Analytica Chimica Acta 570 (2006) 283290

Digital image-based titrations

Edvaldo da Nobrega Gaiao, Valdomiro Lacerda Martins, Wellington da Silva Lyra,
Luciano Farias de Almeida, Edvan Cirino da Silva, Mario Cesar Ugulino Araujo
Universidade Federal da Paraba, CCEN, Departamento de Qumica, P. Box: 5093, 58051-970 Joao Pessoa, PB, Brazil
Received 6 October 2005; received in revised form 8 April 2006; accepted 11 April 2006
Available online 29 April 2006

Abstract
The exploitation of digital images obtained from a CCD camera (WebCam) as a novel instrumental detection technique for titration is proposed
for the first time. Named of digital image-based (DIB) titration, it also requires, as a traditional titration (for example, spectrophotometric,
potentiometric, conductimetric), a discontinuity in titration curves where there is an end point, which is associated to the chemical equivalence
condition. The monitored signal in the DIB titration is a RGB-based value that is calculated, for each digital image, by using a proposed procedure
based on the red, green, and blue colour system. The DIB titration was applied to determine HCl and H3 PO4 in aqueous solutions and total
alkalinity in mineral and tap waters. Its results were compared to the spectrophotometric (SPEC) titration and, by applying the paired t-test, no
statistic difference between the results of both methods was verified at the 95% confidence level. Identical standard deviations were obtained by
both titrations in the determinations of HCl and H3 PO4 , with a slightly better precision for DIB titration in the determinations of total alkalinity.
The DIB titration shows to be an efficient and promising tool for quantitative chemical analysis and, as it employs an inexpensive device (WebCam)
as analytical detector, it offers an economically viable alternative to titrations that need instrumental detection.
2006 Elsevier B.V. All rights reserved.
Keywords: Digital images; RGB colour system; DIB titration; Alkalinity; Waters analysis

1. Introduction
Several instrumental techniques have been widely used for
implementing quantitative chemical analysis based on titration
[1]. In order to find an end point, these titrations employ plots that
are built by using the measured signal after the addition of each
increment of titrant. The shape of the titration curves depend
on some factors such as the reaction of titration, the monitored
specie (indicator, titrant, analyte or formed product), as well as
the chosen instrumental technique (spectrophotometry, conductimetry or potentiometry, for instance).
Recent advances in digital image acquisition technology
have offered video cameras (WebCam) based on charge-coupled
devices (CCD), which are capable to capture digital images with
up to 24 bits (16.7 million colours). In fact, by using the RGB
colour system [2,3], the primary colours are combined in different intensities with values varying in the range 0255 (8 bits)
per colour.

Corresponding author. Tel.: +55 83 32167438; fax: +55 83 3216 7437.

E-mail address: laqa@quimica.ufpb.br (M.C.U. Araujo).

0003-2670/$ see front matter 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.aca.2006.04.048

By using a CCD digital camera, the images obtained with

high resolution have been exploited for different purposes, such
as in studies involving multivariate image regression [4], classification of inhomogeneous food matrices [5], spectral image
analysis for measuring ripeness of tomatoes [6], etc. However, the exploitation of digital images for the quantification
of analyte concentrations has not been found in the literature,
with exception of the work recently proposed by Maleki et al
[7].
Maleki et al. [7] employed a digital camera as detection system for simultaneous determination of Al(III) and Fe(III) in
alloys using the chrome azurol S (CAS) as chromogenic reagent.
The RGB values associated to digital images from Al(III)-CAS
and Fe(III)-CAS complexes were used in the construction of an
artificial neural network (ANN) model. It should be also emphasized that due to the complex relationship between RGB values
and analyte concentrations, the ANN modelling was chosen
by Maleki et al. [7] for multivariate calibration because there
is no need to know the exact form of the analytical function
on which the model should be built. On the other hand, such
modelling requires training and testing procedure of the network, which can be considered laborious and time consuming

