) 147152

et al., 2009; Gonzlez-Martn, Hernndez-Hierro, Vivar-Quintana,

Revilla, & Gonzlez-Prez, 2009).
In the present work we study the use of NIRS technology together with the use of a remote reectance bre-optic probe (with
a 5  5 cm quartz window) for the determination of mineral composition (Ca, P, Mg, K, Na) in 227 samples of cheeses elaborated
with 16 varying mixtures of milk from cows, ewes and goats with
different ripening times over 6 months. The NIR spectra were recorded in reectance mode, applying the bre-optic probe directly
onto the cheese sample, with no sample preparation or manipulation. The reference mineral content was determined by ICP-optic
spectroscopy.
2. Material and methods
2.1. Samples and cheese-making procedure
To perform the present study a total of 227 cheeses of known
composition were elaborated and controlled (Gonzalez-Martin
et al., 2007). Cheeses were prepared in the laboratory following
this procedure: raw milk (40 L), not standardised, was incubated
with 15 mg L 1 direct-vat-set starters made of Streptococcus lactis,
cremoris and Streptococcus diacetilactis, (MA400, Arroyo Laboratories, Santander, Spain) at 30 C. After 10 min at 32 C, 12.5 mg L 1
of calf rennet (90% chymosin, 10% trypsin, 1:150,000 strength)
was added to each vat. Coagulation was allowed to take place over
3540 min. When the curds had developed the desired rmness,
evaluated subjectively, they were cut with a cheese harp until
pieces similar in size to a grain of rice (6 mm) were obtained.
The curd was then stirred for 30 min and heated for 1020 min
at 37 C until it had reached the desired consistency to improve
its drainage with sieves. The curd was packed in round hoops
(1 kg) and pressed for 6 h at 1.5 kg cm 2 at 20 C, with the inversion of the pieces of cheese at the mid-time of the pressed manufacturing. After pressing, the cheeses were salted by soaking them
in sodium chloride brine (18% w/v) at 18 C for 6 h. The cheeses
were then moved to a drying chamber, where temperature
(15 C) and relative humidity (70%) were controlled. They were
made of milk collected directly from farms in winter and summer.
Bovine, ovine and caprine raw milk were obtained directly from
the producers in Zamora (Spain). Cheeses with different compositions were elaborated, prepared with known, varying amounts of
milk from cows, ewes and goats, with percentages ranging between 0% and 100% (Gonzalez-Martin et al., 2007; Revilla et al.,
2009). These were cylindrical, with an initial diameter of 10 cm
and a thickness of 5 cm and they were monitored over 6 months
(at 0.2, 1, 2, 3, 4, 5 and 6 months), using one of the pieces at each
time. Each time, a cheese was monitored, the piece was cut in half
transversally and a rectangular slice of approximately 2 cm in
thickness was collected.
Of all the 227 samples, 170 were employed for the denominated
calibration set, and the other 57 were used for the external
validation.
2.2. Plasma ICP-optic spectroscopy
The chemical measures of reference were performed by ICPOES, after mineralisation in a system of microwaves Model Ethos
Plus (Milestone). The samples of cheese were ground and dried
with a heater at 105 C for 24 h and subjected to mineralisation
in a microwave system. About 0.5 g of dry, ground sample was
later introduced into a high pressure capsule. In a rst stage
5 mL of HNO3 were added (c) and a potential of 300 W was applied
for 5 min. Once the sample was cold a further 5 mL of HNO3 were
added (c) and 1 mL of H2O2 30%, applying a potential of 300 W for

