Analytical Methods

Chemometric approach to evaluate trace metal concentrations in some spices

and herbs
Cennet Karadas , Derya Kara

Department of Chemistry, Art and Science Faculty, Balikesir University, 10100 Balikesir, Turkey
a r t i c l e i n f o
Article history:
Received 12 December 2010
Received in revised form 2 June 2011
Accepted 5 July 2011
Available online 12 July 2011
Keywords:
Classication
Herbs
Spices
Trace elements
Principal component analysis
Linear discriminant analysis
Cluster analysis
a b s t r a c t
Herbs (mint, thyme and rosemary) and spices (black pepper, chili pepper, cinnamon, cumin, sweet red
pepper and turmeric) were analysed using atomic spectrometry and then subjected to chemometric eval-
uation in an attempt to classify them using their trace metallic analyte concentrations (As, Ba, Ca, Cd, Co,
Cr, Cu, Fe, Mg, Mn, Ni, Sr and Zn). Trace metals in acid digests of these materials were determined using
both inductively coupled plasma-atomic emission spectrometry and inductively coupled plasma-mass
spectrometry. The chemometric techniques of principal component analysis (PCA), linear discriminant
analysis (LDA) and cluster analysis (CA) were used for the classication studies. These herbs and spices
were classied into ve groups by PCA and CA. When the results of these techniques were compared with
those from LDA, it was found that all group members determined by PCA and CA are in the predicted
group that 100.0% of original grouped cases correctly classied by LDA.
2011 Elsevier Ltd. All rights reserved.
1. Introduction
Spices and herbal plants contain metal ions over a wide range of
concentrations. Metals can have important positive or negative
roles in human life. Twenty-three elements have known physiolog-
ical activities in humans and other mammals. Some of these ele-
ments, including iron, zinc and copper, are considered essential.
Other metal ions such as cadmium, lead and mercury have toxic
roles in biochemical reactions on our body. The metals vanadium,
chromium, manganese, iron, cobalt, copper, zinc and molybdenum;
and the non-metals selenium, uorine, and iodine are trace ele-
ments that belong to the category of micronutrients, which are
needed by the human body in very small quantities (generally less
than 100 mg/day). Other elements are considered to be macronutri-
ents, e.g. sodium, calcium, magnesium, potassium, chlorine, etc.,
which are required in larger quantities. Some trace elements are
essential components of biological structures, but they can also
be toxic at concentrations beyond those necessary for their biolog-
ical functions. Examples of these metals include cobalt, copper,
chromium and nickel (Fraga, 2005). For example, at trace levels,
nickel is benecial for the activation of some enzyme systems how-
ever, its toxicity at higher levels is more prominent. Fortunately,
nickel toxicity in humans is not a very common occurrence because
the absorption of nickel is very low (Onianwa, Lawal, Ogunkeye, &
Orejimi, 2000). In biological systems, trace elements are mostly
conjugated or bound to proteins forming metalloproteins, or to
smaller molecules, such as phosphates, phytates, polyphenols and
other chelating compounds. Most of the metals in metalloproteins
are part of enzymatic systems, have structural functions, or use
the protein to be transported to their target site in the organism
(Fraga, 2005). For example copper and zinc serve either as a cofac-
tor, as an activator for biochemical reactions and are integral com-
ponents of some enzyme/substrate metal complexes (Mildvan,
1970).
There are many herbs and spices widely consumed in our daily
diet in Turkey and, indeed, worldwide. Amongst the many benets
of these products, rosemary is known for its anti-inammatory
power, cumin for its dementia-ghting power, cinnamon for
combating obesity and sugar regulating powers, turmeric for its
anti-cancer properties, thyme for its skin-saving power. Since these
herbs and spices are being consumed in daily diets or as supple-
ments, it is important to determine their nutrient composition so
that their effect onhumanhealthcanbe understood. It is alsoimpor-
tant totrytoelucidate whether or not there are any relationships be-
tween different plant types and the uptake of metals from the soils.