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E. da Nobrega Gaiao et al. / Analytica Chimica Acta 570 (2006) 283290

especially when optimized parameters for ANN modelling are

aimed.
A titration technique based on digital image exploitation is
proposed in this paper for the first time. Similarly, to spectrophotometric, condutimetric or potentiometric titrations, for
instance, the DIB titrations were always accomplished in order
to make sure the titrant was added in excess, yielding a discontinuity in the titration curve where there is an end point,
which is associated to the chemical equivalence condition.
However, as it can be observed in fundamentals of analytical
chemistry text books, end points are not so precisely determined in original titration curves. This problem has been minimized by using derivative titration curves [8,9]. Initially, two
model titrations (HCl/NaOH and H3 PO4 /NaOH) were studied to evaluate the performance of the DIB titrations. Thereafter, an analytical application involving the determination of
the total alkalinity in mineral and tap waters was carried
out.
2. Experimental
2.1. Reagents, samples, and solutions
All solutions were prepared with freshly distilled-deionised
water and analytical grade chemicals.
About 100.0 mmol l1 NaOH was used for titrations of HCl
and H3 PO4 , whereas for determination of total alkalinity in mineral and tap water samples was employed 10.0 mmol l1 H2 SO4
as titrant. These titrant solutions were standardised employing
conventional methodologies [10,11].
About 0.1% (w/v) phenol red, 0.1% (w/v) bromocresol green
and 0.2% (w/v) methyl orange were used as indicators in the
titrations of HCl, H3 PO4 and total alkalinity in mineral and tap
water samples, respectively.
Mineral water samples were purchased in a local supermarket
and the tap water samples were collected from three points in the
city of Joao Pessoa, Paraba, Brazil. All samples were analyzed
without any further treatment.
2.2. Apparatus
A schematic diagram and a photograph of the system assembled for digital images acquisition is depicted in Fig. 1. A model
CTWCII Creative WebCam (WC) [12], which was positioned in
front of the model QS 137 Hellma flow cell (FC) with inner volume 2.6 ml and optical path 1.0 cm, was used for digital images
acquisition. The coloured solutions were only illuminated by
light from a 5 W Avante fluorescent lamp.
In order to avoid interferences of ambient light, the WebCam
and the flow cell were put into a plastic box with approximately
11 cm 14 cm 9 cm. To provide a uniform illumination and
to reduce glare and specular reflection the internal walls of the
box were covered with a layer of white paper.
The WebCam was connected to the parallel port of a Pentium III 650 MHz microcomputer (PC) and configured to capture 24-bits digital images (16.7 million colours) at a rate of
7.5 images/s with a sensitivity of 15 lx (luminance unit) and a

Fig. 1. Schematic diagram (a) and photograph (b) of the system for digital
images acquisition. B: burette, SB: stirring bar; MS: magnetic stirrer; PP: peristaltic pump; WC: WebCam, SPEC: spectrophotometer; PC: microcomputer;
FC: flow cell and PS: power supply. Arrows indicate directions of pumping. For
further details, see the text.

320 pixels 240 pixels spatial resolution [12]. The images were
captured and stored as jpg compressed files using the software
provided by manufacturer of the WebCam.
A model 8453 Hewlett-Packard (HP) diode array UVvis
spectrophotometer, equipped with a model QS 178.010 Hellma
flow cell (inner volume 100 l and optical path 1.0 cm) was used
for spectrophotometric measurements.
A model 190M Hanna Instruments magnetic stirrer (MS)
driven a stirring bar (SB) inside the Erlenmeyer.
An 8 channel-12 rolls Gilson, peristaltic pump, model Miniplus 3, employing Tygon pumping tubes, and Teflon line
tubes of 1.85 and 0.8 mm inner diameter was used to propel
the solutions.
2.3. The procedure for the RGB-based value calculation
Initially, the user selects, with the computer mouse, the region
of the first titration image that is used for the RGB-based value
calculation. This region must be selected where the light beam
crosses the flow cell (Fig. 2). Right after, the software performs
a scan (column by column) of the pixels on the delimited region,
calculates the RGB-based value of each pixel and then the average of all RGB-based values. This average value is used to build
the DIB titration curves. The software written in Kylix (version
3.0) automatically uses the same coordinates of the delimited

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285

2.4. DIB and SPEC titrations

Fig. 2. Delimited region of a first digital image with n m pixels that is captured
during the DIB titration. x1 , x2 , y1 and y2 are the coordinates of the delimited
region.

