Inorganic Chemistry Communications 188 (2026) 116550

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Inorganic Chemistry Communications

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Research Article

A turn-off fluorescent sensor for selective detection of toxic Hg2+ ions using
acridinedione-pyridine framework
S.P. Ragavi a , D. Thirumalai b , Jebiti Haribabu c , Tanay Debnath a , I.V. Asharani a,*
a
Department of Chemistry, School of Advanced Sciences, Vellore Institute of Technology, Vellore 632 014, Tamil Nadu, India
b
Department of Chemistry, Thiruvalluvar University, Vellore 632 115, Tamil Nadu, India
c
Facultad de Medicina, Universidad de Atacama, Los Carreras 1579, 1532502 Copiapo, Chile

A R T I C L E I N F O A B S T R A C T

Keywords: Metal ions are essential for numerous chemical and biological processes; however, Hg2+ is highly toxic, even at
Mercury ion trace concentrations. Therefore, efficient detection of Hg2+ is crucial. Fluorometric detection, using receptor-
Turn-off based sensors, presents a promising approach. In this study, we developed an acridinedione-pyridine-3,5-
Bio-imaging
dicarbonitrile-based receptor, ARP, for the sensitive and specific detection of Hg2+. ARP was characterized
Test kit
using FTIR, NMR (1H and 13C), and HRMS, respectively. When compared to other metal ions, the receptor
DFT study
showed significant selectivity for Hg2+, exhibiting a noticeable shift in color from yellowish-brown to light
yellow in acetonitrile solution. 1H NMR titrations, HRMS analysis, and DFT calculations were conducted to
determine the interaction between ARP and Hg2+ ions. The binding stoichiometry between ARP and Hg2+ was
observed to be 2:1, as verified by Job's plot. The limit of detection and quantification for Hg2+ were found to be
19.8 nM and 66.6 nM, respectively. The receptor exhibited a quick response to Hg2+ ions, and EDTA titration
studies confirmed its reversibility. ARP successfully detected Hg2+ in real samples, including water, paper strip,
and living HeLa cells, demonstrating a reliable and versatile sensor for monitoring mercury contamination in
environmental and biological systems.

1. Introduction skin rashes, reproductive abnormalities, and neurological disorders,

including sperm and ovary damage, as well as miscarriages [9–13]. The
Mercury (Hg2+) is a pervasive and intensely hazardous, worldwide Minamata disaster in Japan acts as a clear example of the devastating
contaminant that presents significant threats to human well-being and effects of mercury poisoning resulting from contaminated seafood [14].
the environment, even at trace concentrations [1]. Among heavy and In response to the global health threat posed by mercury, the U.S.
transition metal ions, Hg2+ is considered the most hazardous. Mercury Environmental Protection Agency (USEPA) strictly regulates the allow­
ions released due to industrial and human actions are converted into a able level of Hg2+ in drinking water and food to 10 nM (2 ppb) [15,16].
neurotoxic compound, methylmercury, by bacterial and chemical pro­ This underscores the need to advance selective and specific sensing
cesses [2,3]. Hg2+ contamination of water and soil results from the technologies for detecting Hg2+ in biological and ecological matrices.
oxidation of atmospheric mercury vapor [4], whereas aquatic bacteria Although traditional detection methods are widely used, they generally
further convert inorganic mercury into organic forms such as methyl­ involve complex sample preparation and costly equipment [17–20].
mercury, which bioaccumulates in fish and marine organisms [5,6]. This Fluorescence spectroscopy is a simpler, more cost-effective alternative,
bioaccumulation represents a serious threat to human health as mercury providing high sensitivity, rapid response, and real-time detection ca­
enters the food chain, leading to mercury poisoning and related diseases. pabilities. Fluorescent chemosensors, which interact with metal ions to
Coal-fired power stations, gold mining, and mercury-containing devices produce measurable changes in fluorescence, have emerged as prom­
such as lamps, thermometers, and barometers all contribute to the ising tools for the discriminative identification of metal ions [21–24].
widespread distribution of this toxic metal in air, water, and soil [7,8]. Although much literature is available on the surveillance of mercury
Mercury exposure can result in major health problems, such as DNA ions in the atmosphere, there remains a need for improved detection
damage and liver and kidney dysfunction, immune system suppression, strategies (Table S2), the design of a new chemosensor for detecting

* Corresponding author.
E-mail address: [Link]@[Link] (I.V. Asharani).