The mineral andtrace element contents have beendeterminedin
herbs and herbal teas (Basgel & Erdemoglu, 2006; Lozak, Soltyk,
Ostapczuk, & Fijalek, 2002; zcan & Akbulut, 2008; zcan, nver,
Uar, & Arslan, 2008; Sun, Qiao, Suo, Li, & Liang, 2004) and also in
spices (Chizzola, Michitsch, & Franz, 2003; Ozkutlu, 2008; Ozkutlu,
Kara, & Sekeroglu, 2007; Soylak, Tzen, Narin, & Sar, 2004) in
several previous publications. Some of these studies compared the
concentration of one element with another or compared one
0308-8146/$ - see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.07.006

Corresponding author. Tel.: +90 266 612 10 00; fax: +90 266 612 12 15.
E-mail addresses: dkara72@gmail.com, dkara@balikesir.edu.tr (D. Kara).
Food Chemistry 130 (2012) 196202
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sample with another; usually using univariate methods such as
analysis of variance (ANOVA). However, multivariate methods such
as principal component analysis (PCA), correlation analyses, linear
discriminant analysis (LDA) and cluster analysis (CA) can provide
further interpretation and classication between trace metal ions.
In particular, PCA is a very powerful data reduction technique that
aims to explain most of the variance in the data whilst reducing
the number of variables to a few uncorrelated components
(Anderson, 2003). This technique is often used in data reduction
to identify a small number of factors that explain most of the
variance observed in a much larger number of variables. Principal
component analysis may also be used to generate hypotheses
regarding causal mechanisms or to screen variables for subsequent
analysis. For example, it can be used to identify collinearity prior to
performing a linear regression analysis (SPSS 10, 19891999).
Therefore each variable and object is associated with a loading
and a score for each principal component (Brereton, 1990). When
plotted, these scores show the locations of the samples along each
model component, and can be used to detect sample patterns,
groupings, similarities or differences. Principal component analysis
may also be used to plot loadings that are estimated in bilinear
modelling methods where information carried by several variables
is concentrated onto a few components. Each variable has a loading
along each model component. The loadings show how well a
variable is taken into account by the model components. They can
be used to understand how much each variable contributes to the
meaningful variation in the data, and to interpret variable relation-
ships. They are also useful for interpreting the meaning of each
model component (CAMO Software AS, 1998). Cluster analysis is
an unsupervised pattern recognition that involves trying to deter-
mine relationships between objects (samples) without using any
prior information about these relationships. Conversely, when
supervised pattern recognition techniques are used, certain objects
are assumed to fall into pre-determined groups or classes, which are
then used to produce class models. Linear discriminant analysis is
one of the most commonly used supervised pattern recognition
methods (Brereton, 1990).
Principal component analysis has been used to evaluate
whether or not there is a relationship between the metal contents
in different herbal teas (Kara, 2009). Principal component analysis
was also used to evaluate teas (green and black tea) collected from
different parts of the world and their metal contents (Fernndez-
Cceres, Martn, Pablos, & Gonzlez, 2001; Marcos, Fisher, Rea, &
Hill, 1998; Moreda-Pieiro, Fisher, & Hill, 2003).
Principal component analysis has been used to evaluate
whether or not there is a relationship between the metal contents
in different herbal teas (Kara, 2009). Principal component analysis
was also used to evaluate teas (green and black tea) collected from
different parts of the world and their metal contents (Fernndez-
Cceres, Martn, Pablos, & Gonzlez, 2001; Marcos, Fisher, Rea, &
Hill, 1998; Moreda-Pieiro, Fisher, & Hill, 2003).