The titrations of HCl and H3 PO4 in aqueous solutions and

total alkalinity in tap and mineral waters were carried out using
the WebCam and spectrophotometer (Fig. 1) and standardized
NaOH (for HCl or H3 PO4 ) or H2 SO4 (for the total alkalinity).
H2 SO4 was used as titrant because all water samples analyzed
presented very low hardness (<10.0 mg l1 of CaCO3 ). Aliquots
of 20, 25 or 100 ml (HCl, H3 PO4 or water samples) and three
drops of indicator (phenol red, bromocresol green or methyl
orange), respectively, were transferred to the Erlenmeyer. During the titration, the solution inside Erlenmeyer was always kept
under stirring and a closed-loop pumping between the detectors (WebCam and spectrophotometer) and the Erlenmeyer. The
measurements of RGB-based values and absorbance signals
were always carried out with the stopped flow (peristaltic pump
off) and stored in the microcomputer for posterior treatment.
The absorbance measurements were carried out at 558, 617 and
480 nm for titration of HCl, H3 PO4 and total alkalinity in water
samples, respectively.
3. Results and discussion

region (Fig. 2) to calculate RGB-based values of all images of

the titration. The computer code is available as supplementary
information.
Each sampled image has been a matrix with 320 pixels
240 pixels and the delimited region by the user has been a matrix
with about 40 pixels 50 pixels. The pixel resolution has been
87 87 dots/in. The RGB-based value calculations are based on
the product 2R .2G .2B , where R, G and B are the red, green and
blue colour components, respectively. These components may
assume integer values in a range from 0 to 7, reaching up to
16,777,216 colours.

3.1. Digital images and visible spectra of the HCl, H3 PO4

and total alkalinity titrations
To simplify and to make clear the discussions bellow, only
10 digital images and its corresponding visible spectra of HCl,
H3 PO4 and total alkalinity titrations are shown in Figs. 3 and 4,
respectively.
As can be seen in Figs. 3a and 4a, the colours and the corresponding spectral profiles of the images 16 and 710 are
different. Thus, it is feasible to state that between the images 6

Fig. 3. Sequence of the digital images captured during the titration of HCl/NaOH (a), H3 PO4 /NaOH (b) and total alkalinity in a mineral water sample (c) (for
interpretation of the references to colour in this figure legend, the reader is referred to the web version of the article).

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E. da Nobrega Gaiao et al. / Analytica Chimica Acta 570 (2006) 283290

colours; the yellow and magenta are secondary colours (resulting

from the mixture of primary colours green + red and blue + red,
respectively); and the orange is a tertiary colour (resulting from
the mixture of the red + yellow) [2,3].
In H3 PO4 /NaOH titration, the images 15 in Fig. 3b are yellow because the indicator in its acid form absorbs predominantly
blue radiation (see the spectra 15 in Fig. 4b), while the images
610 in Fig. 3b present the cyan secondary colour (mixture of
green + blue) due to the absorption of red light by the indicator
in its basic form (see the spectra 610 in Fig. 4b). As the spectra
and the images 5 and 6 are different, it can be inferred that there
is an end point between them and that it was originated due to
the addition of titrant in excess and the change of the indicator
from its acid to basic form.
A method recommended by the American Public Health
Association [11] to determine total alkalinity of water samples with very low hardness is acid-base titration using the
H2 SO4 titrant, the methyl orange indicator and the visual detection. In general, the analyst may have difficulty to determine
the end point with good precision when this method is used.
This fact is corroborated by the visual examination of the
images 110 (Fig. 3c). As this titration was surely accomplished with the addition of titrant excess, there must be an
end point among the images 110. However, it is not visually located because a colour change is not easily realized
among these images. On the other hand, taking into account
that the spectra 15 (Fig. 4c) are different of the spectra 610,
it may be inferred that there is an end point between the images
5 and 6.
3.2. DIB and SPEC titration curves
To a more accurate evaluation of the variations observed both
in the RGB-based and in the absorbance values and, consequently, to a better comparison of the DIB and SPEC titrations
(Figs. 37), a collection of points with a greater number of added
titrant volumes, especially close to the end point, are shown in
Table 1. The following equations are used by the data treatment
software to build the second-derivative curves.
Equations for the calculation of the first derivatives:
Fig. 4. Absorption spectra in the visible region obtained during the titration of
HCl/NaOH (a) H3 PO4 /NaOH (b) and total alkalinity in a mineral water sample
(c).

