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Syntheses, crystal structures and photophysical

Cite this: DOI: 10.1039/c6dt04039j
properties of Cu(II) complexes: fine tuning of a
coordination sphere for selective binding of
azamethiphos†
Pushap Raj,a Amanpreet Singh,a Ajnesh Singhb and Narinder Singh*a

Received 20th October 2016,
Accepted 7th December 2016
DOI: 10.1039/c6dt04039j
www.rsc.org/dalton
Two copper complexes C1 and C2 have been designed and developed for selective sensing of organo-
phosphates. It is important to develop an efficient method for the detection of these agents for environ-
mental analysis because the overuse of these agents in the environment causes harmful effects on living
systems. Our attempts to utilize the copper complexes for the detection of organophosphates remained
successful: the C1 complex has shown selective binding for the azamethiphos with a detection limit of
19 nM; while the C2 complex has not revealed any selectivity for any of the tested organophosphates.
The results indicated that the coordination sphere of the C1 complex is proficiently engineered in such a
way that it offers judicial binding sites for guest molecules.

Introduction cholinase in cholinergic synapses and damages the central

Organophosphates (OPs) are the esters of phosphoric acid
with C–P and C–PvO bonds and they were first synthesized in
the 1800s through the reaction of alcohols with phosphoric
acid.1 The stability of the C–P bond is high enough that
organophosphates remain stable at moderate temperature.2,3
The use of these organophosphates is extensible in agriculture,
chemical warfare and therapeutics, and in academic research
for organic synthesis.4 In agriculture, OPs are the prime choice
of farmers to eradicate insects, pests and unwanted herbs
from their crops.5,6 The organophosphate-based pesticide
market is extensively growing as OPs are inexpensive and easy
to formulate.7–9 Contrary to the widespread use of OPs in agri-
culture and other applications, OPs are highly neurotoxic and
disrupt the normal functioning of the cholinesterase enzyme.
Cholinesterase hydrolyses acetylcholine into choline and the
acetyl group which are required for proper functioning of the
nervous system.10,11 OPs acquire acute toxicity from the
nucleophilic reaction of the hydroxyl group of the serine
moiety of the cholinesterase enzyme with the electron deficient
phosphorus.12 The esterification of serine generates acetyl-
a
Department of Chemistry, Indian Institute of Technology Ropar, Punjab, 140001,
India. E-mail: nsingh@iitrpr.ac.in; Tel: +91-1881242176
b
Department of Applied Sciences and Humanities, Jawaharlal Nehru Govt.
Engineering College, Sundernagar, Mandi(H.P.)-175018, India
† Electronic supplementary information (ESI) available. CCDC 1462845 and
1462846. For ESI and crystallographic data in CIF or other electronic format see
DOI: 10.1039/c6dt04039j
nervous system.13,14 A careful analysis of the scientific litera-
ture revealed that sincere efforts have been put forward for
the investigation of these analytes through chromatography,
NMR spectroscopy, mass spectrometry, potentiometric
methods, colorimetric methods, and surface acoustic wave
spectroscopy.15–21 However, these techniques are expensive
and time consuming and require experts to handle the instru-
mentation. Nowadays, the organophosphate pesticide kit is
available in the market to investigate organophosphates.
These pesticide kits have several advantages, including cost
effectiveness, less time consumption and easy transportation.
However, the major limitations involve the precision and sensi-
tivity ( ppm level) in the detection of OPs.21 In continuation
with the development of analysis kits, recently optical
methods (colorimetric and fluorometric) for analysis of
OPs have gained tremendous attention because of their high
sensitivity, operativity, and immediate response.22–24 Optical
methods involve the judicial design of a chemosensor in such
a way that the method must offer a change in the absorption/
emission profile of the chemosensor in response to the target
organophosphates.25,26 The resultant optical signal has been
used for the development of a chemosensor for organo-
phosphates.27 In the present manuscript, we have used naphthal-
imide ensemble copper complexes as a chemosensor for OPs,
based on the cation displacement approach.28–30 The displace-
ment assay is the novel approach in the field of supramolecu-
lar chemistry.31 The binding unit and signalling unit are con-
nected through covalent bonding in a coordination complex.
The addition of a targeted analyte results in the ejection of a

