Analytica Chimica Acta 470 (2002) 277288

pH-stat techniques in titrimetric analysis

IV. pH-stat monitoring of chelatometric titrations
Carlo Macc a, , Lidia Sold a , Mirella Zancato b
a Department of Inorganic, Metallorganic and Analytical Chemistry, University of Padua, Via Marzolo 1, I-35131 Padova, Italy
b Department of Pharmaceutical Sciences, University of Padua, Via Marzolo 5, I-35131 Padova, Italy

Received 11 April 2002; accepted 18 July 2002

Abstract
The theory of pH-stat chelatometric titrations recently developed [Anal. Chim. Acta 456 (2002) 313] is experimentally
substantiated here. The titrations of four representative doubly charged cations having different behaviour (copper, zinc,
calcium, magnesium) are taken as examples. Copper is titratable between pH 3 and 5, zinc between 3 and 6, calcium
between 6 and 10, and magnesium between 7 and 10. The shapes of the titration plots agree well with the theory, accounting
for simultaneous equilibria involving proton exchange. The technique yields accurate and precise results, which compare
favourably with those of other instrumental techniques, in particular photometric titrations.
2002 Elsevier Science B.V. All rights reserved.
Keywords: Titrimetric analysis; Chelatometric titrations; pH-stat titrations; Calcium(II); Copper(II); Magnesium(II); Zinc(II)

1. Introduction studies of reaction rates or of enzyme kinetics [2,3].

Automatic pH-metric titrators frequently include
the pH-stat or Stat operating function. In this mode,
the titrator measures the pH of a test solution and
simultaneously counteracts any spontaneous or in-
duced variation (either an increase or a decrease) of
this quantity with respect to a pre-selected operating
control value by adding the appropriate amount of a
suitable reagent (a strong acid or a strong base, re-
spectively). The pH-stat technique [1] is widely used
in the chemical and biochemical laboratory when it is
necessary to keep under strict control that most impor-
tant experimental parameter for instance in the course
of organic reactions or of biological processes, and in
Corresponding author. Tel.: +39-0498275205;
fax: +39-0498275161.
E-mail address: cmacca@chin.unipd.it (C. Macca).
Moreover, the measurement of the reactants delivery
rate provides a direct means for determining reaction
rates and performing kinetic analyses [4,5]. In con-
trast, the use of the pH-stat technique for monitoring
volumetric titrations has been very seldom investi-
gated, and is quite unknown in current practice in the
analytical laboratory. This paper is part of a series in-
tended to study in more depth the theoretical and prac-
tical aspects of this technique in volumetric analysis.
The theory and applications of pH-stat monitoring
of acidbase titrations [6], respectively, are the object
of two recent papers [7,8]. In chelatometric titrations,
the use of pH-stat is currently suggested for the mere
purpose of keeping the pH at an optimal value, when
concentrated buffers could alter the reaction course
[9]. At our knowledge, only a single procedure using
the pH-stat technique for detecting the titration end
point (determination of calcium and magnesium in