[Link]
Received 5 January 2026; Received in revised form 18 March 2026; Accepted 19 March 2026
Available online 20 March 2026
1387-7003/© 2026 Elsevier B.V. All rights are reserved, including those for text and data mining, AI training, and similar technologies.
S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Scheme 1. Synthesis of the receptor ARP

mercury ions at trace levels with improved selectivity and sensitivity 2. Experimental section
remains a critical area of research.
In this work, we developed ARP (2-amino-4-(4-(3,3,6,6,10-pentam­ 2.1. Chemicals and equipment
ethyl-1,8-dioxo-1,2,3,4,5,6,7,8,9,10-decahydroacridin-9-yl)phenyl)-6-
(pyrrolidin-1-yl)pyridine-3,5-dicarbonitrile), a new chemosensor for the Every chemical was obtained from SD Fine Chemicals, Sigma-
selective identification of Hg2+. The ARP, synthesized through a simple Aldrich, and TCI. Analytical-grade solvents from commercial vendors
condensation reaction, demonstrated exceptional fluorescence-based were utilized without additional purification. Every part of the prepa­
sensing properties. It exhibited high specificity and sensitivity for ration was performed using double-distilled water. Infrared (IR) spectra,
Hg2+, effectively distinguishing even in the occurrence of various metal spanning the 4000–400 cm− 1 range, were recorded by the KBr pellet
ions. ARP showed an excellent LOD, LOQ, and rapid response time, method on a Shimadzu FTIR spectrometer. Using a Bruker Avance
making it ideal for real-time detection. Furthermore, ARP enables the spectrometer, 1H and 13C NMR spectra were measured in DMSO‑d₆ and
identification of Hg2+ ions discernible to the naked eye through the CDCl₃ at 400 and 100 MHz, respectively. Mass spectrometry was per­
visualization study and paper strip test. In addition, fluorescence im­ formed on a WATER-XEVO G2XS-QT ESI mass spectrometer to analyze
aging in live HeLa cells demonstrates its applicability in biological the synthesized compounds. A JASCO V-750 spectrophotometer was
monitoring. Compared to previously reported fluorescent receptors, the used for recording UV–visible absorption spectra, while fluorescence
novel structural design of the ARP receptor significantly improves its study were carried out on a HITACHI F-7000. Bioimaging was carried
binding affinity and fluorescence response toward Hg2+ ions, out using an Olympus IX71 fluorescence microscope (Japan). The pre­
completing a notably lower LOD of 19.8 nM. Furthermore, the ARP viously described method [25] was used to produce the precursor
receptor successfully integrates real-time detection, visual recognition, chemical 4, and the supplementary material contains detailed synthetic
and bioimaging capabilities within a single platform. methods.