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Scheme 1 The structure of ligands L1–2 and complexes C1–2; the
potential cation binding sites of ligands are highlighted in red color.
metal ion from the coordination sphere of receptors. The co-
ordinated and free receptors have different photophysical pro-
perties and the analyte induced modulations of photophysical
properties are the basis of analyte estimation.32,33 The main
reason for using the naphthalimide moiety lies in the fact that
this moiety has superior optical and electrochemical pro-
perties and it is easy to engineer the required sensor.34 The
ligand L1 is designed in such a way that the chelate ring for-
mation may prevail with the copper ion and the design of
ligand L2 is devoid of any option for chelate formation;
however, it has a potential binding site for metal coordination
through the sp2 hybridized nitrogen of pyridine. The complex
C1 has distorted octahedral geometry and the coordination
sphere of the copper metal ion was satisfied through coordi-
nation of ligand L1 and nitrate ions as investigated from their
single crystal structure. Similarly, the C2 complex has also
exhibited distorted octahedral geometry and the coordination
sphere of copper was satisfied with ligand L2 and bidentate
nitrate ions as shown in Scheme 1. The chemosensor behav-
iour of C1/C2 was tested with a library of organophosphates
and the original emission of ligand L1 was restored in the
presence of azamethiphos. This shows that the cation displace-
ment assay works in the sensing of azamethiphos. C2 has not
shown any selectivity on addition of a library of organo-
phosphates, which revealed that the system lacks the specific
displacement assay. The possible mechanism of cation displace-
ment behaviour of C1 with azamethiphos was authenticated
through 31P NMR and mass spectrometry.
Results and discussion
Synthesis and characterization of ligands and their metal
complexes
The ligand L1 was synthesized via a condensation reaction
between hydrazine hydrate and 1,8-naphthalic anhydride in
methanol with 93% yield. Similarly, the ligand L2 was syn-
thesized (with 87% yield) through the reaction between 4-amino-
methyl pyridine and 1,8-naphthalic anhydride, however, in
the presence of 2 drops of triethyleneamine. The structural for-
mulae of both the ligands have been elucidated with spectro-
scopic methods: the FTIR (cm−1) ν 3180 (–N–H), the 1H NMR
spectrum with a signal at 5.55 ppm (s, 2H, NH2) and ESI-Mass
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at 212 corresponding to the molecular ion peak confirmed the
formation of the ligand L1. Similarly, the spectroscopic data
recorded for L2 support the formula of the ligand: FTIR (cm−1)
ν 3075 (–Ar–H), 1692 (–CvO), 1552 (Ar–H), the 1H NMR signal
at 5.34 ppm (s, 2H, CH2), the 13C NMR signal at 42.33 ppm
(CH2), and ESI-Mass at 288 confirmed the molecular ion peak.
In order to obtain the crystals of the Cu2+ metal complex of
both the ligands, the reactions were carried out between
Cu(NO3)·3H2O and the respective ligand in a CH3OH : THF
solvent system.
Upon completion of the reaction, the solution was kept in
the dark for slow evaporation of the solvent, which generated
single crystals suitable for diffraction studies. Single crystal
X-ray structure determination of the Cu2+ metal complex of L1
has revealed that the complex crystallizes in the triclinic crystal
system with the space group P1 ˉ and the asymmetric unit con-
sists of one octahedral Cu(II) complex, [Cu(C12H8O2N2)2(NO3)2]
where the Cu2+ ion is located at the centre of inversion and
coordination around the central metal ion is octahedral, with
tetragonal elongation, due to Jahn–Teller distortion (selected
bond length and bond angles are given in Table S2†), and the
complex is named C1. The ORTEP view of complex C1 along
with atom numbering is shown in Fig. 1A. The coordination
sphere of C1 is satisfied with two chelating naphthalimide
ligands while the remaining two coordination sites are
satisfied with oxygen atoms of two nitrate ions. The octahedral
geometry around Cu2+ is slightly distorted with a maximum
deviation of angle as shown by N(2)–Cu(1)–O(2) = 80.30(7)°
from an ideal angle of 90°. The packing analyses of C1
down the b axis have shown the layered arrangements of
moieties, where two adjacent layers are interlocked with
each other through the N–H⋯O/C–H⋯O hydrogen bonding
interaction. The packing diagram of complex salt is shown
in Fig. 1B. The hydrogen bonding parameters are shown in
Table S2.†
The complex [Cu(L2)2(NO3)2] crystallizes in the ortho-
rhombic crystal system with the Pbcn space group and this
complex is named C2. The asymmetric unit consists of one
octahedral Cu2+ in complex C2 where the Cu2+ ion is also
located at the centre of inversion. The ORTEP view of com-
pound C2 along with atom numbering is shown in Fig. 1C.
The distorted octahedral coordination around Cu2+ is com-
pleted with two bidentate nitrato ligands and two mono-
dentate L2 ligands. The coordination around the central metal
ion is distorted octahedrally which deviates as high as 33.29°
(from the ideal value of 90°) as revealed from the geometrical
parameters (Table S3†). Such a distortion can be explained,
considering the spatial juxtaposition of the ligands around the
metal atom in which the O–O bite distance of the nitrato
group is much lower than 2.131 Å. The packing analyses of C2
have revealed that different moieties are interacting with each
other through the C–H⋯O hydrogen bonding interaction
(hydrogen bonding parameters are shown in Table S4†) and
π–π stacking interactions (distance between centroids of two
different naphthalimide rings is 3.758 Å and the shortest
C9⋯C12 distance is 3.495 Å (Fig. 1D)).
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Fig. 1 (A) ORTEP diagram along with the atom numbering scheme of complex C1 with 40% probability thermal ellipsoids. (B) Packing diagram of
complex C1 down the b-axis (N–H⋯O/C–H⋯O hydrogen bonding interactions are shown by green dotted lines). (C) ORTEP diagram along with the
atom numbering scheme of complex C2 with 40% probability thermal ellipsoids. (D) Packing diagram of complex C2 down the c-axis.