0003-2670/02/$ see front matter 2002 Elsevier Science B.V. All rights reserved.
PII: S 0 0 0 3 - 2 6 7 0 ( 0 2 ) 0 0 6 9 1 - 8
278 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
skim milk) has so far been described [10]. In the third
paper of this series [11], the fundamentals of pH-stat
chelatometric titrations have been developed, and the
criteria for optimising the experimental conditions
have been discussed. Accordingly, such titrations are
expected to attain good accuracy and high precision.
The purpose of the present work is to substanti-
ate those favourable anticipations by implementing
experimental procedures for some representative
determinations.
2. Experimental
2.1. Apparatus
2.1.1. pH-stat titrations
The apparatus required must allow the simultane-
ous addition of two reactants: the primary titrant (the
chelating agent), by stepwise addition of equal, small
volume increments, or by continuous addition at a con-
stant volume rate, and the auxiliary titrant (a strong
base) under pH-stat control. Single instruments auto-
matically performing all the required operations are
not currently available. However, a partly automatic
apparatus can easily be assembled by combining a
potentiometric (pH-metric) titrator having the pH-stat
function with a system for controlled delivery of the
primary titrant.
The experiments with stepwise delivery of primary
titrant here reported were performed with a syringe
burette Microbur 2030 (Crison, Barcelona) fitted
with a 1 cm3 syringe 1001 TLL (Hamilton, Bonaduz,
Switzerland). The resolution of delivered volumes is,
4104 and 1103 cm3 , respectively; accuracy was
determined to be better than resolution. The burette
was driven by a PC through the proprietary Bureta
program, modified to repeat additions of constant
volume aliquots at fixed time intervals. Experiments
with continuous delivery of the primary titrant were
performed with the apparatus described in [8].
For delivering the auxiliary titrant under pH-stat
control, a microprocessor-controlled programmable
potentiometric titrator Crison MicroTT 2050 was
used. It was equipped with a Crison Microbur 2030
syringe burette fitted with a 2.5 cm3 1002 TLL sy-
ringe (Hamilton) and was connected to an Epson
LX-400 printer. The signal from a combination glass
electrode with encapsulated Ag/AgCl reference elec-
trode in 3 M KCl and sleeve junction (Ingold, Urdorf,
Switzerland, type 405.60.88.E) was measured with a
resolution of 0.1 mV or 0.01 pH. The Stat program
allowed choosing either mV or pH as the controlled
operational parameter.
The measured solution was contained in a 20
90 cm3 jacketed cell Metrohm (Herisau, Switzerland)
type 6.1418.220 and stirred with a PTFE-coated mag-
netic bar. It was thermostated at 25.0 0.1 C and
fluxed with carbon dioxide-free nitrogen.
2.1.2. Photometric titrations
Photometric titrations were monitored with a
Metrohm 662 Photometer equipped with a light-guide
measuring cell. The photometric probe could be ac-
commodated in the titration cell through the cover.
The stepless wavelength setting is 400700 nm with a
10 nm resolution. The analogue output is 01 V with
a 1 mV resolution. The MicroTT 2050 programmable
potentiometric titrator was used for adding the titrant
EDTA solution and for recording the titration plot
by taking the voltage signal from the photometers
output.
2.2. Reagents
All chemicals of analytical grade or better were used
as received. The 0.1 or 0.01 M standard solution of
metal cations were prepared from high purity metals
(Cu), oxides (MgO, ZnO) or salts (CaCO3 ) with the
usual procedures [1215].
The 0.1 M solutions of primary titrant were prepared
by weighing analytical grade disodium ethylendini-
trilotetraacetate (Na2 H2 Y, EDTA) after desiccation at
80 C.
A 0.1000 M solution of sodium hydroxide low
in carbonate (sodium carbonate <0.2%) BDH cat.
191503E was used as the auxiliary titrant.
All solutions were prepared with ultrapure water
produced with a Milli-Q Plus apparatus (Millipore,
Bedford, MA).
2.3. Procedure
2.3.1. pH-stat titrations
The measuring electrochemical cell was calibrated
separately with pH standards at 4.00 and 7.00. The
 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
titration cell was filled with 20 cm3 of 0.1 M sodium
perchlorate solution containing a suitable buffer at
low concentration (generally (12) 103 M) and a
volume of standard solution of the metal cation con-
taining approximately 50 mol of cation was added.
A preliminary adjustment of the solution pH to a value
slightly more acidic than the operating value was
made either by addition of some drops of diluted acid
or (if the standard solution of cation was too acidic)
by a preliminary end-point titration with strong base.
The exact operating pH value was reached by tem-
porarily activating the Stat function. Finally, the Stat
