Analytica Chimica Acta 396 (1999) 9197

A flow-batch titrator exploiting a one-dimensional optimisation

algorithm for end point search
Ricardo S. Honorato a , Mrio Csar U. Arajo , Ricardo A.C. Lima b , Elias A.G. Zagatto c ,
Rui A.S. Lapa d , Jos L.F. Costa Lima d
b

a Departamento de Qumica Fundamental, Universidade Federal de Pernambuco, Recife PE, Brazil

Departamento de Qumica, Universidade Federal da Paraba, P.O. Box 5093, Zip code 58051-970 Joo Pessoa PB, Brazil
c Centro de Energia Nuclear na Agricultura, Universidade de So Paulo, Piracicaba SP, Brazil
d Faculdade de Farmcia, Universidade do Porto, Porto, Portugal

Received 8 January 1999 ; received in revised form 7 April 1999 ; accepted 21 April 1999

Abstract
A flow-batch hybrid system is proposed as a novel approach to automated titration. Sampling and signal processing are
done as in usual flow systems, whereas chemical reactions take place in a reaction chamber similar to those found in batch
systems. End point is found by means of the Fibonacci method, adapted to act as a one-dimensional optimisation algorithm for
fast titration. A model system involving the spectrophotometric NaOH/HCl titration was initially studied, and the feasibility
of the approach was further demonstrated in the titration of white wines with NaOH using m-cresol purple as indicator. About
20 samples are run per hour and ca. 0.6 ml of sample and titrant are consumed per determination. Within the 5.27.3 g l1
acidity (expressed as tartaric acid) range, relative standard deviations of results were estimated as about 0.7% after six-fold
processing of typical wine samples, and results are in agreement with those obtained by a classical potentiometric titration,
with relative errors <0.8%. 1999 Elsevier Science B.V. All rights reserved.
Keywords: Flow-batch titrator; Fibonacci method; Wines; Flow analysis; Spectrophotometry

1. Introduction
Manually performed titrations are usually laborious
and time consuming. Considering its relevance, automated procedures carried out in batch or flow systems
have been proposed for large-scale analysis.
Batchwise automated titrations are known for at
least 80 years [1] and still subject to studies [2]. They
involve a titration vessel for each sample, addition of
Corresponding
author.
Tel.:
+55-83-216-7438;
+55-83-216-7437
E-mail address: laqa@quimica.ufpb.br (M.C.U. Araujo)

fax:

titrant by means of either an automatic burette or a syringe, and stirring of the reaction medium. Although
simplification of the analytical procedure is achieved,
batch titrators are usually characterised by a low sample throughput, a high consumption of sample and
titrant, and a high implementation cost.
These problems have been circumvented after the
pioneer work of Blaedel and Laessig [3]that led to
the development of ingenious approaches in the field
of flow titrations. Initially, unsegmented flows were
exploited [311] and at least one point with titrant and
analyte in stoichiometry was considered. This point
was usually attained by maintaining the flow rate of

0003-2670/99/$ see front matter 1999 Elsevier Science B.V. All rights reserved.
PII: S 0 0 0 3 - 2 6 7 0 ( 9 9 ) 0 0 3 6 6 - 9

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R.S. Honorato et al. / Analytica Chimica Acta 396 (1999) 9197

one solution (sample or titrant) and linearly increasing

the other flow rate [3,5,79], or by maintaining the
sample flow rate and exploiting a linear concentration
gradient of titrant generated into it. For this purpose,
an external chamber [4] or a coulometric device [6]
were used. Alternatively, mechanically driven cams
linked to syringe pumps [10,11], pulsed flows [12] or
computer-controlled valves [13] have been used for
attaining variations in sample and titrant flow rates.
Development of flow-injection titration systems
[14]started in the seventies. A concentration gradient
was established into the titrant stream as a consequence of the sample injection into this stream. Therefore, two sites with titrantanalyte stoichiometry were
obtained. The time interval 1t elapsed between passage of these sites through the detector was ideally
proportional to the logarithm of analyte concentration. A calibration step requiring several standard
solutions was needed to achieve the (1t versus ln C)
relationship. For sensitivity enhancement, a reverse
flow-injection system was further proposed [15].
An alternative flow-injection procedure, single-point
titration [16], exploited the linear relationship between recorded peak height and acid (or base) concentration when a potentiometric sensor such as the
glass electrode was employed. This feature has been
also observed in acid/base spectrophotometric titrations when the indicators are properly elected [17].
Recently, a sequential injection system for spectrophotometric titrations was proposed [18].
The above mentioned flow systems required analytical curves based on standard solutions. Difficulties in
preparing these solutions, such as in the determination
of hardness, oxidability or acidity/alkalinity in natural
waters [19] might limit the applicability of these systems. Results may be dependent on more than one analyte concentration, and different analytes may present
different reaction stoichiometries towards the titrant,
so that preparation of the standard solutions may become cumbersome.
To solve this drawback, flow-injection titration
procedures requiring only one standard solution (the
titrant) have been proposed [20,21]. In contrast with
earlier systems, the titrant concentration is directly
used for calculating the analyte concentration without the need for an analytical curve. The first system
[20] was based on the calibration of the concentration gradient related to the injected species (sample