and 7 there is an end point and that such fact happens when the
titrant is added in excess and the indicator changes from acid
to basic form. It is worth pointing out that the images 16 are
of orange colour because the indicator in its acid form absorbs
the blue and green-blue lights (see the spectra 16 in Fig. 4a),
while the images 710 present the magenta colour due to the
yellow-green light absorption (see the spectra 710 in Fig. 4a)
by the indicator in its basic form.
In order to understand the complementary colours of the
images obtained during DIB titrations (Fig. 3), it is worth
reminding that the red, green and blue are considered primary

R
R(i+1) Ri
=

V
V(i+1) Vi

and

Vaverage =

V(i+1) + Vi
2

Equations for the calculation of the second derivatives:

2 R

((
R)/(
V ))
=

Vaverage

Vaverage
=
V average =

((
R)/(
V ))i+1 ((
R)/(
V ))i

(Vaverage )i+1 (Vaverage )i

and

(Vaverage )i+1 + (Vaverage )i

2

where R is the analytical response and V is the added titrant

volume. i = 0, 1, 2, . . ., n, and n is the number of titration

E. da Nobrega Gaiao et al. / Analytica Chimica Acta 570 (2006) 283290

points that were used in derivative calculations. The secondderivative curves were built by using V average .
Initially, it is worth arguing that an abrupt variation of the
RGB-based value does not necessarily imply in a association with a region of the DIB titration curve where the end
point may be located. For example, in the HCl/NaOH titration, the abrupt variation of the RGB-based value between the

287

points 67 (Fig. 5a) is associated with a region where an end

point can be found. This fact is corroborated by the difference
of colour (Fig. 3a) and of the corresponding spectral profile
(Fig. 4a) between these images. In addition, an abrupt variation
of absorbance between the points 6 and 7 is also observed in
the SPEC titration curve (Fig. 5c). On the other hand, according to the SPEC titration curve (Fig. 6c) and the differences of
colour (Fig. 3b) and of corresponding spectral profile (Fig. 4b),

Table 1
A collection of points of three DIB and SPEC titrations with a greater number of added titrant volumes close to the end point

2 absorbance

Vaverage

2 RGBbased value

Vaverage

Volume (ml)

Absorbance

RGB-based value (107 )

HCl/NaOH titration
0
4.0
8.0
12.0
16.0
19.4
19.6
19.7
19.8
19.9
20.0
20.1
20.2
22.0
24.0
26.0

0.0053
0.0003
0.0012
0.0031
0.0011
0.0007
0.0028
0.0005
0.3658
0.3503
0.3914
0.3875
0.3901
0.3554
0.3539
0.3529

1.4994
1.5308
1.5842
1.6294
1.7031
1.7449
1.7531
1.8695
1.8753
1.5988
1.5984
1.5971
1.5964
1.5869
1.5816
1.5646

0.0045
0.0023
0.0014
0.0008
0.0700
0.4364
36.7572
38.0781
5.6680
4.5110
2.4580
1.7090
0.0002
0.0009

