View Article Online

View Journal

Analyst
Accepted Manuscript

This article can be cited before page numbers have been issued, to do this please use: J. Zhai and E.
Bakker, Analyst, 2016, DOI: 10.1039/C6AN00538A.

This is an Accepted Manuscript, which has been through the

Royal Society of Chemistry peer review process and has been
accepted for publication.

Accepted Manuscripts are published online shortly after

acceptance, before technical editing, formatting and proof reading.
Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available.

You can find more information about Accepted Manuscripts in the

Information for Authors.

Please note that technical editing may introduce minor changes

to the text and/or graphics, which may alter content. The journal’s
standard Terms & Conditions and the Ethical guidelines still
apply. In no event shall the Royal Society of Chemistry be held
responsible for any errors or omissions in this Accepted Manuscript
or any consequences arising from the use of any information it
contains.

www.rsc.org/analyst
 Page 1 of 10 Analyst
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3
4 Complexometric Titrations: New Reagents and Concepts to Overcome Old
5 Limitations
6
7 Jingying Zhai and Eric Bakker*
8 Department of Inorganic and Analytical Chemistry, University of Geneva, Quai Ernest-Ansermet 30, CH-1211 Geneva,
9 Switzerland
10
11
12 Abstract rapid and quantitative chemical analysis of metal ions. The
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13 Chelators and end point indicators are the most important ions of interest are titrated with the chelator of choice
14 parts of complexometric titrations. The most widely used through a coordination complexation reaction and rapidly
15 universal chelators ethylenediamine tetraacetic acid (EDTA) form stable monodentate or multidentate complexes. The
16 and its derivatives can strongly coordinate with different chelator is sometimes called the complexing reagent or
17 more simply, titrant. The end point can be identified by a

Analyst Accepted Manuscript

metal ions. Their limited selectivity often requires the use
18 of masking agents, and the multiple pKa values of the metallochromic indicating dye, which shows a color change,
19 chelators necessitate a careful adjustment of pH during the or by other instrumental indicators, such as ion-selective
20 procedure. electrodes.
21 Real world requirements for pH independent, selective and The earliest example for this type of titration reactions is
22 sensitive chelators and indicators call for a new design of the determination of cyanide ion concentration by silver
23 these reagents. New concepts and structures of chelators nitrate, proposed by the German chemist Justus Liebig in
24 and indicators have indeed recently emerged. We present the 1850s.1 In 1945, Schwarzenbach, who made a
25 here recent developments of chelators and indicators for significant contribution to this field, formally introduced
26 complexometric titrations. Much of these advances were the complexometric titration method to quantify metal ions,
27 made possible only recently by moving the titration from a mainly using EDTA as chelator.2 The discovery of EDTA
28 homogenous to a heterogeneous phase, using a new class of dramatically pushed the field forward. Since 1950,
29 chelators and indicators based on highly selective complexometric titration has spread all over the world, for
30 example to measure water hardness.3 A comprehensive
ionophores embedded in ion-selective nanosphere
31 theory of complexometry was put forward by
emulsions.
32 Schwarzenbach in his book, published 10 years after the
In view of achieving titrations in situ by complete
33 introduction of the method.4, 5 Almost at the same time,
instrumental control, thin layer electrochemistry has
34 various indicating dyes started to appear to visualize the
recently shown to be an attractive concept that replaces the
35 end point by the naked eye or by spectrophotometric
traditional cumbersome titration protocol with a direct
36 instrumentation. Murexide and Eriochrome Black T were
reagent free sensing tool.
37 established as indicators for water hardness.6, 7 A number of
38 researchers including Reilley, Hildebrand, Patton, Reeder
39 1. Introduction and Tsien contributed to the synthesis of new indicators to
40 Titrimetry is a general and powerful method which is used improve their selectivity.8-11 Powerful instrumental analysis
41 to quantify a wide range of analytes. The high accuracy of methods such as potentiometry, conductometry,
42 the results and maturity of the procedure have made it a thermometry, coulometry and chronopotentiometry were
43 routine method in various fields such as environmental also developed to provide improved choices for
44 monitoring, bioanalytical chemistry and clinic analysis. quantitative analyses.12-19 Although EDTA has always been
45 Compared with quantitative instrumental measurements the most widely recognized chelator in complexometric
46 that depend on a readout using methods as ion titration, good chelators and indicators as well as new
47 chromatography, ICP-MS or AAS, titrimetry is the most concepts have been continuously emerging. In addition,
48 simple and accurate because it relies on an exhaustive various titration protocols have been developed.20, 21 This
49 consumption of the analyte at the end point. Today, review summarizes the most recent developments in
50 titrimetry can be easily automated and commercially complexometric titration reagents and methods.
51 available standardized reagents provide more convenience
52 to the end users. These key advantages have made
53 titrimetric methods an indispensible part of analytical
2. Chelators
54 chemistry, even with the arrival and establishment of newer 2.1 The classical chelator EDTA
55 instrumental techniques. Chelators are today widely applied in chemical industry,
56 Complexometric titration (complexometry or chelatometry) therapy, agriculture, biochemistry and other fields. Most
57 is one of the classical titrimetric methods developed for the chelators contain N, O or S atoms in their molecular
58
59
60 1
 Analyst Page 2 of 10
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3
4
5
6
7
8
9
10
Fig. 1 Chemical structure of fully deprotonated EDTA
11
and its complexation with a metal ion M in 1:1
12
stoichiometry.
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13
14 structure to provide lone-paired electrons available for
15 coordination.4, 22, 23 Functional groups such as carboxylates,
16 amines, hydroxyls and sulfhydryls are very commonly
17 found. One of the structurally simple monodentate ligands