The aim of this study was to apply the chemometric techniques
of principal component analysis (PCA), linear discriminant analysis
(LDA) and cluster analysis (CA) to results obtained from the atomic
spectrometric analysis of acid digests of herbs and spices to evalu-
ate whether or not there is a similarity with respect to their min-
eral and trace metal contents. Since these spices and herbs have
been used to give avour to dishes and as medicinal effects for
health issues, it is important to know their elemental content since
some of these elements have either toxic effects or essential prop-
erties on human health. It is therefore important to classify these
spices or herbs using different statistical methods to try and under-
stand which of them have similar effects for very important essen-
tial elements such as Fe, Mg, Ca, Mn, Zn or toxic elements such as
Cd, Co and As human health. Before any of these chemometric
techniques can be used, it is important to ensure that accurate ana-
lytical data are being obtained since the chemometric techniques
and the conclusions drawn from them can only be as reliable as
the data. Consequently, all sample types were analysed under strict
analytical control with the use of regular check standards and the
analysis of a certied reference material being used routinely.
2. Experimental
2.1. Reagents and solutions
All chemicals used were of analytical grade. Double-deionized
water was used for preparing the solutions and for all dilutions.
All stock solutions of metals (1000 or 100 mg L
1
) were prepared
from the nitrate salts of metals. Plant digests were prepared using
HNO
3
(Merck, Germany). A certied reference material (GBW
07604 Poplar Leaves obtained from National Institute of Standards
and Technology, Beijing, China) was used to verify the accuracy of
the results of analyses.
2.2. Instrumentation
AnICP-MS instrument (VGPlasmaQuad, PQ2+Turbo, ThermoEle-
mental, Winsford, Cheshire, UK) was used for the determination of
Cd, As, Ni, CoandCr. Operating conditions for the ICP-MS instrument
were: forward power 1.35 kW, coolant gas ow rate 12 L min
1
,
auxiliarygas owrate 1 L min
1
, nebulizer gas owrate 0.9 L min
1
.
An ICP-OES instrument (Varian 725-ES, Melbourne, Australia) was
used for the determination of Ba, Cu, Fe, Mg, Mn, Sr, Zn and Ca in
the plant digests. Operating conditions for the ICP-OES instrument
were: forward power 1.4 kW, coolant gas owrate 15 L min
1
, aux-
iliary gas owrate 1.5 L min
1
; nebulizer gas owrate 0.68 L min
1
and the viewing height was 8 mm above the load coil.
2.3. Procedure
2.3.1. Sample preparations
The spices and herbs, thyme (Thymus vulgaris, Labiatae), cumin
(Cuminum cyminum, Apiaceae), mint (Mentha spicata, Lamiaceae),
cinnamon (Cinnamomum verum, Lauraceae), turmeric (Curcuma
longa, Zingiberaceae), chili pepper (Capsicum annuum, Nightshade
family (Solanaceae)), rosemary (Rosemaryinus ofcinalis, Lamia-
ceae), sweet red pepper (Capsicum annuum, Nightshade family
(Solanaceae)), and black pepper (Piper nigrum L., Piperaceae) were
purchased from a supermarket in Balikesir, Turkey. These samples
have been sold as air dried and no further drying was applied to the
samples, because it is in this form in which they are ingested. The
samples were ground using a pestle and mortar. The pulverised
and powdered spice and herb samples were transferred into plastic
bags. All spices and herbs were treated in an identical manner. For
acid digestion, approximately 0.25 g of spices or herbs was
weighed accurately into pre-cleaned beakers. Concentrated nitric
acid (10 mL) was added, the beakers were then covered with
watch-glasses and the samples boiled gently on a laboratory hot-
plate until digestion was complete. This process took approxi-
mately 2 h. The digested samples were then allowed to cool before
being transferred quantitatively into pre-cleaned 25 mL volumetric
asks. The samples were then diluted to volume by the addition of
ultrapure water. Three replicate digestions were made for each
spice and herb type. The accuracy of the results obtained for the
analyses were shown using a certied reference material.
