Author's personal copy

Sensors and Actuators B 145 (2010) 32–38

Contents lists available at ScienceDirect

Sensors and Actuators B: Chemical

journal homepage: www.elsevier.com/locate/snb

A chemosensor for heavy-transition metal ions in mixed

aqueous–organic media
Amal Kumar Mandal, Moorthy Suresh, E. Suresh,
Sanjiv K. Mishra, S. Mishra, Amitava Das ∗
Central Salt & Marine Chemicals Research Institute (CSIR), Bhavnagar 364002, Gujarat, India

a r t i c l e i n f o a b s t r a c t

Article history: A new chemosensor (L1 ) is synthesized and its spectral response was screened with most alkali, alkaline
Received 9 July 2009 earth, transition and lanthanide metal ions in THF–water (2:3, v/v, pH 7.2, 20 mM HEPES buffer) medium.
Received in revised form 29 October 2009 This is found to bind four different transition metal ions such as Hg2+ , Cu2+ , Ag+ and Ni2+ in mixed aqueous
Accepted 6 November 2009
solution at pH 7.2; while binding affinity towards the Hg2+ is found to be an order of magnitude higher
Available online 13 November 2009
as compared to other three cations mentioned. Experimental results reveal that the L1 could be used for
colorimetric detection of Hg2+ , present in the lower organism like Pseudomonas putida.
Keywords:
© 2009 Elsevier B.V. All rights reserved.
Chemosensor
Pyrazole
Mercury
Fluorescence quenching

1. Introduction Fluorescence quenching by heavy metal ions is a well-

documented phenomenon and occurs by a number of pathways
Studies parenting to recognition and selective binding of desired that include spin–orbit coupling, energy transfer, and elec-
metal ions by tailored made designer receptors have gained impor- tron transfer [4]. For Hg2+ ion, efficient spin–orbit coupling is
tance over last few decades. In recent time, these studies have the most common one and forms the basis of the develop-
gained more significance owing to the awareness of the involve- ment of the fluorophore emission-based turn-off detection. Some
ment of certain metal ions in health hazards and carcinogenic activ- recent references illustrate such examples [5], where various
ities. Among various metal ions, recognition and sensing of Hg2+ ion derivatives of bipyridyls or polypyrroles are used. Intricate syn-
in aqueous media is one of the major targets for researchers active thetic methodologies of such receptors and non-aqueous medium
in this area of research for its obvious health related hazards [1–3]. limit their use in practice. More recent reports, which describe
Traditional quantitative approaches to Hg(II) analysis in water sam- receptor molecules that can be used either in mixed aque-
ples generally employ a number of analytical techniques and many ous/organic medium or in pure aqueous solution are relatively
of these methods require complicated, multistep sample prepa- fewer [6].
ration and/or sophisticated instrumentation. These instrumental However, in most such instances receptor molecules are derived
techniques often provide direct and quantitative information about from various macrocycles or polyenes and generally involve multi-
Hg(II) concentrations, but are not well-suited for quick detection step synthetic procedures—barring two examples where relatively
of Hg(II) in the field or for in vivo studies of Hg(II) in biological easily synthesizable receptors are used for recognition studies
samples. All these led to the search for the mercury-responsive [6h,n]. This led us to synthesize a easily synthesizable pyrazole-
small-molecule ligands that could provide immediate optical feed- based receptor molecule (L1 ) having an anthracenyl unit as the
back and can overcome such limitations. Owing to its closed-shell signalling moiety, that could bind preferentially to Hg2+ in mixed
d10 configuration, Hg2+ ion has no optical spectroscopic signature. aqueous/organic medium and be use for in vivo detection of this
Thus, optical detection, following changes in solution fluorescence ion in Gram−ve bacteria.
or UV–vis spectroscopy resulting from a Hg2+ binding induced per- In this present article, we have reported a new sensor molecule
turbation of a chromophore, is commonly used for monitoring Hg2+ (L1 ) that has an anthracenyl unit as the signalling moiety, cova-
in either environmental or biological contexts. lently coupled to an N–N donor receptor fragment. Fluorescence
based emission read-out signal of the anthracenyl unit is governed
by different photoinduced electron transfer (PET) processes in the
∗ Corresponding author. Tel.: +91 278 2567760; fax: +91 278 2567562. absence and presence of the coordinated metal ion. Experimen-
E-mail address: [email protected] (A. Das). tal studies revealed that pyridyl pyrazole, used as the receptor

0925-4005/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.snb.2009.11.016
 Author's personal copy