Photophysical properties of metal complexes (C1–2)
The copper complexes C1–2 were soluble in methanol and
their photophysical properties were evaluated with UV-visible
absorption and emission spectroscopy; the results are com-
pared with the respective ligands (Fig. 2A and B). The absorp-
tion spectrum recorded with 10 μM concentration of L1 in
methanol revealed bands at 332 (εo = 8.0 × 104 L mol−1 cm−1)
due to internal charge transitions (ICT) and a high energy
band at 248 (εo = 1.9 × 104 L mol−1 cm−1) on account of π → π*
transitions.35 Similarly, the absorption spectrum of L2
recorded with the same concentration and the same solvent as

Fig. 2 (A) A comparison of UV-Vis absorption spectra of C1–2 with respect to L1–2, each spectrum was recorded with 10 µM concentration of
ligand/complex in methanol. (B) A comparison of the emission profile of C1–2 with respect to L1–2, each spectrum was recorded with 10 µM con-
centration of ligand/complex in methanol.

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those used for L1 revealed the ICT band at 339 (εo = 5.8 × 104
L mol−1 cm−1) and a band due to π → π* transitions at 254
(εo = 2.1 × 104 L mol−1 cm−1). However, the complex formation
of L2 leads to the bathochromic and hypochromic shift in
both high and low energy bands, which is due to the modu-
lation of the energy gap upon coordination of the metal ion
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The copper ion is paramagnetic and the reduction potential of
copper lies in between the HOMO–LUMO gap of the ligand.
Therefore the excited state energy of the ligand transfers to the
unpaired d orbital of the copper ion, as a result quenching in
the fluorescence intensity of the ligand was observed. It is
already documented in the literature that metal ions such
as Cu2+, Pb2+, Hg2+ and Cd2+ can cause quenching in the

