C H A P T E R

2
Data Analysis and Chemometrics
Paolo Oliveri, Michele Forina
Department of Drug and Food Chemistry and Technology, University of Genoa,
Via Brigata Salerno, 13, Genoa, Italy

O U T L I N E

2.1. Introduction 25 2.3.1. Principal-Component Analysis 34

2.1.1. From Data to Information 25 2.3.2. Signal Pre-Processing 37
2.3.3. Supervised Data Analysis
2.2. From Univariate to Multivariate 27
and Validation 38
2.2.1. Histograms 27
2.3.4. Supervised Qualitative Modeling 39
2.2.2. Normality Tests 29
2.3.5. Supervised Quantitative Modeling 45
2.2.3. ANOVA 30
2.3.6. Artificial Neural Networks 48
2.2.4. Radar Charts 33
2.3. Multivariate Data Analysis 34

2.1. INTRODUCTION generally of the same nature. For instance, gas

2.1.1. From Data to Information
Advances in technology and the increasing
availability of powerful instrumentation now
offer analytical food chemists the possibility for
obtaining high amounts of data on each sample
analyzed, in a reasonable e often negligible e
time frame (Valcárcel and Cárdenas, 2005).
Often, in fact, a single analysis may provide
a considerable number of measured quantities,
chromatographic (GC) analysis of fatty acid
methyl esters allows us to quantify, with a single
chromatogram, the fatty acid composition of
a vegetable oil sample (American Oil Chemist’s
Society, 1998). Spectroscopic techniques as well
may supply, with a single and rapid analysis
on a sample, multiple data of homogeneous
nature: in fact, a spectrum can be considered as
a data vector, in which the order of the variables
(e.g., absorbances at consecutive wavelengths)
has a physical meaning (Oliveri et al., 2011).

Chemical Analysis of Food: Techniques and Applications

DOI: 10.1016/B978-0-12-384862-8.00002-9 25 Copyright Ó 2012 Elsevier Inc. All rights reserved.
26 2. DATA ANALYSIS AND CHEMOMETRICS
In other cases, a set of samples can be
described by a number of heterogeneous chem-
ical and physical parameters at the same time.
For example, a global analytical characteriza-
tion of a tomato sauce may involve the quanti-
fication of color and rheological parameters as
well as pH and chemical composition and e
possibly e a number of sensorial responses
(Sharoba et al., 2005). Also in such cases, each
sample may be described by a data vector,
but without any implication with respect to
the order of the variables. Instead, differences
in variable magnitude and scale between
different variables may affect data analysis
if a proper pre-processing approach is not
followed.
The availability of large sets of data does not
mean at the immediate time availability of infor-
mation promptly accessible to the sample
analyzed: usually, in fact, a number of steps
are required to extract and properly interpret
the potential information embodied within the
data (Martens and Kohler, 2008).
A deep understanding of the nature of
analytical data is the first basic step for any
proper data treatment, because different data
types usually require different processing strat-
egies, which closely depend on their nature and
origin. For this reason, the data analyst should
always have a complete awareness of the
problem under study and about the whole
analytical process from which data derive e
from the sampling to the instrumental analysis.
Such knowledge is fundamental: it makes the
difference between a chemometrician and
a mathematician. A chemometrician is, first of
all, a chemist, who is acquainted with his
data, and utilizes mathematical methods for
the conversion of numerical records into rele-
vant chemical information.
The analytical food chemist William Sealy
Gosset (1876e1937), who worked at the
Arthur Guinness & Son brewery of Dublin,
can be considered as one of the fathers of
chemometrics. In fact, he studied a number
I. ANALYTICAL TECHNIQUES
of statistical tools and adapted them to better
solve actual chemical problems. He had to
present his studies using a pseudonym, since
his company did not permit him to publish
any data. Considering himself as a modest
contributor in the field, rather than a statisti-
cian, he adopted the pen name Student. His
most famous work was on the definition of
the probability distribution that is commonly
referred to as the Student’s t distribution
(Student, 1908).
The term chemometrics was used for the first
time by Svante Wold, in 1972, for identifying the
discipline that performs the extraction of useful
chemical information from complex experi-
mental systems (Wold, 1972).
Statistics offers a number of helpful tools that
can be used for converting data into informa-
tion. Univariate methods, which consider one
variable at a time, independently of the others,
have been and are still extensively used for
such purposes. Nonetheless, they usually
supply just partial answers to the problems
under study, since they underutilize the
potential for discovering global information
embodied in the data. For instance, they are
not able to take into account inter-correlation
between variables e a feature that can be very
informative, if recognized and properly
interpreted.
Multivariate strategies are able to take into
account such an aspect, allowing a more
complete interpretation of data structures.
However, in spite of their big potential, multi-
variate methods are generally less used than
univariate tools.
On the other hand, a number of people try
multivariate analysis as the last-ditch resort,
when nothing seems to provide the desired
results, pretending that chemometrics provide
valuable information from data that do not
contain any informative feature at all.
Such demeanor is very hazardous especially
when complex methods are being used, because
there may be the risk of employing chance
 2.2. FROM UNIVARIATE TO MULTIVARIATE
correlations to develop models with good
performances only on appearance e namely,
on the same samples used for model building e
but with very poor prediction ability on new
samples: this is the so-called overfitting. To over-
come such a possibility, a proper validation of
models is always required. In particular, the
more complex the technique applied, the deeper
the validation recommended.
For these reasons, a good understanding of
the characteristics of the methods employed
for data processing is always advantageous
as well.
In this chapter, an overview of the chemomet-
ric techniques most commonly used for data
analysis in analytical food chemistry will be pre-
sented, highlighting potentials and limits of
each one.
2.2. FROM UNIVARIATE
TO MULTIVARIATE
A bidimensional table is probably the most
typical way to arrange, present, and store
analytical data: conventionally, in chemomet-
rics, each row usually represents one of the
samples analyzed, while each column corre-
sponds to one of the variables measured.
As an example, Table 2A.1 reports the red-
wine data set, which consists of 27 chemical
and physical parameters measured on 90 wine
samples, belonging to three Italian denomina-
tions of origin from the same region (Piedmont):
Barolo, Grignolino, and Barbera. The original
data set was composed of 178 samples (Forina
et al., 1986).
Table 2A.1 contains also additional informa-
tion, which is usually not processed but which
may be extremely helpful in the final under-
standing and interpretation of the results. In
particular, the two heading lines contain the
numbers and the names of the variables, which
are additional information for the columns,
while the two heading columns include the
I. ANALYTICAL TECHNIQUES
27
names identifying the samples and their class,
which represent additional information for the
rows.
It is easy to guess that such data enclose
a great deal of potential information. Anyway,
the simple visual inspection of the table, which
contains a considerable number of records,
does not provide directly any valuable informa-
tion about the samples analyzed. A conversion
from data into information is necessary.
Univariate methods are still the most used
in many cases, although they generally offer
only a very limitative vision of the global
situation.
2.2.1. Histograms
A good way to extract information from data
is to use graphical tools. Among them, histo-
grams are probably the most widely employed
(Chambers et al., 1983).
To build a histogram, the range of interest
of the variable under study is divided into
a number of regular adjacent intervals. For
each interval, the contribution of the measured
samples is graphically displayed by a vertical
rectangle, whose area is proportional to the
frequency (i.e., the number of observations)
within that interval. Consequently, the height
of each rectangle is equal to the frequency
divided by the interval width, so that it has
the dimension of a frequency density.
Frequently, such frequency values are
normalized, dividing each of them by the total
number of observations, thus obtaining relative
values. It follows that, in such cases, the sum of
the areas of all the rectangles e i.e., the sum of
all the relative frequencies e is equal to 1.
The frequency distribution visualized by
a histogram can be used to estimate the proba-
bility distribution of the variable under study
and to make deductions about the samples.
Figure 2.1 shows examples of histograms for
a portion of the data given in Table 2A.1, namely
for variables number 13, 21, and 26.
28 2. DATA ANALYSIS AND CHEMOMETRICS

(a) -3
x 10 (b)
6 0.7

5 0.6

Relative Frequency
0.5
4
Relative Frequency

0.4
3
0.3
2
0.2
1
0.1

0 0
150 200 250 300 350 400 450 500 550 600 1 1.5 2 2.5 3 3.5 4
Phosphate
OD280/OD315 of diluted wines

(c)
-3
x 10
2
1.8
1.6
1.4
Relative Frequency

1.2
1
0.8
0.6
0.4
0.2
0
200 400 600 800 1000 1200 1400 1600 1800
Proline

FIGURE 2.1 Histograms for three variables of Table 2A.1: phosphate (a), OD280/OD315 of diluted wines (b), and proline (c).

Three typical patterns are noticeable. In histograms could be drawn for each class sepa-
particular, variable 13 (phosphate) shows rately, to verify the trend of the within-class
a unimodal and almost symmetric shape, which distributions.
may suggest that such variable follows a normal Instead, the histogram shape for variable
probability distribution (Fig. 2.1a). 26 (proline) reveals an underlying asymmetric
Conversely, variable 21 (OD280/OD315 of distribution (Fig. 2.1c). It is possible to
diluted wines) presents a bimodal distribution, convert such behavior into an almost normal
which may suggest that this variable to be charac- one, simply by applying a logarithmic trans-
terized by different average values for diverse formation to the variable, as it is shown in
sample classes (Fig. 2.1b). In such cases, Fig. 2.2.

I. ANALYTICAL TECHNIQUES
 2.2. FROM UNIVARIATE TO MULTIVARIATE
1.4
1.2
Relative Frequency
1
0.8
0.6
0.4
0.2
0 equal to 1.
5.5 6 6.5 7 7.5
Log Proline
FIGURE 2.2 Histogram for the log-transformed variable
proline of Table 2A.1.
2.2.2. Normality Tests
Assessing for compatibility with a normal
distribution is a basic issue in data analysis,
because many methods require variables to be
normally distributed. As observed, frequency
distributions may be employed for this purpose.
Visual examination of histogram shapes may
supply a preliminary evaluation. Besides, the
cumulative empirical frequency distributions
(EFDs) constitute the basis for a family of statis-
tical normality tests, which are usually referred
to as KolmogoroveSmirnov tests (Kolmogorov,
1933; Smirnov, 1939).
One of the most effective and employed
among them is the Lilliefors test, which may
be used for generally assessing how well an
empirical distribution fits with a theoretical
one (Lilliefors, 1970). In the case for normality
verification, the null hypothesis (H0) is that the
observed empirical frequency distribution for
a given variable is not significantly different
from the theoretical normal probability distribu-
tion, at a given significance level. The alternative
hypothesis (H1) is that the observed EFD is not
compatible with the theoretical normal distribu-
tion, at that significance level.
I. ANALYTICAL TECHNIQUES
29
The test procedure consists in ordering the
values of the variable to be tested and normal-
izing them by means of a Student’s transforma-
tion (or autoscaling):
xi;v  xv
x
i;v ¼ (2.1)
sv
The variable is corrected by subtracting its
mean (xv ) from each of its values and then
dividing by its standard deviation (sv). The
autoscaled variable is dimensionless and
present mean equal to 0 and standard deviation
Then, the corresponding cumulative theoret-
ical probability distribution is estimated from
the statistical parameters computed, and the
maximum distance from such hypothesized
distribution and the empirical one is calcu-
lated. This value is compared with a critical
distance value, at a predetermined significance
level, and such comparison determines the
acceptance/rejection of the null hypothesis.
The critical values, which depend on the
sample size, were obtained by Monte Carlo
simulations and are available on tables or
statistical software.
The Lilliefors test can be performed also in
a graphical way (Iman, 1982), as it is illustrated
in Figs. 2.3 and 2.4 for the same cases of Figs. 2.1
and 2.2.
Charts for the Lilliefors test report the cumula-
tive empirical frequency distributions (EFDs) for
variables number 13, 21, and 26 of Table 2A.1,
after column autoscaling (polygonal curves in
Figs. 2.3 and 2.4), together with the cumulative
theoretical probability distribution (sigmoid
solid curves), and the distance limits according
to the Lilliefors test, at a 5% significance (sigmoid
dot curves). When the EFD curve intersects at
least one of the limits individuated by
the critical distance, the null hypothesis is
rejected. As for the examples reported in
Fig. 2.3, the null hypothesis is accepted only for
the variable phosphate, while for both the other
variables examined, it is rejected at the same
30 2. DATA ANALYSIS AND CHEMOMETRICS

