A Journal of

Accepted Article
Title: Luminescent Eu(III) and Tb(III) Complexes Containing Dopamine
Neurotransmitter: Biological Interactions, Antioxidant Activity and
Cellular Imaging Studies

Authors: Khushbu Singh, Anshika Goenka, Subramaniam Ganesh, and

Ashis K. Patra

This manuscript has been accepted after peer review and appears as an
Accepted Article online prior to editing, proofing, and formal publication
of the final Version of Record (VoR). This work is currently citable by
using the Digital Object Identifier (DOI) given below. The VoR will be
published online in Early View as soon as possible and may be different
to this Accepted Article as a result of editing. Readers should obtain
the VoR from the journal website shown below when it is published
to ensure accuracy of information. The authors are responsible for the
content of this Accepted Article.

To be cited as: Eur. J. Inorg. Chem. 10.1002/ejic.201800732

Link to VoR: http://dx.doi.org/10.1002/ejic.201800732

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
Luminescent Eu(III) and Tb(III) Complexes Containing Dopamine
Neurotransmitter: Biological Interactions, Antioxidant Activity
and Cellular Imaging Studies
Khushbu Singh[a], Anshika Goenka[b], Subramaniam Ganesh[b] and Ashis K. Patra*[a]

Abstract: Two luminescent water soluble lanthanide(III) complexes, for time-resolved live cell or in-vivo imaging application. Such
[Ln(DTPA-DOPA)(H2O)] (1-2) where Ln = Eu(III) (1), Tb(III) (2) formed time-gated techniques will eliminate interfering background

Accepted Manuscript
by reaction with diethylenetriaminepentaacetic acid (DTPA) carrying autofluorescence, scattering in biological milieu and
neurotransmitter dopamine with catechol functional groups i.e. DTPA- enhancing signal-to-noise ratios. Chelation of lanthanides to
bis(3-hydroxytyramide) are characterized and their photophysical multidentate chelating ligands forms stable coordinatively
properties were studied. The strong characteristic red and green
saturated Ln(III) complexes which protects it from quenching
luminescence in the visible region was observed for 1 and 2 upon
through non-radiative vibrational energy dissipation for
photo-excitation, followed by efficient energy transfer from dopamine
application in luminescence imaging or sensing. Thus,
to Ln(III) leading to intraconfigurational f-f transitions. They show
design of the Ln(III) complexes with polydentate ligands like
distinguishable sharp emission bands due to intraconfigurational
5
D07FJ transitions in Eu(III) and 5D47FJ transitions in Tb(III). They DTPA, DOTA appended with a sensitizing antenna
exhibit long excited-state lifetimes: = 0.52 ms (1) and 0.33 ms (2) established as an important approach for designing highly
and calculated quantum yields (Фoverall) = 0.022 (1) and 0.038 (2). The emissive lanthanide based bioprobes for cellular imaging
luminescent complexes were studied for their antioxidant activity, and sensing.[2,3] Recently, K.-L. Wong et al. strategically
ability to interact with calf thymus-DNA (CT-DNA) and bovine serum designed unique lanthanide probes for selective
albumin (BSA), hydrolytic cleavage of pUC19 supercoiled plasmid photodynamic therapeutic (PDT) applications,[4] photo-
DNA and as cellular imaging probes for both cancer cells and triggered controlled delivery and tracking of anticancer
neuroblastoma cells. Both Eu(III) and Tb(III) complexes exhibit potent drugs[5], imaging probe for early prognosis of disease
antioxidant activity. These complexes showed moderate binding biomarkers. [6]
affinity towards CT–DNA (Kb~104 M-1) and BSA (KBSA =103 M-1). Dopamine is an important catecholamine
Complexes at 200 µM exhibit good hydrolytic cleavage of supercoiled neurotransmitter and regulates cognitive abilities and various
pUC19 DNA. The non-cytotoxic nature of these complexes makes
motor functions in humans.[7-10] Several pathobiological
them efficient candidates for cellular imaging probes. Cytosolic
studies reveal an intricate relationship between the
localization is observed for both the complexes in HeLa cells and
imbalance of dopamine and several neurodegenerative
mouse neuroblastoma (Neuro-2a) cells. The present systems with
diseases: viz. Perkinson’s disease (PD), Alzheimer disease,
advantageous antioxidant activity may have potential application in
tracking and delivery of dopamine, which is important for dopamine Huntington’s disease, multiple sclerorosis etc. Dopamine
replacement therapy in various neurodegenerative disorders. mediates its action through binding with dopamine receptors,
thus expression, distribution and mapping of various
dopamine receptor subtypes by bioimaging intended to be
an effective tool towards diagnosis and early prognosis of
Introduction various neurodegenerative diseases. Moreover, metabolism
of various catecholamine-based neurotransmitters like
Luminescent lanthanide probes have certain distinct dopamine, norepinephrine, serotonin provides an antioxidant
incentives in designing and development of traceable defense in brain against oxidative stress damage induced by
delivery vehicles for therapeutic or diagnostic loads. This is free radicals or iron independent or dependent lipid
due to their unique photophysical properties like sharp peroxidation.[11-13] Certain biological characteristics such as
distinguishable emission bands, long excited state lifetimes water solubility, efficient cellular internalization and non-
and large ligand-induced pseudo–Stokes shift.[1-3] The f-f toxicity at working concentration are important criteria for
transitions are Laporte and spin-forbidden leading to long lanthanide complexes to be used as a cellular probe. This
excited state lifetimes in the ms - µs range. These favourable work stems from our current interest in developing
optical properties of lanthanides leads to many opportunities luminescent Ln(III) complexes for cellular imaging and
delivering therapeutic loads.[14]
Here we have conjugated dopamine as an antenna and
[a] Department of Chemistry, Indian Institute of Technology Kanpur,
as potential antioxidant to diethylenetriaminepentaacetic
Kanpur 208016, Uttar Pradesh, India. E-mail: akpatra@iitk.ac.in
https://www.iitk.ac.in/new/ashis-k-patra acid (DTPA) and prepared respective luminescent probes:
[b] Department of Biological Sciences and Bioengineering, Indian [Ln(DTPA-DOPA)(H2O)] [Ln(III) = Eu(III) (1), Tb(III) (2)]
Institute of Technology Kanpur, Kanpur 208016, Uttar Pradesh,
(Scheme 1). Eu(III) and Tb(III) were used for their longer
India.