92

or titrant), and the other [21] exploited the binary

search concept, where variations in sample and titrant
volumetric fractions were considered.
Titration procedures using the titrant as the only
standard solution could be also accomplished by
adding this solution together with the sample aliquot
and eventually the indicator solution to a reaction
chamber analogous to those inherent to batch analysis.
Although some favourable analytical characteristics
were reported [22,23], sampling throughput was low
(612 h1 ) even for sample lots with low variability in analyte concentration. In addition, the systems
were somewhat complex, requiring two pumps.
The aim of the present work was then to develop a
novel automated titrator, which operates with a single
standard solution (titrant) and does not require analytical curves. Sampling and measurements are carried
out in a continuous way, and the involved reactions
occur inside a reaction chamber, as in batch titrations.
The approach is then a flow-batch hybrid system combining the intrinsic advantages of flow systems such
as high sampling rate, low sample and reagent consumption, low cost, easy of automation, etc., with the
wide application range inherent to batch systems. End
point is found by applying the Fibonacci method [24]
adapted for fast titrations.

2. The Fibonacci method

This one-dimensional optimisation method [24]
always minimises a given interval where the optimised value of the considered parameter is located.
The method starts by defining limits for this interval, the so-called initial interval of uncertainty, L[1].
It undergoes successive reductions and converges to
a very narrow one, the final interval of uncertainty,
L[f]. The method permits a given ratio between L[1]
and L[f] to be quickly attained, and its application
results in an optimised range instead of a specific
value. In the present titration procedure, the variable
to be optimised is the titrant volume and its value at
the equivalence point is aimed.
hi [1]) for
The lower and higher limits (Vtitlo [1] andVtit
L[1] are selected according to the expected sample
concentrations in the lot to be assayed. It is good practice to consider a suitable safety margin. When the
variability in analyte concentration is unknown, Vtitlo [1]

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R.S. Honorato et al. / Analytica Chimica Acta 396 (1999) 9197

Table 1
L[1]/L[f] ratio for different n[1] values
N[1]

L[1]/L[f]

0
1
2
3
4
5
6
7
8
9
10

0
1
1
2
3
5
8
13
21
34
55

n[1]

L[1]/L[f]

11
12
13
14
15
16
17
18
19
20
21

89
144
233
377
610
987
1597
2584
4181
6765
10946

should be zero and Vtithi [1] equal to Vtot , a constant

value equivalent to the sum of sample plus titrant volumes added to the reaction chamber. End point is then
related to any value between zero and Vtot , this later
value relating to the most concentrated sample.
In the present titration procedure, the valve timing
courses define the volumes added to the reaction chamber. So, the Fibonacci method was adapted in terms
of time rather than volume and t lo [0], t hi [1] and ttot
tit

tit

lo
hi
were used instead of and V tit
[1], V tit
[1] and Vtot .
For any sampling step (sample, indicator and titrant
addition to the reaction chamber), the sample time,
ts [m], is equal to (ttot ttit [m], the indicator time, ti , is
constant and the titrant time, ttit [m]), is expressed as:
lo
ttit [m] = ttit
[m] +

F [n[m]1] hi
lo
ttit [m] t tit
[m]
F [n[m]]

(1)

or
max
ttit [m] = ttit
[m]
tit

F [n[m] 1]
F [n[m]]

[m] ttit [m]

min

(2)

t max
where: m is the index of the sampling step; F[n[m]] and
(F[n[m] 1]) are integers of the Fibonacci sequence,
and n[m] is the index of the Fibonacci number which
undergoes continuous lowering as the convergence
proceeds. The F[0] = 0, F[1] = 1, F[2] = 1, F[3] = 2,
F[4] = 3, F[5] = 5, F[6] = 8,... Fibonacci sequence follows equation: F[n + 2] = F[n + 1] + F[n], (see also Table 1).
For the first sampling step (m = 1), Eq. (1) or Eq.
(2) can be considered. Integer n[1] is related to the
number of sampling steps and expressed as nhi + 2; or