(a) 1 (b) 1
0.9 0.9
0.8
Cumulative distribution

0.8

Cumulative distribution
0.7 0.7
0.6 0.6
0.5 0.5
0.4 0.4
0.3 0.3
0.2 0.2
0.1 0.1
0 0
-3 -2 -1 0 1 2 3 -3 -2 -1 0 1 2 3
Phosphate (autoscaled) OD280/OD315 of diluted wines (autoscaled)

(c) 1
0.9
0.8
Cumulative distribution

0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
-3 -2 -1 0 1 2 3
Proline (autoscaled)

FIGURE 2.3 Graphical Lilliefors normality test for three variables of Table 2A.1: phosphate (a), OD280/OD315 of diluted
wines (b), and proline (c). The polygonal curves represent the empirical frequency distributions (EFDs) of variables after
column autoscaling, the solid sigmoid curves represent the cumulative theoretical probability distributions and the dot
sigmoid curves represent the distance limits according to the Lilliefors test, at a 5% significance. (For color version of the
figures, refer to the online version)

significance level. In fact, only in the first case

2.2.3. ANOVA
(Fig. 2.3a), the polygonal EFD curve does not Analysis of variance (ANOVA) is the name of
intersect the critical distance lines in any point. a group of statistical methods based on Fisher’s
In addition, it can be easily verified e con- F tests, generally aimed at verifying the
firming the deductions made by looking at the existence/absence of significant differences
histogram of Fig. 2.2 e that the logarithmic between groups of data. The null hypothesis
transformation applied to the variable proline H0 is that all the data derive from the same
makes it compatible with the normal distribu- stochastic population, i.e., there is no significant
tion (see Fig. 2.4). difference between the groups considered. In

I. ANALYTICAL TECHNIQUES
 2.2. FROM UNIVARIATE TO MULTIVARIATE
1
0.9
0.8
Cumulative distribution
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
-3
FIGURE 2.4 Graphical Lilliefors normality test for the log-transformed variable proline of Table 2A.1. The polygonal
curve represents the empirical frequency distributions (EFDs) of variables after column autoscaling, the solid sigmoid curve
represents the cumulative theoretical probability distributions and the dot sigmoid curves represent the distance limits
according to the Lilliefors test, at a 5% significance.
order to verify this hypothesis, the final F test
compares the variability between groups with
the variability within groups (Box et al., 1978).
The simplest case is the one-way ANOVA,
whose procedure is described with a real
numerical example. The two columns of Table
2.1 report the values of the alcoholic degree for
Barolo and Barbera wine samples of the red-
wine data set (Table 2A.1), respectively, and
some basic descriptive parameters. A summary
of all the parameters computed for the ANOVA
test is given in Table 2.2. The aim is to assess
whether there is a significant difference between
the alcohol content of the two wines or not. In
fact, although the mean Barbera alcoholic
percentage (13.07% abv) is noticeably less than
the corresponding Barolo value (13.83% abv),
the two respective ranges overlap, so that it
might be suspected the observed difference to
be due to chance variations.
The within-columns variance can be computed
as a pool variance, under the hypothesis that the
variances of the different groups are
31
-2 -1 0 1 2 3
Log proline (autoscaled)
homogeneous. When only two groups e and,
consequently, two variances e are being
compared, a Fisher’s F test is suitable to verify
this preliminary hypothesis. In the given numer-
ical example, the test value is computed as
s2Barolo 0:274
Ft ¼ 2
¼ ¼ 1:08 (2.2)
sBarbera 0:254
The F critical value, at a 5% right significance level
and for 29 degrees of freedom (d.o.f.) both at the
numerator and at the denominator, is 1.86. So it
is possible to conclude that the variances of the
two groups considered are not significantly
different, at a 5% right significance level.
In problems involving more than two
groups, the comparison among variances can
be performed with multiple F tests on all the
possible pairs, or by means of the Cochran’s
test or the Bartlett’s tests (Snedecor and
Cochran, 1989). The former is valid when there
is an equal numbers of data in each group,
while the latter has a wider applicability.
I. ANALYTICAL TECHNIQUES
32 2. DATA ANALYSIS AND CHEMOMETRICS

TABLE 2.1 Alcohol Content (% abv) for Barolo and TABLE 2.2 Full ANOVA Parameters for the Data
Barbera Samples of Red-Wine Data Set, given in Table 2.1. Computed F ratio
and Basic Statistical Parameters (from variances of columns Barolo and
Barbera) ¼ 1.08. Critical F value (at 5%
Barolo Barbera significance) ¼ 1.86. F test on variances
14.23 12.86 of columns Barolo and Barbera:
13.20 12.88
significance ¼ 41.8%

13.16 12.81 Source of variation d.o.f. Sum of squares Variance

14.37 12.70
Total 60 10874.485
13.24 12.51
Mean 1 10850.384
14.20 12.60
Between columns 1 8.786 8.786
14.39 12.25
Within columns 58 15.314 0.264
14.06 12.53
14.83 13.49 Computed F ratio ¼ 33.28; Critical F value (at 5% significance) ¼ 4.01;
ANOVA F test: significance ¼ 0.0%.
13.86 12.84
14.10 12.93
14.12 13.36
In the numerical example discussed, the
within-columns variance e computed as pooled
13.75 13.52
variance e corresponds to
14.75 13.62
PC PNC 2
14.38 12.25 c¼1 n ¼ 1 ðxnc  xc Þ
s2within ¼
13.63 13.16 NC (2.3)
15:314
14.30 13.88 ¼ ¼ 0:264
13.83 12.87 58
14.19 13.32 Conversely, the between-columns variance is
13.64 13.08 computed as
14.06 13.50 PC
c ¼ 1 NC ðxc  xÞ
12.93 12.79 s2between ¼
C1 (2.4)
13.71 13.11
8:786
12.85 13.23 ¼ ¼ 8:786
1
13.50 12.58
13.05 13.17
Finally, the ANOVA F test value is computed
as between-columns variance:within-columns
13.39 13.84
variance ratio, to perform the final test which
13.30 12.45 compares these variations, for the verification
13.87 14.34 of the ANOVA null hypothesis:
14.02 13.48
s2between 8:786
Min 12.85 12.25 FANOVA ¼ 2
¼ ¼ 32:28 (2.5)
swithin 0:264
Max 14.83 14.34
Mean 13.83 13.07 The F critical value, at a 5% right signifi-
Variance 0.274 0.254 cance level, for 1 degree of freedom at the
numerator and 58 degrees of freedom at the
Standard deviation 0.524 0.504
denominator, is 4.01. From the comparison

I. ANALYTICAL TECHNIQUES
 2.2. FROM UNIVARIATE TO MULTIVARIATE
with the computed test value, it follows that
the null hypothesis is rejected at a 5% signif-
icance level. The conclusion is that the differ-
ence between the alcoholic content of Barolo
and Barbera samples is significantly larger
than the variability within each of the two
groups.
ANOVA tests can be applied also when the
effect of two variability sources (e.g., type of
wine and vintage year) is to be verified: such
a scheme is usually called a two-way ANOVA.
When a number of replicate measurements are
available for each level combination of the two
factors (nested two-way ANOVA), the model
obtained also allows an estimation of the inter-
action between the factors, together with its
significance.
2.2.4. Radar Charts
Radar charts e also known as web charts,
spider charts, star charts, cobweb charts, polar
charts, star plots, or Kiviat diagrams e are
a data display tool that can be considered as
a sort of link between univariate and multivar-
iate graphical representations (Chambers et al.,
1983).
They consist of circular graphs divided into
a number of equiangular spokes, called radii.
Each radium represents one of the variables.
A point is individuated on it, whose distance
from the center is proportional to the magnitude
of the related variable for that datum. Finally, all
the data points e corresponding to all the vari-
ables measured on a sample e are connected
with a line, which represent a sort of sample
profile.
Usually, each plot represents a single
sample, and multiple observations are
compared by examining different plots. It is
also possible to overdraw several lines on the
same chart, although the outcome will be
legible only for small data sets. As a matter of
fact, when the number of samples is large,
such graphical representation is generally not
very functional.
I. ANALYTICAL TECHNIQUES
33
Within radar charts, variables can be repre-
sented without any previous scaling, revealing
what variables are dominant for a given data
set. Nonetheless, when variables are character-
ized by considerably different scales (as in the
case for red-wine data of Table 2A.1), a prelimi-
nary transformation may be helpful in order to
make visible within the graph the contribution
of all of them, by assuring the same a priori
importance.
For instance, by looking at Fig. 2.5, it clearly
appears that, without any scaling, four
features are dominating, corresponding to
the variables number 10, 13, 24, and 16, which
are characterized by the highest mean values
(see Table 2.1). The contribution of the remain-
ing 23 variables is not recognizable within
these graphs. Furthermore, it is not possible
to draw many valuable considerations about
the sample profiles. In particular, it can be
noticed that Grignolino wines (Fig. 2.5a) are
characterized, on average, by smaller values
for the four observable variables. It can also
be deduced that Barolo has a higher contribu-
tion from variable 26, while Barbera (Fig. 2.5c)
has higher contributions from variables 24
and 10.
On the other hand, Fig. 2.6 illustrates that,
after application of column autoscaling (see
Eqn (2.1)), the a priori differences in location
and dispersion among the original variables
are eliminated, thus showing the contribution
of all of them and highlighting the differences
among the observations. In fact, in this second
graph, the profiles of the three wines appear
much more dissimilar than in the previous
one. By a joint examination of the three radar
charts of Fig. 2.6, it can be deduced that Barolo
and Barbera samples present two rather
complementary profiles, while the Grignolino
profile is somewhat intermediate. In particular,
Barolo (Fig. 2.6a) is characterized by higher
average values of variables 1, 2, 13, 15, 16, 18,
21, 22, 23, and 26. Instead, Grignolino
(Fig. 2.6b) presents average lower values of all
the variables, except for the number 20. Finally,
34 2. DATA ANALYSIS AND CHEMOMETRICS

1 1 1
(a) 26
27 1200 2
3 (b) 26
27 1200 2
3 (c) 26
27 1200 2
3
25 1000 4 25 1000 4 25 1000 4
800 800 800
24 5 24 5 24 5
600 600 600
23 6 23 6 23 6
400 400 400
22 200 7 22 200 7 22 200 7
0 0 0
21 8 21 8 21 8

20 9 20 9 20 9

19 10 19 10 19 10

18 11 18 11 18 11
17 12 17 12 17 12
16 13 16 13 16 13
15 14 15 14 15 14

FIGURE 2.5 Radar charts of average profiles of Barolo (a), Grignolino (b), and Barbera (c) samples. Numbers from 1 to 27
correspond to the original variables listed in Table 2A.1.