(ligand to metal charge transfer band). The transition was

recognized from nonbonding electrons of pyridine nitrogen emission spectrum through the charge or energy transfer
(low energy LUMO) to the unpaired HOMO of the copper ion, process.39,40 Furthermore, the solid state emission spectra of
which is already reported by Zaleski et al. in a Cu(II)–pyridine complexes C1 and C2 were studied. The solid state emission of
complex in the range of 300–375 nm.36,37 Similarly, C1 has both complexes matches almost with the solution state emis-
revealed the bathochromic and hypochromic shift in both the sion as shown in Fig. S12.† The XRD pattern of complexes C1
bands; noticeably, the low energy band of C1 has shown a and C2 has been recorded and compared with the simulated
shoulder. This implies that all the binding sites of the chromo- data as shown in Fig. S11.† The result of the powder XRD
phore are not uniformly bound to the metal ion and the same pattern almost matches with the simulated data (Mercury
was already confirmed from the crystal structure. The d–d software).
bands are too weak to show in comparison to the ICT bands.
Upon excitation of methanol solutions containing 10 µM con- Electrochemical properties of metal complexes (C1–2)
centrations of L1 and L2 at 332 nm and 339 nm respectively, The electrochemical properties of both complexes C1–2 were
the corresponding emission was observed at 436 nm and studied by cyclic voltammetry in the potential range 1.0 to
429 nm. As per the expectations, the fluorescence intensity of −1.2 V using 0.1 M tetrabutyl ammonium perchlorate in
both the ligands was quenched upon coordination of Cu2+, methanol solution and data are summarized in Table 3. Both
possibly due to the open shell of the copper ion (Table 2).38 the complexes C1–2 have demonstrated a quasi-reversible
reduction wave in the range of −0.13 to −1.2 V. Complex C1
has shown a cathodic peak at (Epc/V) −0.66 and an anodic
Table 1 Crystal data and refinement parameters of [Cu(L1)2(NO3)2] and peak (Epa/V) at −0.13 and C2 displayed a cathodic peak (Epc/V)
[Cu(L2)2(NO3)2] at −0.77 and an anodic peak at −0.18 V (Fig. 3A);41 the redox
potential of both complexes with a negative value is attributed
Compound C1 C2
to the presence of a conjugated naphthalimide moiety. To
Empirical formula C24H16CuN6O10 C36H24CuN6O10 confirm the number of electron transferred in the redox reac-
Mw 611.97 764.15 tion of the copper complexes, the coulometric experiment by
Temperature [K] 293(2) 293(2)
Crystal system Triclinic Orthorhombic potentiometric exhaustive electrolysis was performed at
Space group P1ˉ Pbcn 100 mV less than the reduction peak of the complexes C1 and
a/[Å] 7.3858(4) 27.0677(13) C2.42 The results of the potentiometric experiment show the
b/[Å] 7.9511(4) 7.5081(3)
c/[Å] 9.9894(5) 16.0875(8) consumption of one electron (n = 0.93, n = 0.89 for C1 and C2
α/[°] 75.700(2) 90° respectively) in the redox couple of both the copper complexes.
β/[°] 78.644(2) 90° Furthermore, to verify the quasi-reversible nature of metal
γ/[°] 84.546(2) 90°
V [Å3] 556.66(5) 3269.4(3) complexes, scan rate experiment has been carried out from 20
Z 1 4 to 100 scan rate (Fig. 3B and C).43 The observations made from
Dc [Mg m−3] 1.826 1.552
μ/[mm−1] 1.061 0.740
Reflections collected 12 175 68 677
Data/restraints/parameters 2745/0/194 4076/0/240
Unique reflections, [Rint] 2745[0.0446] 4076[0.0930] Table 3 Electrochemical data of copper complexes
GOF = Sall 1.046 0.966
Final R indices S. no. Metal complexes Epc/V Epa/V E1/2/V ΔE/mV
R1, wR2 [I > 2σI] 0.0386, 0.0814 0.0458, 0.1233
R1, wR2 (all data) 0.0557, 0.0871 0.0895, 0.1475 1 C1 −0.66 −0.13 −0.39 535
Δρmax/Δρmin [Å3] 0.430 and −0.397 0.240 and −0.559 2 C2 −0.77 −0.18 −0.48 593

Table 2 Photophysical parameters of ligands and metal complexes

Ligand/metal Absorption Quantum

S. no. complex λabs (nm) ε0 (L mol−1 cm−1) transition λem (nm) yield (Φs)

1 L1 332, 248 8.0 × 104, 1.9 × 104 ICT, π → π* 436 0.75

2 L2 339, 254 5.8 × 104, 2.1 × 104 ICT, π → π* 429 0.73
3 C1 344, 269, 236 3.5 × 104, 2.3 × 104, 1.6 × 104 ICT, π → π* 429 0.30
4 C2 360, 275, 239 1.6 × 104, 2.1 × 104, 1.0 × 104 ICT, π → π* 410 0.31

