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Talanta
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art ic l e i nf o
a b s t r a c t
Article history:
Received 10 October 2013
Received in revised form
11 April 2014
Accepted 16 April 2014
Available online 24 April 2014
A multielemental analytical method has been proposed to determine the contents of Al, B, Ca, Cu, Fe, K,
Mg, Mn, Na, Ni, P, Pb, Sr and Zn in paprika samples from the two Protected Designations of Origin
recognized in Spain, such as Murcia and La Vera (Extremadura). The samples are mineralized by acid wet
digestion using a mixture of perchloric and nitric acids and analyzed by means of inductively coupled
plasma atomic emission spectroscopy. The method performance has been checked studying the absence
of matrix effect, trueness, precision, linearity, limit of detection and limit of quantication. The proposed
method has been applied to analyze samples of sweet, hot and hot/sweet paprika from the considered
production areas. Differences between paprika samples from Murcia and Extremadura were found and
pattern recognition methods, such as linear discriminant analysis, linear support vector machines, soft
independent modeling of class analogy and multilayer perceptrons articial neural networks, has been
used to obtain classication models. Sweet and hot/sweet paprika types were differentiated by means of
linear models and hot paprika was differentiated by using articial neural networks. A model based on
articial neural networks is proposed to differentiate the geographical origin of paprika, with
independence of the type, leading to an overall classication performance of 99%.
& 2014 Elsevier B.V. All rights reserved.
Keywords:
Paprika
Inductively coupled plasma atomic emission
spectrometry
Pattern recognition
Linear discriminant analysis
Support vector machines
Articial neural network
Soft independent modeling of class analogy
1. Introduction
Paprika is a red powder made from grinding the dried pepper
pods of some varieties of Capsicum annuum L. [1]. This natural food
product is commonly used as spice and natural colorant in cookery
and to provide redness to meat products and commercial sauces
[2]. There are different types of paprika according to its pungency,
named sweet, sweet/hot and hot. This variety of tastes makes this
product very common in different traditional cuisines. The adequate consumption of pepper fruits is also related to potential
health effects due to the presence of antioxidant compounds [3].
Although paprika is original from America, it is also produced in
Europe, particularly in Hungary, Turkey and Spain. In Spain,
paprika is called pimentn and it is cultivated and produced in
two main areas namely Murcia and La Vera [4]. Paprika from both
regions is recognized under Protected Designation of Origin (PDO)
by the European Commission of Agriculture and Rural Development [5], being these products of great importance for the local
economies. The production region of the paprika protected by La
Vera PDO is located in the centre-west of Spain. This PDO includes
http://dx.doi.org/10.1016/j.talanta.2014.04.025
0039-9140/& 2014 Elsevier B.V. All rights reserved.
some towns of the province of Cceres in the region of Extremadura. La Vera PDO uses peppers belonging to the variety Bola and
Ocales from the species C. annuum L. and C. longum L., respectively
[6]. The other pepper growing region which is currently protected
under the Murcia PDO comprises the homonymous region, located
to the south east of Spain. In this case, the peppers used belong to
the variety Bola from C. annuum L. [7].
Nowadays, consumers relate quality of foodstuff to certain
characteristics inuenced by the production areas, the harvesting
practices and the raw materials. For this reason, it is of great
importance the development of suitable methodologies allowing
the characterization of different products. Within this context,
several studies have been focused on the compositional proling
of paprika samples. Mateo et al. [4] identied volatile compounds
inuencing the overall avor of paprika from La Vera, whilst
volatile composition of paprika from Murcia has been studied by
Guadayol et al. [8]. Kocsis et al. [9] researched on the volatile
component composition of Hungarian red paprika as an important
parameter of quality and identity. Due to the use of red paprika as
a source of pigments to enhance or change food color, the study of
constituents related to this characteristics, such as carotenoids, is
valuable. In fact, color is used as a parameter to monitoring the
quality changes during the elaboration or storage process. Techniques based on the spectrophotometric measurement of color
16
local stores. Within these classes there are three different types
of paprika: sweet, hot/sweet and hot, being 24 the number of
samples of each pair of type-origin.
2.3. Apparatus and methods
ICP-OES analysis was performed using an ULTIMA 2 atomic
emission spectrometer (Horiba Jobin Yvon, Kyoto, Japan). The ICP
operating conditions are shown in the electronic Supplementary
material (Table S1).
Three mixtures of acids were tested to carry out wet ashing
mineralization of paprika samples: M1 (2 ml of H2SO4 and 15 ml of
HNO3), M2 (5 ml of H2O2 and 20 ml of HNO3) and M3 (2 ml of
HClO4 and 20 ml of HNO3). The acids were gradually added to 1 g
of paprika (weighted with precision of 0.1 mg) placed in a beaker
and the mixture was heated till complete mineralization. All
obtained solutions were cooled to room temperature, ltered
(0.45 mm) and transferred to 50 ml volumetric asks. Adequate
blanks were prepared for each method.