1 1 1
(a) 26
27 2
1.5
2
3 (b) 26
27 2
1.5
2
3 (c) 26
27 2
1.5
2
3
25 1 4 25 1 4 25 1 4
24 0.5 5 24 0.5 5 24 0.5 5
0 0 0
23 -0.5 6 23 -0.5 6 23 -0.5 6
-1 -1 -1
22 7 22 7 22 7
-1.5 -1.5 -1.5
-2 -2 -2
21 8 21 8 21 8

20 9 20 9 20 9

19 10 19 10 19 10

18 11 18 11 18 11
17 12 17 12 17 12
16 13 16 13 16 13
15 14 15 14 15 14

FIGURE 2.6 Radar charts of average profiles of Barolo (a), Grignolino (b), and Barbera (c) samples. Numbers from 1 to 27
correspond to the variables listed in Table 2A.1, reprocessed by application of column autoscaling.

Barbera (Fig. 2.6c) has a bigger contribution one of the basic and most useful tools in the
from variables 3, 4, 5, 9, 17, 19, and 24. branch of multivariate analysis. It is an explor-
Such deductions may be useful for character- atory method, which always offers an overview
ization purposes. of the problem studied and often allows the
drawing of significant conclusions and for deci-
sions to be made on the basis of the observed
2.3. MULTIVARIATE DATA results. Furthermore, PCA can be employed
ANALYSIS for feature and noise reduction purposes and
constitutes the basis for other more complex
pattern recognition techniques.
2.3.1. Principal-Component Analysis
PCA is based on the assumption that a high
Principal-component analysis (PCA), which variability (i.e., a high variance value) is synon-
originates in the work of K. Pearson (1901), is ymous with a high amount of information.

I. ANALYTICAL TECHNIQUES
 2.3. MULTIVARIATE DATA ANALYSIS
For this reason, PCA algorithms search for
the maximum variance direction, in the multidi-
mensional space of the original data, preferably
passing through the data centroid, which means
that data have to be at least mean centered
column-wise. The maximum variance direction
represents the first principal component (PC).
The second PC is the direction which keeps
the maximum variance among all directions
orthogonal to the first PC. It follows that the
second PC explains the maximum information
not explained by the first one or, in other words,
these two new variables are not inter-correlated.
The process continues with the identification of
the subsequent PCs: it may stop at reaching
a variance cutoff value or continue until all the
variability enclosed in the original data has
been explained (Jolliffe, 2002).
Since the variance values depend on the
scale of the variables, it becomes difficult to
compare and impossible to combine informa-
tion from variables of different nature, unless
properly normalized: column autoscaling (see
Eqn (2.1)) is the most commonly applied
transform.
Each sample can be projected in the space
defined by the new variables: the coordinate
values obtained are called scores.
The PCs are expressible as linear combina-
tions of the original variables: the coefficients
which multiply each variable are called load-
ings. They represent the cosine values (director
cosines) of the angles between the PCs and the
original variables. These values may vary
between e1 and þ1, indicating the importance
in defining a given PC: the larger the cosine
absolute value, the closer the two directions,
thus the larger the contribution of the original
variable to the PC.
In terms of matrix algebra, the rotation from
the space of the original variables to the PC
space is performed by means of the loading
orthogonal matrix, L:
SNV ¼ XNV LVV (2.6)
I. ANALYTICAL TECHNIQUES
35
where S is the score matrix and X is the original
matrix, constituted by N objects (rows)
described by V variables (columns).
One of the key features of PCA is its high
capability for representing large amounts of
complex information by way of simple bidimen-
sional or tridimensional plots.
In fact, the space described by two or three
PCs can be used to represent the objects (score
plot), the original variables (loading plot), or
both objects and variables (biplot) (Geladi
et al., 2003; Kjeldahl and Bro, 2010). Since prin-
cipal components are not inter-correlated vari-
ables, no duplicate information is shown in PC
plots.
Figure 2.7 represents an example of a highly
informative biplot, which derives from PCA
performed on the red-wine data set given in
Table 2A.1. Data have been previously auto-
scaled in order to eliminate the magnitude
differences among the variables. The two Carte-
sian axes correspond to the first (meaning low-
order) two PCs, which together show a 44.2%
of the information (defined as explained vari-
ance) enclosed in the original multidimensional
data space.
The plot clearly shows the interrelations
existing among samples, among variables, and
between samples and variables. Moreover,
considering also the additional row information
given in Table 2A.1 (namely, the class of each
sample, graphically represented by different
colors), it is possible to get information about
the discrimination among the three wine cate-
gories (Barolo, Grignolino, and Barbera), the
dispersion of samples within each class, and
the discriminatory importance of the variables
measured.
In particular, it appears that PC1 e which
accounts for the 27.4% of the total variance,
i.e., information e is a direction effective in dis-
tinguishing among the three wine classes, espe-
cially between Barolo (circles) and Barbera
(triangles) samples. Instead, PC2 (explaining
the 16.8% of total variance) is useful in
36 2. DATA ANALYSIS AND CHEMOMETRICS

20

14
22
21 11
PC2 (16.8%)

16
25
7 4
15 12
18
13 27 9 17
26

6 5 3

1
2 23 81024
19
PC1 (27.4%)

FIGURE 2.7 Example of PCA biplot for the autoscaled red-wine data. The scores (symbols) correspond to the wine
samples of classes Barolo (circles), Grignolino (squares), and Barbera (triangles), respectively. The loadings (line segments
and numbers from 1 to 27) represent the contribution of the original variables e as listed in Table 2A.1 e to the information
visualized in the plot. (For color version of the figure, refer to the online version)

differentiating mainly Grignolino samples
(squares) from the other two groups.
The variables which present the highest
loading absolute value on PC1 are the numbers
15, 16, 21, 22, 3, 4, 5, 6, 9, and 17. It means that
such variables are the most important in
defining PC1 and, consequently, in discrimi-
nating among the three wine classes, particu-
larly Barolo from Barbera. In more detail,
looking at the correspondences between scores
and loadings, it can be deduced by this plot
that the samples that present on average the
highest values for variables 3, 4, 5, 6, 9, and 17
and the lowest values for variables 15, 16, 21,
and 22 belong to class Barbera. Just opposite
considerations are applicable for the samples
of class Barolo (low values for the variables of
the first group and high values for the variables
of the second group). Instead, Grignolino wines
lay in a halfway position, meaning that they are
characterized by intermediate values for all
these variables. Conversely, variable number
20 has the highest loading value on PC2 and it
is clearly in the same direction of the Grignolino
cluster. This means that Grignolino wines have,
on average, high values of variable 20. Opposite
considerations are valid for variables 1, 2, 8, 10,
19, 23, and 24.
All of these inferences are absolutely in
accord with all the deductions already made
by inspection of the radar charts in Fig. 2.6.
Moreover, the information that variable 21
(OD280/OD315 of diluted wines) has a highly
discriminant power is in perfect agreement
with the bimodal distribution observed in the
histogram of Fig. 2.1b for the same variable.