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
excited state lifetimes () in µs-ms range and emission in Physiochemical [Eu(DTPA-DOPA)(H2O)] [Tb(DTPA-DOPA)(H2O)]
visible region. The photophysical properties, interactions with Data (1) (2)
CT-DNA and BSA, anti-oxidant activity, DNA hydrolysis, and ESI-MS in H2O 814.17 820.18
potential cellular imaging applications were studied. (m/z) [M−H2O+H]+
FT-IR[a] (KBr 1631 (s, νasym CO2−) 1587 (s, νasym CO2−)
−1
pellet, νmax, cm ) 1527(s, νCO of CONH) 1528(s, νCO of CONH)
1384 (s, νsymCO2−) 1384 (s, νsymCO2−)
λmax[b]/ nm 282 (8325) 282 (7134)
(ε/M−1cm−1)
H2O and D2O[c] 0.52 and 1.43 0.33 and 0.36
qEu and qTb[d] 1.2 1.1
ϕEu and ϕTb[e] 0.038 0.022
[a]
In KBr phase. [b]UV-visible spectra in MQ water. [c] Lifetime measured in H2O

Accepted Manuscript
and D2O (τ ± 15%). [d]Hydration number (q =10%).[e]Overall quantum yield (ϕ ±
10%) using quinine sulphate as standard.

This strong coordinative property led the compounds 1 and 2 to

be stable in aqueous medium for prolonged time at room
temperature. The structural integrity of the complexes were also
evidenced by ESI-MS which showed [M-H2O+H]+ peak and
Scheme 1. Simplified scheme for [Ln(DTPA-DOPA)(H2O)] (Ln= Eu(III) (1) and
Tb(III) (2)) complexes showing the energy transfer to Ln(III) through dopamine
matching isotopic distribution pattern in mass spectral profile
moiety and multiple biological importance of the systems. clearly showing the formation of [Ln(DTPA-DOPA)(H2O)] in 1:1
stoichiometry.

Results and Discussion

Synthesis and Characterization

The H3DTPA-DOPA ligand was synthesized from acylation
reaction of DTPA bis(anhydride) with dopamine in dry DMF
at 70 C with 90% yield. The luminescent complexes 1 and
2 are synthesized by reacting H3DTPA-DOPA with
Eu(NO3)3·6H2O and TbCl3·6H2O in 1:1 mole ratio in water at
~80% yield and characterized by physicochemical methods, Figure 1. ESI-MS of (a) complex 1 with m/z [1 H2O + H]+ (C32H38EuN5O12) and
(b) complex 2 with m/z [2 H2O + H]+ (C32H38TbN5O12) in aqueous medium. Inset
ESI-MS, FTIR and UV-visible spectroscopy. The selected
figure shows the theoretical isotopic distribution pattern.
physicochemical data are given in Table 1. The ligand based
absorption peaks were observed at 282 nm due to π→π*
Photophysical Properties
transition. The IR data shows strong absorption ~1600 cm-1
Complexes 1 and 2 exhibit characteristic absorption bands at 282
ascribed to C=O of CO2- which shows shifts in the stretching
nm which can be assigned to the ligand based π→π* transition.
frequency by 20-60 cm-1 upon complexation suggesting At λex = 282 nm, Eu(III) and Tb(III) complexes showed
oxygen coordination to the Ln(III). These results suggests characteristic sharp emission peaks due to 5D0→7FJ (J = 0-4) and
that DTPA-bisamide coordinates to the Ln(III) by three 5
D4→7FJ (J = 6-3) transitions respectively (Figure 2). The excited
nitrogens, three carboxylate oxygen atoms, two amide state lifetimes were measured in H2O and D2O at RT and
oxygen and the ninth site is occupied by water molecule to luminescence decay profile has been fitted by single exponential
form water-soluble thermodynamically stable complexes.[17] fitting to obtain H2O = 0.52 ms; D2O = 1.43 ms for complex 1
The Gd(III) complexes of analogous ligands were previously and H2O = 0.33 ms; D2O = 0.36 ms for complex 2 confirming the
reported as potential receptor-specific MRI contrast presence of single luminescent lanthanide species in solution.
agents.[15] Moreover, the redox stability of these trivalent The decay profiles and their mono exponential fittings were shown
Ln(III) complexes is crucial towards intracellular applications in Figure 2 (c, d). In D2O, the emissive lifetime increases due to
in presence of various reducing agents such as thiols, GSH non-radiative deactivation generated by O-D vibration is lower in
comparison to O-H vibration.[18] The hydration numbers, qEu =1.2
and ascorbates.[16]
and qTb = 1.2 denotes the number of coordinated water
Table 1. Selected physiochemical data for complexes 1 and 2
molecule(s) in the coordination sites calculated through modified
Horrock's equation.[19] The q values are in consistent with eight
coordination sites that are bonded to DTPA-bisamide derivative
and the ninth site is occupied by one water molecule.

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
The luminescence quantum yields measured for the complexes 1 observed in the absorption spectral titration of complexes
and 2 were 0.038 and 0.022 respectively which is indicative of the with CT-DNA attributed to the possible perturbation of π -
fact that the triplet state (3T) of dopamine transfers sufficient orbitals of DNA base pairs by the complexes. The intrinsic
energy to the emissive 5D0 or 5D4 excited state of Eu(III) and Tb(III) binding constants (Kb) of complexes towards CT-DNA: 0.78
respectively. x 104 M-1 (1) and 1.5 x 104 M-1 (2) respectively, suggesting
The luminescence intensity of the probes were sensitive to
moderately strong binding propensity towards CT-DNA.
changes in pH. We observed significant decrease in emission
Ethidium bromide (EthB) is a well-defined emissive probe
intensity of complex 1 with lowering pH due to protonation of
that shows intense emission upon binding to CT-DNA due to
carboxyl moieties of DTPA-DOPA ligand and possible breaking
its strong intercalation between the adjacent DNA base pairs.
down of the complex (Figure 2 (e, f) . Loss of structural integrity
and exposure of Ln3+ core of the complexes leads to significant The complexes exhibit competitive binding to DNA and
quenching by vibrational energy transfer to solvent molecules. displaces EthB and thereby decreasing emission intensity at
605 nm (Table 2). The displacement of EthB by these

Accepted Manuscript
complexes suggestive towards an intercalative or minor
groove binding.[21] The apparent binding constant (Kapp)
calculated are 9.8 x 104 M-1 for (1) and 7.2 x 104 M-1 for (2)
suggesting intercalative or minor grove binding mode for
complexes with moderate affinity.[22]

Table 2. Binding parameters for complexes with CT-DNA and BSA.

Physiochemical [Eu(DTPA-DOPA)(H2O)] [Tb(DTPA-DOPA)(H2O)]

Data (1) (2)
Kb [a]/ M-1 0.78 (±0.2) x 104 1.5 (±0.2) x 104
Kapp [b]/ M-1 9.8 x 104 7.2 x 104
KITC[c]/ M-1
K1 = 1.81 x 10 4
K1 = 9.94 x 104
K2 = 1.10 x 103 K2 = 8.47 x 103
[d] -1 3
KBSA /M 9.8 (±0.3) x 10 8.7 (±0.1) x 103
[e] -1 [f] 11
Kq /M [n] 9.8 x 10 [1.1] 8.7 x 1011 [0.98]
[a] [b]
Kb, intrinsic equilibrium DNA binding constant. Kapp, apparent DNA binding
[c] [d]
constant. KITC, Affinity constant from ITC titration. KBSA , Stern-Volmer
quenching constant for BSA fluorescence. [e] Kq, quenching rate constant. [f] n,
number of binding sites.