2.4. Chemometrics
A data matrix consisting of 14 columns (the determined elements)
and 144 rows (the paprika samples) was created for the chemometrics
calculations. Two-way nested analysis of variance (ANOVA) was used
to compare recoveries of the mineralization methods. Non-parametric
comparison test such as KruskalWallis and MannWhitney tests
were applied in order to draw attention to signicant difference in
elemental content between the paprika types and origins, respectively.
PCA was used to visualize data trends and to get a rst evaluation
of the discriminant power of the variables. LDA, SVM, SIMCA and MLPANN were applied to obtain classication models. The STATISTICA
8.0 software package (StatSoft, 2007) was used for the statistical
analysis.
Table 1
Two-way nested ANOVA results for method comparison.
ANOVA calculations
Element
S2method
Fexp
Al
B
Ca
Cu
27.4
139.7
42.7
162.4
5.8
96
59
8.5
4.72
1.46
0.72
19.11
Fe
K
Mg
Mn
Na
Ni
P
Pb
1423
3.6
193.8
108.9
558.4
74.4
88.3
270.6
24.6
9.4
6.6
74.4
5.8
85.6
21.7
15.1
57.85
0.38
29.36
1.46
96.28
0.87
4.07
17.92
28.96
Sr
489.5
16.9
Zn
815.6
123.5
M1M2
M1M3
6.6
Table 2
Recoveries and expanded uncertainties obtained for the studied elements for the
compared mineralization methods.
Al
B
Ca
Cu
Fe
K
Mg
Mn
Na
Ni
P
Pb
Sr
Zn
M3M2
S2method, variance due to use of different oxidant mixtures; S2pe, pure error variance;
F(0.05,2,18) 3.55.
Element
17
Recovery, %
RM1 7 URM1
RM2 7 URM2
RM3 7 URM3
99 74
98 713
102 715
91 76
82 76
100 74
108 76
93 723
98 75
93 717
96 710
90 78
92 714
110 713
1027 5
1047 15
1037 11
897 8
1067 14
1017 10
1007 5
917 11
847 11
937 15
1027 8
927 13
827 9
1147 24
997 1
977 3
997 2
977 3
1017 3
1007 2
1017 2
987 4
987 3
987 4
1007 1
1017 3
977 3
967 4
corresponding to Ri values is computed from two terms, combined uncertainty and pure repetition uncertainty [19]. Uncertainties (u) are converted to expanded one (U) multiplying by a
coverage factor k 2 [22]. Recoveries can be considered free from
bias when the inequality jRi 100jr URi holds true. As can be
seen in Table 2 mixture M3 leads to recoveries whose condence
interval includes the 100% for all the elements. In addition, the
uncertainties of M3 are generally lower than those of M1 and M2.
Furthermore, there are some practical considerations to take
into account to select the oxidant mixture to be used for paprika
mineralization. The presence of high concentrations of acids
during the measurement could lead to a decrease of sensitivity
due to the higher viscosity of the solution. For this reason an
evaporation step is needed in order to eliminate the excess of
acids. The vapor pressure of sulfuric acid is very low in comparison
to those for perchloric acid, nitric acid and hydrogen peroxide
aqueous solutions. This means that the elimination of sulfuric acid
requires higher temperatures and longer heating times. In addition
the mixture of nitric and sulfuric acids may yield a precipitate of
compounds like lead sulfate. Perchloric acid may react explosively
with organic compounds but it can be used safely after a rst
action of nitric acid. When hydrogen peroxide is used, longer
Table 3
Repeatability, intermediate precision, linearity, limit of detection and limit of
quantication obtained for the analyzed elements.
Element RSDrepeat (%) RSDip (%) % L
Al
B
Ca
Cu
Fe
K
Mg
Mn
Na
Ni
P
Pb
Sr
Zn
0.1
3.2
0.1
0.45
0.05
2.25
0.05
0.05
0.4
0.3
0.55
0.4
0.05
0.05
0.88
6.37
2.44
3.07
2.25
2.10
2.04
8.26
4.34
7.93
0.72
4.19
1.57
10.21
1.86
8.38
3.50
3.97
3.89
2.79
3.16
11.43
4.69
10.00
1.17
4.74
8.30
11.69
98.8
96.78
99.1
99.0
98.87
98.6
95.3
97.2
98.4
98.7
99.1
96.9
98.4
98.3
0.35
10.5
0.35
1.5
0.15
7.5
0.15
0.15
1.35
1
1.85
1.5
0.15
0.15
18
Table 4
Median, minimum and maximum (in brackets) metal content in Spanish paprika. Concentrations expressed in mg kg 1 (dry weight).