I. ANALYTICAL TECHNIQUES
 2.3. MULTIVARIATE DATA ANALYSIS
Variables 7, 12, 25, and 27 have very small
loading values on both the PCs, meaning that
such variables give a negligible contribution to
the portion of information visualized in this
plot.
Further considerations may be drawn by
looking at the distribution of samples inside
each class. For instance, it can be easily seen
that Barolo wines are characterized by the
lowest within-class variability. In the case of
quality control, a low sample variability about
a target value is an index of high quality
(Taguchi, 1986).
It is worth noticing that a single and simple
biplot is able to report a considerable amount
of information that would require a large
number of univariate plots and tests to be
extracted: PCA is, without any doubt, the most
efficient way to account for the information
enclosed in a data table.
2.3.2. Signal Pre-Processing
Instrumental analytical techniques com-
monly provide information in the form of digital
signals. Spectra, chromatograms, and voltam-
mograms are typical examples. Such signals
generally require to be suitably pretreated, since
the analytical information is not the exclusive
component. A number of different variations,
from sources other than the analytical system
under investigation, generally affect signals.
They may be related either with the electric
instrumentation components or with the
surroundings. In particular, unwanted signal
variations may be random or systematic. The
former are generally due to sporadic interfer-
ences or associated with random phenomena
(e.g., Brownian motions of particles and
thermal motion of electrons e the so-called
JohnsoneNyquist noise) which usually follow
a standard normal or a Poisson probability
distribution. This type of noise, also called white
noise, is characterized by frequency values
higher than those of the useful signal.
I. ANALYTICAL TECHNIQUES
37
Instead, systematic unwanted variations are
commonly due to instrumental trends or to
external influences. They may affect the signal
with baseline shifts and/or drifts, which can
be considered as a low-frequency contribution.
Signal processing is generally aimed at mini-
mizing the unwanted variations, thus
improving the quality of signals and, conse-
quently, the conversion of data to valuable
information.
In particular, it is possible to individuate
three main objectives: reduction of random
noise, reduction of systematic unwanted varia-
tions, and reduction of data size.
Several pre-processing techniques accom-
plish this with more than one point. Further-
more, in some cases, the transformation itself
facilitates the interpretation of complex signals,
as in the case for derivatives.
When several digital signals are structured
into a data matrix, each of them corresponding
to a row e following the chemometric conven-
tion e signal pre-processing is also known as
row pre-processing. The mathematical trans-
forms act on each single signal independently
of the others.
Techniques for reduction of random noise
include the moving average e or boxcar e
filters, the SavitzkyeGolay smoothing (Savitzky
and Golay, 1964), and the Fourier transform
(FT)-based filters (Reis et al., 2009).
Regarding the elimination or minimization
of unwanted systematic effects, a number of
mathematical methods for signal transforma-
tion are widely employed, such as the
standard normal variate (SNV) transform and
derivatives.
2.3.2.1. Standard Normal Variate (SNV)
Transform
The SNV transform, or row autoscaling, is
particularly applied in spectroscopy, since it is
useful to correct for both baseline shifts and
global intensity variations (Barnes et al., 1989).
Each signal (xi) is row-centered, by subtracting
38 2. DATA ANALYSIS AND CHEMOMETRICS
its mean (xi ) from each single value (xi,v), and
then scaled by dividing by the signal standard
deviation (si):
xi;v  xi
x
i;v ¼ (2.7)
si
After transformation, each signal presents
mean equal to 0 and standard deviation equal
to 1.
SNV has the peculiarity of possibly shifting
informative regions along the signal range, so
that the interpretation of the results referring
to the original signals should be performed
with caution (Fearn, 2009).
2.3.2.2. Derivatives
The numerical differentiation of digitized
signals may correct for baseline shifts and drifts,
depending on the derivation order. Further-
more, derivative profiles often exhibit an
increased apparent resolution of overlapping
peaks and may accentuate small structural
differences between nearly identical signals
(Taavitsainen, 2009).
The first derivative of a signal y ¼ f(x) is the
rate of change of y with x (i.e., y0 ¼ dy/dx),
which can be interpreted e at the single
points e as the slope of the line tangent to
the signal. It provides a correction for baseline
shifts.
The second derivative can be considered as
a further derivation of the first derivative
(y00 ¼ d2y/dx2); it represents a measure of the
curvature of the original signal, i.e., the rate of
change of its slope. Such transform provides
a correction for both baseline shifts and drifts.
A disadvantageous consequence of deriva-
tion may be an enhancement of the random
noise, characterized by high-frequency slope
variations. To overcome this hurdle, signals
are firstly smoothed, often by using the
SavitzkyeGolay algorithm (Savitzky and Golay,
1964) with a third-order polynomial.
I. ANALYTICAL TECHNIQUES
2.3.2.3. Horizontal Alignment
When a series of chromatograms are used
as vectors to build a data matrix, a typical
problem arises with the horizontal shifts
that commonly characterizes such type of
data.
The most common methods for peak align-
ment, such as the correlation optimized
warping (COW), search for the maximum corre-
lation between a selected reference profile and
a series of piecewise modified (shifted and
warped) versions of the signals to be aligned
(Nielsen et al., 1998; Jellema, 2009).
2.3.3. Supervised Data Analysis
and Validation
Exploratory techniques for data analysis,
such as PCA, are unsupervised, meaning that
they just show the data as they are. Conversely,
supervised chemometric methods look for
determined features within data, explicitly
oriented to address particular issues.
In particular, when a model is developed
with the purpose of predicting a qualitative or
quantitative property of interest, its reliability
in prediction should be assessed prior to using
the model in practice. Prediction ability values
should be presented together with their confi-
dence interval (Forina et al., 2001; Forina et al.,
2007), which depends on the number of samples
used for the validation. The estimation of the
predictive ability on new samples e not used
for building the models e is a fundamental
step in any modeling process and several proce-
dures have been deployed for this purpose. The
most common validation strategies divide the
available samples into two subsets: a training
(or calibration) set used for calculating the
model and an evaluation set used for assessing
its reliability. A key feature for an honest valida-
tion is that the test samples have to be absolutely
extraneous to the model: no information from
them can be used in building the model in the
 2.3. MULTIVARIATE DATA ANALYSIS
pre-processing stages, otherwise the prediction
ability may be overestimated.
In many modeling techniques, some parame-
ters are optimized looking for a setting that
provides the maximum predictive ability for
the model for a given sample subset.
In such cases, a correct validation strategy
would involve three sample subsets: a training
set, an optimization set, and an evaluation set.
The optimization set is used to find the best
modeling settings, while the actual reliability
of the final model is estimated by way of a real
prediction on the third subset, formed by objects
that have never influenced the model.
The evaluation of the predictive ability of
a model can be performed in a unique step or
many times with different evaluation sets,
depending on the strategy adopted.
2.3.3.1. Single Evaluation Set
A single evaluation set is the simplest and
most rapid validation scheme. A fraction e
usually between 50% and 90% of the total
number e of the available samples constitutes
the training set, while the remaining objects
form the evaluation set. The subdivision may
be arbitrary, random, or performed by way of
a uniform design, such as the Kennard and
Stone and the duplex algorithm (Kennard and
Stone, 1969; Snee, 1977), which allows two
subsets that are uniformly distributed and
representative of the total sample variability to
be obtained.
2.3.3.2. Cross-Validation (CV)
Cross-validation is probably the most com-
mon validation procedure. The N available
samples are divided into G cancellation groups
following a predetermined scheme (e.g., contig-
uous blocks or Venetian blinds). The model is
computed G times: each time, one of the cancel-
lation groups is used as the evaluation set, while
the other groups constitute the training set. At
the end of the procedure, each sample has been
used G e 1 times for building a model and
I. ANALYTICAL TECHNIQUES
39
once for evaluation. The number of cancellation
groups usually ranges from 3 to N. Cross-valida-
tion with N cancellation groups is generally
known as the leave-one-out procedure (LOO).
LOO has the advantage of being unique for
a given data set, whereas, when G < N, different
orders of the samples and different subdivision
schemes generally supply different outcomes.
However, especially when the total number of
samples is considerable, predictions made on
a unique object, although repeated many times,
may yield an overly optimistic result. An exten-
sive evaluation strategy consists in performing
cross-validation many times, with different
numbers of cancellation groups, from 3 up to
N. Another possibility is to repeat the validation,
for a given number G < N of cancellation groups,
each time permuting the order of the samples,
thus obtaining a different group composition
each time.
2.3.3.3. Repeated Evaluation Set
This procedure, also called Monte Carlo vali-
dation, computes many models (often many
thousands), each time creating a different evalu-
ation set, with a variable number of samples, by
random selection. Each sample may fall many
times, or even no times at all, in the evaluation
set. The main drawback of this validation
strategy is the longer computational time.
2.3.4. Supervised Qualitative Modeling
2.3.4.1. Classification and Class-Modeling
A wide number of issues within food science
require qualitative answers. It is the case for the
characterization of ingredients or finished
products, the verification of the geographical
origin, and e more generally e the quality
control, the control on food adulterations, and
so on.
Discriminant classification and class-modeling
techniques represent the most commonchemo-
metric tools for addressing such aims (Oliveri
and Downey, 2012). In fact, they build
40 2. DATA ANALYSIS AND CHEMOMETRICS
mathematical rules or models able to characterize
a sample with respect to a qualitative property,
namely the class to which it belongs.
A class (or category) is defined as a group of
samples having in common the same values of
discrete variables or proximate values of contin-
uous variables. Frequently, such variables are
qualitative factors that cannot be determined
experimentally, so that they have to be esti-
mated from the values of some experimentally
measurable predictors, by way of suitable math-
ematical tools.
In more detail, discriminant classification
techniques are able to determine to which class
a sample more probably belongs, among
a number of predefined classes. They work by
building a delimiter between the classes and,
then, each new object is always assigned to the
category to which it more probably belongs,
even in the case of objects which are not perti-
nent to any class studied.
Instead, class-modeling techniques verify
whether a sample is compatible or not with the
characteristics of a given class of interest. In fact,
they provide an answer to the general question:
“Is sample X, claimed to belong to class A, actu-
ally compatible with the class A model?” This is
essentially the question to be answered in most
of the real qualitative problems studied within
the food sciences. Such an approach is also
capable of detecting outliers (Forina et al., 2008).
2.3.4.2. Evaluation Parameters
The effectiveness of a classification rule is
usually evaluated by the classification rate, i.e.,
the percentage of objects correctly classified.
This parameter is often indicated as prediction
rate, when it is estimated by means of an evalu-
ation sample set. A class rule can be considered
valuable when the prediction rate should be
significantly bigger than the null-classification
rate, which is defined as the probability
percentage of chance correct assignments, and
corresponds to 100% divided by the number of
categories.
I. ANALYTICAL TECHNIQUES
A class model is characterized by two param-
eters: sensitivity and specificity. Sensitivity is
defined as the percentage of objects belonging
to the modeled class which are rightly accepted
by the model. Specificity is the percentage of
objects not belonging to the modeled class
which is rightly rejected by the model. A class-
modeling technique builds a class space, whose
wideness corresponds to the confidence
interval, at a pre-selected confidence level, for
the class objects: sensitivity is an experimental
measure of this confidence level. A decrease in
the confidence level for the modeled class gener-
ally reduces the sensitivity and increases the
specificity of the model. Frequently, in order to
evaluate the model performance, taking into
account these features, an efficiency parameter
is computed as the geometric mean of sensi-
tivity and specificity.
When at least two classes are modeled, the
results of the class-modeling analysis can be
visualized by way of the Coomans’ plots
(Coomans et al., 1984). Such graphs represent
the samples in relation to the distances from
the models of two given classes. Often, the
distances are normalized dividing by the critical
distance value that characterizes the corre-
sponding model.
In the example given in Fig. 2.8, the two
Cartesian axes correspond to the distances
from the model of class Barolo and from the
model of class Grignolino (red-wine data set),
respectively, while two straight lines parallel to
the axes describe the limits of the corresponding
class spaces at a 95% confidence level. The plot
area is divided into four regions, which contain
respectively: the samples accepted by the model
of class Barolo (upper left rectangle), the
samples accepted by the model of class Grigno-
lino (lower right rectangle), the samples
accepted by both the models (lower left square),
and the objects rejected by both the models
(upper right square). All the samples belonging
to the class Barbera correctly lay inside such
last area.

Normalised distance from the model of class Grignolino
FIGURE 2.8 Example of a Coomans’ plot for the data set
red-wine given in Table 2A.1. The samples are represented
by class symbols: circles (Barolo), squares (Grignolino), and
triangles (Barbera). (For color version of the figure, refer to
the online version)
Classification and class-modeling techniques
belong to three main families:
• distance-based techniques
• probabilistic techniques
• experience-based techniques.
2.3.4.3. Distance-Based Techniques
2.3.4.3.1. K NEAREST NEIGHBORS (K-NN)
k-NN is one of the simplest approaches for
classification (Vandeginste et al., 1998). As the
first step, k-NN computes the distances of the
test sample from each of the samples of
a training set, whose class membership is
known. Usually, the multivariate Euclidean
distance is employed:
Di;j
2.3. MULTIVARIATE DATA ANALYSIS 41
with xi and xj being two sample data vectors,
respectively, and xi,v the value of variable v for
sample i.
In some cases, the Mahalanobis distance can
be used. It can be considered as a Euclidean
distance modified for taking into account the
dispersion and the correlation of all of the
variables:
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Di;j ¼ ðxi  xj Þ0 V1 ðxi  xj Þ (2.9)
where V is the covariance matrix.
Once the matrix of distances between objects
has been computed, the k samples nearest to
the test sample are then taken into consider-
ation to perform the classification: generally,
Normalised distance from the model of class Barolo a majority vote is employed, meaning that the
new object is classified into the class mostly
represented within the k selected objects. Being
a distance-based method, it is sensitive to the
measurement units and to the scaling proce-
dures applied.
The method provides a nonlinear delimiter
between categories, generally expressible as
a piecewise linear function (see Fig. 2.9). The
delimiter usually becomes more smoothed for
elevated values of k. When the parameter k is
optimized to obtain the highest prediction
ability for a given data set, validation should
be performed by way of a three-set procedure.
Being a nonprobabilistic method, k-NN is
free from statistical assumptions e such as
normality of variable distributions e and
requirements from limitations on the number
of variables. This assures a wide applicability.
Furthermore, in many applications, it has been
shown to perform as well as or better than
more complex methods (Vandeginste et al.,
1998; Dudoit et al., 2002).
vffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
u V qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uX 2.3.4.3.2. A NONPARAMETRIC CLASS-
¼ t ðxi;v  xj;v Þ2 ¼ ðxi  xj Þ0 ðxi  xj Þ MODELING TECHNIQUE
v¼1
Derde et al. (1986) presented a simple and effi-
(2.8) cient nonparametric class-modeling technique,
I. ANALYTICAL TECHNIQUES
42 2. DATA ANALYSIS AND CHEMOMETRICS