Analyst Accepted Manuscript

18 is ammonia. It is able to strongly coordinate with metals
19 including Cu2+, Ni2+, Co3+, Ag+ and is useful to extract or
20 dissolve metals.23, 24 However, it is not suited as a chelator
21 in complexometric titrations because ammonia only
22 exhibits one bond that can be used to coordinate the analyte.
23 The resulting stepwise formation of metal complexes
24 makes it difficult to observe the end point. Cyanide suffers
25 from the same drawback.23, 24
26 It has therefore been proposed early on that for a reagent to
Fig. 2 Structural formulates of the diglycolamide based
27 act as a chelator in complexometric titrations, (i) the
ligands for actinide and lanthanide ions extraction.
28 reaction should be kinetically fast, (ii) it should proceed
29 stoichiometrically; (iii) the change in free energy must be based device contained various amounts of known EDTA
30 sufficiently large. and a small amount of indicator. The device is able to
31 The introduction of EDTA was a revolution in the field of rapidly and quantitatively determine the Ca2+ and Mg2+ in
32 complexometric titrations because it fulfills the above- mineral water, river water and seawater samples.
33 mentioned conditions. As shown in Fig. 1, EDTA exhibits As an effective chelator, EDTA can also be used to remove
34 multiple coordinating groups and forms 1:1 metal-chelator the toxic transitional metals such as Pb2+, Ni2+, Hg2+, Cd2+
35 complexes. EDTA is able to form complexes with various and As3+ from wastewater, contaminated soil and lake
36 metal ions. Since its introduction, a large number of water.43-45
37 elements have been measured with this method. Early Unfortunately, many chelaters are not biodegradable and
38 pioneering works have been done by Schwarzenbach and will be persistent in the environment.46, 47 Therefore,
39 co-workers.4, 25-28 EDTA has been used to analyze almost recyclable/separable EDTA-functionalized compounds or
40 half of the elements in the periodic table while its materials have also been reported for these applications,
41 derivatives such as diethylene triamine pentaacetic acid such as EDTA bonded polymers, particles or natural
42 (DTPA) and ethylene glycol tetraacetic acid (EGTA) materials.43, 48
43 provide similar usage.4, 29-37 EDTA and its derivatives Stark and co-workers reported that EDTA-like chelator
44 belong to the aminopolycarboxylic acid family. It is able to modified nanomagnets can remove Cd2+, Pb2+ and Cu2+
45 dissociate into several protonation states. The effective from contaminated water.43 In a very short time, the
46 formation constant of the metal-EDTA complexes therefore concentrations of transition metal ions may decrease to the
47 ug/L range, which is often acceptable. The nanomagnets
depends on pH, which makes titrations with EDTA pH
48 must be separated from the treated water.
dependent. Because EDTA and its derivatives and the
49 Until today, 95% of the publications in the complexometric
above-mentioned pharmaceutical chelators exhibit high
50 titration area are still based on EDTA and its derivatives.
binding constants to many metals ions, they lack tunable
51 EDTA as an important conventional chelator has also been
selectivity and often require the use of masking reagents.4,
52 23 used medicinally for the treatment of human intoxication
53 with heavy metals. For this type of medical treatment, three
The total amount of Ca2+ and Mg2+ is normally titrated with
54 commonly other commonly used chelators also exist,
EDTA at pH 10 while Ca2+ alone is titrated at pH 13 upon
55 including British Anti-Lewisite (BAL), dimercaptosuccinic
masking the Mg2+ ions with OH-.38-41 Recently, EDTA
56
titration was demonstrated by Kaneta and co-workers in acid (DMSA) and dimercaptopropane sulfonate (DMPS).49
57
58 microfluidic paper-based analytical devices.42 This paper-
59
60 2
 Page 3 of 10 Analyst
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3 These pharmaceutical chelators can in principle equally be
4 used in titrations.
5
6 2.2 Extractant based on diglycoamides
7 Diglycolamides as a new class of extractants for actinide
8 and lanthanides ions have been extensively studied during
9 the past decades.50 Separation, recycling and storage of
10 these long-lived radioactive elements from high level waste
11 generated from nuclear fuel are very important for the
12 environment and human health. Diglycolamides and its
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13
analogs (see figure 2) are found to be very effective and
14
selective for the extraction of trivalent actinides compared
15
with other extractants such as malomide and octyl-
16
(phenyl)-N, N-diisobutyl carbamoyl methyl phosphine
17