2.3.2. Sample analysis
In the spice and herb acid extracts, Ba, Ca, Cu, Fe, Mg, Mn, Sr and
Zn were determined using ICP-OES and As, Cd, Co, Cr and Ni were
determined using ICP-MS. Indium (as an internal standard for
C. Karadas , D. Kara / Food Chemistry 130 (2012) 196202 197
ICP-MS measurements) was added to each digest to give a
concentration of 100 lg L
1
after dilution to 25 mL. All results were
the mean of the three replicates and are quoted on a dry weight ba-
sis. All statistical calculations were made using SPSS 10 (SPSS 10 &
Release 10.0.1, 19891999).
3. Results and discussion
The average results and standard deviations of three replicate
analyses obtained using ICP-OES and ICP-MS are shown in Table 1.
As shown in Table 1, the results obtained for the analysis of the cer-
tied reference material GBW 07604, Poplar Leaves were, in gen-
eral, in close agreement with reference values, with only Fe, Mg
and Sr being signicantly lower than the certied value ( the
uncertainty value). Although the nitric acid attack was particularly
efcient at destroying the organic matter of the samples, the Poplar
Leaves CRM, in particular, also contains signicant proportion of
inorganic material; perhaps a silicaceous backbone. This left a res-
idue that no amount of boiling with nitric or perchloric acids could
destroy, even in the presence of hydrogen peroxide. The elements
that gave slightly lower concentrations than the certied values
are renowned for being associated with silicaceous material.
Therefore, they would not be expected to be extracted completely.
The data given in Table 1 show that, for all samples, the analytes
are below the maximum permissible limits in foods; which are
1 mg kg
1
, 0.3 mg kg
1
, 40 mg kg
1
and 60 mg kg
1
for As, Cd, Cu
and Zn, respectively (WHO, 1999). There are no maximumpermissi-
ble levels for the other analytes whose data are presented in Table 1.
Using the data in Table 1, metals were classied using correla-
tion analysis and principal component analysis. The plants were
classied using principal component analysis, cluster analysis
and linear discriminant analysis.
3.1. Correlation analysis
The correlation coefcient is a measure of the linear relation-
ship between two attributes or columns of data. The correlation
coefcients can range from 1 to +1 and is independent of the
units of measurement. A value near 0 indicates virtually no corre-
lation exists between two attributes; whereas a value near +1 or
1 indicates a high level of correlation. Correlation analysis of total
element contents in spices and herbs (Table 2) shows moderate to
strong correlations in ve groups of elements. The correlations gi-
ven in Table 2 indicate that the relationships between the elements
are complex and are difcult to explain individually. Correlation
coefcient values higher than 0.5 were used for the interpretation
of the correlation analysis. However, some values close to 0.5 were
also included to produce grouping such as 0.45 for As and Ni; 0.40
for Ca and Ba and 0.43 for Sr and Mn. Interpretation of correlation
analyses enabled the groupings below to be obtained:
Group 1: Ca, Sr, Ba
Group 2: Mg, Co, Cu, Fe
Group 3: Ba, Mn, Sr
Group 4: Cd, Zn
Group 5: Ni, Cr, Co, Fe, As
More interpretations between elements and spices and herbs
may be obtained using more powerful chemometric techniques
such as PCA.
3.2. Principal component analysis
Principal component analysis provides information on the most
meaningful parameters that describe the whole data set enabling
data reduction with only a minimum loss of original information.
It is a powerful technique for pattern recognition that attempts
to explain the variance of a large set of inter-correlated variables
and transform them into a smaller set of independent (uncorre-
lated) variables (principal components). The PCs generated by
PCA are sometimes not readily interpreted. A rotation is used to
further analyse initial PCA results with the aim of making the pat-
tern of loadings clearer. This process is designed to reveal the sim-
ple structure (Brown, 2009).
Principal component analysis was applied to the entire data set
using the SPSS programme. The principal components that have
Eigenvalues higher than 1 were extracted. The data were subjected
rst to direct oblimin rotation that is one of the oblique rotation.