93

alternatively, as 2nlo + 1 (odd n[1]) or 2nlo + 2 (even

n[1]). Symbols nhi and nlo correspond to the highest and lowest numbers of sampling steps required to
achieve a given ratio between the initial and final intervals of uncertainty. Table 1 shows the L[1]/L[f] ratios
for different n[1] values from where one can perceive
that L[1]/L[f] = F[n[1]].
The n[1] value is predefined by the analyst as a compromise between aimed sampling throughput (number
of sampling steps) and result precision (L[1]/L[f] ratio). For e.g. n[1] = 10, a complete titration requires
48 sampling steps, and L[f] is 55 times lower than
L[1]. The number of sampling steps is related to the
analyte concentration. It should be emphasised that for
sample lots with low variability in analyte concentration, L[1] can be reduced, lowering the final interval of
uncertainty, thus improving the precision of the result.
After the second sampling step, either Eq. (1) or
Eq. (2) can be used to calculate ttit [m], depending on
whether the signal related to the previous measurement
indicates a titrant or an analyte excess. Therefore, the
program must recognise the species in excess through
the measured signal. For this task, a reference signal
is needed. In spectrophotometric acid/base titrations,
it corresponds to the indicator colour change interval.
Regarding ttit determination for the mthsampling
step, if titrant excess is observed, ttit [m] should be
lower than ttit [m 1], related to previous sampling
step. It is calculated by using Eq. (1) with t lo
tit[m] and
hi
ttit
[m] values used as ttitlo[m 1] and ttit [m 1]. If
hi [m 1] t [m 1]) < (t [m 1] t lo[m 1]),
(t tit
tit
tit
tit
the n[m] integer is used as (n[m 1] 1), otherwise
n[m] is used as (n[m 1] 2). On the other hand, a
higher ttit [m] value relative to the previous sampling
step is required when analyte excess is noted. In this
situation, ttit [m] is determined by using Eq. (2) with
t lo

hi

hi

tit [m] and ttit [m] used as ttit [m 1] and ttit [m 1]. If
hi [m 1] t [m 1]) > (t [m 1] t lo [m 1]),
(ttit
tit
tit
tit

the n[m] integer is used as (n[m 1] 1), otherwise

n[m] is used as (n[m 1] 2).
Search of the equivalence point ends when n[m] < 3
and the final interval of uncertainty in terms of concentration, still containing the analyte concentration
is:
"
#
hi
h
i
ttitlo [f ]
ttit
[f ]
lo
hi
Cs , Cs t
C R, t t hi [f ] Ctit R (3)
tot ttit
lo [f ] tit
tot tit

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R.S. Honorato et al. / Analytica Chimica Acta 396 (1999) 9197

where C slo and C shi are the lower and higher limits
of L[f], in terms of concentration, Ctit is the titrant
lo [f ] and t hi [f ] are the limits of the L[f]
concentration, t tit
tit
and R is the titrant-to-sample flow rate ratio.
When the measured signal falls within the reference
signal range, i.e., near the equivalence point, the titration is completed earlier. Eq. (3) is then applied with
lo
hi
ttit
[f ] = ttit
[f ] = ttit [f], yielding a specific value instead
of a final interval of uncertainty.

94

An IBM/PC compatible microcomputer furnished

with a home made interface card was used to control
the system and for data treatment. The solenoid valves
and the magnetic stirrer were controlled through an
electronic actuator, that provided the required increase
in power (potential and/or current) for signal sent by
the
microcomputer.