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Fig. 3 (A) The cyclic-voltammogram of C1–2 in methanol (scan rate 20 mV s−1) at a silver electrode; supporting electrolyte: [nBu4N][ClO4]. The
effect of scan rate on the cyclic-voltammogram studies of: (B) C1 and (C) C2. (D) TGA graph of C1–2 in the range from 20 °C to 700 °C at a heating
rate of 10 °C per minute.
the analysis of the scan rate are as follows: (1) the Epc and Epa
values change with the increase in scan rate, (2) the ΔEp value
increases with the increase in the scan rate, (3) the cathodic
and anodic peak current ratio (ipc/ipa) is more than 1, (4) the
ΔEp value is more than 59 mV.44 These observations strongly
confirmed the quasi-reversible nature of metal complexes and
involved the reduction of Cu2+ ions to Cu+ ions in the reaction.
Thermal studies of metal complexes (C1–2)
The thermal studies of complexes C1–C2 were analyzed
through TGA analysis in the range of 20°–700 °C under a nitro-
gen atmosphere at the heating rate of 10 °C per minute.
Complex C1 showed adequate crystallinity and thermal sta-
bility at room temperature as confirmed from the TGA result.
The TGA analysis shows that the 63% mass of complex C1 is
lost at 450 °C, this was due to the complete decomposition of
the naphthalimide moiety. 37% of the mass residue left was
due to CuO. Similarly, it was observed from the TGA analysis
that the C2 complex shows adequate crystallinity and thermal
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stability at room temperature. The result of TGA analysis
shows that 71% mass of complex C2 was lost due to the
organic ligand L2 at 400 °C and 29% mass was found which is
again due to CuO as shown in Fig. 4D.
Chemosensor activities of metal complexes (C1–2)
To scrutinize the chemosensor activity of C1 (10 μM, HEPES,
pH 7.4) in methanol towards a library of organophosphates
NADH, NAD, ATP, NADH, azamethiphos, parathion, chlor-
pyrifos, phosmet, azinophos-methyl, and fenitrothion were
employed. The emission profile of C1 (10 μM, HEPES, 7.4) had
showed an emission intensity at 430 nm with a quantum yield
(Φs) of 0.30 which was half of the intensity as represented by
the naphthalimide ligand (Φs = 0.75).45 The quenching in the
fluorescence intensity of C1 was due to the open shell effect of
copper. Upon addition of various organophosphates (30 μM)
to the solution of C1, no significant change in emission
spectra was observed except in the case of azamethiphos, in
which the emission intensity of C1 was enhanced by 4 fold
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Fig. 4 (A) Change in fluorescence spectra of C1 in methanol solution on addition of various organophosphates (0–30 μM). (B) Fluorescence titra-
tion of C1 (10 μM) with azamethiphos (0–30 μM). (C) The nonlinear plot between fluorescence intensity versus concentration of azamethiphos. (D)
Interference studies of complex C1 for determination of azamethiphos in the presence of other organophosphates.

(Fig. 4A). Furthermore, to check the reproducibility of experi-
ment, titration was performed by stepwise addition of aza-
methiphos (0–30 μM) to C1 (10 μM) and the corresponding
enhancement in fluorescence intensity was observed as shown
in Fig. 4B. The calibration plot showed that the fluorescence
intensity increases with the addition of an incremental
amount of azamethiphos as shown in Fig. 4C. To corroborate
the practical utility of complex C1 towards azamethiphos the
interference studies were carried out. In interference studies
competitive organophosphates such as NADH, NAD, ATP,
NADH, parathion, chlorpyrifos, phosmet, azinophos-methyl,
and fenitrothion (30 μM) were added to the solution of C1
along with azamethiphos (30 μM). It was observed from the
fluorescence analysis that none of the competing analytes
interfere in the detection of azamethiphos. This confirmed the
selective binding of azamethiphos with C1 in the presence of
other competitive organophosphates. Nonlinear regression
analysis was used to calculate the binding constant having a
value of 2.453 × 104. The stoichiometry of complexation has
been confirmed through the Job plot.46 The Job plot showed
the maximum [HG] at a mole fraction of 0.5, which confirmed
the formation of 1 : 1 stoichiometry (Fig. S14†). The limit of
detection was calculated by the 3σ method that was achieved
to be 19 nM, far better than the existing sensor (Table S5†).
Similarly, the recognition properties of C2 were tested for
various organophosphates viz. as NADH, NAD, ATP, NADH,
azamethiphos, parathion, chlorpyrifos, phosmet, azinophos-
methyl and fenitrothion. The emission spectrum of C2 (10 μM,
HEPES, pH = 7.4) did not show any significant change in the
emission profile on addition of various organophosphates
(30 μM) as shown in Fig. S13.† This indicates that C2 cannot
show any interaction with the tested organophosphates.