12 12

10 10

8 8

6 6

X2
X2

4 4

2
2

0
0

-2
-2 -2 0 2 4 6 8 10 12
-2 0 2 4 6 8 10 12 X1
X1
FIGURE 2.10 Example of nonparametric class space
FIGURE 2.9 Example of k-NN class delimiter for k ¼ 1 (shadowed region) for the class of interest (artificial data).
(artificial data). (For color version of the figures, refer to the online version)

closely related to k-NN, which defines the class
space on the basis of a critical distance from the
objects of the training set (see Fig. 2.10). Several
settings can be varied, such as the type of distance
(Euclidean or Mahalanobis), and the strategies
adopted to determine the critical distance value.
Unfortunately, this promising class-modeling
technique has not been used anymore, and it
would merit a thorough reconsideration.
2.3.4.3.3. SOFT INDEPENDENT MODELING OF
CLASS ANALOGY (SIMCA)
Soft independent modeling of class analogy
(SIMCA) (Wold and Sjöström, 1977) was the
first class-modeling technique introduced into
chemometrics. This method builds class
models based on PCA performed using only
the samples of the category studied, generally
after within-class autoscaling or centering. In
more detail, SIMCA models are defined by
the range of the sample scores on a selected
number of low-order principal components
(PCs) e ideally the significant ones e and
models therefore correspond to rectangles
(two PCs), parallelepipeds (three PCs), or
hyper-parallelepipeds (more than three PCs)
referred to as the multidimensional boxes of
SIMCA inner space. Conversely, the principal
components not used to describe the model
define the outer space, which represents the
space of uninformative variations, often due
to noise. The score range can be enlarged or
reduced, mainly depending on the number of
samples, to avoid the possibility of under- or
overestimation of the true variability (Forina
and Lanteri, 1984). The standard deviation of
the distance of the objects in the training set
from the model corresponds to the class stan-
dard deviation. The boundaries of the SIMCA
space around the model are determined (as
shown in Fig. 2.11) by a critical distance, which
is obtained by means of the Fisher statistics.
SIMCA is a very flexible technique since it
allows variation in a large number of parame-
ters such as scaling or weighting of the

I. ANALYTICAL TECHNIQUES
 2.3. MULTIVARIATE DATA ANALYSIS 43
12 12

10 10

8 8

6 6
X2

X2
4 4

2 2

0 0

-2 -2
-2 0 2 4 6 8 10 12 -2 0 2 4 6 8 10 12
X1 X1

FIGURE 2.11 SIMCA normal-range model (segment)
and class space (shadowed region) for the class of interest
(artificial data). (For color version of the figure, refer to the
online version)
original variables, number of components, and
expanded or contracted score range.
2.3.4.4. Probabilistic Techniques
2.3.4.4.1. LINEAR DISCRIMINANT ANALYSIS
(LDA)
Linear discriminant analysis (LDA) is
the first multivariate classification technique,
introduced by Fisher (1936). It is a probabilistic
parametric technique, i.e., it is based on the
estimation of multivariate probability density
functions, which are entirely described by
a minimum number of parameters: means, vari-
ances, and covariances. LDA is based on the
hypotheses that the probability density distribu-
tions are multivariate normal and that the disper-
sion is the same for all the categories. This means
that the varianceecovariance matrix is the same
for all of the categories, while the centroids are
different (different location). In the case of two
variables, the probability density function is bell-
shaped and its elliptic section lines correspond
FIGURE 2.12 Iso-probability ellipses under LDA
hypotheses and the resultant linear class delimiter (artificial
data).
to equal probability density values and to the
same Mahalanobis distance from the centroid.
Because of the above-mentioned LDA hypoth-
eses, the ellipses of different categories present
equal eccentricity and axis orientation: they
only differ for their location in the plane. By con-
necting the intersection points of each couple of
corresponding ellipses, a straight line is identi-
fied which corresponds to the delimiter between
the two classes (see Fig. 2.12). For this reason, this
technique is called linear discriminant analysis.
The directions which maximize the separation
between pairs of classes are the so-called canon-
ical variables.
2.3.4.4.2. QUADRATIC DISCRIMINANT
ANALYSIS (QDA)
Quadratic discriminant analysis (QDA) is
a probabilistic parametric classification technique
which represents an evolution of LDA for
nonlinear class separations. Also QDA, like
LDA, is based on the hypothesis that the

I. ANALYTICAL TECHNIQUES
44 2. DATA ANALYSIS AND CHEMOMETRICS

probability density distributions are multivariate
normal but, in this case, the dispersion is not the
same for all of the categories. It follows that the
categories differ not only for the position of their
centroid but also for the varianceecovariance
matrix (different location and dispersion). Conse-
quently, the ellipses of different categories differ
also for eccentricity and axis orientation (Geisser,
1964). By connecting the intersection points of
each couple of corresponding ellipses (at the
same Mahalanobis distance from the respective
centroids), a quadratic delimiter is identified,
which is a parabola in the bidimensional case, as
represented in Fig. 2.13.
2.3.4.4.3. UNEQUAL CLASS MODELS (UNEQ)
UNEQ is a powerful class-modeling tech-
nique, which originated in the work of Hotelling
(1947) and was launched in chemometrics by
Derde and Massart (1986). The method, closely
related to QDA, is based on the hypothesis of
a multivariate normal distribution in each cate-
gory studied and on the use of Hotelling’s T2
statistics to define a class space, whose boundary
is an ellipse (two variables), an ellipsoid (three
variables), or a hyper-ellipsoid (more than three
variables). The dispersion of the class space is
defined by the critical value of the T2 statistics
at a selected confidence level (see Fig. 2.14). The
eccentricity and the orientation of the ellipse
depend on the correlation between the variables
and on their dispersion.
These probabilistic techniques present some
restrictions on the number of objects that can
be used. From a strictly mathematical point of
view, objects have to be one more than the
number of variables measured. Nevertheless,
in order to obtain reliable results, these tech-
niques should be applied in cases when the ratio
between the number of objects in a given cate-
gory and the number of variables is at least
three. Furthermore, the number of objects in
each class should be nearly balanced: it is not
advisable to work when ratios between number
of objects in different categories are greater than
three (Derde and Massart, 1989).

12 12

10 10

8 8

6 6
X2

X2

4 4

2 2

0 0

-2 -2
-2 0 2 4 6 8 10 12 -2 0 2 4 6 8 10 12
X1 X1

FIGURE 2.13 Iso-probability ellipses under QDA FIGURE 2.14 UNEQ model (cross) and class space
hypotheses and the resultant quadratic class delimiter (shadowed region) for the class of interest (artificial data).
(artificial data). (For color version of the figure, refer to the online version)