operation and the record of the titration plot were
simultaneously started and, after a known time delay
(30 or 60 s which allowed to control the stability of
pH), the primary titrant addition was begun. Constant
volume (2448 l) stepwise additions at fixed time
intervals (30 s2 min), or continuous additions (min-
imum rate 0.05 cm3 min1 ) were carried on until the
added primary titrant was 1.52 times the equiva-
lence volume (usually corresponding to 0.5 cm3 ). The
numerical titration report (displaying time, measured
pH or mV, auxiliary titrant volume, single volume
increments, temperature) was printed during opera-
tion at 10 s intervals. The graphical report (auxiliary
titrant volume versus time) was printed after ending
the titration. The true titration plot was recalculated
by using as the abscissa the primary titrant volume VY
and as the ordinate the volume of auxiliary titrant VOH
required to reach stationary conditions (pH constant
in time at the operating value within the resolution of
the pH-stat system, i.e. 0.01 units).
The equivalence volume of the primary titrant, Ve ,
was determined by least-square analysis as the ab-
scissa of the intersection between two straight-line
segments. Data was treated with SigmaPlot scien-
tific graphing software (Jandel Scientific, Erkrath,
Germany).
2.3.2. Photometric titrations
Photometric titrations were performed on a mea-
sured volume of standard solution of the analyte cation
diluted with 25 ml of buffer at the required pH and con-
taining a suitable indicator. The programmable titrator
was operated in the inflection point titration mode
with constant volume additions. This mode allowed
printing and plotting the complete titration record. The
input signal was taken from the photometric probe, op-
279
erated either in the absorbance or transmittance mode.
Very small (10 l) additions were made, and a strict
stability criterion (signal drifting less than 1 mV in
10 s) was chosen, in order to reach the best precision
and accuracy even when the signal change at the end
point was abrupt or the complexation reaction was
slow.
3. Results and discussion
3.1. The pH-stat titration plot and the choice of
operating pH
In pH-stat chelatometric titrations of metal cations,
the addition of primary titrant (EDTA or another suit-
able chelating agent) is accompanied by the addition
of the auxiliary titrant (a strong base solution) at a rate
suitable to maintain the pH constant at a pre-selected
value by neutralising the acidity delivered by EDTA.
The titration plot shows the added volume of strong
base, VOH , against the volume of EDTA, VY . When
the chelation of the titrand cation is complete, the
delivery of acidity by EDTA does stop or change
rate, depending on the operating pH. Therefore, in
the correspondence of the equivalence point the titra-
tion plots slope also changes. If the conditions are
suitable, it is expected that the change of slope is easily
appreciable.
The dependence of the titration plots shape on the
nature of the reactants and on pH has been discussed
in detail [11]. Two main aspects of the titration plot
do condition the feasibility of a titration: the extent of
slope change and its sharpness. The extent of slope
change depends on the difference between the net
number of protons delivered by EDTA before and af-
ter the equivalence point. Its sharpness, in turn, de-
pends on the quantitativity of the chelation reaction.
Both are strongly influenced by pH [11].
According to the primary reaction:
Mm+ + H2 Y2 MYm4 + 2H+ (1)
the slope of the titration plot before the equivalence
point ideally corresponds to 2 mol of strong base per
mole of EDTA. However, competing reactions in-
volving proton exchange, whose extent depends on
pH, can decrease this ratio [11]. The slope after the
equivalence point, ideally null after the completion of
280 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
reaction (1), is dictated by the degree of protonation
of the unreacted ligand at the operating pH. At pH
increasing over about 5, the prevailing form is HY3 ;
at still higher pH it is Y4 . Therefore, a finite and
increasing amount of base is consumed to deproto-
nate the excess titrant, H2 Y2 ; the relative amount
gradually approaches the maximum value of 2 mol of
auxiliary titrant per mole of primary titrant in excess.
In consequence, the change of slope at the equivalence
point vanishes at a pH is larger than about 11.5 [11].
In turn, the quantitativity of the chelation reaction
depends on the conditional stability constant of the
chelate complex KMY  in the reaction medium. This
quantity is heavily conditioned by the dependence of
the degree of protonation of EDTA anion on pH [16].
Competing ligands decreasing the free-cation fraction
of the unreacted metal may also have an appreciable