Correlations were lower than 0.32 for four components at the com-
ponent correlation matrix. It was therefore decided that the data
Table 1
Concentrations of elements in different plants (mean and standard deviations).
Plants Cd (ng g
1
) As (ng g
1
) Co (ng g
1
) Ni (lg g
1
) Cr (lg g
1
) Ba (lg g
1
) Cu (lg g
1
)
Reference plant (certied value) 320 70 370 90 420 30 1.9 0.3 0.55 0.07 26 4 9.3 1.0
Reference plant (found value) 260 10 360 10 470 10 1.68 0.06 0.45 0.02 22.9 0.5 9.92 0.78
Thyme 26 6 277 5 193 19 2.34 0.09 0.97 0.11 18.06 1.27 12.17 0.52
Cumin 45 9 174 14 234 4 2.56 0.09 1.79 0.21 17.12 1.06 8.33 0.54
Mint 20 6 104 9 403 29 2.40 0.27 1.01 0.05 31.78 0.93 10.04 0.79
Cinnamon 118 9 38 4 60 2 0.32 0.05 0.52 0.04 39.22 2.52 2.78 0.20
Turmeric 8 2 39 5 79 19 0.36 0.06 0.44 0.01 2.02 0.04 2.90 0.41
Chili pepper 39 4 151 9 522 28 6.50 0.46 2.22 0.06 3.86 0.44 11.70 0.65
Rosemary 5 1 173 27 268 38 8.05 0.07 3.14 0.23 11.42 1.51 3.41 0.43
Sweet red pepper 79 8 621 64 460 28 6.08 0.73 1.56 0.27 6.05 0.63 9.54 0.29
Black pepper <LOD 28 3 131 3 4.04 0.60 0.71 0.05 29.77 1.02 10.19 0.88
Plants Fe (lg g
1
) Mn (lg g
1
) Sr (lg g
1
) Zn (lg g
1
) Mg (mg g
1
) Ca (mg g
1
)
Reference plant (certied value) 274 17 45 4 154 9 37 3 6.5 0.5 18.1 1.3
Reference plant (found value) 217 13 39.1 1.8 113 3 32.8 1.5 5.8 0.1 18.1 0.2
Thyme 301 26 44.5 1.6 27.60 0.82 20.4 1.7 1.67 0.06 13.81 0.60
Cumin 269 14 29.3 1.4 21.51 1.32 45.7 4.8 2.59 0.09 8.27 0.60
Mint 281 19 98.0 0.9 111.5 1.7 23.6 2.2 3.63 0.02 11.94 0.57
Cinnamon 53 4.9 156.2 12.6 45.49 3.25 38.3 3.2 0.40 0.02 9.60 0.53
Turmeric 71 9.8 14.2 1.6 7.31 0.97 16.5 6.9 0.95 0.08 8.97 0.49
Chili pepper 339 17 17.6 0.4 6.22 0.32 32.7 2.5 1.78 0.03 2.55 0.62
Rosemary 346 2 56.2 4.3 4.44 0.49 20.0 1.4 1.79 0.06 3.54 0.10
Sweet red pepper 409 17 21.5 0.6 8.70 0.15 25.9 3.6 1.95 0.04 2.51 0.15
Black pepper 158 19 164.7 4.0 19.49 0.63 22.0 1.1 1.38 0.04 3.14 0.13
198 C. Karadas , D. Kara / Food Chemistry 130 (2012) 196202
should be subjected to Varimax rotation; i.e. one of the best known
approaches for orthogonal rotation. The principal components are
rotated so that the total sum of squares of the loadings along each
new axis is maximised (Brereton, 1990). The goal of all of these
strategies is to obtain a clear pattern of loadings, i.e. factors that
are somehow clearly marked by high loadings for some variables
and low loadings for others. This general pattern is also sometimes
referred to as simple structure (a more formalised denition can be
found in most standard textbooks). The higher the loading of a var-
iable implies a larger contribution to the variation, accounting for
the Varimax rotated principal components (Brereton, 1990). This
led to the formation of four principal components. The rst compo-
nent accounted for 39.7%, the second for 19.5%, the third for 13.7%
and the fourth for 10.6% of the total variation of the data. The rst
four components account for 83.5% of variances for all of the data.