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

2.2. Synthesis of receptor ARP

Compound 5 (0.5 g, 1.2 mmol) was dissolved in ethanol (7 mL), and

pyrrolidine (2–3 drops) was added, resulting in an immediate color
change from light brown to dark brown. The reaction mixture was
refluxed for 30 min. Malononitrile (0.161 mL, 2.5 mmol) was then added
dropwise, and the mixture was further refluxed for 4 h. Upon comple­
tion, a reddish-brown solid was obtained, which was isolated as ARP
(C35H38N6O2, 75% yield, m. p. 272–274 ◦ C). 1H NMR δ (ppm) (400 MHz,
CDCl3): 6.90-6.77 (m, 4H), 6.75 (s, 2H, -NH2), 4.91 (s, 1H), 3.16 (s,
broad, 8H), 3.00 (s, 3H), 2.37-2.09 (2d, 4H), 1.91 (s, 4H), 0.80-0.74 (2 s,
12H). 13C NMR δ (ppm) (100 MHz, CDCl3): 196.2 (C=O), 168.2 (C–N),
166.5 (C-NH2), 162.5 (Aromatic C–C), 159.1 (C-N-C), 149.1, 137.4
(Aromatic C–C), 129.4-128.4 (Ar–C), 115.1 (-CN), 113.2 (Ar–C), 86.0
(C-CN), 53.0 (H2C-N-CH2), 50.6 (Aromatic -CH2), 40.8 (Aromatic -CH2),
32.1 (N-CH3), 32.1 (C-C-C), 29.2 (CH3), 27.2 (H2C-N-CH2). HRMS (m/z):
calculated mass: 574.3056 and found mass: 574.3031 [M+] (Figs. S1-
S3).

3. Results and discussion

5
Fig. 1. Absorption spectra of receptor ARP (2 × 10− M) upon adding different
metal ions (1 × 10− 3 M) in ACN solution 3.1. Design and synthesis of receptor ARP

Compounds 4 and 5 were synthesized according to the procedures

described in the Supporting Information [26]. The general synthetic
route for the preparation of receptor ARP is shown in Scheme 1. The
structure of the target compound was confirmed by spectral analyses,
including 1H NMR, 13C NMR and HRMS (Figs. S4-S9).

3.2. UV–vis absorption spectral studies of receptor ARP

The interaction of receptor ARP with different cations, like Al3+,

Ag+, Cr6+, Co2+, Cd2+, Cu2+, Mn2+, Ni2+, Fe2+, Fe3+, Hg2+, Pb2+, K+,
and Zn2+, was examined using absorption spectroscopy. As shown in
Fig. 1, the free receptor solution in acetonitrile (ACN) exhibited two
separate bands of absorption at 198 nm and 245 nm. When Hg2+ (1 ×
10− 3 M) aqueous solution is involved, a distinct absorption band
emerges at 250 and 350 nm, accompanied by a redshift [27]. In contrast,
the spectra did not significantly change when other metal ions were
present. These findings show that receptor ARP specifically interacts
with Hg2+, demonstrating its selective complexation behavior.

3.3. Fluorescence and visualization study of receptor ARP

The fluorescence response of receptor ARP after the addition of

several metal ions was also examined. When excited at 245 nm, receptor
ARP displayed an intense fluorescence peak at 445 nm. However, when
exposed to Hg2+, the emission intensity was completely quenched
(Fig. 2a). This selective quenching was further illustrated by a bar graph
(Fig. 2b), suggesting that the quenching mechanism may involve
chelation-enhanced quenching (CHEQ). In the fluorescence studies,
ARP displayed yellowish-brown fluorescence, which was entirely
quenched upon interaction with Hg2+. These results indicate that Hg2+
ions specifically bind to ARP, causing both a color change and a
reduction in emission intensity. The other metal ions tested showed no
discernible variations in fluorescence (Fig. 3). The quantum yields (Φ) of
ARP and the ARP–Hg2þ complex were calculated using quinine sulfate
(Φ = 0.546 in 0.1 M H₂SO₄) as a standard sample analyzed under
indistinguishable conditions. The quantum yields for the ARP receptor
and the ARP–Hg2þ complex were 38.1% and 2.04%, respectively. The
significant decrease in quantum yield upon Hg2+ binding indicates
strong fluorescence quenching, which can be ascribed to the develop­
Fig. 2. (a) ARP emission spectrum response with and without various metal ment of a stable ARP–Hg2þ complex that restricts molecular motion
ions (b) Bar diagram illustrating the relative fluorescence emission response of and promotes non-radiative decay pathways. These results confirm the
ARP at 445 nm following the addition of several cations.
efficient interaction between ARP and Hg2+ and are consistent with the
observed fluorescence quenching.