0.0246
0.0164
0.0148
0.0008
0.2505
5.8260
11.0670
28.2220
27.6030
0.0820
0.0550
0.0390
0.0067
0.0004

H3 PO4 /NaOH titration

0
2.0
3.0
5.0
5.8
5.9
6.0
6.1
6.2
6.3
6.4
6.6
7.0
8.0
10.0

0.0023
0.0061
0.0079
0.0150
0.0719
0.2384
0.4049
0.8221
0.8987
0.9753
0.9875
0.9966
1.0893
1.0825
1.0457

1.5382
1.5991
1.6028
1.6234
1.6308
1.6327
1.6346
1.6272
1.5632
1.4991
1.5314
1.5637
1.8600
1.8778
1.9174

0.0016
0.0005
0.0430
15.7190
20.1200
25.0710
34.0630
16.0100
6.4310
0.3170
0.3150
0.2563
0.0095

0.0178
0.0075
0.0099
0.0010
0.0010
0.0926
0.5668
0.0006
0.9640
0.9219
0.8798
0.3714
0.0093

2 absorbance

Vaverage

Alkalinity titration
0
2.0
4.0
6.0
7.8
7.9
8.0
8.1
8.2
8.3
8.4
8.5
8.6
8.8
9.0

0.1435
0.1438
0.1483
0.1470
0.1529
0.1613
0.1691
0.1807
0.1850
0.1967
0.2049
0.2128
0.2314
0.2369
0.2289

8.7500
8.6400
8.6900
8.7100
8.7600
8.9600
9.0600
8.8500
8.7600
8.6600
8.5500
8.4400
8.3800
8.2300
8.1700

0.0130
0.0017
0.0015
0.0004
0.0938
0.3220
0.0650
0.3820
0.7340
0.7460
0.3510
0.0310
0.1072

2 RGBbased value

Vaverage

0.0600
0.1400
0.0500
0.0100
1.2000
26.0000
10.0000
8.8000
0.7000
0.6000
0.5000
0.2000
0.1000

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Fig. 5. Zero-order and second-derivative curves of the HCl/NaOH model titration with WebCam (a and b) and spectrophotometer (c and d) detectors.

the end point of the H3 PO4 /NaOH titration is located between

the points 5 and 6. However, the variation of the RGB-based
value is much greater between the points 7 and 8 (Fig. 6a).
This abrupt change of RGB-based value between these points
occurs due to the abrupt increase of the contribution of the blue
emerging component (B) regarding to the red (R) and green
(G) components, resulting from the increase (in the red spectral
region) and decrease (in the blue and green region) of the absorption by the indicator in its basic form. In contrast, the change
of RGB-based value between points 5 and 6 occurs due to the
decrease of the contribution of R component, regarding to G and
B components.

The volume of titrant associated to the end point (Vep ) has

always been determined by the data treatment software adopting the zero crossing criteria in the second-derivative curves.
Independent of the particular titration considered, the software
uses all points of titration (points 2030) to build such curves.
About 10 points with increments of 0.1 ml of titrant have always
been measured around the end point. As in a traditional SPEC
titration, the increment of 0.1 ml of titrant in DIB titration has
been started when the RGB-based value (displayed on computer
screen by the software) presented a significant change.
It can be noted that the zero crossing, indicated by an arrow in
the second-derivative curves of HCl, H3 PO4 and total alkalinity

Fig. 6. Zero-order and second-derivative curves of the H3 PO4 /NaOH model titration with WebCam (a and b) and spectrophotometer (c and d) detectors.

E. da Nobrega Gaiao et al. / Analytica Chimica Acta 570 (2006) 283290

289

Fig. 7. Zero-order and second-derivative curves of the total alkalinity/H2 SO4 titration of a mineral water sample with WebCam (a and b) and spectrophotometer (c
and d) detectors.