effect. The overall trend is towards a decrease of KMY
on decreasing pH. As the reaction becomes less and
less quantitative, the slope change occurs in a larger
and larger interval around the equivalence point, thus
impairing the sharpness of the equivalence point.
For every analyte cation, a compromise value of pH
for the two contrasting effects must be found. This op-
timum value can be accurately forecast if the stability
constant of the chelate complex and the equilibrium
constants of the competing reactions are known with
sufficient accuracy [11].
It must be noted that pH does not need to be known
with the greatest accuracy, as long as it lies in the
wanted range and is kept rigorously constant during
each single titration. For quantitative determinations,
calibration of the measuring cell with usual pH stan-
dards is quite acceptable.
3.2. Choice of the instrumental and other
experimental parameters
The experimental parameters to be set at the be-
ginning of each titration are either of chemical or
of instrumental nature. The chemical parameters are
operating pH and buffer power of the supporting
solution. The instrumental ones are addition rate,
equilibration time and control band [8].
The choice of pH is dictated by the criteria dis-
cussed in Section 3.1. The buffering power of the mea-
sured solution regulates the sensitivity of the pH-static
response, i.e. the deviation of pH from the operat-
ing value caused by unbalanced titrants ratio. In the
absence of any buffer, EDTA addition, particularly
when discontinuous, could cause sudden and large pH
changes, hindering the pH-static control. A too high
buffering power would instead cover the titrants ac-
tion. Both effects cause large random errors [8]. With
synthetic samples, optimum conditions are realised by
adding a suitable buffer pair.
The approach to the choice of the instrumental pa-
rameters is similar to the one discussed in detail for
acidbase pH-stat titrations [8]; in the present instance,
however, it is much more dependent on the nature of
the chemical system involved. Indeed, an important
peculiarity of chelation reactions is that they are more
or less appreciably slow, at variance with neutralisa-
tion, which is generally fast.
When the reaction is fast, the practical rate-
determining stage can be, alternatively, the mixing of
reactants, the electrode response, or the ability of the
pH-stat device to keep pace with the acidity change.
This last is mostly conditioned by the instrumental
parameters set. By acidbase titrations, it was quite
generally possible to operate with continuous, al-
though moderately slow, addition of titrand (which in
acidbase titrations operationally takes the place of
the primary titrant). In contrast, preliminary experi-
ments did show that complexometric reactions with
continuous additions, although yielding apparently
satisfactory linear segmented plots, were frequently
affected by appreciable systematic errors. These were
removed by operating with constant additions at
fixed time intervals. With this procedure, the plot of
auxiliary titrant volume against time together with
the numerical record of both quantities and pH (see
Section 2.3) allowed to verify point by point the time
necessary to reach equilibrium conditions (Fig. 1).
The size of the added aliquots (1/20 to 1/10 of the
equivalence volume), and thus their number, was cho-
sen to satisfy the somewhat contrasting requirement
of efficient pH-stat control, good least-squares evalu-
ation of data and reasonable total titration time. The
time between two subsequent additions, set after the
results of a preliminary titration, was just long enough
to allow the reaction to reach equilibrium after each
addition (Fig. 1).
The size of additions also has a role in dictating the
instrumental parameters specific of the pH-stat oper-
ational mode, i.e. equilibration time and control band
 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
Fig. 1. (A) Recorded plot of the volume of auxiliary titrant
(0.1000 M NaOH) against time for a pH-stat titration at pH 3.00
of 48.64 mol of copper(II) in approximately 20 ml of supporting
solution (0.1 M sodium perchlorate containing 2 103 M formate
buffer). Aliquots of 48 l of 0.1 M of EDTA were added at 1 min
intervals. Equilibration time 2 s; control band 2 pH units. (B) The
corresponding plot of measured pH against time.
[8]. The equilibration time is the time after which in
the Stat mode the meter is made to accept the mea-
sured signal as stable and ideally corresponding to the
equilibrium emf of the measuring cell when stationary
conditions are reached. Given the fast response of the
physical measuring system, an equilibration time of
2 s was sufficient. The choice of the control band [8]
is possibly less critical than in acidbase titrations,
because the reaction and, consequently, the delivery
of acidity, are moderately slow and do not require
fast compensation of large pH changes if the size of
added primary titrant aliquots is small. The purposely
added pH buffer (see above) helped to damp pH from
swinging.
281
Fig. 1A shows a typical graphical titration record
(volume of auxiliary titrant against time) for an ap-
preciably slow reaction (copper(II) at pH 3). Fig. 1B
shows the corresponding pattern of pH as recorded in
the numerical titration report (see Section 2.3). Step-