A.K. Mandal et al. / Sensors and Actuators B 145 (2010) 32–38 33

fragment for Hg2+ showed a higher affinity for this ion in aque- (1H-pyrazole-3yl)-pyridine (Fig. 1). 2-(1H-pyrazole-3yl)-pyridine
ous/organic media. (0.557 g, 0.0038 mol) was dissolved in 50 ml of THF and was
taken in a 100 ml two neck round bottom flask. To this 6.63 ml
2. Experimental of 10 M NaOH solutions and 9-chloromethyl anthracene (1.13 g,
0.00498 mol) were added. Then the reaction mixture was stirred
2.1. Materials and methods for 30 h at ∼65 ◦ C. After this, the solvent was removed under
reduced pressure. Then desired compound was extracted in CHCl3
9-chloromethyl anthracene, Hg(ClO4 )2 ·xH2 O, Cu(ClO4 )2 ·6H2 O, layer from an aqueous suspension and this process was repeated
Ni(ClO4 )2 ·5H2 O, Ag(ClO4 )·xH2 O, Mg(ClO4 )2 , Ca(ClO4 )2 ·4H2 O, thrice. Then all organic fractions were collected and dried over
Sr(ClO4 )2 ·xH2 O, Li(ClO4 ), Na(ClO4 ), K(ClO4 ), Cs(ClO4 ), Na2 SO4 . The crude product was obtained by removing solvents
Fe(ClO4 )3 ·xH2 O, Cr(ClO4 )3 ·6H2 O, Co(ClO4 )2 ·6H2 O, Zn(ClO4 )2 ·6H2 O, under reduced pressure. L1 with desired purity was achieved
Ba(ClO4 )2 ·xH2 O, Cd(ClO4 )2 ·xH2 O, Eu(NO3 )3 ·5H2 O, through crystallization from hot methanol (Yield: 0.629 g (49%)).
Er(NO3 )3 ·5H2 O, Yb(NO3 )3 ·5H2 O, La(OCOCH3 )3 ·H2 O, Pr(NO3 )3 · 1 H NMR (CD Cl , ı ppm): 8.57 (1H, s), 8.55 (1H, d, J = 7 Hz), 8.39
2 2
6H2 O, Nd(NO3 )3 ·6H2 O, Tb(NO3 )3 ·5H2 O, Ce(NO3 )3 ·6H2 O, were (2H, d, J = 9 Hz), 8.08 (2H, d, J = 8.5 Hz), 8.03 (1H, d, J = 8 Hz), 7.72
obtained from Sigma–Aldrich and were used as received. All the (1H, t, J = 7.5 Hz), 7.58 (2H, t, J = 6.5 Hz), 7.51 (2H, t, J = 7.5 Hz), 7.18
other reagents used were procured from S.D. fine chemical, India. (1H, t, J = 6 Hz), 7.09 (1H, d, J = 6.5 Hz), 6.72 (1H, d, J = 6.5 Hz), 6.35
2-(1H-pyrazole-3yl)-pyridine was synthesized following a known (2H, s). IR (KBr, cm−1 ): 3644, 3381, 3107, 3048, 2356, 1589, 1447,
procedure [7a]. All solvents were dried using standard procedure 1317, 1216, and 1043. Es-Ms (m/z): 335.94 (M+ ). The structure
prior to use. of L1 was further established by single crystal X-ray data (Fig. 1)
[12].
2.2. Instrumentation

Electronic spectra were recorded using a Shimadzu UV-3101 2.4. Spectrophotometric titrations
PC or a Cary 500 Scan UV–Vis–NIR spectrometer; while steady-
state luminescence spectra and time resolved fluorescence decay A stock solution of L1 with the concentration of 1.07 × 10−4 M
experiments were carried out with a HORIBA JOBIN YVON spec- was prepared in THF–water (2:3, v/v) mixed solvent and was stored
trofluorometer. 1 H NMR spectra were recorded on a Bruker in dark. This stock solution was used for all spectrophotometric and
500 MHz FT NMR (model: Avance-DPX 200) using CD2 Cl2 as the spectrofluoremetric titrations after appropriate dilution. Similarly,
solvent and tetramethylsilane (TMS) as an internal standard. X-ray 1.0 × 10−3 M solutions of individual metal ions were also prepared
single crystal data for L1 was collected using Mo K␣ ( = 0.7107 Å) in THF–water (2:3, v/v) mixed solvent. The fluorescence titration
radiation on a SMART APEX diffractometer equipped with a CCD was carried out using the above referred solution after maintaining
area detector. The data integration and reduction were processed the pH of the solution to 7.2 using 20 mM HEPES buffer. For fluores-
with SAINT [8] software. An empirical absorption correction was cence studies samples were excited at 367 nm with an excitation
applied to the collected reflections with SADABS [9]. The struc- and emission slit width of 1 nm. Binding constants are evaluated
tures were solved by direct methods using SHELXTL [10] and were based on the binding induced changes in fluorescence intensity at
refined on F2 by the full-matrix least-squares technique using the 416 nm using the standard equation [13].
SHELXL-97 [11] program. All hydrogen atoms were geometrically
fixed at their idealized position. The FT-IR spectra were recorded 3. Result and discussion
using PerkinElmer Spectra GX 2000 spectrometer.
To demonstrate the sensing nature of the newly synthesized
2.3. Synthesis chemosensor L1 , we carried out absorption and fluorescence study
in an optimized THF–water (2:3, v/v) mixed solvent medium.
L1 (2-(1-(anthracen-9-ylmethyl)-1H-pyrazol-3-yl)-pyridine) Absorption spectrum for 2-(1H-pyrazole-3yl)-pyridine, recorded
was synthesized by reacting 9-chloromethyl anthracene and 2- in THF–water (2:3, v/v) mixed solvent medium reveals two strong

Fig. 1. (a) Synthetic route of L1 ; (b) ORTEP diagram of the L1 (40% probability factor for the thermal ellipsoids).
 Author's personal copy

34 A.K. Mandal et al. / Sensors and Actuators B 145 (2010) 32–38

Fig. 2. Changes in (a) absorption (L1 = 1.4 × 10−4 M) with varying [Hg2+ ] (0–3 × 10−4 M) and (b) fluorescence (L1 = 1.0 × 10−5 M) spectra of L1 with varying [Hg2+ ]
(0–4.5 × 10−4 M) in THF/water (2:3, v/v; 20 mM pH 7.2, HEPES buffer); exc = 367 nm.