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Fig. 3. Visual fluorescence detection of ARP with various metal ions using a UV–vis chamber set at 365 nm

Fig. 5. Job's plot for ARP–Hg2þ.

Fig. 4. (a) FL titration of ARP with Hg2+ at several concentrations (b) Cali­
bration plot of ARP.

3.4. Study of fluorescence spectral titration

Fig. 6. B–H plot of the receptor ARP–Hg2+ complex.

The solution was excited at 245 nm at room temperature to perform a

fluorescence spectrum titration of the ARP receptor with Hg2+ ions in illustrating the fluorescence intensity of solutions containing different
acetonitrile. Interestingly, the intensity of the fluorescence at 445 nm mole fractions of Hg2+ ions, between 0.1 and 0.9. A significant drop in
gradually dropped as the concentration of Hg2+ ions raised from 0 to fluorescence intensity is noted at a mole fraction of 0.7 for Hg2+,
100 μM (Fig. 4a). The LOD for Hg2+ was calculated using the 3σ/s implying a likely 2:1 binding interaction between ARP and Hg2+ ions
standard formula from the titration data, resulting in a detection limit of [28]. The binding constant was calculated from the titration results
19.8 nM (1.980 × 10− 8 M) (Fig. 4b). Fig. 5 presents Job's plots using the Benesi–Hildebrand (B–H) method, which came out to be
3.362 × 105 M− 1 (Fig. 6). These outcomes demonstrate that ARP is an

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Fig. 8. (a) Interference analysis of ARP–Hg2+ complex with various metal ions
Fig. 7. (a) Reversibility study of ARP þ Hg2þ complex with EDTA (b) Recy­ (b) Time-dependent interaction of the ARP receptor to Hg2+ ions.
clability study of ARP þ Hg2þ complex.

active chemical tool for sensing Hg2+ ions, with a highly linear standard
plot (R2 = 0.9983) suitable for quantitative mercury analysis. Further
confirmation of the ARP–Hg2þ interaction was obtained using LC-
HRMS analysis. The spectrum exhibited a peak at m/z 1348.5648 cor­
responding to [M]+, which closely matches the calculated value of
1348.5651 (Fig. S10), thereby supporting the proposed molecular
composition of the ARP–Hg2þ complex.

3.5. Study of reversibility and recyclability

The reversibility of receptor ARP is a crucial factor in evaluating its

potential as a reusable fluorescent chemosensor. To assess this property,
the reversible behavior of the ARP–Hg2þ complex was investigated
through the sequential addition of EDTA (50 μL). Upon the addition of
EDTA, the fluorescence emission intensity at 445 nm was restored,
indicating the dissociation of Hg2+ from the receptor. Subsequent
addition of Hg2+ (50 μL) resulted in quenching of the emission intensity
once again (Fig. 7a). These observations confirm the reversible coordi­
nation between ARP and Hg2+, enabling its function as an “on–off”
fluorescent chemosensor in acetonitrile. The reversibility and repro­
ducibility of the system were further demonstrated through multiple Fig. 9. The pH plot on the receptor ARP and the complex ARP þ Hg2þ.
switching cycles achieved by the alternate addition of Hg2+ and EDTA
(50 μL each), with no significant loss in fluorescence response (Fig. 7b).

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Fig. 10. Optimized structure of receptor ARP and ARP–Hg2þ complex.

Fig. 11. ARP and ARP þ Hg2þ Frontier Molecular Orbital.