The rst component represents the maximum variation of the data
set. Table 3 gives the rotated loadings and communality for each
element. The loadings were large for Ni, Cr and Ca on the rst com-
ponent, for Co, Cu, Fe, Mg and Sr on the second component, for Mn,
Ba and Sr on the third component and for Cd and Zn on the fourth
component. On the rst component, Ni and Cr have opposite prop-
erties with Ca because Ni and Cr have highest positive loadings
while Ca has the highest negative loading value. A very similar
classication of the analytes was achieved using classication
analysis, described previously. Table 3 also gives the score values
for each principal component for each spice or herb sample. From
the scores on the rst principal component it can be interpreted
that the concentrations of Ni and Cr on the rst principal compo-
nent loadings are higher but the concentrations of Ca are lower
for rosemary, chili pepper and sweet red pepper than the other
plants. The concentrations of Ni and Cr on the rst principal com-
ponent loadings are lower but the concentration of calciumis high-
er for turmeric, cinnamon, thyme and mint than the other plants.
When the second principal component is interpreted, Co, Cu, Fe,
Mg concentrations are higher for mint, sweet red pepper and cu-
min and are lower for turmeric and cinnamon than for the other
plants investigated. On the third principal component, Ba, Mn
and Sr concentrations are higher for cinnamon, black pepper and
mint and are lower for turmeric, sweet red pepper and chili pepper
than for the other plants. Arsenic concentrations have the opposite
properties in these samples compared with Ba, Mn and Sr. This is
demonstrated by the loading values of Ba, Mn and Sr having highly
positive values while the loading for As has the highest negative
value. Zinc and Cd concentrations in the fourth principal compo-
nent are higher for cinnamon, cumin and sweet red and lower
for turmeric, black pepper and rosemary.
Fig. 1 shows the two way loadings and score plots. Every prin-
cipal component was plotted against PC1 to show high percentage
of the total variance (59.1150.22). Fig. 1b shows the behaviour of
variables on PC1 and PC2. As can be seen, there is an association
between Ni and Cr that have the highest positive loading values
while Ca has highest negative value. There is also association be-
tween Co, Cu, Fe and Mg on the PC2 whereas the rest of the metals
appear more dispersed into the components space, showing a more
individualised behaviour.
The superposition of the loading (Fig. 1b) and score (Fig. 1a)
plots for PC1 and PC2 show calcium concentrations are higher
for thyme, mint and turmeric. Nickel and Cr concentrations are
highest for rosemary and are also higher for chili pepper and
sweet red pepper whereas the concentrations of Co, Cu, Fe and
Table 2
Correlation matrix for the element concentrations in plants.
Cd As Ni Co Cr Ba Cu Fe Mg Mn Sr Zn Ca
Cd 1.00
As 0.30 1.00
Ni 0.23 0.45 1.00
Co 0.01 0.55 0.66 1.00
Cr 0.18 0.29 0.86 0.55 1.00
Ba 0.29 0.43 0.44 0.40 0.45 1.00
Cu 0.18 0.35 0.25 0.57 0.03 0.04 1.00
Fe 0.11 0.74 0.78 0.84 0.72 0.43 0.56 1.00
Mg 0.31 0.21 0.23 0.62 0.30 0.02 0.51 0.60 1.00
Mn 0.16 0.49 -0.27 0.46 0.43 0.90 0.13 0.52 0.22 1.00
Sr 0.07 0.25 0.41 0.08 0.36 0.68 0.15 0.11 0.56 0.43 1.00
Zn 0.62 0.03 0.14 0.07 0.11 0.26 0.02 0.03 0.06 0.06 0.04 1.00
Ca 0.04 0.25 -0.76 0.39 0.52 0.40 0.00 0.33 0.15 0.08 0.60 0.02 1.00
Table 3
The loading and the scores of the rst four rotated principal components.