Fig. 3. Changes in (a) absorption (L1 = 1.4 × 10−4 M) with varying [Cu2+ ] (0–7.2 × 10−4 M) and (b) fluorescence (L1 = 1.0 × 10−5 M) spectra of L1 with varying [Cu2+ ]
(0–8 × 10−3 M) in THF/water (2:3, v/v, 20 mM pH 7.2, HEPES buffer). exc = 367 nm.

bands at 246 and 279 nm. Thus, the observed absorption spectrum be 6214.3 M−1 L at abs
max of 367 nm and emission spectrum showed
of L1 (Figs. 2a, 3a and 4a) is actually a summation of the elec- an emission spectrum with typical emission maximum at 394, 416
tronic spectrum of two individual chromophores, where absorption and 440 nm, characteristic for anthracene moiety.
bands in the wavelength range of 240–290 nm are predominantly Electronic spectra recorded for L1 in the absence and pres-
due to the 2-(1H-pyrazole-3yl)-pyridine-based ␲–␲* transitions ence of twenty four different metal ions in 2:3, v/v THF–water (pH
(Supporting information) alongwith some contribution from S0 7.2, 20 mM HEPES buffer) mixed solvent media. In 2:3 THF–water
to S2 based transition involving anthracene moiety. Absorption mixed solvent media L1 was found to have two major absorption
bands in the wavelength range of 320–390 nm could be attributed bands around 367 and 280 nm. A red shifted absorption band was
to the anthracence-based ones owing to the presence of S0 –S1 evident when spectra for L1 was recorded in presence of any one of
based transition [14]. The molar absorptivity value was found to the four added metal ions like Hg2+ , Cu2+ , Ag+ or Ni2+ . Upon addition

Fig. 4. Changes in (a) absorption (L1 = 1.4 × 10−4 M) with varying [Ni2+ ] (0–4.5 × 10−3 M) and (b) fluorescence (L1 = 1.0 × 10−5 M) spectra of L1 with varying [Ni2+ ] (0–4 × 10−3 M)
in THF/water (2:3, v/v; 20 mM pH 7.2, HEPES buffer). exc = 367 nm.
 Author's personal copy

A.K. Mandal et al. / Sensors and Actuators B 145 (2010) 32–38 35

Table 1
Binding constants of L1 for Hg2+ , Cu2+ , Ag+ and Ni2+ .

Metal ions Binding constant (Kf ) M−1

Hg 2+ 11256 (±20)a,c
10876 (±20)b,c

Cu2+ 4178 (±10)a,c

4325 (±15)b,c

Ag+ 3472 (±10)a,c

5950 (±15)b,d

Ni2+ 2654 (±20)a,c

2454 (±25)b,c

Binding constants were evaluated from a fluorescence and

b
spectrophotometric titration either in c THF/water (2:3, v/v)
medium or d acetonitrile medium. Each stability constant value
reported is an average of at least five individual evaluations and
associated errors are shown in parentheses. For Ag+ , specropho-
tometric titration was carried out in CH3 CN medium (Table 1).
Fig. 5. Bar diagram represents the luminescence intensity of L1 at [2.0 × 10−6 (M)] Higher binding constant value evaluated in acetonitrile medium
for different metal salts at [8 × 10−4 (M)]. exc of 367 nm was used as the excitation is understandable if one considers that the competing aquation
wavelength. process is absent in this case.

of increasing amount of Hg2+ , Cu2+ , Ag+ and Ni2+ electronic spec- constant for Hg2+ was found to be 11256 ± 20 M−1 and was more
tral changes were observed with a characteristics isobestic points at than those for Ni2+ , Cu2+ and Ag+ (Table 1). Observed luminescence
287, 354, 372 and 391 nm, respectively (Figs. 2a, 3a and 4a). These change for Hg2+ was more appreciable than the other three metal
spectral changes signified a coordinative binding of any of these ions for a comparable concentration of the respective metal ion.
four metal ions to the pyrazole-based receptor unit in L1 . Red shift Linear detection range for Hg2+ was determined based on the fluo-
of the absorption band could be accounted based on the favoured rescence spectral studies with varying [Hg2+ ] and lowest detection
intra molecular charge transfer process [15]. We have also exam- limit was found to be 3 ppm (Supporting information).
ined the affinity of the 2-(1H-pyrazole-3yl)-pyridine towards Hg2+ , Compared to the other receptors, which work in the aque-
Cu2+ , Ni2+ and Ag+ using UV–vis spectroscopy. As it was observed in ous/organic medium, the formation constants for L1 ·Hg2+ is either
case of L1 , binding induced shifts in absorption spectra (Supporting comparable or an order of magnitude lower—while switching off of
information) is most significant for Hg2+ . This also confirms the the observed fluorescence on binding to Hg2+ ion is not a common
pyrazole–pyridine functionality as the binding site for these metal phenomena among these sensor molecules [6b–e,g–o]. However,
ions. two specific receptors, derived either from calyx[4]arene or aza-
Spectral changes were restricted mostly in the UV-region and macrocycle derivative showed much higher selectivity towards
thus binding induced changes in solution colour could not be Hg2+ [6a,f]. However, in most cases, Cu2+ and Pb2+ interfere with
detected in naked eyes; however, significant changes in lumines- the Hg2+ ion detection and in some instances complete quench-
cence intensity for L1 were observed on addition of these four metal ing of fluorescence could not be achieved. Binding constants for
ions (Figs. 2b, 3b, 4b and 5). A complete quenching of lumines- the turn-off sensors for Hg2+ , which work in pure aqueous medium
cence was observed in presence of added Hg2+ ; while only a partial are generally modest and comparable to the one we have observed
quenching was observed for Cu2+ , Ag+ , Ni2+ and Fe3+ (Fig. 5). [2j].
Addition of all other metal ions like Ca2+ , Mg2+ , Li+ , Na+ , K+ , Relative difference in the receptor (L1 )–metal ion binding affin-
Ba2+ , Sr2+ , Cs+, Fe3+ , Co2+ , Zn2+ , Cd2+ , Cr3+ , Pb2+ , Eu3+ , Er3+ , Yb3+ , ity was also evident in the relative spectral shifts in the 1 H NMR
Pr3+ , La3+ , Tb3+, and Ce3+ did not show any observable changes in spectra for L1 in the presence of comparable concentrations of Hg2+
luminescence intensity. Further, luminescence for L1 (6.9 × 10−6 M) and Ag+ (Fig. 6). Up-field shift in 1 H NMR signals for most pro-
was found to remain quenched by Hg2+ (2.0 × 10−4 M) even in tons was observed, which is not an unusual phenomena [7a,b]. As
presence of 20 molar excess of other metal ions (4.0 × 10−3 M) expected no sensible 1 H NMR spectra for L1 was observed in the
like Ca2+ , Mg2+ , Li+ , Na+ , K+ , Ba2+ , Sr2+ , Cs+ , Ni2+ , Fe3+ , Ag+ , Co2+ , presence of added Cu2+ and Ni2+ .
Zn2+ , Cd2+ , Cr3+ , Pb2+ , Eu3+ , Er3+ , Yb3+ , Pr3+ , La3+ , Tb3+ and Ce3+ Binding of the pyridyl pyrazole functionality of L1 to the respec-
(Supporting information). This indicates the possibility of the quan- tive metal ions do not actually involve the lone pair of electron
titative detection of Hg2+ in the presence of other metal ions present of the N-atom that links the anthryl unit through a –CH2 – group.
in excess. Thus, coordination to the metal ions do not actually interfere with
To evaluate the affinity of these four metal ions (Hg2+ , Cu2+ , the PET mechanism involving the electron transfer from the N-
Ag and Ni2+ ) towards receptor L1 , we have carried out system-
+ atom based HOMO to the partially filled HOMO of the excited state
atic UV–vis, fluorescence and 1 H NMR titrations studies. Respective
binding constants, evaluated for these four individual metal ions,
using these three spectroscopic techniques were actually agreed
very well and are given in Table 1. In all cases, 1:1 receptor–metal
ion binding stoichiometry was established from spectrophotomet-
ric titration profile; as well as ESI–mass spectral studies (Supporting
information).
Thus, luminescence studies corroborated our optical spectral
observation of selective binding of L1 to either one of these
(Hg2+ /Cu2+ /Ni2+ or Ag+ ) metal ions. The fluorescence quenching of
these metal ions could be explained based on the spin–orbit cou- Fig. 6. Partial 1 H NMR spectra for L1 (3.0 × 10−3 M) in CD3 CN in the absence and
pling phenomena [16]. From the luminescence study, the binding presence of Ag+ (3.0 × 10−3 M) and Hg2+ (3.0 × 10−3 M).
 Author's personal copy