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Possible mechanistic insights of chemosensor activities
To study the possible binding mechanism of azamethiphos
with the C1 complex, 31P NMR spectroscopy was employed in
DMSO-d6. The 31P NMR spectrum was analyzed by adding
equal molar concentrations of azamethiphos to the solution of
C1. The 31P NMR spectrum involves the broadening and sig-
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nificant change in the chemical shift value of azamethiphos
(originally δ at 26.72) on complex formation with C1 (δ at
30.44) as shown in Fig. S15.† The broadening in the 31P NMR
peak of azamethiphos is due to the formation of the copper :
azamethiphos complex in solution. To further confirm the for-
mation of the copper : azamethiphos complex in the solution,
ESI-Mass has been recorded. The result of the mass spectrum
displays a peak at 512 due to the copper : azamethiphos
complex as shown in Fig. S16.† The 31P NMR spectra and mass
spectrum confirmed the dissociation of C1 into a free ligand
and copper and simultaneously formation of the copper :
azamethiphos complex. On the basis of fluorescence, 31P NMR
and mass spectrometry, the possible mechanism has been
drawn as shown in Scheme 2. The selective binding of C1 with
azamethiphos was due to hydrogen bonding between the
nitrogen of azamethiphos and the NH2 group of naphthal-
imide moieties which stabilised the transition state followed
by the formation of the copper : azamethiphos complex. Such
binding sites are not available in the C2 complex and did not
interact with any of the tested organophosphates as a result
the emission profile remains the same as that of the C2
complex. Furthermore, we have added an excess aliquot of
Cu2+ ions in the C1 : azamethiphos solution and a decrease in
the intensity of the emission profile was observed as shown in
Fig. S17.† These results confirm that the excess of copper
further reacted with the free naphthalimide moieties forming
the C1 complex again. This clearly confirmed the cation dis-
placement mechanism work in the sensing of azamethiphos
as shown in Scheme 2. The reversibility in the response of
complex C1 with azamethiphos has been recorded during its
five cycles of titration by alternate addition of azamethiphos
and copper to C1 solution. The fluorescence intensity of the
tested solution shows alternate enhancement and quenching
as shown in Fig. S18.† Upon addition of azamethiphos to C1
Scheme 2 The possible mechanism of interaction of azamethiphos
with C1 complex.
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solution the observed enhancement in emission intensity and
further addition of copper cause quenching in the emission
profile. This process is repeated 5 times and little fluorescence
efficiency loss was observed.
Experimental details
General information
All chemicals were purchased from Sigma Aldrich Co. and SD
Fine India, and used without purification. 1H NMR and
13
C NMR were recorded on a JEOL instrument operated at
400 MHz for 1H NMR, 100 MHz for 13C NMR and 31P NMR at
160 MHz. The FTIR spectra of the dried sample of receptors L1
and L2 and their complex with copper were recorded on a
Bruker Tensor 27 spectrophotometer using a solid cell tech-
nique. Elemental analysis was monitored through a Fisons
instrument (Model EA 1108 CHN). The photophysical properties
and recognition properties of the metal complexes were deter-
mined with a Shimadzu UV-2400 spectrophotometer and
through a Perkin Elmer L55 fluorescence spectrophotometer.
The monitoring of UV-visible absorption and the fluorescence
range was fixed at 200–900 nm and the measurement was per-
formed at room temperature using a 1 cm path length quartz
cuvette.
Synthesis and characterization of L1. Ligand L1 was syn-
thesized via a condensation reaction between 1,8-naphthalic
anhydride (1.98 g, 10 mmol) and hydrazine (0.500 g, 10 mmol)
in 50 ml methanol. A light yellow color solid was separated out
after 8 hours. The precipitate was filtered out and recrystallized
in methanol to afford a pure product. The yield of the product