7 min. The sample was cooled to room temperature, made up to

100 mL with distilled water and maintained at 4 C until analysis.
A Jobin Yvon ICP OES model Ultima2 powered by a radiofrequency generator at 1100 W was used for elemental determination.
The following conditions were used to carry out the elemental analysis: speed of the bomb: 20 rpm; plasma argon gas ow PL1: 12 L/
min; pod gas ow G1: 0.2 L/min; nebulizer ow: 1.0 L/min; nebulizer pressure: 2.95 bars; with the use of an argon humidicator.
The analytical lines used for the different elements were Ca:
317.9333 nm; K: 766.490 nm; Mg: 279.553 nm; Na: 589.592 nm;
and P: 177.440 nm. The range of the standards concentrations has
been used for Ca, Na and P: 51020 ppm; for K and Mg: 0.25
0.501 ppm (concentrations of the standard dissolution which in
the equivalence in cheese were: Ca, Na and P: 10002000
4000 mg/kg; and K and Mg: 100200400 mg/kg). In those samples
which the concentration was not found in its respective lineal margin were diluted with ultrapure water and were re-analysed until
they were in the lineal response margin. The detection limits were:
Ca: 10 mg/kg; K: 22 mg/kg; Mg: 10 mg/kg; Na: 36 mg/kg; P: 28 mg/
kg.
The mineralisation and the ICP measurements were carried
out in the Servicio de Anlisis Qumico of the Universidad de
Salamanca, Spain.
2.3. NIR spectroscopy
A Foss NIRSystem 5000 with a standard 1.5 m 210/7210 bundle
bre-optic probe, Ref. no. R6539-A, was used. The probe employs a
remote reectance system and uses a ceramic plate as reference.
The window is of quartz with a 5  5 cm surface area, measuring
reectance in the IR zone close to 11002000 nm. The measurement of the spectra was carried out using NIRS technology and a
remote reectance bre-optic probe that was applied directly to
the cheese samples with no prior treatment or manipulation. The
spectra were recorded at intervals of 2 nm, performing 32 scans
for both the reference and samples. To minimise sampling error,
all the samples were analysed in triplicate. The software used
was Win ISI 1.50 installed on a HewlettPackard Pentium III
computer.
2.4. Statistical analyses
Calibrations were developed using WinISI II version 1.5 (Infrasoft International). The samples were divided into calibration
(n = 170) and validation (n = 57) sets. All types of cheeses were represented in both the validation and calibration sets. The modied
partial least squares (MPLS) regression method was used to obtain
the NIR equations for all the parameters studied (Shenk &
Westhaus, 1995). Partial least squares (PLS) regression is similar
to principal component regression (PCR), but uses both reference
data (chemical, physical, etc.) and spectral information to obtain
the factors useful for tting purposes (Martens & Naes, 2001).
MPLS is often more stable and accurate than the standard PLS algorithm. In MPLS, the NIRS residuals at each wavelength obtained
after each factor has been calculated are standardised (dividing
by the standard deviations of the residuals at each wavelength) before calculating the next factor. When developing MPLS equations,
cross-validation is recommended in order to select the optimal
number of factors and to avoid overtting. For cross-validation,
the calibration set is partitioned into several groups. Each group
is then validated using a calibration developed on the other samples. Finally, validation errors are combined into a standard error
of cross-validation (SECV) (Davies & Williams, 1996). It has been
reported that the SECV is the best single estimate of the prediction
capability of the equation and that this statistic is similar to
the average standard error of prediction (SEP) from 8 to 10

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I. Gonzlez-Martn et al. / Food Chemistry 127 (2011) 147152

3.1. Chemical composition

Table 1 shows the minimum, maximum and mean concentrations and standard deviations of the mineral composition in calcium, phosphorus, magnesium, potassium and sodium in the
samples of cheese studied obtained by the method of reference.
The total of 227 samples was divided into two groups, calibration
(170 samples) and external validation (57) sets and the Table 1
presents the values for both groups.
The average levels of calcium (8.1 g/kg) are generally up to 10
times higher in hard cheeses and 4 times more in cheeses with
mould than in milk (Scott, 1989). The average concentration of
phosphorus, 3.73 g/kg, is in agreement with those found in cured
cheeses of goat (3.60 g/kg) (Almenara et al., 2007). The levels of
calcium and phosphorus are higher in cheeses of cow in comparison to those found by (Raynal-Ljutovac, Lagriffoul, Paccard, Guillet,
& Chilliard, 2008), which shows that these two elements are found
in similar quantities in the milk of goat and cow; they are the most
abundant in ewes milk.
The relationship Ca: P is 2.2, a high value compared with those
obtained in goats cheeses (Park, 2000) and in cows cheeses
(Jenness, 1980; Prieto et al., 2002). The average content of potassium, 1.26 g/kg is in agreement with other works (Herrera et al.,
2006). The quantity of potassium depends on the animal origin
of the milk, the highest values being presented in cows cheeses
and the lowest in ewes. In addition, the concentration of this mineral diminishes throughout the process of ripening, which suggests
that a considerable quantity is lost during the process of elaboration of the cheese, as a consequence of the acidication of the curds
(Almenara et al., 2007).
The high concentration of sodium throughout the whole process of ripening, with an average value of 7.99 g/kg, is principally
due to the addition of salt during the process of elaboration and
the period of ripening (Gambelli et al., 1999). Moreover, the seasonal variability is observed, thus the cows cheeses elaborated in
winter present lower concentrations of sodium than the cheeses
elaborated from goats milk; the opposite of what occurs for the