Element
Al
B
Ca
Cu
Fe
K
Mg
Mn
Na
Ni
P
Pb
Sr
Zn
MURCIA
EXTREMADURA
Sweet
Hot
140 (95276)
14.2 (7.420.8)
1946 (14982442)
20.3 (9.322.2)
229 (183403)
15687 (916920158)
2012 (17582657)
24.9 (10.735.8)
509 (419881)
0.93 (0.811.43)
3396 (30813575)
0.49 (0.0310.7)
29.8 (9.638.5)
43.3 (14.962.2)
136
12.2
2191
12.9
201
14538
2256
20.5
441
1.06
3555
0.40
15.1
15.1
(43353)
(0.2622.7)
(10122790)
(9.122.6)
(123418)
(922619765)
(17032718)
(12.353.1)
(168784)
(0.831.56)
(26854093)
(0.0911.1)
(8.538.4)
(14.260.8)
Hot/Sweet
Sweet
42
7.9
1494
11.1
106
14387
1785
10.2
546
0.61
3644
0.10
12.2
30.5
132
14.8
1895
11.6
178
14734
2189
26.9
328
1.00
3613
0.40
8.7
35.7
(26137)
(5.523.6)
(10312174)
(9.222.6)
(85264)
(769117547)
(16422190)
(9.127.7)
(278948)
(0.540.88)
(31544226)
(0.019.1)
(9.032.1)
(23.969.0)
Hot
(80246)
(7.327.9)
(10632465)
(6.020.0)
(84295)
(1117517581)
(14832461)
(19.797.6)
(155531)
(0.172.94)
(24994266)
(0.0319.0)
(5.729.1)
(18.869.6)
117
13.5
2044
12.7
168
14261
2174
26.1
333
1.00
3568
1.12
9.4
32.0
Hot/Sweet
(83160)
(7.023.5)
(8332437)
(7.420.6)
(123236)
(1064023416)
(18672390)
(15.355.6)
(255497)
(0.522.13)
(31703950)
(0.1016.8)
(5.521.3)
(18.962.2)
128
10.5
1862
9.8
183
15206
2194
28.7
377
1.02
3619
0.42
9.2
33.4
(91227)
(5.823.8)
(11132422)
(6.520.7)
(150247)
(1031820213)
(19512346)
(19.168.5)
(21560)
(0.493.40)
(32634140)
(0.0421.3)
(6.616.3)
(18.360.5)
19
Fig. 1. Scatterplot of paprika in the space formed by the three rst PCs for (A) global comparison; (B) sweet paprika; (C) hot paprika and (D) hot/sweet paprika.
20
Fig. 2. Distribution of the samples according to their scores for the computed discriminant function for (A) global comparison; (B) sweet paprika; (C) hot paprika and
(D) hot/sweet paprika.
Table 5
SENS and SPEC results (%) obtained by the built differentiation models for (1) global
comparison, (2) sweet paprika, (3) hot paprika and (4) hot/sweet paprika.
Model
LDA1
LDA2
LDA3
LDA4
SVM1
SVM2
SVM3
SVM4
SIMCA1
SIMCA2
SIMCA3
SIMCA4
MLP-ANN1
MLP-ANN2
MLP-ANN3
MLP-ANN4
Extremadura
Murcia
Overall
SENS
SPEC
SENS
SPEC
SENS
SPEC
987 3
100
937 12
100
947 4
100
837 12
100
847 6
767 15
787 20
91 715
997 2
100
967 7
100
91 77
100
897 13
100
877 9
987 7
877 18
100
847 6
100
887 20
100
997 4
100
997 4
100
917 7
100
897 13
100
877 9
987 7
877 18
100
877 6
897 15
897 16
73 724
997 4
100
997 4
100
987 3
100
937 12
100
947 4
100
837 12
100
64 77
100
357 21
100
997 2
100
967 7
100
957 4
100
91 710
100
91 73
997 3
857 9
100
867 3
837 10
857 11
867 12
997 2
100
987 4
100
947 5
100
91 710
100
91 74
997 4
857 8
100
747 5
100
597 19
100
997 2
100
977 4
100
21
Fig. 3. Cooman's plots obtained from SIMCA models for (A) global comparison; (B) sweet paprika; (C) hot paprika and (D) hot/sweet paprika.
4. Conclusions
A method to perform the determination of Al, B, Ca, Cu, Fe, K,
Mg, Mn, Na, Ni, P, Pb, Sr and Zn in paprika samples has been
proposed. Different mixtures of acids to perform the mineralization have been tested, being the mixture of perchloric and nitric
acids which leads to adequate recoveries for all the considered
elements and also presents practical advantages. The method
22