3.3. Procedure
3. Experimental
3.1. Solutions
All solutions were prepared with freshly distilleddeionized water and analytical reagent grade
chemicals.
For NaOH and wine analysis 24.2 mmol l1 HCl or
83.4 mmol l1 NaOH standardised [25] solutions were
used. NaOH solutions were daily prepared.
Wine samples were purchased in a local supermarket and analysed without any further treatment.
3.2. Apparatus
The diagram of the flow-batch hybrid system is
shown in Fig. 1. A model B342II Micronal spectrophotometer furnished with a tubular (inner volume 100 ml,
optical path 2 mm) flow cell [26] was employed. A debubbler in the analytical path was not used because the
flow cell did not retain air bubbles. A model 78002-50
Ismatec peristaltic pump was used with Tygon (i.d.
1.02 mm) pumping tubes. The manifold was build-up
with 0.8 mm i.d. Teflon tubing.
The reaction chamber, CR, was a 0.4 ml Teflon
cylinder with an inner diameter of 7 mm. A 5 mm
2 mm magnetic bar was used inside CR to improve
mixing conditions of the added solutions. In order to
avoid electricmagnetic fields altering the valves operation, it was driven by a homemade low-power magnetic stirrer.
Four Cole Parmer three-way solenoid valves, Vs ,
Vt , Vw , Vi , were used to direct the sample, titrant,
water and indicator solutions. Another valve, Vd , was
added to select the stream flowing through the flow
cell.

End point search required successive variations in

the sample and titrant volumes added to the reaction
chamber, CR, which were proportional to the valves
switching time. Flow rates were tested within 1.0 and
5.0 ml min1 .
Initially, CR was washed by adding and removing
water from it. This was accomplished by successively
ON/OFF switching of Vw and Vd valves in an opposite
way. When Vd was switched ON, the solution leaving
CR was aspirated through the flow cell towards waste;
otherwise a water stream was aspirated through the
flow cell. During the washing step, no electric pulse
was sent to Vt , Vs or Vi valves, which were then
maintained OFF. The corresponding solutions were
allowed to recycle.
After the washing step, Vt , Vs and Vi valves were
switched ON and sample, titrant and indicator solutions were pumped towards CR during specific time
intervals. A short (0.5 s) pre-defined time interval was
further required to improve conditions for mixing and
reaction completeness. Thereafter, Vd valve was operated to direct the processed sample towards the detector. In the present procedure, 3200 measurements
could be performed per second and the analytical signal corresponded to the averaged signal related to
the central portion of the processed sample. Measurements related to the front and tailing portion of the
solution aspirated from CR were however, not considered in view of their instability (airliquid interface). After measurement, the cycle was repeated for
the next sample/titrant volumetric ratio defined by the
Fibonacci algorithm. Titration ended when L[f] was
obtained.
There are always differences in flow rates among
pumping tubes with the same nominal flow rate. Tube
ageing may also lead to variations in flow rate. In order
to compensate for these experimental variations, the

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R.S. Honorato et al. / Analytica Chimica Acta 396 (1999) 9197

95

Fig. 1. Flow diagram of the proposed flow-batch hybrid system. S, T, W, I represent sample, titrant, water, indicator solutions, respectively.
V: valves with alternative position specified by dotted lines; CR: reaction chamber with a stirring bar; D: detector; l1 = 3 cm; l2 = 10 cm,
arrows: sites where pumping is applied.

R-value (Eq. (3)) should be periodically determined.

This was accomplished by pumping a coloured solution and water through the sample and titrant channels
during a given time interval. The solution leaving RC
was aspirated through the detector and an absorbance
value As was obtained. Provided that a linear relationship between absorbance and dye concentration
holded,
As =

qtit
Vtit /t
Vtit
Atit
=
=
=
qs
Vam /t
Vam
As

Initially, the model system was studied by using

NaOH titration by HCl. A suitable concentration
for the bromothymol blue indicator (6.0 < pH < 7.6,
yellowblue transition) was found to be 40 mg l1
and the wavelength was set as 615 nm. It was realised
that pumping rates (1.04.0 ml min1 ) were not critical. In this way, the sample, titrant and indicator
1

Vs
Adye
V

flow rates were selected as 1.6 ml min and the wa

ter stream as 3.4 ml min1 . In order to speed up the
procedure, the aspiration rate through the flow cell
was chosen as 3.4 ml min1 . In this situation, R (Eq.
(3)) was periodically determined and only slight variations, usually <1%, were observed during extended
(8 h) operation periods. This robustness comes from
the batch fashion of the approach.
The volume of the reaction chamber shoul
modate the
added solutions and to provide suitable conditions

as low as to
possible,
acc
yet enough om

R =

4. Results and discussion

(4)

In the present procedure, 1.0 mmol l1 KMnO4 was

used as the dye solution and the wavelength was set
as 525 nm. Linearity between absorbance and permanganate concentration was then observed.

for mixing and reaction development. For smaller

chamber volumes, however, ttot (Eq. (3)) should be
reduced, impairing the measurement precision. Transmission lines between CR and flow cell were kept
as short as possible (10 cm) in order to speed up the
procedure.
Results obtained for synthetic NaOH solutions are
shown in Table 2. For comparative purposes, results
based on a potentiometric titration are also shown.
Analysis of these data reveals the reliability of results
obtained with the proposed system. The number of