Figure 2. (a) UV – vis spectra of ligand H3DTPA-DOPA, complexes 1 and 2

(100 µM) in aqueous medium at 298 K. (b) Excitation and time-resolved
luminescence spectra for complexes 1 and 2 in aqueous medium at 298 K.
Lifetime measurement of (c) complex 1 and (d) 2 (100 µM) in H2O and D2O
medium and fitted by single-exponential decay curves. (e) The emission
intensity changes of complex 1 in aqueous medium with variation of pH at ex =
282 nm. (f) The changes in emission intensity of 5D07F2 transition at 616 nm
as a function of pH.

DNA Binding Studies

DNA is a critical pharmacological target for metallodrugs like
cisplatin, carboplatin or oxaliplatin or Pt(IV) prodrugs where
they cross-link with nitrogen of nucleobases of DNA, thereby
arresting transcription or replication and trigger cell-death Figure 3. (a) Electronic spectral traces of 1 with increasing [CT-DNA] in saline
pathways.[20] Considering potential biological activity of Tris–HCl buffer (5 mM, pH 7.2) at 25 C. Inset: plot of Δεaf/Δεbf vs. [DNA] and
[DNA] / εaf vs. [DNA] for 1. (b) Quenching of emission intensity of EthB bound
[Ln(DTPA-DOPA)(H2O)] (1, 2), the mode of interaction of the
CT-DNA adduct with increasing [1] in 5 mM saline Tris–HCl buffer (pH 7.2) at
complexes with CT-DNA at physiological condition were 25 C; ex = 546 nm, em= 605 nm, [EthB] =3.0 μM, [CT-DNA] = 280 μM. Inset:
studied using different spectroscopic techniques. The plot of relative fluorescence intensity (I/I0) at λem = 605 nm vs. concentration of
bathocromic shift (2-4 nm) and hypochromism (72-75%) complexes 1 and 2. (c) ITC plot showing the interaction of CT-DNA (0.1 mM)

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
with complex 1 (1 mM) in 5 mM Tris–HCl/NaCl buffer (pH 7.2) at 25C. The top in structural conformation of BSA. The quenching constants
plot shows the raw ITC data for 20 injections of 2 μL each. The bottom panel
represents extent of evolved heat vs. mole ratio of [1]/[CT-DNA].
(KSV) for the complexes was determined from Stern-Volmer
equation[27] The KSV values obtained for the complexes are:
9.8 x 103 M-1 (1) and 8.7 x 103 M-1 (2) respectively. The
Isothermal titration calorimetry (ITC) experiments, designed
binding constants (K) and number of binding sites (n) were
to investigate the DNA-binding thermodynamics of the
obtained from curve fitting of Scatchard equation: log(I0-I)/I =
complexes with CT-DNA were performed in Tris-HCl/NaCl
logK + nlog[Q].[28] The kq values for the complexes 1 and 2
buffer (5 mM, pH 7.2) at 30 C using MicroCal iTC200 system.
are ∼1011 M−1s−1 suggesting existence of static quenching
The binding data were fitted by two mode of binding
mechanism (Table 2).[29]
interaction with CT-DNA. The ITC fitting reveals that
interaction of these complexes with DNA is exothermic and
biphasic at two binding sites with varying affinities attributed Hydrolytic Cleavage of DNA
for such hairpin shaped complexes.[23] The calculated The high Lewis acidity and higher effective charge

Accepted Manuscript
binding constant values are K1 = 1.81 x 104 M-1, K2 = 1.10 x associated with the trivalent lanthanide complexes makes
103 M-1 (1) and K1 = 9.94 x 104 M-1, K2 = 8.47 x 103 M-1 (2). them effective catalysts for phosphodiester bonds of DNA or
The ITC plot and best-fitting model are shown in Figure 3 (c). RNA under physiological conditions.[30,31] The ability of Ln-
The affinity constants obtained are consistent with the Kb and complexes to readily catalyze the DNA hydrolysis is highly
Kapp values measured through UV-vis and emission titration. desirable to develop lanthanide-based artificial nucleases
important for various clinical applications including DNA-
Interaction with Serum Protein finger printing, sequence-specific DNA lesions etc.[31] The
BSA is the most extensively studied serum transport protein complexes 1 and 2 displayed efficient cleavage of plasmid
due to its structural homology with human serum albumin supercoiled DNA (SC, form I) to nicked circular (NC, form II)
(HSA) and serve important roles in drug transport, storage form in dark.[32] Both the complexes at 100 µM
and metabolism.[24,25] The fluorescence of BSA originates concentrations effectively promoted cleavage of SC-DNA to
from the intrinsic fluorescence of the tryptophan residues, 80% NC form when incubated for 5 h (Figure 5 a). The
Trp-134 and Trp-212 located at the surface and within a gradual disappearance of SC-DNA form and the appearance
hydrophobic binding pocket respectively. The interaction of of NC form of DNA with increasing time is observed. The
the tested complexes and BSA was investigated using linear curve fitting of ln(%SC-DNA) vs. time plot suggesting
tryptophan fluorescence quenching. The intrinsic BSA pseudo-first-order reaction profile. The rate constant (k) for
fluorescence at 342 nm gets quenched in presence of these this conversion obtained was 5.9 x 10-3 s-1 and 6.0 x 10-3 s-1
complexes, indicating their effective binding interaction with in presence of 100 M complexes 1 and 2 respectively. To
BSA (Figure 4). The fluorescence quenching possibly investigate the mechanism of DNA phosphodiester cleavage
induced from a variety of molecular interactions like by the complexes, hydroxyl radical scavengers (DMSO and
collisional encounters between the fluorophores and glycerol), singlet oxygen scavengers (NaN3) were added
quenchers or ground-state complex formation. prior to addition of complex 1 and it was observed that DNA
cleavage was not inhibited. The results showed that DNA
cleavage by complex 1 is not inhibited by any of the classical
radical scavengers (OH, 1O2).[31]

Figure 4. (a) Fluorescence spectral traces of BSA showing the quenching effect
upon addition of complex 1 in saline Tris-buffer (5 mM, pH 7.2) at 25 C. Inset:
the plot of I0/I vs. [Complex] for 1 and 2. (b) Scatchard plots between log (I0-I)/I
vs. log[complex] ex=295 nm, [BSA]=5 M.