Analyst Accepted Manuscript

oxide (CMPO) based extractants.51, 52
18
The lipophilicity of diglycolamide compounds can be tuned
19
with different alkyl chains on N atoms. Sasaki and co-
20
workers reported that water soluble diglycolamines, N, N,
21
N’, N’-tetramethyldiglycolamide (TMDGA), N,N,N,N’-
22
23 tetraethyldiglycolamide (TEDGA), N,N,N’,N’-
24 tetrapropyldiglycolamide (TPDGA) can be used as
25 complexing agents for Pu(IV) and Am(III).53 As neutral
26 complexing reagents, these diglycolamines display high
27 affinity to Pu(IV) and Am(III) and form more stable
Fig. 3 Ion-selective emulsions containing nanospheres as
28 complexes with Pu(IV) and Am(III) than EDTA in highly
complexing agent and structures of the compounds used
29 concentrated HNO3 solution.
to form nanospheres. The nanosphere core is made of
30 Water insoluble diglycolamides with long alkyl chains can
dodecyl 2-nitrophenyl ether (D-NPOE) and the
31 be dissolved in solvents to perform liquid-liquid extraction
hydophobic sub-structure of Pluronic F-127. The
32 of actinides and lanthanides.50, 54 Many groups have studied
complexation reaction comprises (1) ion exchange
33 the synthesis and the characteristics of different
between target ion (Ca2+ or Pb2+) in aqueous phase and the
34 diglycolamides extractants. Among these extractants,
counter ion of R- (K+) in the organic phase and (2) the
35 N,N,N’,N’-tetraoctyl diglycolamide (TODGA) was found
complexation reaction between target ion and the
36 to be a promising extractant for trivalent actinides.55, 56 The
receptor, which lowers the solvation energy for the target
37 metal ions will be extracted into the solvent together with a
ion and provide the driving force for its uptake into the
38 counteranion (NO3-) in high concentrated HNO3 solution or
nanospheres.
39 its salt. If the concentration of the counter anion (NO3-) is
40 not sufficiently high, the extractants could not function three functional groups at C-pivot and trialkylphenyl
41 properly. The extraction process can also be based on ion platforms.62 These ligands also showed high affinity for
42 exchange. Naganawa and co-workers reported on the role Am(III) and Eu(III) with a 1:1 metal to ligand
43 of the hydrophobic counteranions (TFPB-) in the extraction stoichiometry. In general, ligands based on diglycoamide
44 of lanthanides(III) with TODGA.57 The metal ions may be with various platforms have demonstrated satisfactory
45 exchanged into the solvent by the counter ion of TFPB-. extraction performance.
46 The introduction of ion exchanger is greatly helpful to The water soluble diglycoamides based compounds are
47 improve the extraction ability and selectivity with low promising chelators for the titration and extraction of
48 background of ionic strength. On the other hand, without actinides and lanthanides in homogenous titrations.
49 addition of hydrophobic cationic exchanger, dissolving the However, for the use of hydrophobic titrants/extractants,
50 extractant into the ionic liquid can also result in a high the extraction system should best contain an ion exchanger,
51 extraction efficiency for lanthanides, still based on the which is further explained below with the nanospheres.
52 cation-exchange mechanism.58, 59
Diglycolamide-
53 functionalized task specific ionic liquids with functional 2.3 Ion selective nanospheres as a new generation of
54 groups attached to the cationic part of the ionic liquid chelators
55 showed high extraction efficiency to actinides and Conventional titration reactions occur directly in the
56 lanthanides.59-61 Verboom and co-workers also reported on aqueous sample. Recently, Bakker and co-workers
57 a series of new ligands based on diglycolamides that have proposed ion selective nanospheres as a novel class of
58
59
60 3
 Analyst Page 4 of 10
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3
4
5
6
7
8
9
10
11
12
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

2
13
14 Fig. 4 Comparing calcium selective emulsion with
15 EDTA as complexing agents for the potentiometric
titrations of 4 µM calcium. NS 1: titration in non- c)
16
17 buffered water by calcium selective emulsion; NS

Analyst Accepted Manuscript

18 2: titration in 1mM pH 7.0 Tris-HCl by emulsion;
19 EDTA 1: titration in non-buffered water by EDTA;
20 EDTA 2: titration in 1 mM pH 7.0 Tris-HCl by
21 EDTA. The dashed vertical line marks the end
22 point.
23 complexometric titration reagents, which moved the
24
titration process from the homogeneous to the
25
heterogeneous phase.63-65 One key advantage of using this
26
new toolbox is that the titration reagents no longer need to
27
be water soluble. Other lipophilic chelators with high
28
selectivity and high affinity to the analyte can be used, such
29
as ionophores for Pb2+, Ca2+, Cu2+, Na+, K+ and Cl-.66-68
30
These classical ionophores have originally been introduced Fig. 5 Illustration of coulometric release concept
31
as active reagents in ion selective electrode membranes for (labeled as ion pump) into a thin layer sample. (a) Flat
32
many decades. As shown in Fig. 3, the chelating sheet configuration with potentiometric readout
33
nanospheres for calcium titration contain a lipophilic (detector). E1 and E2 correspond to two different
34
calcium ionophore II and cation exchanger in the core of signals as a consequence of different excitation times.
35
emulsified organic nanodroplets, which is made of (b) Tubular configuration with a coulometric readout.
36
37 surfactant Pluronic F-127 and plasticizer.63 Based on the The Ag/AgCl served as working electrode. The
38 principle of ion exchange, the analyte calcium readily integrated charge, q1 and q2 correspond to two
39 exchanges into the nanospheres for the original counter different signals obtained at two excitation times. IFS,
40 ions (K+ or Na+, which would only interference at internal filling solution; R–, cation exchanger; L,
41 extremely high concentration) of the ion exchanger. Every ionophore.
42 Ca2+ exchanges with two monovalent counter ion so that (c) Complexometric titration by using the calcium
43 the core of the nanospheres remains neutral. In the pump plus potentiometric detection for three EDTA
44 chelating nanospheres the ionophore is chosen at molar concentrations (0.25, 0.50, and 0.75 mM). Solid lines
45 excess compared to the ion exchanger. It is therefore the correspond to the calculated concentrations from
46 ion-exchanger that defines the extraction capacity, not the equilibrium theory.
47 ionophore, and various ion–ionophore stoichiometries can Flat sheet configuration indicates that the membrane of
48 be tolerated. working electrode (calcium pump) use a flat porous
49 Fig. 4 shows a comparison between titrations effected with polypropylene sheet. Tubular configuration means that
50 calcium selective nanospheres and the chelator EDTA.63 the working electrode is a silver/silver chloride wire
51 Because the calcium ionophore exhibits no protonatable placed inside a hollow fiber doped with the lipophilic
52 groups, it was not necessary to control the sample pH cocktail.
53 during the titration: titrations with the calcium selective
visible endpoint in unbuffered water and the transition was
54 nanospheres showed an almost indistinguishable behavior
not easily observable in samples buffered at pH 7.0. As
55 in pH 7.0 buffered solution than in unbuffered water. The
established, EDTA titrations of calcium must be performed
56 titration curves were nearly identical, with sharp transitions
above pH 10.
57 at the endpoint. Titrations with EDTA did not show a
58
59
60 4
 Page 5 of 10 Analyst
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3 The chelating nanospheres exhibit attractive versatility. By
4 simply replacing the ionophore in the nanospheres, it is
5 possible to create a palette of reagents of different
6 selectivity. A potential limitation is that the nanospheres
7 tend to coagulate at high concentration, resulting in
8 undesirable light scattering if the end point is observed by
9 optical methods. This method is still young and emulsion
10 based titrations for monovalent metal ions such as Na+ and
11 K+ still wait to be demonstrated.
12
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13
14 3. Coulometric titration
15 Coulometry conversion has also been considered to directly Fig. 6 The analytical reverse reaction between squaraine
16 generate titrants for the purpose of effecting titrations. This derivatives and Hg2+.
17 principle works on the basis of Faraday’s law, which