36 A.K. Mandal et al. / Sensors and Actuators B 145 (2010) 32–38

Fig. 7. Fluorescence decay profile of chemosensor (L1 ) with Cu2+ , Ni2+ , Ag+ , and Hg2+
in THF/water (2:3, v/v) using exc = 340 nm and mon = 367 nm.

of the fluorophore. On the contrary, LUMO for L1 is expected to

be lower on binding to the cationic metal center and thereby the Fig. 8. Fluorescence microscope images recorded using TRITC filter at 25 ◦ C for (a)
HOMO-LUMO gap. This effect is most prominent for Hg2+ (Figs. 2–4) Pseudomonas putida, (b) bacterial cells exposed to Hg2+ (10 ␮M) for 10 min at 25 ◦ C,
(c) pretreated cells further exposed to 10 ␮M of L2 (10 ␮M) for 15 min, (d) when the
owing to the most efficient binding. According to the energy gap
slide (c) exposed on 200 ␮M of L1 .
law, lower HOMO-LUMO gap favours shortening of the life time of
the excited state due to deactivation of the excited state through
cessively exposed to 10.0 ␮M solution of L2 (a Rhodamine based
preferred radiative pathways. This presumably along with the pos-
dye [3p], Supporting information) and L1 (200 ␮M). These treated
sibility of the spin–orbit coupling could account for the observed
bacterial cells were monitored through a fluorescence microscope
luminescence quenching. In case of Cu2+ and Ni2+ ions, associated
(AXIO IMAGER, Carl Zeiss; 40×) using TRITC filter (ext = 541 nm;
paramagnetic effect might also have contributed to the resultant
ems = 572 nm). Fluorescence microscopic images of bacterial cell
luminescence quenching.
in the absence and presence of Hg2+ are shown as (a) and (b) in
Fluorescence spectra recorded after addition of excess of dis-
Fig. 8. Fig. 8c revealed that bacterial slide, pre-exposed to Hg2+
odium salt of ethylene diamine tetraacetic acid (THF–water; 2:3,
solution turned red fluorescent, when exposed further to L2 . This
v/v) to the solution of Mn+ –L1 (Mn+ is Hg2+ /Cu2+ /Ni2+ /Ag+ ) revealed
slide was further exposed to 200 ␮M solution of L1 and images
restoration of the original fluorescence for free L1 (Supporting
were viewed after 15 min. Disappearance of the red fluorescence
information). This confirmed the reversibility in binding of respec-
of Hg2+ –L2 , when exposed to large excess of L1 , supports the bind-
tive metal ion to L1 .
ing of L1 to the Hg2+ ion adsorbed in bacterial cells (Fig. 8). These
Time resolved fluorescence decay experiments for chemosensor
results demonstrate that L1 could bind and be used for detecting
(L1 ) were measured using 340 nm nano LED as a light excitation
Hg2+ adsorbed on the cell wall of these bacteria. Examples of using
source in air-equilibrated THF–water (2:3, v/v) medium. Fluores-
a sensor molecule for detection of Hg2+ in lower organism are not
cence decay traces for L1 showed single exponential life time
common in literature.
 1 = 8.23 ± 0.02 ns (2 = 1.09).
However, fluorescence decay traces in the presence of different
4. Conclusion
metal ions were best fitted with different time constants (Fig. 7).
In the presence of Hg2+ , L1 showed bi-exponential decay with
In conclusion, a new fluorescence-based chemosensor (L1 ) is
 1 = 0.141 ± 0.029 ns and  2 = 6.902 ± 0.1 ns (2 = 1.104). The pres-
reported. This shows preferential binding to Hg2+ ion in aqueous
ence of other metal ions like Cu2+ , Ni2+ and Ag+ the respective
solution at pH 7.2 as compared to those of Cu2+ , Ag+ and Ni2+ . We
life time are  1 = 0.3789 ± 0.014 ns,  2 = 7.54 ± 0.036 ns (2 = 1.08)
could also demonstrate that L1 could bind and detect Hg2+ ion,
(Cu2+ );  1 = 6.55 ± 0.04 ns (2 = 1.13) (Ni2+ ) and  1 = 3.49 ± 1 ns,
adsorbed on the cell wall of the bacterial cells, through quenching
 2 = 13.4 ± 6 ns,  3 = 0.433 ± 0.02 ns (2 = 1.103) (Ag+ ). In all cases,
of the fluorescence of L2 .
longer life time presumably reflects the life time of the free receptor
present in the equilibrium mixture.
One of the most common pollutants that are present in vari- Acknowledgments
ous effluents and soil irrigated with contaminated water is Hg2+
ion. There are several microorganisms that exist in soil and various Authors wish to acknowledge DST and CSIR for financial support.
effluents. Pseudomonas putida, a Gram−ve bacteria is one such bac- A.D. and S.M. thank P.K. Ghosh for his keen interest in this work.
teria [17,18] and is known to have a high affinity to absorb the Hg2+ A.K.M, M.S and S.K.M acknowledge CSIR for a research fellowship.
ion.
To examine whether this newly developed chemosensor was Appendix A. Supplementary data
capable of binding to Hg2+ ion present in lower organism, we have
used this bacterial cell for detection of Hg2+ adsorbed when exposed Supplementary data associated with this article can be found, in
to the Hg2+ solution. P. putida was cultured in the King’s B (KB) the online version, at doi:10.1016/j.snb.2009.11.016.
medium (Peptone 20 g, glycerol 15 g, K2 HPO4 1.5 g, MgSO4 ·7H2 O
1.5 g, distilled water 1000 ml, pH 7.2). Cells were harvested and References
vortexed for making the homogenous suspension in sterile dis-
[1] (a) A.P. de Silva, D.P. Fox, A.J.M. Huxley, T.S. Moody, Combining luminescence,
tilled water. These bacterial cells were exposed to Hg2+ (10.0 ␮M) coordination and electron transfer for signalling purposes, Coord. Chem. Rev.
in water–ethanol (7:3, v/v) for 10 min at 25 ◦ C and were then suc- 205 (2000) 41–57;
 Author's personal copy