The other possible factors include perturbation of secondary

structure of BSA, subunit association, conformational
changes induced by these quenchers.[26] The observed small
blue shift in em (~15 nm) possibly resulted from the changes

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
Figure 5. (a) Agarose gel electrophoresis of pUC19 DNA (0.2 μg, 30 μM) in
saline Tris–buffer (50 mM, pH 7.2) with complex 1 (100 µM) on increasing the
incubation time showing the disappearance of SC DNA and appearance of NC
DNA. Inset: Gel-electrophoresis picture snapped for both complex 1 and 2 with
increasing time. (b) Plot of ln(%SCDNA) vs. time for complex 1 (100 µM). (c)
Gel electrophoresis of pUC19 DNA incubated for 5 h at 37 °C and pH 7.2
(50 mM Tris–HCl) with increasing [complex]: L1, DNA control; L2, Eu(NO3)3,
200 μM; L3–L7, complex 1 of 10, 20, 40, 100, 200 μM concentrations
respectively. (d) gel electrophoresis of pUC19 DNA of 1 with different additive: Figure 6. (a) DPPH radical scavenging ability of tested compounds expressed
L1, DNA control; L2, DNA + 1; L3, DMSO (0.4 M); L4, glycerol (0.4 M); L5, NaN3 as percentage reduction of absorbance of DPPH measured at max = 517 nm in
(2 mM); L6, H2O2 (50 μM) after incubation at 37 C for 5 h. aqueous medium of pH 7.2 at 298 K. (b) ABTS cation radical scavenging ability
of tested compounds measured as percentage inhibition of ABTS absorbance
at λmax = 734 nm in aqueous medium of pH 7.2 at 298 K.
When oxidant (50 µM H2O2) was added to the reacting
mixture, it could not enhance the DNA cleavage induced by
the complexes. Therefore, the DNA cleavage promoted by The antioxidant activity of the complexes 1 and 2 was further

Accepted Manuscript
the complexes might not occur via an oxidative pathway. The studied by scavenging of cationic ABTS+ radicals by
hydrolytic cleavage of phosphodiester bonds possibly measuring percentage inhibition of the initial ABTS+
involve occurred via readily oxidizable appended catechol absorbance at λmax= 734 nm. The Eu(III) and Tb(III)
group forming dopamine semiquinone or quinone radicals. complexes showed 53.3% and 59.1% scavenging ability
Antioxidant activity compared to Trolox (89.1%) used as reference antioxidant
(Fig. 6b). Both the tested Ln-compounds are more active
Overproduction of free radicals due to imbalance of defence
than free H3DTPA-DOPA ligand (41%) and in consistent with
or immune mechanism eventually leads to a wide range of
the DPPH scavenging assay. Enhanced radical scavenging
diseases such as chronic inflammation, cancer, rheumatoid
by Ln-complexes compared to the free ligand clearly show
arthritis, cardiovascular disease, diabetes, aging and
significant role of Ln(III) centre.
especially neurodegenerative diseases.[33,34] The role of
Intracellular localization studies
antioxidants are to be antagonistic towards the free radicals
thereby protecting us from such diseases. The oxidative In order to understand the potential ability of the emissive Eu(III)
and Tb(III) compounds as visible bioimaging probes, the cellular
stress originated from ROS and free radicals contributes to
imaging capabilities of the complexes were studied using HeLa
the progression of several neurodegenerative disorders like
cells and mouse neuroblastoma cells (Neuro-2a) by fluorescence
Alzheimer’s disease, Parkinson’s disease etc.[34,35]
microscopy (Figure 7). DAPI was used as nucleus stain to
Therefore, considerable attention has been directed for the
ascertain the intracellular localization of the complexes. The
development of anti-oxidant based therapies for limiting conjugation of dopamine with DTPA is a possible promising
neuronal cell damage.[36,37] Dopamine and related strategy in visualizing delivery or understanding mechanism of the
catecholamine based neurotransmitters (viz. L-DOPA, dopamine action pathways for neurodegenerative diseases by
epinephrine, norepinephrine) or phenolic-OH containing utilizing the intrinsic luminescence of Eu(III) and Tb(III) in visible
tyramine, -tocopherol, tyrosine reported to show antioxidant region avoiding background autofluorescence.[38]
activity due to presence of catechol or phenolic-OH The complexes showed intrinsic lanthanide luminescence
moiety.[11-13] upon direct excitation of Ln(III) within HeLa cells and mouse
The presence of appended dopamine moieties in neuroblastoma cells depicting their cellular localization within 24
[Ln(DTPA-DOPA)(H2O)] (1, 2) prompted us to study their h of incubation. The Eu(III) and Tb(III) complexes showed their
ability to scavenge free radicals to evaluate promising characteristic red and green luminescence upon internalization.
antioxidant capacity. The ability of the complexes 1 and 2 to The cells remained healthy and viable during complete
experiment tenure suggesting their non-cytotoxic nature (Figure
scavenge DPPH and ABTS (ABTS+) radicals was assessed
7a). Counterstaining with DAPI indicated cytoplasmic localization
and compared with the well-established antioxidant agents
of the complexes inside the HeLa and mouse neuroblastoma cells
(e.g. butylatedhydroxytoluene (BHT), 6-hydroxy-2,5,7,8-
(Figure 7b,c) suggesting towards effective application of these
tetramethylchromane-2-carboxylic acid (Trolox)) used as
complexes as cellular imaging agents.
reference standard to evaluate potential antioxidant activity.
DPPH free radical exhibit a strong absorption peak at max =
517 nm which on gaining protons from antioxidants
decreases its intensity. The quantification was done after 20
and 60 minutes, which did not show any notable change
demonstrates that the scavenging action is time
independent. The effectiveness of the complexes to
scavenge DPPH radical is quite promising compared to BHT
as reference (60% for complexes, 30.4% for H3DTPA-
DOPA and 73.6% for BHT) (Figure 6a).

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
thus promising candidates for cellular imaging studies. The
fluorescence imaging study with HeLa and neuroblastoma cell
lines revealed that these complexes showed cytosolic localization
inside the cell convincing towards their good candidature for cell
imaging application. This strategy developed here may have
potential application towards delivery and in situ tracking of
dopamine, other neurotransmitters or therapeutic loads important
for the treatment of neurodegenerative diseases.

Experimental

Materials: The A.R. grade chemicals used were purchased

Accepted Manuscript
commercially and used with no further purifications. The solvents
were of spectroscopic grade or were purified by standard
procedures.[39] Eu(NO3)3·6H2O and TbCl3·6H2O were purchased
from Sigma-Aldrich. 2,2-diphenyl-1-picrylhydrazyl (DPPH) and
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)
(ABTS) were purchased from Sigma (U.S.A). 3-(4,5-
Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT),
trypsin–EDTA, Dulbecco’s modified eagle’s medium (DMEM,
Gibco® Life Technologies, Bengaluru, India), penicillin–
streptomycin antibiotic, bisBenzimide H33258 and gelatin (from
cold water fish skin) were purchased from Sigma-Aldrich
(Bengaluru, India) and used as received. Tris-(hydroxymethyl)-
aminomethane-HCl (Tris-HCl) buffer solution was prepared using
Milli-Q water (18.2 MΩ). Calf thymus DNA (CT-DNA), bovine
serum albumin (BSA), ethidium bromide (EthB) were purchased
from Sigma (USA).