3.6. Interference and time response study higher temperatures, increased molecular motion weakens the
ARP–Hg2þ interaction, resulting in partial dissociation of the complex
To evaluate the receptor ARP's selectivity for the Hg2+ ion in the and a relative increase in fluorescence intensity [29] (Fig. S11).
presence of several cations, 150 μL of every ion was mixed with the pre-
formed ARP–Hg2+ complex. The emission intensity of ARP at 445 nm 3.7. Effect of pH on receptor ARP
persisted almost unchanged in the occurrence of these competing ions,
indicating that none of these metal ions interfered with the detection of The pH-dependent fluorescence behavior of receptor ARP and the
Hg2+ (Fig. 8a). The time-dependent response of the receptor ARP to­ ARP–Hg2þ complex was investigated over a pH range of 1.0–12.0. A
ward Hg2+ in ACN was investigated. When Hg2+ is added, a sharp significant increase in fluorescence intensity was observed for ARP at
decline in fluorescence intensity was detected, reaching a stable value pH 2, which remained stable up to pH 11. In contrast, pronounced
within 30 s (Fig. 8b). This behavior indicates fast binding kinetics and fluorescence quenching of the ARP–Hg2þ complex was observed from
complete complexation at room temperature, ensuring efficient and pH 3.0 and remained consistent up to pH 10.0 (Fig. 9). These results
rapid detection. indicate that the ARP–Hg2þ complex is stable and functionally active
Temperature-dependent fluorescence spectra further reveal that the over a broad pH range (pH 3–10), making it suitable for applications in
quenching of ARP by Hg2+ follows a static (chelation-enhanced environmental and biological sample analysis, where pH conditions may
quenching) mechanism. Upon Hg2+ binding, the fluorescence of ARP is vary.
strongly quenched at room temperature, reflecting the formation of a
stable ground-state ARP–Hg2þ complex. At lower temperatures,
3.8. DFT studies
reduced molecular motion further stabilizes this complex, leading to
enhanced quenching and lower fluorescence intensity. Conversely, at
To refine the structural properties and electronic configurations of

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Fig. 12. Tau's plot of ARP and ARP + Hg2+.

the receptor ARP and its Hg2+ complex, DFT study was performed in the and explaining the experimentally observed fluorescence quenching.
gas phase using the Gaussian 16 package. The optimized geometries and In addition, the stability of the ARP–Hg2þ complex was corrobo­
frontier molecular orbitals (FMOs) of ARP and the ARP–Hg2þ (Figs. 10 rated by ESP, DFT-D calculations, and Monte Carlo simulations were
and 11). Gas-phase computational methods were used to elucidate the performed to optimize adsorption configurations and evaluate the
intrinsic electronic structure and binding-induced changes in orbital adsorption energies of cations on the ARP surface. These simulations
distribution. Analysis of the FMOs reveals that the HOMO of ARP is explicitly incorporated water molecules to account for solvent effects,
predominantly localized on the acridinedione electron-donor frame­ providing a realistic description of ARP–Hg2þ interactions under
work, while the LUMO shows significant electron density shifted toward aqueous conditions (Figs. S12–S16 and Table S1).
the pyrrolidine moiety, indicating efficient intramolecular charge
transfer. The calculated HOMO and LUMO energies of ARP are 5.90 eV
3.9. Binding behavior of ARP toward Hg2+
and 1.79 eV, respectively, corresponding to a theoretical bandgap of
4.11 eV, which is in reasonable agreement with the experimentally
The 1H NMR titration study was accomplished to examine the
determined optical bandgap of 5.02 eV.
binding interaction between ARP and Hg2+ (Fig. 13). In the 1H NMR
Upon coordination with Hg2+, the ARP molecule adopts a more
spectrum of ARP (DMSO‑d₆), the -NH₂ proton signal is present at δ 6.7
planar conformation, resulting in stabilization of both frontier orbitals.
ppm. Upon gradual addition of Hg2+ (0.5–2 equivalents), this –NH₂
The HOMO and LUMO energies of the ARP–Hg2þ complex decrease to
signal progressively decreases and eventually disappears, while a new
6.10 eV and 2.30 eV, respectively, resulting in a reduced theoretical
-NH signal appears at δ 4.4 ppm, indicating a slight deshielding effect.
bandgap of 3.80 eV. This bandgap narrowing accounts for the red shift
The appearance of this -NH peak after adding 0.5 equivalents of Hg2+,
detected in the absorption spectrum of the ARP–Hg2þ complex relative
along with an increase in its intensity, clearly suggests coordination of
to the free receptor. The experimentally obtained bandgap of the com­
Hg2+ with the nitrogen atom of the -NH₂ group. These results are
plex (2.86 eV) is consistent with this theoretical trend [28] (Fig. 12).
consistent with a 2:1 binding stoichiometry, as indicated by the Job plot
Furthermore, strong coordination of Hg2+ to the electron-rich donor
analysis, demonstrating the establishment of a stable ARP–Hg2þ
sites of ARP facilitates efficient nonradiative decay pathways through
complex.
enhanced spin-orbit coupling, thereby suppressing radiative emission
Following complexation, an increase in the charge density within the