The loading The scores
Element PC1 PC2 PC3 PC4 Plants PC1 PC2 PC3 PC4
Cd 0.116 0.141 0.004 0.947 Thyme 1.09 0.46 0.68 0.41
As 0.224 0.395 0.624 0.332 Cumin 0.18 0.32 0.25 0.86
Ni 0.918 0.295 0.181 0.118 Mint 0.90 1.80 1.09 0.51
Co 0.446 0.75 0.30 0.13 Cinnamon 0.58 1.27 1.13 1.70
Cr 0.797 0.26 0.23 0.03 Turmeric 1.09 1.40 1.04 1.18
Ba 0.313 0.04 0.89 0.26 Chili pepper 1.06 0.40 0.50 0.23
Cu 0.026 0.73 0.08 0.02 Rosemary 1.62 0.34 0.12 0.87
Fe 0.513 0.721 0.405 0.032 Sweet red pepper 0.59 0.53 1.27 1.01
Mg 0.014 0.913 0.056 0.144 Black pepper 0.57 0.49 1.40 0.84
Mn 0.068 0.2 0.922 0.097
Sr 0.513 0.547 0.573 0.036
Zn 0.037 0.047 0.136 0.809
Ca 0.896 0.151 0.099 0.041
Eigenvalue 5.16 2.53 1.78 1.37
Variance (%) 39.66 19.45 13.72 10.56
Cumulative (%) 39.66 59.11 72.84 83.40
C. Karadas , D. Kara / Food Chemistry 130 (2012) 196202 199
Mg are higher for mint, sweet red pepper and thyme. It can be
interpreted from the score and loading plots for PC1PC3
(Fig. 1c and d) that Ba, Mn and Sr concentrations are higher
for black pepper, cinnamon and mint. The superposition of the
loading (Fig. 1c) and score (Fig. 1d) plots for PC1 and PC3 show
that the arsenic concentration is highest for sweet red pepper
and turmeric. From the score and loading plots for PC1PC4
(Fig. 1e and f) it can be interpreted that Zn and Cd concentra-
tions are higher for cinnamon, sweet red pepper and cumin.
The metal concentrations are the lowest or lower, comparatively
in turmeric because it stays in the negative side of the plots in
all PC score plots.
The classication of the spices from the view point of metal
contents can be made using three way PC score graphs. The highest
percentage of total variance of about 72.84 was observed with PC
123 while PC 124 score graph shows about 69.67% of total
variance. It can be seen from the PC 123 and PC 124 graphs
(Fig. 2) that the spices can be classied into ve groups. These
groups were:
Group 1: Sweet red pepper, chili pepper
Group 2: Rosemary, black pepper
Group 3: Thyme, mint,
Fig. 1. The score and loading plots (a, c and e are the score plots and b, d and f are the loading plots).
200 C. Karadas , D. Kara / Food Chemistry 130 (2012) 196202
Group 4: Cinnamon, turmeric
Group 5: Cumin
From Fig. 2, It can be seen that cinnamon and turmeric have
similar negative PC1 and PC2 scores but that cinnamon has highly
positive PC3 and PC4 scores while turmeric has negative PC3 and
PC4 factors. This means that the concentrations of Ba, Mn, Sr, Cd
and Zn are higher in cinnamon but are lower in turmeric. There-
fore, cinnamon and turmeric appear separate from each other in
PC 123 and PC 124 score graphs but they were put in the same
group because they have similar PC1 and PC2 score values which
show the 59.11% of variance.