3.2. Calibration equation

To obtain the calibrations, a starting set of 170 cheese samples
of varying compositions of cows, ewes and goats milk was used.
Fig. 1 shows the spectra of ten samples. Initially, a principal component analysis was carried out (PCA). In all cases, the spectral variability explained was above 99%, and 12 principal components
were required for calcium; 8 for phosphorus; 10 for potassium,
10 for sodium and 11 for magnesium.
The calibration process was implemented with the spectra of
the resulting samples and their chemical data. The statistical
parameters of the calibration were obtained for each of the components after removing the samples for spectral (H criterion) or
chemical reasons (T criterion). Anomalous spectra were detected
by applying the Mahalanobis distance. Furthermore, the risk of
1.8
1.6

(a)

1.4

Log (1/R)

3. Results

samples elaborated in summer, the higher concentrations of sodium are presented in cows cheeses in comparison to those of
goat.
The content of Mg does not present variations throughout the
process of ripening and its average levels of 401.2 ppm coincide
with those found in other studies (Cichoscki et al., 2002; Herrera
et al., 2006; Moreno-Rojas et al., 1994).

1.2
1
0.8
0.6
0.4
0.2

0
1100

1200

1300

1400

1500

1600

1700

1800

1900

2000

wavelenghts, mn

none / 2 derivate (Log(1/R))

randomly-chosen prediction sets. In all cases, cross-validation was

performed by splitting the population into four groups. The effects
of scattering were removed using MSC (Multiplicative Scatter
Correction), SNV (Standard Normal Variate), DT (DeTrend) or
SNV-DT, (Dhanoa, Lister, & Barnes, 1995). Moreover, the mathematical treatments were tested in the development of the NIRS
calibrations. The following is an example: 1, 4, 4, 1, where the rst
digit is the number of the derivative, the second is the gap over
which the derivative is calculated, the third is the number of data
points in a running average or smoothing and the fourth is the
second smoothing. The statistics used to select the best equations
were RSQ (multiple correlation coefcients) and the standard error
of cross-validation (SECV).

0.06

Calcium

(b)

0.04
0.02
0
-0.02
-0.04
-0.06
-0.08
1100

1200

1300

1400

1500

1600

1700

1800

1900

2000

wavelengths, nm
Fig. 1. (a) NIR spectra of 10 samples of cheese and (b) mathematical treatments
(none/2 derivate) in the spectra.

Table 1
Main mineral composition of the samples of cheese.
Calibration set (N = 170)

External validation set (N = 57)

Element

Minimum

Maximum

Mean

SD

Minimum

Maximum

Mean

SD

Ca (g/kg)
P (g/kg)
K (g/kg)
Na (g/kg)
Mg (ppm)

4.49
2.43
0.62
2.76
257.85

40.38
6.03
2.17
13.92
686.69

8.11
3.73
1.26
7.99
401.19

3.8
0.7
0.4
2.7
67.0

5.12
2.35
0.73
3.83
269.43

12.56
5.68
1.88
12.96
561.96

7.89
3.83
1.23
7.74
405.56

1.6
0.8
0.3
2.2
58.8

N: number of samples; SD: standard deviation.