(Figs. 5b and d, 6b and d, 7b and d), provides Vep values very

close for both DIB and SPEC titrations.
Table 2 displays the results of HCl and H3 PO4 in aqueous
solutions and of total alkalinity in mineral and tap waters as
determined by the DIB and SPEC titration. They show a good
agreement between the obtained values by both methods. Actually, no statistic difference at the 95% confidence level has been
verified between the results by applying the paired t-test.
Table 2
Results of HCl (in mmol l1 ) and H3 PO4 (in mmol l1 ) in aqueous solutions and
of total alkalinity (in mg CaCO3 l1 ) in mineral and tap waters as determined
by the proposed DIB and SPEC titration
Samples

Titrations
DIB

4. Conclusions
SPEC

(1) HCl
(2) HCl
(3) HCl
(4) HCl
(5) HCl
(6) HCl

98.8
50.1
72.6
147.7
198.8
258.7

0.1
0.1
0.1
0.1
0.1
0.1

99.1
50.0
72.6
147.6
199.0
258.5

0.1
0.1
0.1
0.1
0.1
0.1

(1) H3 PO4
(2) H3 PO4
(3) H3 PO4
(4) H3 PO4
(5) H3 PO4
(6) H3 PO4

24.8
64.6
99.8
127.1
175.8
208.6

0.1
0.1
0.1
0.2
0.1
0.1

24.7
64.5
99.8
127.1
175.8
208.7

0.1
0.1
0.1
0.2
0.1
0.1

(1) Mineral water

(2) Mineral water
(3) Mineral water
(1) Tap water
(2) Tap water
(3) Tap water

In terms of precision, the obtained values by the proposed

and SPEC methods are identical for the HCl and H3 PO4 determinations and both values are better than the total alkalinity
determinations. This finding may be ascribed to a clearer identification and determination of the end point for the HCl and
H3 PO4 titrations than total alkalinity titrations. On the other
hand, the slightly better precision of the DIB titration regarding
to SPEC titration may be credited to the multivariate advantage [13]. In fact, the monitored signal in the DIB titration
is of multivariate nature (variation of three RGB components),
while the SPEC titration is univariate (variation of absorbance
in 480 nm).

82.4 0.4
82.7 0.2
16.0 0.1

82.5 0.5
82.7 0.3
15.9 0.2

144.8 0.5
6.0 0.1
4.0 0.1

144.7 0.6
6.0 0.2
4.0 0.2

The feasibility of the exploitation of digital images obtained

from a WebCam as novel instrumental detection technique for
titration was demonstrated. To attain such a goal, the proposed
technique requires a variation of the RGB image components
from the analytical system and a discontinuity in titration curves
where there is an end point that is associated to the chemical
equivalence condition. The DIB titration can be considered a
promising analytical tool for accomplishing quantitative chemical analysis, as well as it offers an economically viable alternative to titrations with a difficult visualization of the end point.
By exploring the spatial-resolution-related advantages inherent to digital images, it is expected to increase considerably
sampling throughput carrying out several DIB titrations at a
same time, by acquiring contemporarily in a single image information on more analytical samples. The DIB titration speed can
also be increased by using the Flow-Batch automatic system
[14,15].

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E. da Nobrega Gaiao et al. / Analytica Chimica Acta 570 (2006) 283290

It also aims at investigating the use of higher resolution CCD

cameras and/or other colour system, such as HIS (hue, saturation and intensity system), which has already provided quite
satisfactory results in colour image analysis [5], to improve precision on titrations with difficult visualization (as seen in the
determinations of total alkalinity).
In order to build parsimonious and robust models of multivariate analysis the use of variable and sample selection algorithms [1618] or a wavelet transform-based strategy [19] in
RGB space will also be investigated, especially when inhomogeneous matrices are involved as exploited by Antonelli et al.
[5]. It is worth noting that the proposed approach was applied in
a homogeneous analytical system (titration of samples in solution).
Acknowledgement
E.N. Gaiao, V.L. Martins, L.F. Almeida, and M.C.U. Araujo
thank the Brazilian agency CNPq for scholarship.
Appendix A. Supplementary data
Supplementary data associated with this article can be found,
in the online version, at doi:10.1016/j.aca.2006.04.048.
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