3.3. Cluster analysis
This technique is an unsupervised classication procedure that
involves a measurement of the similarity between objects to be
clustered. Objects will be grouped in clusters in terms of their
nearness or similarity. The measurement of the similarity is based,
amongst others, on the squared Euclidean distance. The clustering
method used was the Wards method. The cluster analysis was
applied using the Statistica package (Statistica 99 edition, 1984
1999). Similar groupings to those found above were obtained from
cluster analysis (Fig. 3). These groups contain:
Group 1: Black pepper and rosemary
Group 2: Sweet red pepper and chili pepper
Group 3: Turmeric, cinnamon
Group 4: Cumin
Group 5: Mint, thyme
Cluster analyses shown in Fig. 3 indicated that Group 1 (black
pepper and rosemary) and Group 2 (sweet red pepper and chili
pepper) spices are close to each other. The same result was
obtained in the PC 123 and PC 124 score graphs (Fig. 2).
This means that black pepper and rosemary have a similarity in-
spite of metal concentrations data and the nearest spices to
them are sweet red pepper and chili pepper. Turmeric and cin-
namon are in the same cluster and this shows that they have
very similar metal concentrations. In addition, cumin was
grouped separately into a fourth Group that was also a sub-
cluster of the Group 3. It shows that cumin does have some sim-
ilarities to either (or both) turmeric and/or cinnamon. This is
inspite of the trace elements concentrations, whose trends do
not completely match those of the other spices. The next nearest
cluster to the Group 4 (cumin) includes mint and thyme (Group
5). Figs. 1 and 2 also prove that these similarities between herbs
and spices exist.
Fig. 2. Three way PCA scores plot (a) PC123 (b) PC124 (1: thyme; 2: cumin; 3: mint; 4: cinnamon; 5: turmeric; 6: chili pepper; 7: rosemary; 8: sweet red pepper; 9:
black pepper).
Fig. 3. Dendrogram of cluster analysis.
C. Karadas , D. Kara / Food Chemistry 130 (2012) 196202 201
3.4. Linear discriminant analysis
Linear discriminant analysis (LDA) is a supervised classication
tool used to differentiate (discriminate) groups of samples as a
function of one or several combinations of experimental variables
(Massart, 1997). Linear discriminant analysis can be used to show
how these group members may be correctly classied as a percent-
age of the original group. The linear discriminant analysis was per-
formed for the 13 elements on the classied ve groups resulting
from the PCA and CA as discussed above using SPSS 10 statistics
software (SPSS 10, 19891999). The recognition of these groups
was highly satisfactory. All group members determined by PCA
and CA are in the predicted group that 100.0% of original grouped
cases correctly classied. Three canonical discriminant functions
whose Eigenvalues were greater than 1 were obtained from the
data. The rst canonical discriminant function explains 98.7% of
the variance. The discriminant function of the rst function is
Z = 283.761 + 4.026 Cu + 0.029 Fe + 0.03 Mg + 0.027 Ca.
4. Conclusions
The chemometric evaluation shows that there is a relationship
between herbs and spices from the perspective of metal concentra-
tions. The spices and herbs were classied into ve groups by PCA
and cluster analysis. These groupings are correctly classied as
100.0% by using linear discriminant analysis. From the chemomet-
ric evaluation of trace metal concentrations, the rst group con-
tains black pepper and rosemary, the second group contains the
spices sweet red pepper and chili pepper, the third group are tur-
meric, cinnamon, the fourth group contains only cumin and lastly,
the fth group of herbs and spices are mint and thyme.
The rst group of herbs (black pepper and rosemary) contains
comparatively higher concentrations of Cr, Ni, Co, Fe, Ba, Mn and
Sr. The second group of spices (sweet red pepper and chili pepper)
has got higher concentration of Cr, Ni, Co, Fe, Cd and Zn. The third
group of spices (turmeric, cinnamon and cumin) has higher con-
centrations of Ba, Mn, Sr, Cd and Zn. The fourth group of plants
(mint and thyme) has higher concentrations of Ca, Co, Cu, Fe, Mg
and Sr cluster analysis also found a very similar grouping. All group
members determined by principal component analyses and cluster
analyses are in the predicted group that 100.0% of original grouped
cases correctly classied.
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