was 93% and FTIR (cm−1) ν 3180 (–NH2), 1662 (–CvO),
1552 (–Ar–H). 1H NMR (400 MHz, CDCl3) δ, 8.66 (d, 2H, Ar–H,
J = 8 Hz), 8.25 (d, 2H, Ar–H, J = 8 Hz), 7.75 (t, 2H, Ar–H, J =
8 Hz), 5.55 (s, 2H, NH2). 13C NMR (CDCl3, 100 MHz) 163.07,
134.61, 134.70, 131.69, 127.15, 126.69, 121.94, EI-Mass:
212 and Anal calcd for C12H8N2O2: C, 67.92; H, 3.80; N, 13.20;
O, 15.08; found: C, 65.05; H, 3.45; N, 12.90.
Synthesis and characterization of L2. Ligand L2 was syn-
thesized via a condensation reaction between 4-aminomethyl
pyridine (1.08 g, 10 mmol) and 1,8-naphthalic anhydride
(1.98 g, 10 mmol) in 30 ml methanol. In this reaction scheme
1,8-naphthalic anhydride (10 mmol) was taken in 30 ml
of methanol and 4-aminomethyl pyridine (10 mmol) was
added in the presence of 2 drops of triethylene amine as the
catalyst. The reaction mixture was refluxed for 6 hours and
after six hours the product was cooled at room temperature
and a white color solid was separated out after 4 hours. The
white solid was washed with methanol 4–5 times and
dried under vacuum. The yield of the product was 87%,
FTIR (cm−1) ν 3075 (–Ar–H), 1692 (–CvO), 1552 (Ar–H).
1
H NMR (400 MHz, CDCl3) δ = 8.61 (d, 2H, Ar–H, J = 8 Hz),
8.51 (d, 2H, Ar–H, J = 8 Hz), 8.23 (d, 2H, Ar–H, J = 8 Hz),
7.8 (t, 2H, Ar–H, J = 8 Hz), 7.35 (d, 2H, Ar–H, J = 8 Hz),
5.35 (s, 2H, CH2), 13C NMR (CDCl3, 100 MHz) 164.24,
150.08, 146.01, 134.55, 131.82, 128.33, 127.17, 123.33,
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122.34, 42.33, EI-Mass: 288 and Anal Calcd for C18H12N2O2:
C, 74.99; H, 4.20; O, 11.10; N, 9.72; found: C, 74.21; H, 4.00;
N, 8.50.
Synthesis and characterization of copper complex of C1.
24.1 mg of Cu(NO3)·3H2O was dissolved in 1 ml of methanol
in one test tube and in the second test tube 21.2 mg of ligand
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L1 was dissolved in 2 ml CH3OH : THF (1 : 1). Both the solu-
tions were mixed, filtered and kept for slow evaporation in
the dark without any disturbance. After 3 days a blue color
crystal was obtained which was washed with methanol and
was suitable for X-ray studies. Yields: 40%, Anal Calcd for
C24H16CuN6O10: C; 47.10, H; 2.63, N; 13.73; found: C; 47.09, H;
2.19, N; 13.53 preferred FTIR: (cm−1) ν 3300 (–NH2), 1663 (–CvO),
3208 (–Ar–H).
Synthesis and characterization of copper complex of C2.
28.8 mg of ligand L2 was dissolved in 1 ml methanol in one
test tube and heated for 30 minutes, and in the second test
tube 24.1 mg of Cu(NO3)·3H2O was dissolved in 1 ml metha-
nol. Both the solutions were mixed and kept for slow evapo-
ration at room temperature. After 4 days a blue color crystal
was formed which was washed with methanol and was suit-
able for X-ray crystallography. Yields: 60%, Anal Calcd for
C36H24CuN6O10: C; 56.58, H; 3.16, N; 10.99; found: C; 56.45, H;
3.07, N; 10.01 preferred FTIR: (cm−1) ν 3072 (–Ar–H), 1696
(–CvO).
X-ray data collection and refinement
The X-ray diffraction data for C1 and C2 were collected on a
Bruker D8 Venture PHOTON 100 CMOS CCD diffractometer
at 293(2) K using mirror monochromatized Mo-Kα radiation
(λ = 0.71073 Å). Data reduction and multi-scan absorption
were carried out using the APEX II program suite (Bruker,
2007). The structures were solved by direct methods with the
SIR97 program [S1] and refined using full-matrix least
squares with SHELXL-2014 [S2]. Anisotropic thermal para-
meters were used for all non-H-atoms. The hydrogen atoms
of C–H groups were with isotropic parameters equivalent to
1.2 times those of the atoms to which they were attached. All
other calculations were performed using the programs
WinGX [S3] and PARST [S4]. The molecular diagrams were
drawn with DIAMOND [S5]. The final R-values together with
selected refinement details are given in Table 1. Related
crystallographic information is provided for CCDC no. 1462845
and 1462846.
Photophysical properties of metal complexes
The photophysical properties of both metal complexes were
evaluated at room temperature and 10 μM solutions of the
respective complexes are prepared in methanol and shaken