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Fig. 13. Titration of ARP with Hg2+ monitored by 1H NMR spectroscopy.

Fig. 14. FTIR spectrum of the receptor ARP and ARP–Hg2þ complex.

core moiety was observed, which likely contributed to the CHEQ effect. theory [27]. The strong and stable connection between the nitrogen
FTIR spectroscopy provided additional evidence for the binding inter­ atom and Hg2+ is supported by this preference, a finding that is further
action. The spectra of both ARP and the ARP–Hg2+ complex were substantiated by 1H NMR, FTIR, and HRMS data. Based on these
recorded (Fig. 14). Notably, while the stretching frequencies of all experimental results, a 2:1 binding stoichiometry between ARP and
functional groups, except for the -NH₂ moiety, remained largely unaf­ Hg2+ is proposed, as described in Scheme 2.
fected, a distinct shift in the -NH₂ stretching frequency was observed,
from 3348 cm− 1 to 3452 cm− 1, following the addition of Hg2+. This shift
3.10. Paper strip test
confirms the direct interaction between the -NH₂ nitrogen and Hg2+.
The structure of the ARP–Hg2þ complex has been theoretically
A paper strip test was performed to assess the receptor ARP's prac­
confirmed through DFT calculations, which were aligned with the Hard
tical usefulness. First, uniform sections of Whatman filter paper, soaked
and Soft Acids and Bases. Mercury is categorized as a soft acid that
in ARP solution and then dried for five minutes in a hot air oven. After
favorably interacts with soft bases like nitrogen (N) under the HSAB
drying for five minutes, each dried paper strip was separately immersed

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Scheme 2. Proposed sensing mechanism of ARP for Hg2+ ions.

Fig. 15. A paper strip test to assess the ARP receptor's interaction with different metal ions.

Fig. 16. Cytotoxicity test of receptor ARP.

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Fig. 17. Fluorescence bioimaging of HeLa cells using ARP (10 μM) and ARP with various concentrations of Hg2+ ions (50 μM).