Measurements: The elemental microanalyses for C, H and N

were done using a Perkin-Elmer 2400 Series-II elemental
Figure 7. (a) Cytotoxicity profile of the complexes 1 and 2 with HeLa cells on 24 analyzer instrument. Infrared spectra were taken in Perkin-Elmer
h incubation at 37 ᴏC as evaluated by MTT assay. (b) Fluorescent microscopic model 1320 FT-IR spectrometer in KBr pellets in the 4000–400
images of HeLa cells on treatment with complexes 1 and 2 (100 μM) for 24 h at
cm−1 range. Electrospray ionization mass spectral (ESI-MS)
37 ᴏC and DAPI (5 μg mL−1): λex (1) = 405 nm, λex (2)= 488 nm; scale bar = 10
μM. (c) Fluorescence microscopic images of mouse neuroblastoma cells
measurements were achieved using a WATERS Q-TOF Premier
(Neuro-2a) on treatment with complex 1 (100 μM) for 24 h at 37 ᴏC and DAPI
mass spectrometer. Electronic spectra were recorded at room
dye (5 μg mL−1): λex = 405 nm; scale bar = 10 μm for all the panels. temperature using a Perkin-Elmer Lambda 25 UV-vis
spectrophotometer and fluorescence spectra by Agilent Cary
eclipse fluorescence spectrophotometer at 298 K. ITC binding
study were measured using MicroCal iTC200. Lifetime
Conclusions measurements for complex 1 and 2 was conducted by using
pulsed xenon lamp at λex =282 nm and λem = 616 nm and 543 nm
In conclusion, this work focusses on the detailed investigation on for Eu(III) and Tb(III) complexes respectively with a delay time and
nine-coordinated luminescent DTPA-bisamide dopamine gate time of 0.1 ms under ambient condition. Decay curve fitting
was done through non-linear least square method. The
complexes viz. [Ln(DTPA-DOPA)(H2O)] of Eu(III) (1) and Tb(III)
luminescence quantum yield measurement was performed in
(2). The complexes were water soluble and thermodynamically aqueous medium at pH 7.2 at 25 ᴏC by taking quinine sulphate as
stable under experimental condition. Eu(III) complex exhibited red reference with the help of reported procedure using the following
and Tb(III) showed green luminescence on excitation at the ligand equation[40]
centered band at 282 nm which is indicative of efficient energy
transfer from the triplet excited state of the ligand (chromophore) 𝐴ref 𝐼𝑛2
to the Eu(III) (5D0→7Fj) and Tb(III) (5D4→7FJ). DNA binding assay ∅overall = ∅ref 2
𝐴𝐼ref 𝑛ref
of the complexes 1 and 2 were performed by UV–Vis, emission where A, I and n represents the respective absorbance at the
spectroscopy and isothermal titration (ITC) and the results excitation wavelength, area under the emission spectral curve
obtained proves that the complexes interacts to CT-DNA by and refractive index of the solvent respectively.
groove binding or partial intercalation. They are also effective The ϕref represents the quantum yield of the standard quinine
towards the cleavage of pUC19 DNA through the hydrolytic sulfate solution. The lifetime measurements in excited state in
pathway. The antioxidant activity demonstrates their promising H2O and D2O allowed the determination of the coordinated water
free radical scavenging activity than free ligands and reference molecules (q) directly attached to the respective lanthanide center
standards. The complexes were non-cytotoxic upto 100 µM and

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
using the following modified Horrocks' equations for Eu(III) and 822.18 (27.5%). FT-IR (KBr pellet, νmax, cm−1): 3402 (w), 1587 (s,
Tb(III) respectively.[19] νasym CO2−), 1528 (s, νC=O of CONH), 1440 (m), 1384 (s, νsymCO2−),
1 1 1 1 1328(m), 1115 (m), 930 (m), 817 (m), 715 (m). UV-vis (H2O, 298
q Eu =1.2 ( - - 0.25) , q Tb =5.0 ( - - 0.06)
τH2O τD2O τH2O τD2 O K), λmax, nm (ε, M−1 cm−1): 282 (7135 M-1cm-1).

Fluorescence images were captured using an epifluorescence DNA Binding Methods

microscope (Axiovision) attached with the ApoTome module (Carl
Interaction of Eu(III) and Tb(III) complexes were carried out
Zeiss, Bangalore, India).
by UV-visible absorption titration, ethidium bromide (EthB)
Synthesis of N,N’-bis(3-hydroxytyramide)diethylenetriamine displacement assay, and isothermal titration calorimetry
N, N’, N’’-triacetic acid (H3DTPA-DOPA) (ITC) study.
The ligand was prepared following a modified literature The binding study of the complexes 1 and 2 with CT-DNA