A.K. Mandal et al. / Sensors and Actuators B 145 (2010) 32–38 37

(b) A.P. de Silva, H.Q.N. Gunaratne, T. Gunnlaugsson, A.J.M. Huxley, C.P. McCoy, (c) C.N. Burress, M.I. Bodine, O. Elbjeirami, J.H. Reibenspies, M.A. Omary,
J.T. Rademacher, T.E. Rice, Signaling recognition events with fluorescent sensors F.P. Gabbai, Enhancement of external spin–orbit coupling effects caused by
and switches, Chem. Rev. 97 (1997) 1515–1567; metal–metal cooperativity, Inorg. Chem. 46 (2007) 1388.
(c) B. Bag, P.K. Bharadwaj, Attachment of an electron-withdrawing fluorophore [5] (a) A.M. Costero, R. Andreu, E. Monrabal, R. Martínez-Máñez, F. Sancenón,
to a cryptand for modulation of fluorescence signaling, Inorg. Chem. 43 (2004) J. Soto, 4,4-Bis(dimethylamino)biphenyl containing binding sites. A new flu-
4626; orescent subunit for cation sensing, J. Chem. Soc., Dalton Trans. (2002)
(d) J.F. Callan, A.P. de Silva, D.C. Magri, Luminescent sensors and switches in the 1769;
early 21st century, Tetrahedron 61 (2005) 8551–8588. (b) X.-J. Zhu, S.-T. Fu, W.-K. Wong, J.-P. Guo, W.-Y. Wong, A near-infrared-
[2] (a) D.A. Jose, D.K. Kumar, B. Ganguly, A. Das, Efficient and simple colorimetric fluorescent chemodosimeter for mercuric ion based on an expanded porphyrin,
fluoride ion sensor based on receptors having urea and thiourea binding sites, Angew. Chem. Int. Ed. 45 (2006) 3150;
Org. Lett. 6 (2004) 3445; (c) Z. Wu, Y. Zhang, J.S. Ma, G. Yang, Ratiometric Zn2+ sensor and strategy for
(b) D.A. Jose, D.K. Kumar, B. Ganguly, A. Das, Urea and thiourea based efficient Hg2+ selective recognition by central metal ion replacement, Inorg. Chem. 45
colorimetric sensors for oxyanions, Tetrahedron Lett. 46 (2005) 5343; (2006) 3140.
(c) D.A. Jose, A. Singh, B. Ganguly, A. Das, A density functional study towards [6] (a) R. Métivier, I. Leray, B. Valeur, Lead and Mercury sensing by calixarene-
the preferential binding of anions to urea and thiourea, Tetrahedron Lett. 48 based fluoroionophores bearing two or four dansyl fluorophores, Chem. Eur. J.
(2007) 3695; 10 (2004) 4480;
(d) M. Suresh, D.A. Jose, A. Das, [2,2 -Bipyridyl]-3,3 -diol as a molecular half- (b) Q.-Y. Chen, C.-F. Chen, A new Hg2+ -selective fluorescent sensor based
subtractor, Org. Lett. 9 (2007) 441; on a dansyl amide-armed calix[4]-aza-crown, Tetrahedron Lett. 46 (2005)
(e) D.A. Jose, D.K. Kumar, P. Kar, S. Verma, A. Ghosh, B. Ganguly, H.N. Ghosh, 165;
A. Das, Role of positional isomers on receptor–anion binding and evidence for (c) J.H. Kim, A.-R. Hwang, S.-K. Chang, Hg2+ -selective fluoroionophore of p-
resonance energy transfer, Tetrahedron 63 (2007) 12007; tert-butylcalix[4]arene-diaza-crown ether having pyrenylacetamidesubunits,
(f) D.A. Jose, P. Kar, D. Koley, B. Ganguly, W. Thiel, H.N. Ghosh, A. Das, Phenol- Tetrahedron Lett. 45 (2004) 7557;
and catechol-based ruthenium(II) polypyridyl complexes as colorimetric sen- (d) S.H. Kim, K.C. Song, S. Ahn, Y.S. Kang, S.-K. Chang, Hg2+ -selective flu-
sors for fluoride ions, Inorg. Chem. 46 (2007) 5576; oroionophoric behavior of pyrene appended diazatetrathia-crown ether,
(g) D.A. Jose, D. Krishna Kumar, B. Ganguly, A. Das, Rugby-ball-shaped Tetrahedron Lett. 47 (2006) 497;
sulfate–water–sulfate adduct encapsulated in a neutral molecular receptor (e) S.-Y. Moon, N.J. Youn, S.M. Park, S.-K. Chang, Diametrically disubstituted
capsule, Inorg. Chem. 46 (2007) 5817; cyclam derivative having Hg2+ -selective fluoroionophoric behaviors, J. Org.
(h) A. Ghosh, B. Ganguly, A. Das, Urea-based ruthenium(II)–polypyridyl com- Chem. 70 (2005) 2394;
plex as an optical sensor for anions: synthesis, characterization, and binding (f) S.M. Park, M.H. Kim, J.-I. Choe, K.T. No, S.-K. Chang, Cyclams bearing diamet-