wise additions of EDTA were made at 1 min intervals;
pH was measured at 2 s intervals and recorded every
10 s. The patterns of both plots of pH and NaOH vol-
ume against time show that 3045 s were required to
reach stationary conditions after each addition of pri-
mary titrant. After the equivalence point, the measured
solution is no any more under pH-stat control, because
the excess EDTA behaves as a weak base (it hydrol-
yses by about 30%) and slowly increases the solution
pH [11].
3.3. The auxiliary titrant
For determining the metal cation contained in the
sample solution, it is not necessary to know accurately
the titre of the auxiliary titrant solution. Indeed, the
titre of the strong base only decides on the experimen-
tal slopes (cm3 /cm3 ) of the two segments composing
the titration plot without altering the abscissa of their
intersection, i.e. the equivalence volume of the pri-
mary titrant.
In principle, if the reacting strength of the secondary
titrant is accurately determined [7,8] and the real path
of the overall reaction (cation+chelating agent+base)
with its exact stoichiometry [11] is known, the amount
of analyte can also be determined from the end-point
volume of the auxiliary titrant. Much more interesting
is the possibility of determining the stoichiometry of
the overall titration and the stability constants of mixed
complexes from the mole ratio R of strong base to
chelating agent after measuring the slope of the linear
segment preceding the equivalence point [11]. For this
purpose, the reacting strength of the strong base must
be known with the best possible accuracy.
It is known that strong base solutions are gener-
ally contaminated by carbonate, and in consequence
their reacting strength depends on the end-point pH
of the individual titration in which they are used [8].
The effective titre can be conveniently determined
by pH-stat titration of a primary standard acid [7,8]
at the intended operating pH of the pH-stat chelato-
metric titration. By this means, the 0.1000 M solution
of sodium hydroxide low in carbonate used in the
282 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
experiments described here was seen to increase only
negligibly its contamination as an effect of the nec-
essary manipulations. In preliminary experiments,
solutions at the usual level of carbonate contami-
nation (about 2%) were used without appreciable
inconvenience.
3.4. Selected titration systems
When discussing the theory of pH-stat complex-
ometric titrations [11], summarised in Section 3.1,
we took those of copper(II) and magnesium(II) with
EDTA as examples representative of the main features
of the technique. Owing to their diverse properties,
these systems are also very suitable for introducing
the experimental application of the pH-stat technique,
while confirming the theoretical deductions. In addi-
tion, we report here the results of investigations on two
other cations of great analytical relevance, calcium(II)
and zinc(II).
For comparison with other well-established tech-
niques, we have considered only volumetric methods
based on chelatometric reactions. Indeed, the most
common instrumental determination technique of
metals in solution, atomic absorption spectrometry,
measures concentration in a much lower range and
with limited precision, with the inconvenience of re-
quiring a specific lamp for every element. Direct po-
tentiometry also requires a specific electrode for each
determinand cation and has only moderate precision.
When higher precision determinations are required,
the choice is usually chelatometric titration (even in
preference to gravimetry, which is too time consum-
ing). Visual titrations are performed after adding a
suitable indicator. The procedures can be automated
and their precision improved by using photometric
titrators. Therefore, in order to state whether the new
technique offers any advantage in an automated ana-
lytical laboratory, we have compared pH-stat titrations
of each investigated metal cation with corresponding
indicated photometric titrations. However, it must be
stressed that a photometric titration can hardly be
considered an accurate reference method. Indeed, the
shape of the titration curve (let it be absorbance A,
or transmittance T, against titrant volume V), and by
consequence the procedure for locating the equiva-
lence volume, are strongly dependent on the values
of the conditional stability constants of the com-
plexes involved (cation-EDTA and cation-indicator).
In practice, the location of the equivalence point may
become rather subjective, although less than in visual
titrations. With computerised photometric titrators,
the accuracy depends on the algorithm chosen.
An alternative automatable technique, the poten-
tiometric titration, is convenient only if an electrode
selective for the analyte cation is available (proce-
dures involving the use of electrodes responding to an
auxiliary cation, e.g. mercury(II), are not of current
use in the analytical laboratory). Even in this instance,
it can suffer some limitation due to the relatively high
lower response limit of the sensor.
All titrations were performed using primary stan-
dard solutions of cations. In order to favour a direct
comparison among all series of titrations, the results
were expressed by calculating the titre CY of the