Accepted Manuscript
procedure and characterization data shows close resemblance were conducted in saline-Tris buffer (5 mM, pH 7.2). The
with reported values.[15] 3-hydroxytyramine hydrochloride (0.2 g, A260/A280 ratio of 1.9 for CT-DNA in Tris buffer showed DNA
1.05 mmol) was added to DMF solution of is free from proteins. The DNA concentration is calculated
diethylenetriaminepentaacetic acid dianhydride (0.189 g, 0.52 from the A260 value and known 260 = 6600 M-1.[41] In this
mmol) and then stirred at 80 C under N2 atmosphere for 8 h. The assay, the complex concentration was kept constant while
solvent was removed by evaporation, 5 mL of distilled water was varying the concentration of DNA during titration. The
added to the resulting compound, and pH was adjusted to 8.0 with intrinsic interaction constant (Kb) was derived from the
0.1 M NaOH. The solvent was further evaporated under reduced
equation [41]: [DNA]/(af) = [DNA]/(bf) + 1/ Kb(bf)
pressure, washed with ethanol, and dried in vacuo overnight.
where [DNA] is the CT-DNA concentration, εa is the apparent
(Yield: 0.36 g, 92%). ESI-MS in H2O (m/z): [M + H]+ calcd for
extinction coefficient of complexes, εf and εb represent the
C30H41N5O12: 664.27; Found: 664.28.1H-NMR (D2O), δ (ppm):
extinction coefficient of complex in free form and fully bound
2.64 (t, 4 H); 2.98 (t, 4 H); 3.10 (m, 4 H); 3.39 (m, 4 H,); 3.50 (s, 2
H); 3.56 (s, 4 H) ; 3.70 (s, 4 H); 6.56–7.12 (m, 6 H). FT-IR (KBr form to CT-DNA. The intrinsic binding constant is calculated
pellet, νmax, cm-1 ): 3420 (br), 1651 (vs, νC=O of CONH1), 1529 (s, by ratio of slope and intercept of [DNA]/(af) vs. [DNA] plots.
νC=O of CONH2), 1440 (m), 1404 (s), 1286 (m), 1120 (s), 1050 (m), For DNA binding study through emission spectroscopy, ethidium
920 (m), 817 (m), 707 (m) (vs, very strong; s, strong; m, medium; bromide (EthB) was used as emissive spectral probe in saline Tris
br, broad). UV-vis (H2O, 298 K), λmax, nm (ε, M-1cm-1): 282 (5150 buffer (5 mM, pH 7.2) to calculate the apparent binding constant
M-1cm-1). of the complexes with CT-DNA. The fluorescence spectra were
Synthesis of [Ln(DTPA-DOPA)(H2O)] (Ln = EuIII (1), TbIII recorded on excitation at 546 nm with an emission at 605 nm with
(2)): The complexes 1 and 2 were prepared by following general increasing concentration of the complexes. The apparent binding
procedure. H3DTPA-DOPA ( 0.2 g, 0.3 mmol, pH 8) dissolved in constant (Kapp) was calculated from the equation:21 Kapp [C]50 =
5 mL of distilled water was added dropwise to the 5 mL aqueous KEthB  [EthB], where [C]50 is the complex concentration when
solution of respective lanthanide salts, viz. Eu(NO3)3•6H2O (0.101 emission intensity reaches half of the CT-DNA bounded ethidium
g, 0.3 mmol) and TbCl3•6H2O (0.079 g, 0.3 mmol) at 50 C. The bromide emission.
reaction was continued for 6 h to obtain light yellow coloured The isothermal calorimetric titration (ITC) was performed to
solution. The solution was evaporated and the desired product study the CT-DNA interaction with Eu (III) and Tb (III) complexes
was washed with diethyl ether and finally dried in a vacuum over by MicroCal iTC200 system. The titration was done in
P4O10 [yield: ~80%]. homogeneous solvent medium for both CT-DNA and complexes
[Eu(DTPA-DOPA)(H2O)] (1): Yield: 0.213 g (81%). Anal. calcd for at 30 ᴏC in saline tris buffer (5 mM, pH 7.2). The samples were
C30H40EuN5O13: C, 43.38; H, 4.85; N, 8.43. Found: C, 43.62; H, thoroughly degassed before titrating. The sample cell was feed
4.59; N, 8.55. ESI-MS in H2O (m/z): [M − H2O + H]+ calcd for with 0.02 mM CT-DNA and the complex was loaded in 40 µL
C30H38EuN5O12 (relative abundance): 814.17 (100.0%), 812.17 syringe at stirring rate of 1000 rpm. Titration initiated first by
(55%), 813.17 (29%), 815.17 (54%), 816.17 (19.0%). Found: baseline stability. A titration experiment consisted of 20
814.17 (100.0%), 812.17 (89%), 813.17 (41%), 815.17 (54%), consecutive injections of 2 μL volume and 4 sec duration each,
816.18 (14 %). FT-IR (KBr pellet, νmax, cm−1): 3410 (w), 1631 with a filter period of 5 sec. The reference power was set at 5
(s,νasym CO2−), 1527(s, νC=O of CONH), 1441 (m), 1384 μcal/sec with an initial delay of 60 sec. The resulting data were
(s, νsymCO2−), 1328(m), 1285 (m), 1116 (m), 930 (m), 816 (m). UV- fitted by sequential binding site model (number of sites= 2) using
vis (H2O, 298 K), λmax, nm (ε, M−1 cm−1): 282 (8325 M-1 cm-1). MicroCal® ORIGIN software supplied with the instrument.

Protein Binding Assay: The interaction of proteins with complex

[Tb(DTPA-DOPA)(H2O)] (2): Yield: 0.185 g (78%). Anal. calcd for
1 and 2 was studied by quenching of fluorescence of tryptophan
C30H40TbN5O13: C, 43.02; H, 4.81; N, 8.36. Found: C, 43.41; H,
in saline tris buffer (5 mM, pH 7.2). Buffer containing BSA was
4.71; N, 8.65. ESI-MS in H2O (m/z): [M − H2O + H]+ calcd for
excited at λmax 295 to get an emission spectra at 344 nm. The
C30H38TbN5O12 (relative abundance): 820.18 (100.0%), 821.18
emission peak was monitored by adding the complex 1 and 2
(33.4%), 822.18 (8.5%) Found: 820.18 (100.0%), 821.18 (52.4%),
which showed significant quenching on increasing concentration

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
of the complexes. The quenching constants (KSV or KBSA) for the Cellular Viability Assay
complexes was determined from Stern-Volmer equation:[27] I0/I= 1
Cellular Viability assay was done using HeLa cell line
+ kq0[Q] = 1 + KSV[Q]. Here I0 and I denotes the fluorescence
obtained from the national repository, the National Centre of
intensities in the absence and presence of quencher of
concentration [Q], τ0 the average lifetime of the fluorophore in the Cell Science, Pune, India. The cells were grown in
absence of quencher (10−8 s), kq is quenching rate constant, and Dulbecco's modified Eagle's medium (Sigma Aldrich
KSV is the Stern–Volmer quenching constant. For static quenching Chemicals Pvt. Ltd., New Delhi, India) supplemented with
interaction, both the binding constant (K) and the number of 10% (v/v) fetal calf serum, 100 units/ml penicillin and 100
binding sites (n) can be determined according to the Scatchard mg/ml Streptomycin. Cells grown on gelatin coated dishes
equation: log (I0 − I)/ I = log K + n log[Q]. The n and K can be were incubated with increasing concentrations of Eu(III) and
calculated by the slope and the intercept of the double logarithm Tb(III) complexes for a period of 24 hours.
regression curve of log(I0 − I)/ I versus log[Q]. Concentrations of 1, 10, 25, 50 and 100 µM were used for
each of the two compound and these were harvested along