Analyst Accepted Manuscript

thin layer gap. Calcium EDTA titration was demonstrated
18 defines a direct relationship between the charge passing
in the range of 0.25−0.75 mM with a precision of 3%,
19 through the electrode and the molar amount of analyte that
whereas the coulometric readout gave a range of 0.02−0.12
20 has reacted. A quantitative release of titrant can be
mM and a precision of 2%. This method requires only a
21 achieved through precise manipulation of current and
very small amount of sample and is suitable for in-situ
22 duration, if there are no cross-reactions.
measurements. Moreover, it is potentially calibration free
23 Reilley and Porterfield reported earlier on a general method
owing to the coulometric mode. For the latter to be true, the
24 for the coulometric generation of EDTA, where EDTA was
selectivity of the ionophore has to be high (which it
25 indirectly released upon the reduction of mercuric-EDTA.
normally is) and any non-Faradaic processes during the
26 This method has been successfully demonstrated to
coulometric release must be negligible.
27 measure calcium, copper, zinc and lead ions.12 By applying
28 an appropriate potential or current, the direct release of
29 non-redox active ions was demonstrated. The released ions 4. Indicators
30 served directly as the titrant and were also used determine 4.1 Metallochromic dye based indicators
31 the end point of the titration. Compared with traditional An end point detection by the naked eye is sometimes
32 volumetric titrations, this method is able to accurately thought to be the most convenient way to visualize the
33 release the titrant without requiring standardized stock titration end point, and metallochromic indicators can be
34 solution. In addition, the sample is not diluted during the applied for this purpose. As an indicator, it should also
35 titration and the sample volume can remain quite limited. fulfill key criteria that include a high sensitivity to exhibit a
36 However, the lack of selectivity and limited options of drastic change at the end point, a high selectivity to obtain
37 reagents are still to be improved, and the use of mercury is accurate results, and a sufficiently stable complex with the
38 today often no longer acceptable. metal of interest. In the early days, Murexide and
39 More recently, the introduction by our group of ion Eriochrome Black T were the classical dye indicators for
40 selective membranes to achieve coulometric titration has Ca2+, Mg2+ as well as other metal ions.4, 6, 7 However, these
41 aimed to overcome the above mentioned limitations.69 dyes are not very selective and can only be used in a
42 Calcium and barium ions (titrant ions) were chosen as narrow window of pH.
43 initial examples. When a constant current and duration was The development of colorimetric/fluorescent metal sensors
44 applied across the ion selective membrane, Ca2+ or Ba2+ has gradually drawn people’s attention to their wider
45 was released from the membrane to the sample solution applications in biochemical and environmental sciences.
46 with high selectivity and accuracy. A second ion selective With the pursuit of high selective and sensitive metal ion
47 sensors, a great number of specially designed molecules
electrode was used as end point indicator. As above, the
48 have emerged where many proved to be good candidates as
amount of released titrant ions could be accurately
49 indicators for complexometric titration. The
calculated using Faraday’s law.
50 fluorophore/chromophore was usually modified with metal
Bakker and co-workers subsequently introduced the
51 chelating groups and the metal binding would induce a
concept of thin layer coulometric titration (Fig. 5).70 By
52 change of the optical signal. Rhodamine, porphyrin,
applying a constant potential, Ca2+ was electrochemically
53 BODIPY ， fluorescein, spiropyran, coumarin, dansyl
injected into a thin sample layer through transport across a
54
calcium selective membrane (calcium pump). The free derivatives and many others have been used as the
55
calcium ion activity in the thin layer was measured by fluorophore/chromophore71-76 and dipicolylamine,
56
another set of potentiometric ion-selective electrodes glucosamine, Gly-His as receptor.74, 77, 78 pH response
57
placed opposite the pumping electrode, only spaced by the sometimes accompanies the metal response because of
58
59
60 5
 Analyst Page 6 of 10
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3 protonatable groups on the fluorophore/chromophore and its deprotonated state. To serve as an indicator, the metal
4 also on the metal chelating groups. indicator complex should generally be 10 to 100 times less
5 Mártinez-Máñez, Rurack and co-workers designed a series stable than the metal chelator complex so that the chelator
6 of metal triggered dyes formation system for the highly can effectively displace the metal ion from the indicator
7 selective determination of Hg2+.79 The squaraine complex. This is effectively achieved here with the same
8 derivatives were passivated first by a chemical addition chelator/ionophore, since the effective affinity between
9 reaction with thiols (spectroscopic inhibitor) that switch off
10 the colorimetric and fluorescence properties of the indicator.
11 When the target ions are present, they react with the thiols
12 and release the indicator to induce the optical signal
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13
recovery shown in Fig. 6. From the passivated state
14
(colorless) to activated state (blue), the indicators generated
15
dramatic changes in the optical signals, thereby improving
16
sensitivity. The color change was observed within a few
17