EDTA solution (Table 1).
3.5. Copper
The chelatometric titration of copper(II) can be
performed either in acidic solution at pH not exceed-
ing 5, above which precipitation occurs, or at pH
near to 8 [12,13]. We have tested the determination
of Cu(II) in the acid pH range from pH 5 to 3, where
the chelate is still strong enough (log KCuY  8.3)
[16] to form quantitatively even in the proximity of
the equivalence point and assign the titration a good
intrinsic [17] precision.
Typical pH-stat titrations plots of 2.4103 M cop-
per(II) (48.64 mol in approximately 20 cm3 of sup-
porting solution) at various pH values are represented
in Fig. 2A. The buffers were, respectively, 2 103 M
chloroacetate at pH 3 and 2 103 M acetate at pH 4
and 5.
It is known that the chelation of Cu(II) is moder-
ately slow. The speed increases with increasing pH. At
pH 3, reaching stationary conditions required at least
45 s as shown in Fig. 1; with increasing pH, 3015 s
became sufficient. With additions of primary titrant
aliquots corresponding to approximately 1/10 of the
equivalence volume (48 l), titrations lasted not more
than 1215 min and yielded satisfactory results. Some
experiments performed by halving the aliquot volume
(24 l) yielded identical results, with an only moderate
improvement in the statistical evaluation of the linear
parameters and the drawback of longer duration.
C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288 283
284 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
Fig. 2. (A) Titration plots of volume of auxiliary titrant (0.1000 M
NaOH) against volume of primary titrant (0.1002 M EDTA) for
pH-stat titrations of 48.64 mol of copper(II) in approximately
20 ml of supporting solution at pH(): 5.00 (2103 M acetate
buffer); (): 4.00 (2 103 M acetate); (): 3.00 (2 103 M
chloroacetate). Aliquots of 48 l of 0.1 M EDTA were added at
intervals of 1 min (at pH 3) or 30 s (at pH 4). Equilibration
time 2 s; control band 2 pH units. (B) Deviations (residuals) of
the experimental auxiliary titrant volumes from the interpolated
linear segments for the titration at pH 5. Empty symbols after the
equivalence point.
The shape of the plots agrees well with the theoret-
ical expectations (see [11], Fig. 2). With decreasing
pH, the slope of the linear segment at V < Ve de-
creases, being increasingly affected by the formation
of protonated chelate (CuHY ). In contrast, the slope
of the segment at V > Ve , which is null at pH 4 and
5, becomes slightly positive at pH 5, as a result of the
partial reaction of the base with the excess of H2 Y2
to produce H3 Y .
The linearity of the two segments composing the
titration plots was always satisfactory. The standard
deviation of the linear parameters was generally <1 l
for the intercept and 0.002 for the slope (which for
V < Ve changed from about 1.5 at pH 3 to nearly
2 at pH 5). Fig. 2B represents the deviations (resid-
uals) of the experimental volumes of auxiliary titrant
from the interpolated linear segments for the plot at
pH 5. In titrations at pH 4 and 5, the residuals were
generally <1 l and did not show any trend. At pH
3, the residuals of the first segment showed some de-
viation from linearity (a convexity), which however
was not significantly influential on the straight-line pa-
rameters. Only at this pH a single point very near to
the equivalence volume showed a systematic devia-
tion due to nonquantitative reaction, and was omitted
from least-square linearisation. The results, expressed
as the titre of the EDTA solution (mean of five titra-
tions), are summarised in Table 1.
For sake of completeness, pH-stat titrations at pH 2
and 8 were also attempted. In a 0.01 M hydrochloric
acid, the combined effect of slower reaction and higher
buffering power prevented titrations being carried out
in a reasonable time with acceptable accuracy and pre-
cision. The tedious procedure necessary to keep cop-
per(II) dissolved at pH 8 in ammonia buffer without
increasing too much the buffering power made titra-
tion at this pH rather impractical.
Photometric titrations for comparison were made
in 0.1 M acetate buffer at initial pH 5.0 using PAN
(1-(2-pyridylazo)-2-naphthol) as the indicator at the
wavelengths of the absorption peaks both of the free
indicator (460 nm) and of its complex with copper
(545 nm). The end point was located, according to
the customary criteria [9], just after the beginning of
the very sharp transmittance change. The results are
reported in the last column of Table 1.
The results of pH-stat titrations at pH 4 and 5 agree
with their theoretical value as well as the photomet-
ric one. Only at pH 3 is a systematic error, small but
significant, present. It does not depend on imperfect
linearity (partial extrapolation does not change the re-
sult), nor on the nature of the buffer (similar results
were obtained in phthalate buffer). The error corre-
sponds to a consumption of EDTA by copper below
the stoichiometric 1:1 ratio. It could be caused by the
reaction being slower than apparent from the direct
titration plot (Fig. 1); however, longer (2 min) inter-
vals between additions did not change the result. It
is also compatible with the hypothesis of apprecia-
ble formation of binuclear complexes; however, this
is little probable at a low concentration of the metal
 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288 285