Accepted Manuscript
with their experimental controls. Cell survival was assessed
Hydrolytic Cleavage of DNA: DNA cleavage assay was using the MTT method for measuring cellular viability. Cells
performed with SC pUC19 DNA of (30 µM, 0.2 µg) by observing were incubated with 0.5 mgml-1 3-(4,5-dimethylthiazol-2-yl)-
its conversion from supercoiled (SC) form to nicked circular (NC) 2,5-diphenyltetrazolium bromide (MTT) for 2 h at 37 °C prior
and linear DNA forms. The experiment was executed in 50 mM
to harvesting and the absorbance of the metabolic product
saline Tris buffer at pH 7.2 by the treatment of SC pUC19 DNA
was measured at 570 nm using a spectrophotometer. The
with varying concentration of Eu(III) and Tb(III) complexes
absorbance was thus plotted as a measure of percentage of
performed by further dilution upto 20 µL by agarose gel
cell survival.
electrophoresis technique. The treated DNA and complex solution
was kept for incubation for varying time at 37 ᴏC and then analysis
of the cleaved DNA was done by gel electrophoresis. The Intracellular localization Studies
mechanistic study was performed by adding different additives HeLa cells and rat neuroblastoma cells were cultured at 37
DMSO (0.4 M), NaN3 (2 mM), glycerol (0.4 M), H2O2 (50 µM) prior °C and incubated with complexes 1 and 2 (100 µM) for 24 h.
to the addition of complexes. After 24 h, cells were fixed with 4% paraformaldehyde. For
post fixation, the nuclei were stained with 4',6-diamidino-2-
phenylindole (DAPI) reagent. The cells were then mounted
Antioxidant Assay
in 1,4-diazabicyclo[2.2.2]octane (DABCO) mounting medium
DPPH Radical Scavenging Activity: To examine the free
and processed for imaging. Fluorescence images were
radical scavenging activity of the Eu(III) (1) and Tb(III) (2)
captured using an epifluorescence microscope (Axiovision)
complexes, DPPH assay was done using absorption
spectroscopy. DPPH being free radical, give strong absorption attached with the ApoTome module (Carl Zeiss, Bangalore,
peak at 517 nm at room temperature. On interaction of DPPH with India) and the images were assembled using the Adobe
H3DTPA-DOPA (0.1 mM), complexes 1 and 2 in aqueous medium, Photoshop.
change in absorption spectra is observed and depicted as %
reduction of the absorbance of DPPH. H2O was used as a control Acknowledgements
medium. The spectra were recorded after 20 and 60 minutes for We thank Science and Engineering Research Board (SERB),
each tested compounds.BHT is used as a reference compound. Government of India (EMR/2016/00521) and IIT Kanpur for
financial support. K. S. thank the Council of Scientific and
ABTS Radical Scavenging Activity: Stock solution of aqueous Industrial Research (CSIR) for research fellowship.
ABTS (2 mM) on reaction with potassium persulfate (0.17 mM)
produced ABTS•+.The complete mixture was kept in dark for 12-
14 h before being used. The reaction between potassium References
persulfate and ABTS takes place with stoichiometry of 0.5:1; so
[1] a) J.-C.G. Bünzli, Chem. Rev., 2010, 110, 2729–2755; b) J.-
in this case ABTS oxidation remains incomplete. Oxidation
C. G. Bünzli , S. V. Eliseeva, Chem. Sci., 2013, 4, 1939–
process starts immediately but the absorption maxima of the 1949.
solution stabilize after 6 h. So, ABTS•+ can be stored at room [2] a) M. C. Hefferin, L. M. Matosziuk, T. J. Meade, Chem. Rev.,
temperature in dark conditions for more than 2 days. The ABTS•+ 2014, 114, 4496–4539; b) A. J. Amoroso, S. J. A. Pope,
solution on dilution with ethanol gives absorption value at 0.7 at Chem. Soc. Rev., 2015, 44, 4723–4742; c) M. Sy, A. Nonat,
wavelength of 734 nm. 10 µl of reference compound (Trolox) or N. Hildebrandt, L. J. Charbonnière, Chem. Commun., 2016,
the compound to be tested is added and spectra was measured 52, 5080-5095.
after 2 minutes of mixing. The ability of the compounds for [3] a) E. J. New, A. Congreve, D. Parker, Chem. Sci., 2010, 1,
111–118; b) E. G.Moore, A. P. S. Samuel, K. N. Raymond,
scavenging ABTS radicals was plotted as the percentage
Acc. Chem. Res., 2009, 42, 542–552; c) S. Shuvaev, M.
inhibition of the absorbance with respect to the initial ABTS Starck, D. Parker, Chem. Eur. J. 2017, 23, 9974 – 9989.
solution (ABTS %).

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
[4] a) T. Zhang, R. Lan, C.-F. Chan, G.-L. Law, W.-K. Wong, [22] a) M. Lee, A. L. Rhodes, M. D. Wyatt, S. Forrow, A. Hartley,
K.-L. Wong, Proc. Natl. Acad. Sci. 2014, E5492-E5497. Biochemistry, 1993, 32, 4237-4245; b) M . J. Waring, J. Mol.
[5] a) H. Li, R. Lan, C.-F. Chan, L. Jiang, L. Dai, D. W. J. Kwong, Biol. 1965, 13, 269-282.
M. H.-W. Lam, K.-L. Wong, Chem. Commun., 2015, 51, [23] P. B. Glover, P. R. Ashton, L. J. Childs, A. Rodger, M.
14022-14025; b) H. Li, C. Xie, R. Lan, S. Zha, C.-F. Chan, Kercher, R. M. Williams, L. De Cola, Z. Pikramenou, J. Am.
W.-Y. Wong, K.-L. Ho, B. D. Chan, Y. Luo, J.-X. Zhang, G.- Chem. Soc., 2003, 125,9918–9919.
L. Law, W. C. S. Tai, J.-C. G. Bünzli, K.-L. Wong, J. Med. [24] D.C. Carter, J.X. Ho, Adv. Protein Chem., 1994, 45, 153-
Chem. 2017, 60, 8923-8932. 203.
[6] a) H. Li, R. Lan, C.-F. Chan, G. Bao, C. Xie, P.-H. Chu, W. [25] X. M. He, D. C. Carter, Nature, 1992, 358, 209−215.
C. S. Tai, S. Zha, J.-X. Zhang, K.-L. Wong, Chem. Commun., [26] a) U. Kragh-Hansen, Pharmacol. Rev., 1981, 33, 17–53.
2017, 53, 7084-7087; b) Z. Liang, T.-H. Tsoi, C.-F. Chan, L. b)T.Peters, Adv. Protein Chem,1985, 37, 161–245.
Dai, Y. Wu, G. Du, L. Zhu, C.-S. Lee, W.-T. Wong, G.-L. Law, [27] J.R. Lakowicz, Principles of Fluorescence Spectroscopy,
K.-L. Wong, Chem. Sci., 2016, 7, 2151-2156. 2006, 237–265.
[7] a) S. Basu, P. S. Dasgupta, J. Neuroimmunol, [28] D. Voet, Biochemistry, John Wiley & Sons, Inc., 3rd edn.,
2000,102,113–124; b) C. Sarkar, B. Basu, D. Chakroborty, 1995.