Analyst Accepted Manuscript

seconds, making the compounds potential indicators for the
18
titration of mercury ions. Fluorescent titration demonstrated
19
that the indicator may detect less than 2 ppb of Hg2+ in
20
solution. The indicator with a hydrophobic side chain was
21
adsorbed onto powered silica and subsequently coated onto
22
23 a polyethyleneterephthalate film. This film served to
24 analyze Hg2+ and could easily be reused after washing with
25 the thiol inhibitor.
26 Reymond and co-workers reported new types of fluorescent
27 sensors for Cu2+ which are pH independent and exhibit a
28 high selectivity.80 These sensors were quinacridone
29 (fluorophore) derivatives functionalized with a
30 ethylenediamine group (binding site). The
31 ethylenediamines were attached to each nitrogen atom of
32 the symmetrical quinacridone via a linker. The presence of
33 Cu2+ induced the quenching of the fluorescence by
34 formation of the macrocyclic metal-chelator complex,
35 which brought the complex and fluorophore closer to each
36 other. To overcome the pH dependence, the authors
37 modified the chemical structures of the indicator. By
38 making the linker sufficiently long, the
39 protonation/deprotonation of the chelating groups was
40 found to no longer influence the fluorescence of the
41 fluorophore. At the same time, the long linker did not affect
42 the ability for the metal to coordinate with the two
43 ethylenediamines groups and to cause fluorescence
44 quenching. The sensors showed good selectivity to copper Fig. 7 Schematic illustration of ion selective
45 and were independent of pH in the range from 2 to 10. The nanospheres as chelators and indicators in the
46 fluorescent titration showed a 1 to 1 stoichiometry of the complexometric titration of calcium. Chelating
47 complex. nanospheres contain calcium ionophore II and cation
48 Recently, our group has introduced emulsion based ion exchanger. Indicating nanosphere contain calcium
49 exchange nanospheres (discussed above) to serve also as ionophore IV, cation exchanger and solvatochromic
50 optical indicators for complexometric titrations.65 The dye SD.
51 indicating nanospheres contained a lipophilic pH sensitive
52 metal and receptor is dictated by ion-exchange, which is
dye (chromoionophore), an ion exchanger and an ionophore.
53 weakened by the presence of the lipophilic pH indicator.
For cationic analytes, the working principle of the
54 The chromoionophore-based indicating nanospheres may
indicating nanospheres is the exchange between the analyte
55 work in a wide pH range and even at very acidic pH.
and the H+ released from the chromoionophore. The color
56 Unfortunately, however, the transitions become rather
of the indicating nanospheres changes because the
57 difficult to identify with increasing pH because the
chromoionophore transitions from the protonated state to
58
59
60 6
 Page 7 of 10 Analyst
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3 chromoionophore becomes more easily deprotonated at Daniele and co-workers used amperometry to detect the
4 high pH. endpoint of Ca2+ and Mg2+ titrations with EDTA.82 A
5 To overcome this pH dependence, cationic solvatochromic platinum disc microelectrode was used to reduce the H+ to
6 dye based indicating nanospheres were recently introduced. H2 in nonbuffered sample solutions. A second wave in the
7 The solvatochromic dye is not sensitive to pH and changes linear scan voltammogram was observed after the end point
8 color with the solvent environment.64 In the emulsion based due to excess EDTA. The precision of the method was
9 titration, a large amount of the chelating nanospheres and a found to be satisfactory, the relative standard deviation
10 much smaller amount of indicating nanospheres are mixed being not larger than 2% for at least three replicates.
11 together, and the sample solution was gradually added. Besides electrochemical methods, thermometric titration
12 Here, the indicating nanospheres only function as indicator has been found to be an attractive universal method that
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13
to show the color change at the endpoint. Only when the measures the change of temperature with the added volume
14
chelating nanospheres become saturated at the endpoint, the of titrant during a volumetric titration.83 Since heat change
15
cationic solvatochromic dye in the indicating nanopheres is one of the general characteristic of most reactions, a
16
will be exchanged from the nanosphere core to the outside thermometric titration is suited for a wide range of
17