cation and at a pH where protonated forms of the lig-

and prevail, and is not reported in the literature. The
real cause could be some experimental artifact de-
pending on pH, but it was not further investigated.
In practice, titrations can be made at pH 3 if ex-
treme accuracy is not required (however, the high pre-
cision obtained suggests that a blank correction is
possible).

3.6. Zinc

Titration of Zn(II) with EDTA according to tradi-

tional procedures is feasible in basic media up to pH
12, and in acidic media at pH 46 [1214]. The higher
buffering power required to keep zinc dissolved makes
pH-stat titration at basic pH impractical. On the other
hand, the specific properties of the technique prompted
us to test the pH-stat titration at a more acidic pH. The
investigated range was thus pH 36.
The pH-stat titrations plots of 2.0 103 M zinc(II)
(49.57 mol in approximately 25 cm3 of supporting
solution) at pH between 3 and 6 are shown in Fig. 3A.
The solution was buffered with 2 103 M chloroac-
etate at pH 3, 2 103 M acetate at pH 4 and 5,
7 103 M hexamethylene tetramine at pH 6. The
response was acceptably fast at low pH, much faster Fig. 3. (A) Titration plots for pH-stat titrations of 49.57 mol
of zinc(II) in 25 ml of supporting solution at pH():
at pH 4, where 15 s was sufficient for its completion
6.00 (7 103 M hexamethylene tetramine buffer); (): 5.00
after each addition. (2 103 M acetate); (): 4.00 (2 103 M acetate); (): 3.00
The linearity of the two segments of titration plot (2 103 M chloroacetate). Other experimental parameters as in
was satisfactory (Fig. 3B). Even at pH 3, where Fig. 2. (B) Residuals of the plot at pH 6.
log KZnY is as low as 6.0 [16], only points very near to
the equivalence (in Fig. 3A, a single point at 480 l) a sudden slope change, which was taken as the end
showed barely appreciable deviations from quantita- point.
tivity and had to be omitted from the least-squares
fitting. Formation of protonated chelate (ZnHY ) 3.7. Magnesium
strongly affects the slope at V < Ve at pH 3, much
less at pH 4. By contrast, the effect of the reaction of In discussing the theory of complexometric pH-stat
the base with the excess of H2 Y2 to produce H3 Y titration [11], magnesium was taken as an example
becomes very evident at pH 6. of a cation giving a moderately stable chelate, with a
The results, summarised in Table 1, show that the titration pH range restricted to basic values. It is usu-
best results are obtained at pH 46. At pH 3, a small 
ally titrated at pH 10, where log KMgY is 8.2. By con-
but significant error, similar to that found in copper trast, pH-stat titrations are expected to be still feasible
titrations at the same pH, was present. Otherwise, at significantly lower pH values, where the stability
they agree both with the theoretical value and with of the EDTA chelate is so small as to prevent tradi-
that resulting from photometric titrations. These were tional titrations, particularly with a visual end point.
made at pH 10.0 in ammonia buffer with Eriochrome A wide pH range (pH 610) was explored in order to
Black T (EBT) as the indicator at 620 nm (free indi- investigate the experimental feasibility limits with not
cator) and 550 nm (complex with zinc) and showed strictly quantitative reactions. In Fig. 4A, titrations of
286 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
Fig. 4. (A) Titration plots for pH-stat titrations of 50.78 mol
of magnesium(II) in 20 ml of supporting solution (0.1 M sodium
perchlorate) at pH(): 10.00 (2 103 M ammonia buffer);
(): 9.00 (2 103 M ammonia); (): 8.00 (5 103 M tri-
ethanolamine); (): 7.00 (5103 M triethanolamine); (): 6.00
(5103 M hexamethylene tetramine). 48 l of 0.1 M EDTA were
added at intervals of 1 min (at pH 6 and 7) or 30 s (at higher pH).
Other experimental parameters as in Fig. 2. (B) Residuals of the
plot at pH 8.
2.5 103 M magnesium(II) (50.78 mol in approxi-
mately 20 cm3 of supporting solution) are represented.
The buffer was 5 103 M hexamethylene tetramine
at pH 6, 5 103 M triethanolamine at pH 7 and
8, 2 103 M ammonia at pH 9 and 10.
The reaction was quantitative starting from pH 8

(log KMgY = 6.4), where even the experimental points
nearest to equivalence did not show appreciable de-
viations from the linear trend (Fig. 4B). At pH 7

(log KMgY = 5.3), only the points lying at about (1.0
0.1)Ve deviated significantly and had to be neglected
in the linear fitting. The slope of the linear segment
corresponding to an excess of titrand cation was found
to be practically independent of pH, showing that mag-
nesium does not significantly take part in side reac-
tions involving proton exchange [11] in the experi-
mental pH range. Beyond the equivalence point, the
slope increases steadily with increasing pH. At pH 10,
the slope change at the equivalence is only moderate,
because the excess of H2 Y2 reacting with the base
produces an appreciable fraction of H4 Y together with

H3 Y2 . At pH 6 (log KMgY = 3.9), the linear range of
the two segments was too restricted to allow reliable
interpolation of the equivalence volume.
The photometric titrations were made at pH 10.0
in ammonia buffer with EBT as the indicator at
620 nm (free indicator) and 560 nm (complex with
magnesium). The end point was taken at the point
of maximum slope of the sigmoidal titration curve.
The results of both methods (Table 1) agree with the
theoretical value. However, the photometric titrations
show a larger uncertainty.
The experiments confirm that, in agreement with
theoretical expectations [11], pH-stat titrations of mag-
nesium have their optimum operating range around pH
89. At pH 10, the optimum value of traditional titra-
tions, the slope change begins to be critically small.
By contrast, a lower pH can be used, if necessary [10],
although not much under 7.
3.8. Calcium
The chelatometric determination of calcium is usu-
ally performed at pH 912 in ammonia buffer. In vi-
sual titrations, the best precision is usually obtained at
pH 910 by using magnesium as auxiliary cation and
EBT as the indicator.
The plots in Fig. 5 represent pH-stat titrations of
51.55 mol of calcium in 25 cm3 of supporting solu-
tion containing 5 103 M hexamethylene tetramine
(pH 6), 1.0 102 M triethanolamine (pH 7 and 8),
or 2 103 M ammonia (pH 9 and 10). At pH 6,
the points within about 5% of the equivalence vol-
ume showed significant deviations due to nonquanti-
tative reaction and were omitted from the calculations.
The results are summarised in Table 1.
It is seen that for calcium, like magnesium, the
higher limit of the available pH range is conditioned
by the acidbase properties of the chelating agent.
By contrast, the higher stability of the chelate of cal-
cium with respect to magnesium allows a lower pH
to be used. Fig. 5A and Table 1 show that pH 6 (with
 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
Fig. 5. (A) Titration plots for pH-stat titrations of 51.55 mol of
calcium(II) in 25 ml of supporting solution (0.1 M sodium perchlo-
rate) at pH(): 10.00 (2 103 M ammonia buffer); (): 9.00
(5 103 M ammonia); (): 8.00 (1.0 102 M triethanolamine);
(): 7.00 (1.0 102 M triethanolamine); (): 6.00 (5 103 M ture, log KCuHY = 3.0 at 25 C, = 0.1 [16,18]. The
hexamethylene tetramine). Aliquots of 48 l of 0.1 M EDTA were
added at intervals of 1 min. Other experimental parameters as in
Fig. 2. (B) Residuals of the plot at pH 10.