well for recording the experiment. The UV-visible absorption
spectra were recorded in the range of 200–900 nm at room
temperature.
Chemosensor behaviour of metal complexes
The recognition properties of metal complexes towards
a library of organophosphates were determined by using
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fluorescence spectroscopy. For these studies we have
prepared a standard solution (30 μM) of the respective
organophosphates and 10 μM solution of metal complexes in
methanol. The organophosphate recognition was analyzed by
the change in the emission profile on addition of various
organophosphates (30 μM) in the respective host solution. To
further confirm the reproducibility of experiment, titration
experiment was carried out. Interference studies were carried
out to find the practical utility of the sensor. All recognition
experiments were repeated three times and then the final
conclusion has been drawn. The fluorescence quantum yields
were calculated by using the optically matching solution as
the standard at an excited wavelength of 300–340 nm and the
quantum yield can be calculated by using the following
equation as
I ODr ή2
Φs ¼ Φr 
Ir OD ήr 2
where Φs and Φr are the radiative quantum yields of the
sample and reference and ODr and OD are the optical den-
sities of the reference and sample respectively. I and Ir are the
absorbance and ή and ήr are the refractive indexes of the
sample and reference solution respectively (2-aminopyridine
in 0.1 M H2SO4).47 The binding constant was calculated by
using the Benesi–Hildebrand equation.48 The detection limit
was calculated by plotting a graph between fluorescence
intensity vs. concentration and the slope and standard devi-
ation were determined from a nonlinear regression graph.
The limit of detection was calculated by the IUPAC rec-
ommended equation as σ = 3 × SD/slope.49
Conclusion
In conclusion we have synthesized two copper complexes of
naphthalimide and their structures were well characterized
with X-ray crystallography. The X-ray crystal shows that both
the copper complexes are distorted octahedrally in geometry
and their supramolecular frameworks are connected through
a number of interactions between the naphthalic moiety
such as hydrogen bonding, π⋯π, and N⋯O interaction.
These prepared metal complexes were explored as a chemo-
sensor for organophosphates. The C1 complex selectively
binds with azamethiphos having a binding constant value
of 2.4 × 104 with a detection limit of 19 nM. Furthermore,
no interference was observed with other tested organo-
phosphates for the detection of azamethiphos through C1.
The possible interaction between C1 and azamethiphos was
studied by 31P NMR, mass and fluorescence spectroscopy
which confirmed the cation displacement assay governed for
the detection of azamethiphos. The C2 complex did not
show any binding with any of the tested organophosphates
because the coordination framework of the complex func-
tions in such a way that it does not show any binding with
the target analyte.
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Safety consideration
Organophosphates are very toxic and following precautions
were taken for preparation and handling of these solutions. All
the solutions of organophosphates were prepared in a separate
fume hood. Protective clothes, gloves and eyeglasses were
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used. After complete recognition studies, the organophosphate
solution was mixed with sodium hydroxide solution to
maintain pH 12 and refluxed for 1 h at 60 °C. The degradation
study of organophosphates was monitored through gas
chromatography. The hydroxide ion attacks on the electro-
philic phosphorus center and hydrolyses the P–O aryl bond
into its respective component.50
Acknowledgements
We are thankful to DST India [Project no. EMR/2014/000613]
for financial support. Pushap Raj is thankful to UGC
(New Delhi) for a junior research fellowship. We are highly thank-
ful to reviewers for their valuable comments and suggestion.
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