Analyst Accepted Manuscript

solution by the analyte (Fig. 7). Owing to the different
18
polarity between nanosphere core and aqueous solution, the (a)
19
color of the solution will change. Essentially the same
20
sharp titration curves were obtained at pH 5.5 and pH 9,
21
which suggest that this concept can efficiently overcome
22
23 the challenge of pH dependence in such titrations (Fig. 8).
24
25 4.2 Instrument-based indicators
26 While the metallochromic indicators discussed above can
27 directly visualize the end point by a color change,
28 sometimes appropriate indicating dyes are not easily found
29 and instrumental methods are needed to identify the end
30 point.
31 Potentiometry by ion selective electrodes is likely the most
widely applied electrochemical indicator. An ion selective Fig. 8 Optical reverse titration curves for calcium
32
electrode can not only measure the activity of the ions but using solvatochromic dye SD based optical
33
also act as end point indicator to visualize the so-called free nanospheres as indicator at the indicated pH values.
34
activity of the analyte. In potentiometry, the observed End point indicator: SD-based and Ca2+-selective; pH
35
36 signal, the electromotive force, is related to the analyte 9.0: 10-3 M tris-H2SO4; pH 5.5: 10-3M MES-NaOH;
37 activity according to the Nernst equation. With the The dashed vertical line indicates the expected end
38 appropriate selectivity, a large change in the signal is point. Fitting parameters: VT = 2 ml, [TFPB-]is =
39 usually observed at the end point. 3.29×10-6.15 M, Vis = 1.5 µL, [TFPB-]cs =2.35×10-5
40 Pretsch and co-workers suggested ways to improve the M, Vcs = 8 µL, [Ca2+]titrant = 10-3 M,
41 detection limits and sensitivity of potentiometric titrations,
=10-7 M, =10-5.5 M.
42 with lead(II) as example.81 By changing the sensing
43 components of the membrane and adjusting the flux of the
reactions such as acid/base, precipitation, coordination and
44 primary ion, the detection limit of the titration could be
45 redox titrations.83-87 Some factors such as light scattering,
improved by several orders of magnitude, and lower
46 absorption inducing surface blocking, color change or
concentrations of lead could be titrated. To obtain lower
47 overlay, will influence the optical or potentiometric signal
detection limits of the lead selective electrode, a metal
48 but have no significant influence on a thermometric
chelator, nitrilotriacetic acid (NTA) or EDTA, acting as a
49 titration. Thermometric titration has been successfully
metal buffer at low concentrations was added to the inner
50 applied to monitor sulfate, total alkalinity, chlorinity in
solution of the ion selective electrode to keep the
51 seawater,85 and a wide range of metal ions such Ca2+, Mg2+,
concentration of lead low and stable. In this particular case,
52 Fe2+, Pb2+.83, 84, 87 The first thermometric titration research
the net Pb2+ flux was directed from the sample to the inner
53 work was on acid and base neutralization and reported by
solution. With a significant inward flux, the ion selective
54 Bell and Cowell in 1913.88 Jordan successfully applied
electrode may show a super-Nernstian response slope to the
55 thermometric titration to estimate the heat produced by the
analyte, thereby improving the sensitivity of the titration
56 complex formed between divalent metal ions and EDTA.89
quite dramatically.
57 For example, both Ca2+ and Mg2+ could be determined by
58 thermometric titration even if the formation constants differ
59
60 7
 Analyst Page 8 of 10
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3 by less than 2 orders of magnitude.84, 87 By comparison, the same result as volumetric titrations, but without the hassle
4 metallochromic indicator Eriochrome Black T is not of sampling, splitting into aliquots, standardization of
5 sufficiently selective to separate Ca2+ and Mg2+ at the same reagents, and volumetric delivery are potentially very
6 time and require masking reagents or pH control.4, 6 In attractive for a range of applications.
7 addition, different dynamic characteristics may also help A number of the receptors or ionophores have been
8 separate the analytes by monitoring their release of heat in published and applied in different fields. However, there
9 order to obtain a higher selectivity. A key disadvantage of are not sufficient receptors of quality for anions, and highly
10 this method is the relatively high detection limit compared selective and pH independent receptors are still very much
11 with other instrumental indicators. The reason is that one needed.
12 requires a sufficiently large quantity of reaction substrate to Universal methods are also very popular, such as
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13
observe a detectable temperature variance. The thermometric titrations, which measure the temperature
14
development of dedicated instruments encouraged the change to indicate the end point. It is suitable to almost all
15
widespread use of this method. Recently, Barin and co- reactions and not limited to complexation, but to obtain
16
workers introduced a very simple and inexpensive setup for observable measurable temperature changes, it requires