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I. Gonzlez-Martn et al. / Food Chemistry 127 (2011) 147152

there being mistakes in the equations under practical conditions is

very low or almost null when the standardised H statistic (Mahalanobis distance) is used during routine analysis of unknown samples. This tells us how different the spectrum of the unknown
sample is from the average spectrum in the calibration set. Samples with an H-value greater than three may be considered as
not belonging to the population from which the equations are
developed, and in this case the equations should not be used to

make any prediction; 7 samples were removed for calcium; 9 for

phosphorus, 7 for potassium, 6 for sodium and 7 for magnesium.
Calibrations were performed by modied partial least squares
regression (MPLS). Using the T P 2.5 criterion, samples that were
different from the population owing to the chemical criterion were
removed from the set. On the basis of this chemical criterion, 7
samples were removed for calcium, 8 for phosphorus, 5 for potassium, 14 for sodium and 8 for magnesium.

Table 2
Descriptors of NIR calibration.
Element

Mathematical treatment

SD

Estimate

RSQ

SEC

SECV

N fac PLS

Cross-validation groups

RPD

Ca (g/kg)
P (g/kg)
K (g/kg)
Na (g/kg)
Mg (ppm)

156
153
158
150
155

None 2, 4, 4, 1
Standard MSC 1, 4, 4, 1
Standard MSC 2, 10, 10, 1
Standard MSC 2, 8, 6, 1
Detrend only 0, 0, 1, 1

1.3
0.5
0.3
2.7
60.4

3.7511.52
2.035.28
0.202.30
015.88
216.17578.82

0.74
0.69
0.86
0.92
0.72

0.65
0.30
0.13
0.74
31.95

0.82
0.33
0.16
0.80
40.25

8
8
8
10
10

4
4
4
4
4

2.0
1.8
2.7
3.8
2.0

N: number of samples; MSC: Multiplicative Scatter Correction , SD: standard deviation; RSQ: multiple correlation coefcients; SEC: standard error of calibration;
SECV: standard error of cross-validation; PLS: partial least squares; RPD: ratio performance deviation.

11.5

CALCIUM

Phosphorus (g/kg).REF

Calcium (g/kg).REF

12.5

10.5
9.5
8.5
7.5

RSQ: 0.76

6.5

SEP: 0.64

5.5

SEP(c): 0.64

4.5
4.5

5.5

6.5

7.5

8.5

9.5

10.5

PHOSPHORUS

5.5
5
4.5
4

RSQ: 0.70

3.5

SEP: 0.29

SEP(c): 0.29

2.5
2.5

11.5

3.5

Calcium (g/kg).NIR

Potassium (g/kg). REF

Sodium (g/kg).REF

SODIUM

14.5
12.5
10.5
8.5

RSQ: 0.93

6.5

SEP: 0.71

4.5

SEP(c): 0.71

2.5
2.5

4.5

5.5

2.5

18.5
16.5

Phosphorus (g/kg).NIR

7.5

12.5

2.3

POTASSIUM

2.1
1.9
1.7
1.5
1.3
1.1

RSQ: 0.87

0.9

SEP: 0.13

0.7

SEP(c): 0.13

0.5
0.5

17.5

0.7

0.9

Sodium (g/kg).NIR

1.1

1.3

1.5

1.7

1.9

2.1

2.3

Potassium (g/kg).NIR

Magnesium (ppm). REF

700
650

MAGNESIUM

600
550
500
450
400

RSQ: 0.74

350

SEP: 30.80

300

SEP(c): 30.90

250
250

300

350

400

450

500

550

600

Magnesium (ppm ). NIR

Fig. 2. Correlation of the values obtained in the laboratory with respect to those predicted by NIR with a bre-optic probe for the main mineral composition in cheese.

I. Gonzlez-Martn et al. / Food Chemistry 127 (2011) 147152

The best of the different mathematical treatments, concentration range and standard deviation for calcium, phosphorus, potassium, sodium and magnesium are shown in Table 2. The results
obtained indicate that it is possible to determine calcium, phosphorus, potassium, sodium and magnesium in cheeses of varying
compositions of cows, ewes and goats milk by the direct application of the bre-optic probe on the samples of cheese, without
treatment or manipulation.