I. ANALYTICAL TECHNIQUES
 2.3. MULTIVARIATE DATA ANALYSIS
In cases involving many variables, it is
possible to apply LDA and QDA-UNEQ
following a preliminary reduction in the vari-
able number, for instance, by a PCA-based
compression.
2.3.4.4.4. POTENTIAL FUNCTION METHODS
Potential function techniques were intro-
duced into chemometrics by Coomans and
Broeckaert (1986). These methods estimate
a probability density distribution as a sum of
contributions of each single sample in a training
set. A variety of functions can be used to define
the individual contributions. The most
commonly used are Gaussian-like functions,
with a smoothing coefficient that is formally
analogous to the standard deviation of the
Gaussian probability function, thus deter-
mining the shape of the distribution. Such coef-
ficient can be the same for all the samples of
a given class (fixed potential), otherwise it
can be varied as a function of the local density
of samples: such latter strategy, known as
normal variable potential, is useful when the
underlying multivariate distribution is very
asymmetric, with regions characterized by
nonuniform density of samples (Forina et al.,
1991).
The value of the smoothing coefficient can be
optimized by means of a leave-one-out proce-
dure with an optimization sample set.
As represented in Fig. 2.15, the resulting esti-
mated overall probability distribution can be
very complex, capable of effectively describing
nonuniform distributions of samples. From the
probability distribution, the boundary of the
class space can be obtained at a selected confi-
dence level.
2.3.5. Supervised Quantitative
Modeling
Regression defines mathematical relationships
between variables or groups of variables, and
provides models for quantitative predictions.
I. ANALYTICAL TECHNIQUES
45
12
10
8
6
X2
4
2
0
-2
-2 0 2 4 6 8 10 12
X1
FIGURE 2.15 Potential function class space (shadowed
region) for the class of interest (artificial data). (For color
version of the figure, refer to the online version)
Regression techniques can be univariate or
multivariate, depending on the number of
predictors and, eventually, of response variables
involved. Furthermore, they can be linear or
nonlinear, depending on the type of relationship
they are able to model.
Univariate linear regression is a very common
tool in analytical chemistry, generally used to
describe the relation between a chemical quan-
tity (typically, the concentration of an analyte in
a series of standards) and a measured physical
variable (e.g., absorbance values at a given wave-
length). The mathematical model obtained is
used inversely, to compute the chemical quantity
in real samples from the values of the physical
measures performed on them.
2.3.5.1. Ordinary Least Squares (OLS)
The ordinary least-squares (OLS) e or clas-
sical least-squares (CLS) e method is probably
the most widely used and studied historically.
It looks for the combination of parameters of
the linear model (intercept and slope) that
46 2. DATA ANALYSIS AND CHEMOMETRICS
provides the minimum value for the squared
residuals (i.e., the squared differences between
the values estimated by the model and the corre-
sponding true values). The statistical implica-
tions lying on the basis of the method allow us
to calculate the confidence interval for each pre-
dicted value.
OLS can be applied as well to multivariate
data, namely when the predictors are two or
more. In such cases, the method is also known
as multivariate linear regression (MLR) (Draper
and Smith, 1981).
The model can be expressed as a mathemat-
ical relationship between the response y and
the V predictors:
^
y ¼ b 0 þ b 1 x1 þ b 2 x2 þ . þ b V xV (2.10)
that is, in the matrix notation:
^ ¼ X0 b
y (2.11)
where X is the matrix of the predictors
augmented with a column of 1, necessary for
the estimation of the intercept values, and b is
the column vector of the regression coefficients.
The regression coefficients are estimated by
b ¼ ðX0 XÞ1 X0 y (2.12)
The elements of the vector y are the reference
values of the response variable, used for
building the model.
The uncertainty on the coefficient estimation
varies inversely with the determinant of the
information matrix ðX0 XÞ which, in the case of
a unique predictor, corresponds to its variance.
In the multivariate cases, the determinant value
depends on the variance of the predictors and
on their inter-correlation: a high correlation
gives a small determinant of the information
matrix, which means a large uncertainty on
the coefficients, and, consequently, unreliable
regression results.
This is the typical situation when vectors cor-
responding to almost continuous signals (e.g.,
spectra) are used as predictors. In fact, in such
I. ANALYTICAL TECHNIQUES
cases, contiguous variables are considerably
inter-correlated. In a spectrum, for instance,
absorbances evaluated at two consecutive
wavelengths frequently carry almost the same
information, so that their correlation coefficient
is nearly 1. In such cases, standard OLS is abso-
lutely not recommendable.
Furthermore, the number of objects required
for OLS regression must be at least equal to
the number of predictors plus 1. Such a condi-
tion is rarely satisfied in many practical cases.
2.3.5.2. Principal-Component Regression
(PCR)
Principal-component analysis offers a very
simple approach to overcome these hurdles.
The model is obtained by a classical least-
squares approach, which uses a reduced
number of significant principal components,
computed from the original variables, as the
predictors (Jolliffe, 1982). The PCs are, by defini-
tion, orthogonal and, therefore, uncorrelated.
This technique, which is very efficient in
many cases, is known as principal-component
regression (PCR). Since not always the direc-
tions which explain the highest variance
amount (i.e., the lowest-order PCs) are the
most important in predicting a response vari-
able, it is possible to follow a refined approach,
which performs a stepwise selection of the prin-
cipal components to be used in the model on the
basis of their modeling efficiency.
2.3.5.3. Partial Least Squares (PLS)
PLS (partial least squares, or even projections
onto latent structures) is probably the most
widely used multivariate regression technique
(Wold et al., 2001) and represents a better solu-
tion to both of the problems of variable number
and inter-correlation. The latent structures,
more frequently called latent variables (LVs) or
PLS components, are directions in the space of
the predictors. In particular, the first latent
variable is the direction characterized by
 2.3. MULTIVARIATE DATA ANALYSIS
the maximum covariance with the selected
response variable. The information related to
the first latent variable is then subtracted from
both the original predictors and the response.
The second latent variable is orthogonal to the
first one, being the direction of maximum
covariance between the residuals of the predic-
tors and the residuals of the response. This
approach continues for the subsequent LVs.
The optimal complexity of the PLS model,
i.e., the most appropriate number of latent
variables, is determined by evaluating, with
a proper validation strategy, the prediction
error corresponding to models with increasing
complexity. The parameters considered are
usually the standard deviation of the error of
calibration (SDEC), computed with the objects
used for building the model, and the standard
deviation of the error of prediction (SDEP),
computed with objects not used for building
the model:
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
PN
i ¼ 1 ðyi  ^ yi Þ 2
SDECðPÞ ¼
N
3.5
3.0
2.5
2.0
SDE
1.5
1.0
0.5
0.0
0 5 10 15 20
LV number
I. ANALYTICAL TECHNIQUES
47
where yi is the value of the response variable y
for sample i, ^yi is the corresponding value
computed or predicted by the model, and N is
the number of samples.
In general, the calibration error always
decreases when the number of LVs augments,
because the fitting increases (toward an overfit-
ting). On the contrary, the prediction error
generally decreases until a certain model
complexity and then raises: this indicates that
the LVs further introduced are bringing noise,
as shown in the example given in Fig. 2.16.
A simple and practical criterion is the choice
of the LV number corresponding to the SDEP
absolute minimum or e better e to the first local
minimum as the optimal model complexity. In
the example of Fig. 2.16, it corresponds to six
LVs.
When the number of noisy (noninformative)
variables is too large, the performance of PLS
models may be improved by a selection of
useful predictors performed in advance (Forina
et al., 2007).
(2.13) A number of PLS variants have been
deployed, for instance, for developing nonlinear
FIGURE 2.16 Example of calibra-
SDEC tion and prediction error typical
SDEP trends at the increasing of the PLS
model complexity (number of latent
variables).
25 30
48
models and for predicting together two or more
response variables (PLS-2). Furthermore, when
category indices are taken as dummy response
variables, PLS may work as a classification
method which is usually called PLS discrimi-
nant analysis (PLS-DA) or discriminant PLS
(D-PLS).
2.3.6. Artificial Neural Networks
Artificial neural networks (ANNs) are
a family of nonparametric versatile tools that
can be employed both for data exploration and
for qualitative and quantitative predictive
modeling. ANNs offer some advantages. For
instance, they are generally well suited for
nonlinear problems, and the related software
is easily available. Conversely, a number of
important drawbacks should limit ANN use
only to the cases in which other simpler tech-
niques fail and, primarily, when a large number
of samples are available.
Multilayer feedforward neural networks
(MLF) represent the configuration of ANNs
most widely applied to electronic tongue
data. An exemplificative scheme is shown in
Fig. 2.17.
MLF are composed by a number of computa-
tional elements, called neurons, generally orga-
nized in three layers (Zupan, 1994). In the first
OUTPUT
O1 O2
LAYER
Z Z
HIDDEN
LAYER H1 H...
S S S represented by the Kohonen’s self-organizing
INPUT
I1 I2 I3
LAYER
FIGURE 2.17 Exemplificative scheme of general inter-
neuronal connections and transmission/correction mecha-
nisms for a multilayer feedforward neural network.
2. DATA ANALYSIS AND CHEMOMETRICS
one, the input layer, there are usually N neurons
which correspond to the original predictors.
The predictors are scaled (generally range
scaled). When their number is very large, the
principal components are often used, in order
to reduce the data amount and the computa-
tional time.
The first layer transmits the value of the
predictors to the second e hidden e layer. All
the neurons of the input layer are connected
to the J neurons of the second layer by
means of weight coefficients, meaning that the
J elements of the hidden layer receive, as infor-
mation, a weighted sum S of the values from
the input layer. They transform the information
received (S) by means of a suitable transfer
function, frequently a sigmoid.
These neurons transmit information to the
third e output e layer, as a weighted combina-
tion (Z) of their values. The neurons in the output
layer correspond to the response variables which,
in the case for classification, are the coded class
indices. The output neurons transform the infor-
mation Z, from the hidden layer, by means of
a further sigmoid or semilinear function.
After a first random initialization of the
values, a learning procedure modifies the
weights wn,j and wj during several optimization
cycles, in order to improve the performances of
the net. The correction of the weights at each
step is proportional to the prediction error of
the previous cycle. The optimization of many
parameters and the elevated number of learning
cycles considerably increase the risk of overfit-
C or
wj,o ting and, for this reason, a deep validation is
rec
required, with a consistent number of objects.
tion
Hj Another type of widely employed ANN is
wn,j
maps (SOMs), used for unsupervised explor-
atory analysis, and by the counterpropagation
(CP) neural networks, used for nonlinear regres-
I... I4
sion and classification (Kohonen, 2001). In addi-
tion, these tools require a considerable number
of objects to build reliable models, and a severe
validation.
I. ANALYTICAL TECHNIQUES
TABLE 2A.1 Red-Wine Data Set, and Basic Statistical Parameters

Alcalinity
Alcohol Sugar-free Fixed Tartaric Malic Uronic Ash of ash Potassium Calcium Magnesium Phosphate
Name Category (% abv) extract (g/l) acidity (g/l) acid (g/l) acid (g/l) acids (mg/l) pH (g/l) (meq/l) (mg/l) (mg/l) (mg/l) (g/l)
OLO0171 Barolo 14.23 24.82 73.10 1.21 1.71 0.72 3.38 2.43 15.60 950 62 127 320

OLO0271 Barolo 13.20 26.30 72.80 1.84 1.78 0.71 3.30 2.14 11.20 765 75 100 395

OLO0371 Barolo 13.16 26.30 68.50 1.94 2.36 0.84 3.48 2.67 18.60 936 70 101 497

OLO0471 Barolo 14.37 25.85 74.90 1.59 1.95 0.72 3.43 2.50 16.80 985 47 113 580

OLO0571 Barolo 13.24 26.05 83.50 1.30 2.59 1.10 3.42 2.87 21.00 1088 70 118 408

OLO0671 Barolo 14.20 28.40 79.90 2.14 1.76 0.96 3.39 2.45 15.20 868 71 112 418

OLO0771 Barolo 14.39 27.02 64.30 1.64 1.87 0.95 3.42 2.45 14.60 889 67 96 306

OLO0871 Barolo 14.06 26.40 73.50 1.33 2.15 1.14 3.54 2.61 17.60 894 50 121 502

OLO0971 Barolo 14.83 26.80 69.50 1.82 1.64 0.67 3.30 2.17 14.00 765 49 97 440

OLO1071 Barolo 13.86 27.00 68.50 1.92 1.35 0.67 3.27 2.27 16.00 794 51 98 391

OLO1171 Barolo 14.10 26.08 72.50 1.64 2.16 0.62 3.31 2.30 18.00 838 61 105 399

OLO1271 Barolo 14.12 28.35 72.90 1.51 1.48 0.96 3.20 2.32 16.80 827 60 95 424

OLO1371 Barolo 13.75 30.25 75.10 1.92 1.73 0.64 3.18 2.41 16.00 752 65 89 453

OLO1471 Barolo 14.75 30.40 98.90 2.08 1.73 0.72 3.01 2.39 11.40 910 46 91 510

OLO1571 Barolo 14.38 27.10 72.30 1.95 1.87 0.67 3.20 2.38 12.00 927 29 102 523

OLO1671 Barolo 13.63 27.15 69.60 1.48 1.81 0.67 3.47 2.70 17.20 905 28 112 385

OLO1771 Barolo 14.30 27.90 74.90 1.41 1.92 0.82 3.40 2.72 20.00 860 108 120 513

OLO1871 Barolo 13.83 26.30 64.90 1.93 1.57 0.68 3.43 2.62 20.00 905 68 115 419

OLO1971 Barolo 14.19 26.40 72.00 1.85 1.59 0.82 3.38 2.48 16.50 964 86 108 488

OLO0173 Barolo 13.64 27.72 91.50 1.35 3.10 0.82 3.30 2.56 15.20 1038 111 116 402

OLO0273 Barolo 14.06 25.32 71.10 1.34 1.63 1.00 3.47 2.28 16.00 905 79 126 323

OLO0373 Barolo 12.93 28.80 102.10 1.05 3.80 0.89 3.26 2.65 18.60 915 79 102 294

OLO0473 Barolo 13.71 27.63 80.00 2.23 1.86 1.21 3.33 2.36 16.60 815 89 101 476

OLO0573 Barolo 12.85 25.80 69.60 1.54 1.60 0.79 3.45 2.52 17.80 958 101 95 415

OLO0673 Barolo 13.50 25.00 81.60 1.55 1.81 0.95 3.42 2.61 20.00 992 62 96 476

(Continued)
TABLE 2A.1 Red-Wine Data Set, and Basic Statistical ParametersdCont’d