17

Analyst Accepted Manuscript

simultaneous enthalpimetric analysis by using an infrared relatively large amount of substrates.
18
camera as detector to monitor the temperature and
19
disposable microplates to process the enthalpimetric ASSOCIATED CONTENT
20
analysis.90 The noncontact and nondestructive infrared AUTHOR INFORMATION
21 Corresponding Author
thermal imaging technique provided rapid signal
22 *
E-mail : eric.bakker@unige.ch
acquisition with very good quantitative results. Even if
23 Notes
24 some limitations for this new method remain, such as not The authors declare no competing financial interest.
25 being suitable for low reaction rate reactions, it still shows
ACKNOWLEDGMENTS
26 potential to be applied in a range of important applications. The authors thank the Swiss National Science Foundation
27 (SNSF) and the University of Geneva for financial support.
28 5. Conclusion Jingying Zhai gratefully acknowledges the support by the
29 The introduction of EDTA has made complexometric China Scholarship Council.
30 titrations a well known analytical technique. However, the
31 drawbacks of EDTA have until recently not been overcome. REFERENCES
32 As summarized in this work, the advent of new titration 1. J. Liebig, Justus Liebigs Ann. Chem., 1851, 77,
33 reagents and end point indicators have injected new vitality 102-105.
34 into the field and also raised more issues to be resolved in 2. G. Schwarzenbach, E. Kampitsch and R. Steiner,
35 further work. Helv. Chim. Acta., 1945, 28, 1133-1143.
36 3. M. C. Yappert and D. B. DuPré, J. Chem. Educ.,
Recent new concepts for chelators and indicators based on
1997, 74, 1422.
37 heterogeneous reactions involving emulsion based reagents 4. G. Schwarzenbach and H. Flaschka,
38 show very promising characteristics for complexometric Complexometric titrations, London, 1969.
39 titration. Here, ion selective nanospheres really are 5. H. Ackermann, J. E. Prue and G. Schwarzenbach,
40 multicomponent nanoscale solvent reactors that act in Nature, 1949, 163, 723-724.
41 analogy to traditional chelators and indicators and extend 6. H. J. Gitelman, C. Hurt and L. Lutwak, Anal.
42 the usage of lipophilic ionophores and other non-water-
Biochem., 1966, 14, 106-120.
43 7. R. A. C. Lima, S. R. B. Santos, R. S. Costa, G. P.
soluble compounds. This approach exhibits a high S. Marcone, R. S. Honorato, V. B. Nascimento
44 selectivity and sensitivity to a range of analytes, does not and M. C. U. Araujo, Anal. Chim. Acta, 2004,
45 have to limit itself to a unique 1:1 complex stoichiometry, 518, 25-30.
46 and is largely pH independent. Simply changing one or 8. J. Patton and W. Reeder, Anal. Chem., 1956, 28,
47 1026-1028.
more of the components in the droplets, the nanospheres
48 9. G. P. Hildebrand and C. N. Reilley, Anal. Chem.,
can be extended to other ions. This concept is highly
49 1957, 29, 258-264.
suitable for titrating low concentrations of analyte. High
50 10. R. Y. Tsien, Biochemistry, 1980, 19, 2396-2404.
analyte concentrations are more problematic, since 11. G. Grynkiewicz, M. Poenie and R. Y. Tsien, J.
51
nanosphere coagulation normally results in light scattering Biol. Chem., 1985, 260, 3440-3450.
52
that interferes with an optical endpoint detection. 12. C. N. Reilley and W. W. Porterfield, Anal. Chem.,
53
Thin layer coulometric titrations based on ion selective 1956, 28, 443-447.
54 13. C. N. Reilley and R. W. Schmid, Anal. Chem.,
membranes consume only very small amount of sample.
55 1958, 30, 947-953.
With a highly selective release of the titrant, only the
56 14. J. T. Stock, Anal. Chem., 1976, 48, 1R-9R.
analyte of interest is consumed or converted, which is very
57 15. C. N. Reilley, Anal. Chem., 1960, 32, 185-193.
58 attractive for in situ analysis. Direct probes that give the
59
60 8
 Page 9 of 10 Analyst
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3 16. K. R. Williams, V. Y. Young and B. J. Killian, J. 45. C. Chen, T. Tian, M. K. Wang and G. Wang,
4 Chem. Educ., 2011, 88, 315-316. Geoderma, 2016, 275, 74-81.
5 17. K. Granholm, T. Sokalski, A. Lewenstam and A. 46. M. Bucheli-Witschel and T. Egli, FEMS
6 Ivaska, Anal. Chim. Acta, 2015, 888, 36-43. Microbiol. Rev., 2001, 25, 69-106.
7 18. Y. Ni and Y. Wu, Anal. Chim. Acta, 1997, 354, 47. B. Nowack, Environ. Sci. Technol., 2002, 36,
8 233-240. 4009-4016.
19. D. Wilson, S. Alegret and M. d. Valle, 48. M. A. Rahman, M.-S. Won and Y.-B. Shim, Anal.
9
Electroanal., 2015, 27, 336-342. Chem., 2003, 75, 1123-1129.
10 20. M. D. DeGrandpre, T. R. Martz, R. D. Hart, D. M. 49. M. E. Sears, Scientific World J., 2013, 2013, 1-
11 Elison, A. Zhang and A. G. Bahnson, Anal. 13.
12 Chem., 2011, 83, 9217-9220. 50. S. A. Ansari, P. Pathak, P. K. Mohapatra and V.
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13 21. C. Maccà, L. Soldà and M. Zancato, Anal. Chim. K. Manchanda, Chem. Rev., 2012, 112, 1751–
14 Acta, 2002, 470, 277-288. 1772.
15 22. C. N. Reilley and A. Vavoulis, Anal. Chem., 51. S. A. Ansari, P. Pathak, P. K. Mohapatra and V.
16 1959, 31, 243-248. K. Manchanda, Sep. Purif. Rev., 2011, 40, 43-76.
23. C. N. Reilley, R. W. Schmid and F. S. Sadek, J. 52. H. Narita, T. Yaita, K. Tamura and S. Tachimori,
17