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R.S. Honorato et al. / Analytica Chimica Acta 396 (1999) 9197

Table 2
Alkalinity of sodium hydroxide solutions as determined by the
model system (Flow-batch) and by a conventional potentiometric
titration (Reference)a
Sample
1
2
3
4
5
6

Reference
5.03
9.84
20.0
30.2
39.6
50.0

Flow-batchb
4.835.05
9.579.82
19.519.8
30.130.5
40.040.4
49.550.1

96

Table 3
White wine acidities as determined by the proposed method
(Flow-batch) and by a conventional potentiometric titration
(Reference)a

Sampling steps

Sample

Reference

Flow-batchb

Sampling steps

9
7
6
6
7
7

1
2
3
4
5
6
7

6.32
7.25
5.29
5.76
5.29
7.04
5.82

6.326.39
7.187.25
5.225.28
5.785.85
5.285.35
6.987.06
5.865.92

5
5
5
6
6
5
4

Results are in mmol l1 based on six replications.

Relative standard deviations of results estimated as about 0.5%
(N = 6).
a

Fig. 2. Experimental titration curve. ttit [m] is the titrant time for
hi
each sampling steps (m = 1, 2, 3, 4, 5); t lo
tit [f ] and t tit[f ] are the
lower and higher limits of the final interval of the uncertainty.

most frequently required sampling steps is also presented. Only 69 steps are needed even for sample lots
with variability in acidity as high as one decade.
Thereafter, the system was applied to the determination of total acidity in white wines, with a
83.4 mmol l1 NaOH solution used as titrant. Wine
acidity is related to the content of tartaric acid [11]
and is usually determined by potentiometric titration up to pH 8.3. For a spectrophotometric titration,
m-cresol purple (7.4 < pH < 9.0 yellowpurple transition) is usually used as indicator. In the present
work, 8.0 mg l1 concentration was selected for the
indicator and wavelength was adjusted to 578 nm. An
experimental titration curve is shown in Fig. 2 and
the results for seven wine samples are presented in
Table 3.
The proposed system results in a final interval of
uncertainty instead of a specific value. Both for syn-

Results in g l1 tartaric acid [11] based on six replications.

Relative standard deviations of results were estimated as about
0.7% (N = 6).
a

thetic samples and for wines, results were in good

agreement with the expected values. For NaOH titrations, relative standard deviations of results based on
six replications were estimated as about 0.5%. The
mean deviations were always <1.0%. For wines, these
figures were 0.7% and <0.8%, respectively.
For NaOH titrations, L[1] was determined based
on the 4.060.0 mmol l1 concentration range and
n[1] was equal to 11; seven sampling steps were
usually required. For wines, L[1] was based on the
25.055.0 mmol l1 range. As L[1] for wines were
percentile lower in relation to the NaOH samples, a
lower value (8) was used for n[1]. As a consequence,
sampling throughput was increased and consumption of sample and titrant was reduced. Usually, five
sampling steps were required.
With the proposed flow-batch hybrid system, a complete titration of a wine sample requires about 2.4 min
and typical sample and titrant consumption are about
0.6 ml. This means 25 samples run per hour.
5. Conclusions
The proposed system is very stable: after several
months of operation, all system components are in
excellent working conditions. Reproducible selection
of the added aliquots are always found for ON/OFF
valve intervals>1 s.
Concerning sampling throughput and precision of
results, the system is very versatile, as the analyst selects the n[1] index as a compromise between these
parameters. The R-value is efficiently determined, being
independent
on
the
pumping
time.

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R.S. Honorato et al. / Analytica Chimica Acta 396 (1999) 9197

The system is particularly attractive for analysis

of samples where use of standard solutions is cumbersome, such as for acidity determination in wines.
However, application of the proposed system to red
wine analysis was hindered by variations of sample
absorbance with pH. This led to variations in the measured signal near the equivalence point, affecting the
comparison with the reference signal, thus impairing the application of Fibonacci algorithm. This problem can be overcome by the suitable choice of wavelength/indicator combination. Studies focusing on this
point are presently in progress.

Acknowledgements
This work was partially supported by a bi-national
project involving CNPq (Brazil) and JNICT (Portugal)
agencies. R.S.H., M.C.U.A., R.A.C.L and E.A.G.Z.
grants from CNPq are also appreciated.

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