Alcalinity
Alcohol Sugar-free Fixed Tartaric Malic Uronic Ash of ash Potassium Calcium Magnesium Phosphate
Name Category (% abv) extract (g/l) acidity (g/l) acid (g/l) acid (g/l) acids (mg/l) pH (g/l) (meq/l) (mg/l) (mg/l) (mg/l) (g/l)

OLO0773 Barolo 13.05 25.72 78.30 1.15 2.05 1.08 3.57 3.22 25.00 1095 63 124 536

OLO0873 Barolo 13.39 27.10 72.30 1.52 1.77 1.05 3.46 2.62 16.10 936 68 93 395

OLO0973 Barolo 13.30 22.70 68.30 1.74 1.72 1.06 3.44 2.14 17.00 882 52 94 434

OLO1073 Barolo 13.87 29.30 68.30 1.38 1.90 0.75 3.42 2.80 19.40 1085 68 107 396

OLO1173 Barolo 14.02 25.20 69.60 1.71 1.68 0.79 3.26 2.21 16.00 780 62 96 510

GRI0170 Grignolino 12.37 18.30 90.10 2.80 0.94 0.73 3.11 1.36 10.60 580 77 88 296

GRI0270 Grignolino 12.33 22.90 72.20 2.25 1.10 0.69 3.26 2.28 16.00 715 85 101 365

GRI0370 Grignolino 12.64 23.90 95.70 1.93 1.36 1.06 3.19 2.02 16.80 688 83 100 395

GRI0470 Grignolino 13.67 22.20 64.80 2.20 1.25 0.74 3.40 1.92 18.00 725 51 94 301

GRI0570 Grignolino 12.37 23.50 70.00 2.06 1.13 0.72 3.30 2.16 19.00 785 73 87 422

GRI0670 Grignolino 12.17 23.03 65.70 1.84 1.45 0.72 3.35 2.53 19.00 790 62 104 411

GRI0770 Grignolino 12.37 26.80 62.70 1.70 1.21 0.88 3.40 2.56 18.10 978 55 98 310

GRI0870 Grignolino 13.11 23.70 80.00 1.40 1.01 0.77 3.10 1.70 15.00 730 80 78 297

GRI0970 Grignolino 12.37 20.90 63.70 1.94 1.17 0.67 3.40 1.92 19.60 785 40 78 212

GRI0171 Grignolino 13.34 23.72 70.00 2.02 0.94 1.09 3.26 2.36 17.00 760 64 110 451

GRI0271 Grignolino 12.21 22.70 90.70 3.62 1.19 0.94 3.14 1.75 16.80 795 134 151 448

GRI0371 Grignolino 12.29 21.40 55.60 1.43 1.61 0.87 3.54 2.21 20.40 682 102 103 324

GRI0471 Grignolino 13.86 25.25 59.50 1.27 1.51 1.09 3.63 2.67 25.00 785 63 86 383

GRI0571 Grignolino 13.49 22.30 60.90 1.74 1.66 0.67 3.44 2.24 24.00 680 60 87 300

GRI0671 Grignolino 12.99 26.10 50.50 1.42 1.67 1.24 3.52 2.60 30.00 974 55 139 473

GRI0771 Grignolino 11.96 24.50 65.70 2.18 1.09 0.73 3.40 2.30 21.00 681 98 101 366

GRI0871 Grignolino 11.66 20.30 61.70 1.70 1.88 0.60 3.30 1.92 16.00 785 52 97 312

GRI0971 Grignolino 13.03 23.50 78.60 1.90 0.90 0.76 3.30 1.71 16.00 790 57 86 396

GRI0172 Grignolino 11.84 26.40 108.70 1.70 2.89 0.91 3.11 2.23 18.00 790 71 112 350

GRI0272 Grignolino 12.33 20.60 58.70 2.41 0.99 0.84 3.32 1.95 14.80 680 124 136 438

GRI0372 Grignolino 12.70 27.15 93.30 1.46 3.87 1.11 3.19 2.40 23.00 890 110 101 321
GRI0472 Grignolino 12.00 23.20 58.40 1.88 0.92 0.82 3.30 2.00 19.00 680 63 86 408

GRI0572 Grignolino 12.72 22.90 58.40 1.40 1.81 0.81 3.50 2.20 18.80 890 83 86 418

GRI0672 Grignolino 12.08 23.50 56.90 1.33 1.13 0.71 3.65 2.51 24.00 980 85 78 215

GRI0772 Grignolino 13.05 25.50 104.80 1.64 3.86 0.73 3.19 2.32 22.50 938 98 85 195

GRI0173 Grignolino 11.84 23.40 70.80 1.80 0.89 1.00 3.40 2.58 18.00 922 80 94 378

GRI0273 Grignolino 12.67 24.30 74.10 1.70 0.98 0.88 3.35 2.24 18.00 840 81 99 336

GRI0373 Grignolino 12.16 25.80 78.90 1.84 1.61 0.78 3.37 2.31 22.80 845 98 90 285

GRI0473 Grignolino 11.65 22.90 62.90 1.80 1.67 0.64 3.55 2.62 26.00 1045 125 88 281

GRI0573 Grignolino 11.64 24.20 72.40 1.84 2.06 0.89 3.40 2.46 21.60 962 79 84 304

ERA0174 Barbera 12.86 26.80 87.30 0.99 1.35 0.92 3.22 2.32 18.00 830 52 122 266

ERA0274 Barbera 12.88 23.95 78.90 1.85 2.99 0.98 3.50 2.40 20.00 795 55 104 269

ERA0374 Barbera 12.81 24.45 76.20 2.93 2.31 0.87 3.64 2.40 24.00 785 49 98 266

ERA0474 Barbera 12.70 24.75 91.00 1.91 3.55 1.80 3.26 2.36 21.50 805 47 106 356

ERA0574 Barbera 12.51 23.50 104.70 1.34 1.24 0.98 3.50 2.25 17.50 975 60 85 273

ERA0674 Barbera 12.60 23.60 80.60 2.26 2.46 0.97 3.31 2.20 18.50 760 103 94 275

ERA0774 Barbera 12.25 25.30 91.40 1.42 4.72 1.25 3.40 2.54 21.00 995 105 89 262

ERA0874 Barbera 12.53 27.10 99.80 1.88 5.51 1.19 3.30 2.64 25.00 930 100 96 360

ERA0974 Barbera 13.49 25.70 115.50 2.17 3.59 1.47 3.24 2.19 19.50 825 111 88 315

ERA0176 Barbera 12.84 26.20 82.00 1.79 2.96 1.26 3.50 2.61 24.00 925 48 101 398

ERA0276 Barbera 12.93 26.78 80.00 1.69 2.81 1.15 3.31 2.70 21.00 965 40 96 351

ERA0376 Barbera 13.36 24.12 97.80 2.83 2.56 0.77 3.35 2.35 20.00 880 47 89 235

ERA0476 Barbera 13.52 27.90 85.00 1.46 3.17 1.23 3.28 2.72 23.50 880 38 97 325

ERA0576 Barbera 13.62 25.52 93.70 2.70 4.95 1.56 3.41 2.35 20.00 805 57 92 191

ERA0179 Barbera 12.25 23.40 113.50 3.54 3.88 1.04 3.01 2.20 18.50 785 77 112 358

ERA0279 Barbera 13.16 22.90 117.90 3.15 3.57 1.18 3.14 2.15 21.00 805 88 102 456

ERA0379 Barbera 13.88 21.40 99.30 2.81 5.04 1.29 3.28 2.23 20.00 750 43 80 171

ERA0479 Barbera 12.87 24.35 98.90 2.51 4.61 1.25 3.18 2.48 21.50 830 63 86 366

ERA0579 Barbera 13.32 21.46 96.90 2.85 3.24 1.75 3.30 2.38 21.50 790 42 92 306

ERA0178 Barbera 13.08 26.80 120.60 2.90 3.90 1.11 3.16 2.36 21.50 790 73 113 303

(Continued)
TABLE 2A.1 Red-Wine Data Set, and Basic Statistical ParametersdCont’d

Alcalinity
Alcohol Sugar-free Fixed Tartaric Malic Uronic Ash of ash Potassium Calcium Magnesium Phosphate
Name Category (% abv) extract (g/l) acidity (g/l) acid (g/l) acid (g/l) acids (mg/l) pH (g/l) (meq/l) (mg/l) (mg/l) (mg/l) (g/l)

ERA0278 Barbera 13.50 26.50 105.50 2.31 3.12 1.31 3.23 2.62 24.00 980 67 123 338

ERA0378 Barbera 12.79 23.40 117.80 3.12 2.67 0.82 3.21 2.48 22.00 890 53 112 407

ERA0478 Barbera 13.11 25.20 95.40 2.26 1.90 0.86 3.49 2.75 25.50 1140 74 116 289

ERA0578 Barbera 13.23 23.85 120.60 2.80 3.30 0.80 3.20 2.28 18.50 915 68 98 351

ERA0678 Barbera 12.58 21.75 102.70 2.92 1.29 0.79 3.21 2.10 20.00 875 107 103 368

ERA0778 Barbera 13.17 23.20 129.30 2.28 5.19 1.49 3.58 2.32 22.00 1045 102 93 241

ERA0878 Barbera 13.84 24.70 122.90 2.76 4.12 1.07 3.19 2.38 19.50 840 108 89 402

ERA0978 Barbera 12.45 25.35 105.90 2.23 3.03 1.24 3.62 2.64 27.00 1050 118 97 393

ERA1078 Barbera 14.34 29.10 97.50 2.73 1.68 1.60 3.42 2.70 25.00 1095 78 98 462

ERA1178 Barbera 13.48 26.95 102.50 3.75 1.67 1.37 3.41 2.64 22.50 1055 79 89 480

mean 13.13 25.07 82.46 1.97 2.22 0.95 3.35 2.37 19.27 868.7 72.6 100.6 369.5

variance 0.59 5.25 332.34 0.35 1.27 0.07 0.02 0.08 13.24 13102.4 536.4 194.7 7453.5

standard deviation 0.77 2.29 18.23 0.59 1.12 0.26 0.14 0.28 3.64 114.5 23.2 14.0 86.3

OD280/
Total Non- OD315 OD280/ 2-3- Total
Chloride phenols flavanoid Pro- Color of diluted OD315 Glycerol butanediol nitrogen Proline Methanol
(mg/l) (g/l) Flavanoids phenols anthocyanins intensity Hue wines of flavanoids (g/l) (g/l) (mg/l) (mg/l) (% A.A.)