Analyst Accepted Manuscript

Chem. Educ., 1959, 36, 555-564. J. Radioanal. Nucl. Chem., 1999, 239, 381-384.
18 24. A. E. Martell and S. Chaberek, Anal. Chem., 53. Y. Sasaki, H. Suzuki, Y. Sugo, T. Kimura and G.
19 1954, 26, 1692-1696. R. Choppin, Chem. Lett. , 2006, 35, 256-257.
20 25. E. J. Wheelwright, F. H. Spedding and G. 54. Y. Sasaki, Y. Sugo, S. Suzuki and S. Tachimori,
21 Schwarzenbach, J. Am. Chem. Soc., 1953, 75, Solvent Extr. Ion Exch., 2001, 19, 91-103.
22 4196-4201. 55. G. Modolo, H. Asp, C. Schreinemachers and H.
23 26. G. Schwarzenbach and E. Freitag, Helv. Chim. Vijgen, Solvent Extr. Ion Exch., 2007, 25, 703-
24 Acta., 1951, 34, 1503-1508. 721.
27. G. Schwarzenbach, W. Biedermann and F. 56. S. A. Ansari, P. N. Pathak, V. K. Manchanda, M.
25 Bangerter, Helv. Chim. Acta., 1946, 29, 811-818. Husain, A. K. Prasad and V. S. Parmar, Solvent
26 28. G. Schwarzenbach, Helv. Chim. Acta., 1946, 29, Extr. Ion Exch., 2005, 23, 463-479.
27 1338. 57. H. Suzuki, H. Naganawa and S. Tachimori, Phys.
28 29. Y. Ni and Z. Peng, Anal. Chim. Acta, 1995, 304, Chem. Chem. Phys., 2003, 5, 726-733.
29 217-222. 58. K. Shimojo, K. Kurahashi and H. Naganawa,
30 30. C. Maccà, L. Soldà, G. Favaro and P. Pastore, Dalton Trans., 2008, 5083-5088.
31 Talanta, 2007, 72, 655-662. 59. P. K. Mohapatra, Chem. Prod. Process Model.,
32 31. J. Gao, Y. Guo, S. Wang, T. Deng, Y.-W. Chen 2015, 10, 135-145.
and N. Belzile, J. Chem., 2013, 2013, 1-4. 60. P. K. Mohapatra, A. Sengupta, M. Iqbal, J.
33 32. W. Xian-ke, Analyst, 1990, 115, 1611-1612. Huskens and W. Verboom, Chem. Eur. J., 2013,
34 33. M. Vlasák, Z. Luxemburková, V. Sychra and M. 19, 3230-3238.
35 Suchánek, Accred. Qual. Assur. , 2013, 18, 491- 61. A. Sengupta, P. K. Mohapatra, R. M. Kadam, D.
36 499. Manna, T. K. Ghanty, M. Iqbal, J. Huskens and
37 34. S. G. Novick, J. Chem. Educ., 1997, 74, 1463. W. Verboom, RSC Adv., 2014, 4, 46613-46623.
38 35. S.-P. Yang and R.-Y. Tsai, J. Chem. Educ., 2006, 62. D. Jańczewski, D. N. Reinhoudt, W. Verboom, C.
39 83, 906. Hill, C. Allignol and M.-T. Duchesne, New J.
36. M. Romero, V. Guidi, A. Ibarrolaza and C. Chem., 2008, 32, 490-495.
40
Castells, J. Chem. Educ., 2009, 86, 1091. 63. J. Zhai, X. Xie and E. Bakker, Chem. Commun.,
41 37. P.-L. Fabre and O. Reynes, J. Chem. Educ., 2010, 2014, 50, 12659-12661.
42 87, 836-837. 64. J. Zhai, X. Xie and E. Bakker, Anal. Chem., 2015,
43 38. J. S. Fritz, J. P. Sickafoose and M. A. Schmitt, 87, 12318-12323.
44 Anal. Chem., 1969, 41, 1954-1958. 65. J. Zhai, X. Xie and E. Bakker, Anal. Chem., 2015,
45 39. M. C. Yappert and D. B. DuPré, J. Chem. Educ., 87, 2827-2831.
46 1997, 74, 1422-1423. 66. J. D. Rodriguez, T. D. Vaden and J. M. Lisy, J.
47 40. H. V. Malmstadt and T. P. Hadjiioannou, Anal. Am. Chem. Soc., 2009, 131, 17277-17285.
Chim. Acta, 1958, 19, 563-569. 67. E. Bakker, P. Bühlmann and E. Pretsch, Chem.
48
41. P. G. McCormick, J. Chem. Educ., 1973, 50, Rev., 1997, 97, 3083-3132.
49 136-137. 68. P. Bühlmann, E. Pretsch and E. Bakker, Chem.
50 42. S. Karita and T. Kaneta, Anal. Chim. Acta, 2016, Rev., 1998, 98, 1593-1688.
51 924, 60-67. 69. V. Bhakthavatsalam, A. Shvarev and E. Bakker,
52 43. F. M. Koehler, M. Rossier, M. Waelle, E. K. Analyst, 2006, 131, 895-900.
53 Athanassiou, L. K. Limbach, R. N. Grass, D. 70. M. G. Afshar, G. A. Crespo and E. Bakker, Anal.
54 Günther and W. J. Stark, Chem. Commun., 2009, Chem., 2015, 87, 10125-10130.
55 4862-4864. 71. D. B. Papkovsky, G. V. Ponomarev and O. S.
44. E. Vassileva, B. Varimezova and K. Hadjiivanov, Wolfbeis, Spectrochim. Acta, Part A 1996, 52,
56
Anal. Chim. Acta, 1996, 336, 141-150. 1629-1638.
57
58
59
60 9
 Analyst Page 10 of 10
View Article Online
DOI: 10.1039/C6AN00538A

1
2
3 72. R. Sheng, P. Wang, Y. Gao, Y. Wu, W. Liu, J. 81. S. Peper, A. Ceresa, E. Bakker and E. Pretsch,
4 Ma, H. Li and S. Wu, Org. Lett., 2008, 10, 5015- Anal. Chem., 2001, 73, 3768-3775.
5 5018. 82. M. A. Baldo, S. Daniele, C. Bragato and G. A.
6 73. S. B. Maity, S. Banerjee, K. Sunwoo, J. S. Kim Mazzocchin, Analyst, 1999, 124, 1059-1063.
7 and P. K. Bharadwaj, Inorg. Chem., 2015, 54, 83. S. T. Zenchelsky, Anal. Chem., 1960, 32, 289-
8 3929-3936. 292.
74. Y. Zheng, X. Cao, J. Orbulescu, V. Konka, F. M. 84. R. H. Callicott and P. W. Carr, Clin. Chem., 1976,
9
Andreopoulos, S. M. Pham and R. M. Leblanc, 22, 1084-1088.
10 Anal. Chem., 2003, 75, 1706-1712. 85. F. J. Millero, S. R. Schrager and L. D. Hansen,
11 75. R. D. Hancock, Chem. Soc. Rev., 2013, 42, Limnol. Oceanogr., 1974, 19, 711-715.
12 1500-1524. 86. W. L. Everson, Anal. Chem., 1971, 43, 201-205.
Published on 27 May 2016. Downloaded by University of Sussex on 27/05/2016 19:56:49.

13 76. J. D. Winkler, K. Deshayes and B. Shao, J. Am. 87. H. Yoshida, T. Hattori, H. Arai and M. Taga,
14 Chem. Soc., 1989, 111, 769-770. Anal. Chim. Acta, 1983, 152, 257-263.
15 77. J. Hatai and S. Bandyopadhyay, Chem. Commun., 88. J. M. Bell and C. F. Cowell, J. Am. Chem. Soc.,
16 2014, 50, 64-66. 1913, 35, 49-54.
78. A. Mitra, A. K. Mittal and C. P. Rao, Chem. 89. J. Jordan and T. G. Alleman, Anal. Chem., 1957,
17

Analyst Accepted Manuscript

Commun., 2011, 47, 2565-2567. 29, 9-13.
18 79. J. V. Ros-Lis, M. D. Marcos, R. Mártinez-Máñez, 90. J. S. Barin, B. Tischer, A. S. Oliveira, R. Wagner,
19 K. Rurack and J. Soto, Angew. Chem. Int. Ed., A. B. Costa and E. M. M. Flores, Anal. Chem.,
20 2005, 44, 4405-4407. 2015, 87, 12065-12070.
21 80. G. Klein, D. Kaufmann, S. Schürch and J.-L.
22 Reymond, Chem. Commun., 2001, 561-562.
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60 10