studies, Inorg. Chem. 46 (2007) 9912; rically disubstituted pyrenes as Cu2+ - and Hg2+ -selective fluoroionophores, J.
(i) D.A. Jose, S. Mishra, A. Ghosh, A. Shrivastav, S.K. Mishra, A. Das, Colorimetric Org. Chem. 72 (2007) 3550;
sensor for ATP in aqueous solution, Org. Lett. 9 (2007) 1979; (g) N.J. Youn, S.-K. Chang, Dimethylcyclam based fluoroionophore having Hg2+ -
(j) E.M. Nolan, S.J. Lippard, Tools and tactics for the optical detection of mercuric and Cd2+ -selective signaling behaviours, Tetrahedron Lett. 46 (2005) 125;
ion, Chem. Rev. 108 (2008) 3443–3480. (h) Y.-H. Kim, J.S. Youk, S.Y. Moon, J.-I. Choe, S.-K. Chang, Hg2+ -selective flu-
[3] (a) M.C. Aragoni, M. Arca, A. Bencini, A.J. Blake, C. Caltagirone, A. Danesi, F.A. orogenic chemosensor derived from 8-aminoquinoline, Chem. Lett. 33 (2004)
Devillanova, A. Garau, T. Gelbrich, F. Isaia, V. Lippolis, M.B. Hursthouse, B. Val- 702;
tancoli, C. Wilson, New fluorescent chemosensors for heavy metal ions based on (k) J. Yoon, N.E. Ohler, D.H. Vance, W.D. Aumiller, A.W. Czarnik, A fluorescent
functionalized pendant arm derivatives of 7-anthracenylmethyl-1,4,10-trioxa- chemosensor signalling only Hg(II) and Cu(II) in water, Tetrahedron Lett. 38
7,13-diazacyclopentadecane, Inorg. Chem. 46 (2007) 8088–8097; (1997) 3845;
(b) K. Rurack, M. Kollmannsberger, U. Resch-Genger, Daub, A selective and (l) A.B. Descalzo, R. Martínez-Máñez, R. Radeglia, K. Rurack, J. Soto, Coupling
sensitive fluoroionophore for Hg(II), Ag(I), and Cu(II) with virtually decoupled selectivity with sensitivity in an integrated chemosensor framework: design of
fluorophore and receptor units, J. Am. Chem. Soc. 122 (2000) 968–969; a Hg2+ -responsive probe, operating above 500 nm, J. Am. Chem. Soc. 125 (2003)
(c) N. Kaur, S. Kumar, A differential receptor for selective and quantita- 3418;
tive multi-ion analysis for Co2+ and Ni2+ /Cu2+ , Terahedron Lett. 49 (2008) (m) L. Wang, W.-K. Wong, L. Wu, Z.-Y. Li, A highly selective fluorescent
5067–5069; chemosensor for Hg2+ in aqueous solution, Chem. Lett. 34 (2005) 934;
(d) H. Yang, Z. Zhou, K. Huang, M. Yu, F. Li, T. Yi, C. Huang, Multisignaling (n) L. Wang, X.-J. Zhu, W.-Y. Wong, J.-P. Guo, W.-K. Wong, Z.-Y. Li,
optical-electrochemical sensor for Hg2+ based on a rhodamine derivative with Dipyrrolylquinoxaline-bridged Schiff bases: a new class of fluorescent sensors
a ferrocene unit, Org. Lett. 9 (2007) 4729–4732; for mercury(II), J. Chem. Soc., Dalton Trans. (2005) 3235;
(e) A. Coskun, M.D. Yilmaz, E.U. Akkaya, Bis(2-pyridyl)-substituted boratri- (o) Y. Yu, L.-R. Lin, K.-B. Yang, X. Zhong, R.-B. Huang, L.-S. Zheng, p-
azaindacene as an NIR-emitting chemosensor for Hg(II), Org. Lett. 9 (2007) Dimethylaminobenzaldehyde thiosemicarbazone: a simple novel selective and
607–609; sensitive fluorescent sensor for mercury(II) in aqueous solution, Talanta 69
(f) Q. He, E.W. Miller, A.P. Wong, C. Chang, A selective fluorescent sensor for (2006) 103.
detecting lead in living cells, J. Am. Chem. Soc. 128 (2006) 9316–9317; [7] (a) P.S. Argent, H. Adams, R.T. Johannessen, C.J. Jeffery, P.L. Harding, W.
(g) Y.-Q. Weng, F. Yue, Y.-R. Zhong, B.-H. Ye, A copper(II) ion-selective on–off- Clegg, W.R. Harrington, D.M. Ward, Complexes of Ag(I), Hg(I) and Hg(II) with
type fluoroionophore based on zinc porphyrin − dipyridylamino, Inorg. Chem. multidentate pyrazolyl-pyridine ligands: from mononuclear complexes to
46 (2007) 7749–7755; coordination polymers via helicates, a mesocate, a cage and a catenate, Dal-
(h) G. Klein, D. Kaufmann, S. Schürch, J.-L. Reymond, A fluorescent metal sensor ton Trans. (2006) 4996–5013;