Accepted Manuscript
P. S. Dasgupta, Brain Behav. Immun., 2010, 24, 525–528. [29] A. Gong, X. Zhu, Y. Hu, S. Yu, Talanta, 2007, 73, 668–673.
[8] J. S. Fowler, N. D. Volkow, G.-J. Wang, N. Pappas, J. Logan, [30] a) M. Komiyama, N. Takeda, H. Shigekawa, Chem.
R. MacGregor, D. Alexoff, C. Shea, D. Schlyer, A. P. Wolf, Commun., 1999, 1443–1451; b) S. J. Franklin, Curr. Opin.
D. Warner, I. Zezulkova, R. Cilento, Nature, 1996, 379, 733 Chem. Biol., 2001, 5, 201–208.
– 73. [31] M. A. Camargo, A. Neves, A. J. Bortoluzzi, B. Szpoganicz,
[9] K. L. Davis, R. S. Kahn, G. Ko and M. Davidson, Am. J. F. L. Fischer, H. Terenzi, O. A. Serra, V. G. Santos, B. G.
Psychiatry, 1991, 11, 1474-86. Vaz, M. N. Eberlin, Inorg. Chem., 2010, 49, 6013-6025.
[10] E. D. Huey, K. T. Putnam, J. Grafman, Neurology, 2006, 66, [32] L. A. Basile, A. L. Raphael, J. K. Barton, J. Am. Chem. Soc.
17-22. 1987,109,7550-7551.
[11] J. Leu, A. Mori, Arch. Biochem. Biophys., 1993, 302, 118- [33] K. J. Barnham, C. L. Masters, A. I. Bush, Nat. Rev. Drug
127. Discov., 2004, 3, 205-214.
[12] G. C. Yen, C. L. Hsieh, Biosci. Biotech. Biochem., 1997, 61, [34] a) B. Uttara, A. V. Singh, P. Zamboni and R. T. Mahajan,
1646-1649. Curr. Neuropharmacol. 2009, 7, 65-74; b) L. M. Sayre, G.
[13] a) A. D. Stefano, P. Sozio, A. Cocco, A. Lannitelli, E. Perry, M. A. Smith, Chem. Res. Toxicol., 2008, 21, 172–188.
Santucci, M. Costa, L. Pecci, C.Nasuti, F. Cantalamessa, F. [35] S. D. Maleknia, K. M. Downard, Chem. Soc. Rev., 2014, 43,
Pinnen, J. Med. Chem., 2006, 49, 1486–1493; b) J. L. Cadet, 3244-3258.
C. Brannock, Neuro. Chem.Int.1998, 32, 117-131. [36] V. A. Carozzi, P. Marmiroli, G. Cavaletti, Curr. Cancer Drug,
[14] a) K. Singh, S. Singh, P. Srivastava, S. Sivakumar, A. K. 2010, 10, 670-682.
Patra, Chem. Commun., 2017, 53, 6144-6147; b) A. [37] J. E., Slemmer, J. J. Shacka, M. I. Sweeney, J. T.Weber,
Chandra, K. Singh, S. Singh, S. Sivakumar, A. K. Patra, Curr. Med. Chem., 2008, 15, 404-414.
Dalton Trans., 2016, 45, 494-497; c) S. Dasari, S. Singh, S. [38] L. T. Eliassen, G. Berge, A. Leknessund, M.Wikman, I.
Sivakumar, A. K. Patra, Chem. Eur. J., 2016, 22, 17387– Lindin, C. Lokke, F. Ponthan, J. I. Johnsen, B.
17396. Sveinbjornsson, P. Kogner, T. Flaegstad, O. Rekdal, Int. J.
[15] a) S. Laurent, T. N. Parac-Vogt, K. Kimpe, C. Thirifays, K. Cancer, 2006, 119, 493–500.
Binnemans, R. N. Muller and L. V. Elst, Eur. J. Inorg. Chem. [39] D. D. Perrin, W. L. F. Armarego, D. R. Perrin, Purification of
2007, 2061–2067; b) T. N. Parac-Vogt, K. Kimpe, K. Laboratory Chemicals, Pergamon Press, Oxford, 1980.
Binnemans, J. Alloys Compd. 2004, 374, 325-329. [40] J. R. Lakowicz, Principles of Fluorescence Spectroscopy,
[16] a) S. W. Young, K. W. Woodburn, M. Wright, T. D. Mody, Q. Springer, New York, 3rd edn, 2006.
Fan, J. L. Sessler, W. C. Dow, R. A. Miller, Photochem. [41] a) M. E. Reichmann, S. A. Rice, C. A. Thomas, P. Doty, J.
Photobiol. 1996, 63, 892-897; b) D. M.Guldi, T. D. Mody, N. Am. Chem. Soc., 1954, 76, 3047–3053; b) A. Wolfe, G. H.
N. Gerasimchuk, D. Magda, J. L. Sessler, J. Am. Chem. Soc. Shimer ,T. Meehan Jr, Biochemistry, 1987, 26, 6392–6396.
2000, 122, 8289-8298; c) J. L. Sessler, R. A. Miller, Biochem.
Pharmacol. 2000, 59, 733-739.
[17] a) S. Aime , F. Benetollo , G. Bombieri , S. Colla , M. Fasano ,
S. Paoletti, Inorg. Chim. Acta, 1997, 254, 63–70; b)Y.-M.
Wang , Y.-J. Wang , R.-S. Sheu, G.-C. Liu , W.-C. Lin, J.-H.
Liao, Polyhedron,1999, 18 , 1147 –1152.
[18] G. Stein, E. Würzberg, J. Chem. Phys. 1975, 62, 208-213.
[19] a) W. D. W. Horrocks Jr., D. R. Sudnick, J. Am. Chem. Soc.
1979, 101, 334-340; b) A. Beeby, I. M. Clarkson, R. S.
Dickins, S. Faulkner, D. Parker, L. Royle, A. S. de Sousa, J.
A. G. Williams, M. Woods, J. Chem. Soc., Perkin Trans.,
1999, 1, 493–504.
[20] a) A. Pyle, J.K. Barton, in: S.J. Lippard (Ed.), Progress in
Inorganic Chemistry, 1990, 38, 413-442; b) P.J Dandliker,
R.E Holmlin, J.K Barton, Science, 1997, 275,1465-1468.
[21] R. Palchaudhuri, P. J. Hergenrother, Curr. Opin. Biotechnol.,
2007, 18, 407-503.

This article is protected by copyright. All rights reserved.

European Journal of Inorganic Chemistry 10.1002/ejic.201800732

FULL PAPER
Entry for the Table of Contents

FULL PAPER
Luminescent Ln(III) Key Topic*
Probe: Luminescent Khushbu Singh, Anshika
lanthanide(III) complexes Goenka, Subramaniam
of DTPA bisamide Ganesh and Ashis K. Patra*
dopamine: [Ln(DTPA- Page No. – Page No.
DOPA)(H2O)] (Ln= Eu(III) Luminescent Eu(III) and
(1), Tb(III) (2)) were Tb(III) Complexes

Accepted Manuscript
explored for their Containing Dopamine
photophysical properties, Neurotransmitter:
interactions with DNA and Biological Interactions,
BSA, DNA hydrolysis, Antioxidant Activity and
anti-oxidant and neuronal Cellular Imaging Studies
cell imaging properties.
The complexes exhibit
efficient antioxidant activity
and intracellular
luminescence via via f-f
transitions in HeLa and
neuroblastoma cells
showing potential for
bioimaging applications.

This article is protected by copyright. All rights reserved.