log KCaY = 5.9) is still available for sufficiently pre-
cise titrations.
Independent of pH, pH-stat titrations of calcium
seem slightly more precise than photometric ones (last
column of Table 1). These were made at pH 10.0 in
0.1 M ammonia buffer with EBT as the indicator at
620 or 550 nm; the end point was obtained by ex-
trapolating the sloping branch of the absorbance or
transmittance.
3.9. The determination of equilibrium constants
of protonated complexes
It has been theoretically deduced [11] and exper-
imentally confirmed (Sections 3.5 and 3.6) that the
287
slope segment of the titration plot preceding the equiv-
alence depends on proton-exchange side reactions in
which the titrated cation is involved. If only forma-
tion of the protonated chelate occurs as such a side
reaction, its stability constant can be determined from
the slope of that segment, expressed as the mole ratio
R = n(NaOH)/n(EDTA) [11]:
2R
KMHY = (2)
(R 1)[H+ ]
For determining accurately KMHY , accurate mea-
surement of the equilibrium concentration of the hy-
dronium ion and careful control of the ionic strength
are essential. An internal calibration of the electro-
chemical measuring cell in the supporting solution
should be made before each titration [7,8]. In our
experiments, the cell was calibrated externally with
usual pH standards, and only the repeatability of the
operating value of pH during a single experiment
was carefully controlled, the exact value being of
low relevance for titration accuracy (see Section 3.1).
Therefore, their results can be used only for a mere
preliminary check of the theoretical expectations.
The titrations most suitable for this purpose are
those of copper at pH 3.00, where the side-reaction
effect is larger. R was 1.4651 0.0007 (n = 5), yield-
ing KCuHY = 1.150 103 , log KCuHY = 3.061. This
value compares well with the one given in the litera-
very small standard deviation of R shows that KMHY
can be determined with precision possibly much bet-
ter than with traditional methods and with accuracy
limited only by that of the pH measurement. Even the
value R = 1.89220.0006 obtained at pH 4.00, yield-
ing KCuHY = 1.208 103 , log KCuHY = 3.082, is
acceptable, particularly considering that the effect on
R amounts to only 10% of the limit value 2.
4. Conclusions
The experiments here described fulfil satisfactorily
the theoretical expectations [11] about the analytical
suitability of pH-stat chelometric titrations as well as
about the shape of the titration plots. Accuracy and
precision are very good. The experimental parameters
are easily adjusted to fit the properties of the titration
system. The duration of a single titration is acceptable.
288 C. Macca et al. / Analytica Chimica Acta 470 (2002) 277288
In comparison, photometric titrations can reach com-
parable precision only at the cost of longer duration;
indeed, in our experiments, they required smaller ad-
ditions in the proximity of the equivalence both when
the end point was interpolated (with sigmoidal titra-
tion plots) or extrapolated (to identify a discontinuity).
The most serious limit for practical applications of
pH-stat titrations with EDTA is the requirement that
the titrated solution has a moderate buffering power.
The moderately basic maximum operating pH (not
much higher than 10) is largely counterbalanced by
the possibility of exploiting reactions that are not
strictly quantitative, i.e. with the smaller conditional
stability constants of the chelate complex prevailing
at lower pH. A clear advantage of the pH-stat proce-
dure with regularly timed discontinuous addition of
primary titrant is that the direct titration plot of the
auxiliary titrant volume against time generally gives
immediate evidence of slowly occurring reaction
and of reaching equilibrium. Such a direct control is
difficult or impossible, for instance, with indicated
photometric titrations.
Even the suitability of pH-stat procedures for the
determination of some stability constants has been
confirmed. Both applications seem to deserve larger
consideration in the analytical and research laboratory.
Acknowledgements
The work was carried out with the financial support
of CNR. The co-operation of Alberto Doimo in the
preparation of the computer program for timed discrete
additions is gratefully acknowledged.
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