82 2.80 3.06 0.28 2.29 5.64 1.04 3.92 4.77 9.29 757 153 1065 113

90 2.65 2.76 0.26 1.28 4.38 1.05 3.40 3.80 8.93 881 194 1050 94

67 2.80 3.24 0.30 2.81 5.68 1.03 3.17 3.46 11.74 900 206 1185 125

49 3.85 3.49 0.24 2.18 7.80 0.86 3.45 3.54 10.13 1119 292 1480 80

65 2.80 2.69 0.39 1.82 4.32 1.04 2.93 3.22 10.27 799 215 735 73

58 3.27 3.39 0.34 1.97 6.75 1.05 2.85 3.16 10.85 865 364 1450 68

52 2.50 2.52 0.30 1.98 5.25 1.02 3.58 3.94 9.05 931 378 1290 80

64 2.60 2.51 0.31 1.25 5.05 1.06 3.58 3.94 10.13 865 358 1295 100

58 2.80 2.98 0.29 1.98 5.20 1.08 2.85 3.03 9.89 825 438 1045 141

64 2.98 3.15 0.22 1.85 7.22 1.01 3.55 3.75 12.65 788 350 1045 121
61 2.95 3.32 0.22 2.38 5.75 1.25 3.17 3.27 8.59 964 378 1510 123

79 2.20 2.43 0.26 1.57 5.00 1.17 2.82 3.04 11.52 894 294 1280 134

257 2.60 2.76 0.29 1.81 5.60 1.15 2.90 2.92 12.24 784 289 1320 164

50 3.10 3.69 0.43 2.81 5.40 1.25 2.73 2.82 12.29 766 224 1150 105

55 3.30 3.64 0.29 2.96 7.50 1.20 3.00 3.32 9.53 1041 324 1547 114

50 2.85 2.91 0.30 1.46 7.30 1.28 2.88 3.12 7.92 812 229 1310 97

62 2.80 3.14 0.33 1.97 6.20 1.07 2.65 3.10 9.24 836 308 1280 113

58 2.95 3.40 0.40 1.72 6.60 1.13 2.57 2.66 9.41 722 274 1130 99

28 3.30 3.93 0.32 1.86 8.70 1.23 2.82 3.17 9.85 808 230 1680 135

67 2.70 3.03 0.17 1.66 5.10 0.96 3.36 4.00 10.39 726 227 845 119

73 3.00 3.17 0.24 2.10 5.65 1.09 3.71 3.75 10.30 828 225 780 145

62 2.41 2.41 0.25 1.98 4.50 1.03 3.52 3.66 11.88 589 237 770 123

134 2.61 2.88 0.27 1.69 3.80 1.11 4.00 4.31 8.81 715 270 1035 109

73 2.48 2.37 0.26 1.46 3.93 1.09 3.63 3.82 9.14 568 248 1015 102

47 2.53 2.61 0.28 1.66 3.52 1.12 3.82 4.00 9.45 667 210 845 86

82 2.63 2.68 0.47 1.92 3.58 1.13 3.20 3.63 10.34 753 238 830 124

52 2.85 2.94 0.34 1.45 4.80 0.92 3.22 4.44 9.96 854 285 1195 85

46 2.40 2.19 0.27 1.35 3.95 1.02 2.77 3.10 9.70 757 350 1285 84

76 2.95 2.97 0.37 1.76 4.50 1.25 3.40 3.72 9.53 702 280 915 99

53 2.65 2.33 0.26 1.98 4.70 1.04 3.59 3.77 9.94 689 293 1035 100

52 1.98 0.57 0.28 0.42 1.95 1.05 1.82 2.12 5.40 736 287 520 98

108 2.05 1.09 0.63 0.41 3.27 1.25 1.67 1.42 6.90 658 345 680 127

53 2.02 1.41 0.53 0.62 5.75 0.98 1.59 1.86 8.20 691 321 450 60

47 2.10 1.79 0.32 0.73 3.80 1.23 2.46 1.73 8.60 797 262 630 87

306 3.50 3.10 0.19 1.87 4.45 1.22 2.87 3.07 7.20 748 141 420 157

116 1.89 1.75 0.45 1.03 2.95 1.45 2.23 2.73 7.50 627 219 355 58

69 2.42 2.65 0.37 2.08 4.60 1.19 2.30 2.60 7.96 680 259 678 118

148 2.98 3.18 0.26 2.28 5.30 1.12 3.18 3.33 8.20 604 100 502 114

54 2.11 2.00 0.27 1.04 4.68 1.12 3.48 4.07 7.10 554 425 510 98

(Continued)
TABLE 2A.1 Red-Wine Data Set, and Basic Statistical ParametersdCont’d

OD280/
Total Non- OD315 OD280/ 2-3- Total
Chloride phenols flavanoid Pro- Color of diluted OD315 Glycerol butanediol nitrogen Proline Methanol
(mg/l) (g/l) Flavanoids phenols anthocyanins intensity Hue wines of flavanoids (g/l) (g/l) (mg/l) (mg/l) (% A.A.)

111 2.53 1.30 0.55 0.42 3.17 1.02 1.93 1.92 8.10 704 363 750 137

88 1.85 1.28 0.14 2.50 2.85 1.28 3.07 3.23 6.81 714 195 718 116

50 1.10 1.02 0.37 1.46 3.05 0.91 1.82 2.00 6.38 661 301 870 78

59 2.95 2.86 0.21 1.87 3.38 1.36 3.16 3.52 7.62 748 170 410 99

43 1.88 1.84 0.27 1.03 3.74 0.98 2.78 3.50 8.04 614 160 472 64

35 3.30 2.89 0.21 1.96 3.35 1.31 3.50 3.60 8.00 731 293 985 113

48 3.38 2.14 0.13 1.65 3.21 0.99 3.13 3.15 7.70 563 183 886 57

59 1.61 1.57 0.34 1.15 3.80 1.23 2.14 2.35 6.14 596 109 428 129

122 1.95 2.03 0.24 1.46 4.60 1.19 2.48 2.85 8.40 756 167 392 145

58 1.72 1.32 0.43 0.95 2.65 0.96 2.52 3.25 5.22 514 246 500 122

99 1.90 1.85 0.35 2.76 3.40 1.06 2.31 2.70 7.96 654 259 750 59

52 2.83 2.55 0.43 1.95 2.57 1.19 3.13 3.82 8.66 700 227 463 101

27 2.42 2.26 0.30 1.43 2.50 1.38 3.12 3.52 5.80 645 199 278 95

64 2.20 2.53 0.26 1.77 3.90 1.16 3.14 3.33 7.38 664 199 714 111

53 2.00 1.58 0.40 1.40 2.20 1.31 2.72 3.50 8.11 548 203 630 115

48 1.65 1.59 0.61 1.62 4.80 0.84 2.01 2.07 8.64 649 207 515 114

95 2.20 2.21 0.22 2.35 3.05 0.79 3.08 3.81 6.36 586 138 520 141

70 2.20 1.94 0.30 1.46 2.62 1.23 3.16 3.60 7.90 600 217 450 121

54 1.78 1.69 0.43 1.56 2.45 1.33 2.26 2.92 8.04 643 195 495 116

36 1.92 1.61 0.40 1.34 2.60 1.36 3.21 3.27 9.54 608 262 562 120

70 1.95 1.69 0.48 1.35 2.80 1.00 2.75 3.60 7.97 523 223 680 120

46 1.51 1.25 0.21 0.94 4.10 0.76 1.29 1.26 6.43 673 252 630 122

72 1.30 1.22 0.24 0.83 5.40 0.74 1.42 1.34 10.10 918 319 530 102

67 1.15 1.09 0.27 0.83 5.70 0.66 1.36 1.24 10.02 1095 258 560 132

118 1.70 1.20 0.17 0.84 5.00 0.78 1.29 1.23 8.52 1020 238 600 121

29 2.00 0.58 0.60 1.25 5.45 0.75 1.51 1.40 8.32 764 178 650 79
77 1.62 0.66 0.63 0.94 7.10 0.73 1.58 1.37 6.47 573 174 695 100

144 1.38 0.47 0.53 0.80 3.85 0.75 1.27 1.12 8.25 680 217 720 107

6 1.79 0.60 0.63 1.10 5.00 0.82 1.69 1.80 8.35 821 230 515 139

56 1.62 0.48 0.58 0.88 5.70 0.81 1.82 2.23 10.40 700 245 580 150

15 2.32 0.60 0.53 0.81 4.92 0.89 2.15 2.25 10.60 940 269 590 132

25 1.54 0.50 0.53 0.75 4.60 0.77 2.31 2.34 10.62 955 260 600 82

71 1.40 0.50 0.37 0.64 5.60 0.70 2.47 2.60 10.41 814 216 780 106

21 1.55 0.52 0.50 0.55 4.35 0.89 2.06 2.21 10.20 976 201 520 118

16 2.00 0.80 0.47 1.02 4.40 0.91 2.05 2.55 8.90 899 205 550 140

14 1.38 0.78 0.29 1.14 8.21 0.65 2.00 2.23 8.16 521 218 855 97

17 1.50 0.55 0.43 1.30 4.00 0.60 1.68 2.24 5.61 696 252 830 63

10 0.98 0.34 0.40 0.68 4.90 0.58 1.33 1.81 7.94 670 156 415 154

50 1.70 0.65 0.47 0.86 7.65 0.54 1.86 2.10 8.52 806 213 625 122

21 1.93 0.76 0.45 1.25 8.42 0.55 1.62 2.19 6.12 604 219 650 106

50 1.41 1.39 0.34 1.14 9.40 0.57 1.33 1.26 7.36 733 164 550 114

106 1.40 1.57 0.22 1.25 8.60 0.59 1.30 1.29 6.28 568 129 500 107

127 1.48 1.36 0.24 1.26 10.80 0.48 1.47 1.40 7.00 898 154 480 91

55 2.20 1.28 0.26 1.56 7.10 0.61 1.33 1.25 8.57 905 249 425 125

35 1.80 0.83 0.61 1.87 10.52 0.56 1.51 1.42 10.80 915 154 675 84

100 1.48 0.58 0.53 1.40 7.60 0.58 1.55 1.34 7.52 924 142 640 100

84 1.74 0.63 0.61 1.55 7.90 0.60 1.48 1.31 9.50 969 207 725 84

6 1.80 0.83 0.48 1.56 9.01 0.57 1.64 1.92 9.29 902 159 480 132

53 1.90 0.58 0.63 1.14 7.50 0.67 1.73 2.18 10.20 865 252 880 118

49 2.80 1.31 0.53 2.70 13.00 0.57 1.96 2.25 10.82 764 223 660 182

35 2.60 1.10 0.52 2.29 11.75 0.57 1.78 2.09 11.09 1080 250 620 160

66.5 2.24 1.90 0.36 1.51 5.27 0.97 2.51 2.75 8.79 759.7 240.4 779.3 110.2

1977.3 0.40 0.99 0.02 0.35 4.90 0.06 0.62 0.87 2.76 20104.3 4691.8 102493.8 649.9

44.5 0.63 1.00 0.13 0.59 2.21 0.25 0.78 0.93 1.66 141.8 68.5 320.1 25.5
56 2. DATA ANALYSIS AND CHEMOMETRICS
Acknowledgment
The authors wish to thank Mr. Patrick Guerin for the careful
revision of the manuscript.
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