based on macrocyclic chelation, Chem. Commun. (2001) 561–562; (b) T.K. Ronson, H. Adams, M.D. Ward, Mononuclear and polynuclear chain
(i) M. Kerim Gattás-Asfura, R.M. Leblanc, Peptide-coated CdS quantum dots complexes of a series of multinucleating N/S donor ligands, Eur. J. Inorg. Chem.
for the optical detection of copper(II) and silver(I), Chem. Commun. (2003) (2005) 4533–4549;
2684–2685; (c) R.L. Paul, S.M. Couchman, J.C. Jeffery, J.A. Mecleverty, Z.R. Reeves, M.D. Ward,
(j) M. Suresh, A. Ghosh, A. Das, A simple chemosensor for Hg2+ and Cu2+ that Effects of metal co-ordination geometry on self-assembly: a dinuclear double
works as a molecular keypad lock, Chem. Commun. (2008) 3906–3908; helicate complex and a tetranuclear cage complex of a new bis-bidentate bridg-
(k) M.-H. Ha-Thi, M. Penhoat, V. Michelet, I. Leray, Highly selective and sensi- ing ligand, J. Chem. Soc., Dalton Trans. (2000) 845–851;
tive phosphane sulfide derivative for the detection of Hg2+ in an organoaqueous (d) S. James, K.L. Fleming, V. Mann, S.M. Couchman, J.C. Jeffery, J.A. Mecleverty,
medium, Org. Lett. 9 (2007) 1133–1136; M.D. Ward, Double-helical dinuclear copper(I) and mononuclear copper(II)
(l) J. Liu, Y. Lu, A DNAzyme catalytic beacon sensor for paramagnetic Cu2+ ions complexes of a compartmental tetradentate bridging ligand: crystal struc-
in aqueous solution with high sensitivity and selectivity, J. Am. Chem. Soc. 129 tures and spectroscopic properties, J. Chem. Soc., Dalton Trans. (1998)
(2007) 9838–9839; 2047–2052;
(m) X. Guo, X. Qian, L. Jia, A highly selective and sensitive fluorescent chemosen- (e) J. Mukherjee, R. Mukherjee, Reaction with dioxygen of a Cu(I) complex
sor for Hg2+ in neutral buffer aqueous solution, J. Am. Chem. Soc. 126 (2004) of 1-benzyl-[3-(2’-pyridyl)]pyrazole triggers ethyl acetate hydrolysis: acetato-
2272–2273; /pyrazolato-, dihydroxo- and diacetato-bridged Cu(II) complexes, Dalton Trans.
(n) D. Wu, W. Huang, C. Duan, Z. Lin, Q. Meng, Highly sensitive fluorescent probe (2006) 1611–1621.
for selective detection of Hg2+ in DMF aqueous media, Inorg. Chem. 46 (2007) [8] G.M. Sheldrick, SAINT, 5.1 ed., Siemens Industrial Automation Inc., Madison,
1538–1540; WI, 1995.
(o) W. Wen Shi, H. Ma, Rhodamine B thiolactone: a simple chemosensor for [9] SADABS, Empirical Absorption Correction Program, University of Göttingen,
Hg2+ in aqueous media, Chem. Commun. (2008) 1856; Göttingen, Germany, 1997.
(p) M. Suresh, A. Shrivastav, S. Mishra, E. Suresh, A. Das, A rhodamine-based [10] G.M. Sheldrick, SHELXTL Reference Manual: Version 5.1, Bruker AXS, Madison,
chemosensor that works in the biological system, Org. Lett. 10 (2008) 3013. WI, 1997.
[4] (a) P. Svejda, A.H. Maki, R.R. Anderson, Methylmercury as a spin–orbit probe. [11] G.M. Sheldrick, SHELXL-97: Program for Crystal Structure Refinement, Univer-
Effect of complexing on the excited state properties of tryptophan, tryptamine, sity of Göttingen, Göttingen, Germany, 1997.
and benzimidazole, J. Am. Chem. Soc. 100 (1978) 7138; [12] The structural data in CIF format has been deposited for L1 at the Cam-
(b) H. Masuhara, H. Shioyama, T. Saito, K. Hamada, S. Yasoshima, N. Mataga, bridge Crystallographic Data Centre and allocated the deposition number CCDC
Fluorescence quenching mechanism of aromatic hydrocarbons by closed-shell 714417.
heavy metal ions in aqueous and organic solutions, J. Phys. Chem. 88 (1984) [13] (a) A.C. Tedesco, D.M. Oliveira, Z.G.M. Lacava, R.B. Azevedo, E.C.D. Lima, P.C.
5868; Morais, Investigation of the binding constant and stoichiometry of biocom-
 Author's personal copy