151

nicance obtained were between 0.65 for magnesium and 0.15 for
phosphorus, potassium and sodium. There were no differences between the results obtained, so it can be concluded that the method
provides signicantly identical data to the starting reference data.
Table 3 shows the results obtained in the external validation, the
means of the residuals and the Root Mean Standard Error (RMSE)
values.
4. Conclusion

3.3. Validation
3.3.1. Internal validation (prediction)
Models evaluations were performed by cross-validation. In this
method, the set of calibration samples is divided into a series of
subsets, in our case 4. Of these, 3 were taken for the calibration
set and one for the prediction set. The process is repeated as many
times as there are sets, so that all pass through the calibration set
and the prediction set. Using this process, we validated the models
used and checked their prediction capacities. Fig. 2 shows the correlation of the values obtained in the laboratory with respect to
those predicted by NIR with a bre-optic probe for calcium, phosphorus, magnesium, potassium and sodium in cheeses elaborated
with different percentages of cows, ewes and goats milk with ripening times of up to 6 months.
3.3.2. Prediction capacity of the model
The prediction capacity of the model obtained was evaluated
with the ratio performance deviation (RPD) (Williams & Sobering,
1996), a capacity parameter that is dened as the relationship between the standard deviation of the chemical method (SD ref) and
the standard prediction error (SEP) encountered in the NIRS model.
If the RPD value is greater than 2.5 the model is considered suitable. Table 2 shows the values obtained for RPD parameter that
were, comprehended between 3.8 for sodium and 1.8 for phosphorus. These results demonstrate that the capacity for prediction can
be considered excellent for the potassium and sodium, and acceptable for calcium, phosphorus and magnesium, indicating that NIRS
technology presents a capacity for prediction that is interesting for
the determination of mineral composition in samples of unknown
cheese of up to six months of ripening elaborated with milk (cows,
ewes and goats) from different seasons.
3.3.3. External validation
We checked the robustness of the method by applying NIRS
technology to 57 new samples of different compositions and ripening times which did not belong to the calibration group. The procedure was as follows: spectra were recorded in triplicate and the
spectral mean was taken. Then, the calibration equations obtained
during the work were applied and the predicted values were compared with the reference data for calcium, phosphorus, magnesium,
potassium and sodium determined by ICP-optic spectrometry.
The NIRS methodology and the reference data concerning the
calcium, phosphorus, magnesium, potassium and sodium were
compared with those of samples not belonging to the calibration
model using the Students t-test for paired values. The levels of sig-

Table 3
External validation (57 samples).
Element

p (Level of signicance)

Residual mean

RMSE

Ca (g/kg)
P (g/kg)
K (g/kg)
Na (g/kg)
Mg (ppm)

0.40
0.15
0.15
0.15
0.65

0.69
0.34
0.16
1.01
39.81

0.93
0.46
0.22
1.52
49.80

RMSE: Root Mean Standard Error.

The results obtained in this work are comparable to those obtained in fresh, pure cheeses of cow and goat, using ground samples of cheese (Lucas et al., 2008). It should be emphasised that
the capacity of prediction (RPD) for potassium and sodium in our
work are higher, whereas for calcium and magnesium they are
lower due to the greater heterogeneity of the cheeses analysed
here.
Moreover, in this work the determination of phosphorus in the
cheeses in a range of 2.035.28 g/kg is achieved. Phosphorus, as is
known, is the second mineral in importance in this type of samples
and affects the capacity of absorption of the calcium.
The most determinant aspect is that the methodology developed can be applied to any type of unknown cheese, pure or elaborated with percentages of milk from different origins without
previously treating or destroying the sample, which is to say, by
direct application of the bre-optic probe on a slice of cheese, with
results comparable to ICP-optic spectroscopy.
Acknowledgements
This study was made possible by funds from Projects CTQ200604383; SA 139A08. We would like to thank Mr. G.H. Jenkins for his
help with the English version of the manuscript.
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