38 A.K. Mandal et al. / Sensors and Actuators B 145 (2010) 32–38

patible cobalt ferrite-based magnetic fluids to serum albumin, J. Magn. Magn. Biographies
Mater. 272–276 (2004) 2404–2405;
(b) Y. Shiraishi, S. Sumiya, Y. Kohno, T. Hirai, A rhodamine − cyclen conjugate
as a highly sensitive and selective fluorescent chemosensor for Hg(II), J. Org. Amal Kumar Mandal received his BSc and MSc degrees from Vidyasagar Univer-
Chem. 73 (2008) 8571. sity, West Bengal. In November 2007 he joined as the CSIR Jr. Research Fellow in
[14] Molecular Fluorescence, Principle and Application by Prof. B. Valeur, Wiley- the Department of Analytical Science Discipline of Central Salt & Marine Chemi-
VCH, 2005, p. 127. cals Research Institute, Bhavnagar. He is interested in molecular recognition and
[15] (a) B. Valeur, I. Leray, Design principles of fluorescent molecular sensors for supramolecular chemistry.
cation recognition, Coord. Chem. Rev. 205 (2000) 3–40;
(b) G. Grynkiewich, M. Poenie, R.Y. Tsien, A new generation of fluorescent cal- Moorthy Suresh received his MSc degree from Bharathidasan University. He then
cium indicators with greatly improved fluorescence properties, J. Biol. Chem. joined Analytical Science Discipline of Central Salt & Marine Chemicals Research
260 (1985) 3440–3450; Institute, Bhavnagar as the CSIR Jr. Research Fellow. His research interest includes
(c) S. Maruyama, K. Kikuchi, T. Hirano, Y. Urano, T. Nagano, A novel, cell- molecular recognition and molecular electronics.
permeable, fluorescent probe for ratiometric imaging of zinc ion, J. Am. Chem.
E. Suresh obtained his PhD (2001) in chemistry from Bhavnagar University. Cur-
Soc. 124 (2002) 10650–10651;
rently, he is a Senior Scientist at the Analytical Science Discipline of the Central Salt
(d) Z. Xu, Y. Xiao, X. Qian, J. Cui, D. Cui, Ratiometric and selective fluorescent
& Marine Chemicals Research Institute, Bhavnagar. His major research interests are
sensor for Cu(II) based on internal charge transfer (ICT), Org. Lett. 7 (2005)
supramolecular self assembly, anion recognition and metal–organic framework.
889–892;
(e) S. Banthia, A. Samanta, A two-dimensional chromogenic sensor as well as Sanjiv K. Mishra received his MSc degree from Banaras Hindu University in 2005.
fluorescence inverter: selective detection of copper(II) in aqueous medium, Presently he is a Sr. Research Fellow at the Marine Biotechnology and Ecology Disci-
New J. Chem. 29 (2005) 1007. pline of Central Salt & Marine Chemicals Research Institute, Bhavnagar. His research
[16] (a) D.S. McClure, Intersystem conversions in polyatomic molecules within the interest includes cyanobacterial biotechnology and fluorescent proteins.
model of stepwise relaxation of electronic recitation energy, J. Chem. Phys. 20
(1952) 682–686; S. Mishra obtained her PhD degree in 1989 from Bhavnagar University. At present
(b) A.W. Varnes, R.B. Dadson, E.L. Wahry, Interactions of transition-metal ions she is a Senior Scientist at the Marine Biotechnology and Ecology Discipline
with photoexcited states of flavines. Fluorescence quenching studies, J. Am. of Central Salt & Marine Chemicals Research Institute, Bhavnagar. Her research
Chem. Soc. 94 (1972) 946–950; interest includes phycobiliproteins, biodegradable biopolymers, and fluorescent
(c) Y. Zheng, J. Orbulescu, X. Ji, F.M. Andreopoulos, S.M. Pham, R.M. Leblanc, proteins.
Development of fluorescent film sensors for the detection of divalent copper,
J. Am. Chem. Soc. 125 (2003) 2680–2686. Amitava Das obtained his PhD (1989) in chemistry from Jadavpur Univer-
[17] T.J. Beveridge, Biosorption of lead and uranium by Streptomyces sp., Biotechnol. sity. Currently, he is a Senior Scientist at the Central Salt & Marine Chemicals
Bioeng. 16 (1986) 127. Research Institute, Bhavnagar. His research interest includes molecular recognition,
[18] M. Ledin, K. Pedersen, B. Allard, Effects of pH and ionic strength on the adsorp- supramolecular chemistry and electronically active organic and macromolecular
tion of Cs, Sr, Eu, Zn, Cd and Hg by Pseudomonas putida, Water, Air, Soil Pollut. materials.
93 (1997) 367–381.