significantly quenched, with no observable fluorescence (Fig. 17). This

Table 1 observation confirms that ARP functions effectively as a biosensor for
Recovery result of Hg2+ in various water samples. Hg2+ ions in live cells.
Sample Spiked amount of Hg2+ Found amount of Hg2+ Recovery
(μM) (μM) (%)
4.2. Analysis of water samples
DD Water 0 – –
10 9.72 ± 0.04 97.25
20 19.64 ± 0.08 98.21 To assess the practical feasibility of the ARP receptor for the detec­
30 29.54 ± 0.02 98.49 tion of Hg2+ ions, a quantitative analysis was conducted on various
Well 0 – – water samples, such as double-distilled (DD) water, well water, and river
Water 10 9.64 ± 0.06 96.41 water, gathered from different regions within the Vellore district, Tamil
20 19.53 ± 0.09 97.67
Nadu, India. Receptor ARP (20 μM) was added to each water sample,
30 29.38 ± 0.02 97.94
River 0 – – which was spiked with Hg2+ ion concentrations of 0, 10, 20, and 30 μM.
Water 10 9.65 ± 0.03 96.55 The recovery of Hg2+ ions by ARP in each sample ranged from 96.41%
20 19.45 ± 0.08 97.27 to 98.49% (Table 1). A plot of Hg2+ ion concentration versus fluores­
30 29.33 ± 0.04 97.77
cence intensity demonstrated a strong linear correlation for all water
samples (Fig. 18). These results confirm that receptor ARP is effective in
in various metal ions (1 × 10− 3 M). After treatment, the strips were quantifying Hg2+ in conservational water samples.
examined under UV illumination (365 nm). A distinct color shift was
observed exclusively in the strip treated with Hg2+, while no significant 5. Conclusion
color alteration was noted for the strips exposed to multiple metal ions.
These findings reveal the high selectivity of receptor ARP for Hg2+ ions In conclusion, the acridinedione-pyridine-3,5-dicarbonitrile-based
(Fig. 15). ARP receptor was successfully developed and thoroughly described as
an extremely sensitive and selective receptor for Hg2+ ion detection. The
4. Applications receptor showed extraordinary specificity for Hg2+ against other metal
ions, including Ag+, Cu2+, Pb2+, and Al3+, as demonstrated by a distinct
4.1. Cell imaging color shift from yellowish-brown to light yellow in acetonitrile. Binding
studies, including 1H NMR titration and Job's plot analysis, revealed a
To evaluate the ability of receptor ARP to detect Hg2+ ions in bio­ 2:1 stoichiometry for the ARP–Hg2þ complex, providing insight into
logical matrices, a bioimaging study was performed. Before the imaging the sensing mechanism. The receptor demonstrated a low limit of
experiments, the cytotoxicity of ARP was assessed by the MTT assay detection (19.8 nM), along with rapid response and good reversibility, as
against HeLa cancer cells [30,31]. The results indicated that ARP confirmed by EDTA. Furthermore, ARP was successfully applied for the
exhibited negligible cytotoxic effects, with no significant reduction in quantitative detection of Hg2+ in real-world water samples, test strips,
cell viability observed at higher concentrations (Fig. 16). These findings and live HeLa cells, highlighting its practical applicability in environ­
suggest that the ARP receptor is non-toxic and suitable for use in live- mental and biological monitoring. Overall, these results establish ARP
cell imaging applications for Hg2+ ion detection. as a versatile and reliable fluorescent sensor for Hg2+ detection, with
Following the successful toxicity evaluation, a bioimaging study was significant potential for real-world applications.
conducted using a fluorescence microscope to demonstrate the practical
utility of ARP in Hg2+ ion detection. HeLa cells were treated with ARP CRediT authorship contribution statement
(10 μM) and incubated for 1 h, resulting in strong red fluorescence. Upon
subsequent treatment with 50 μM Hg2+ ions, the fluorescence was S.P. Ragavi: Writing – original draft, Methodology, Investigation,

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S.P. Ragavi et al. Inorganic Chemistry Communications 188 (2026) 116550

Fig. 18. Quantitative Hg2+ detection in water samples using ARP at varying concentrations.

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[15] J. Du, M. Liu, X. Lou, T. Zhao, Z. Wang, Y. Xue, Y. Xu, Highly sensitive and S.P. Ragavi received her Bachelor of Science in Chemistry from
selective chip-based fluorescent sensor for mercuric ion: development and Muthurangam Government Arts and Science College, Vellore.
comparison of turn-on and turn-off systems, Anal. Chem. 84 (18) (2012) She did her Master of Science at Thiruvalluvar University,
8060–8066, [Link] Serkkadu, Vellore. Currently, she is pursuing her Ph.D at Vel­
[16] K.E. Kristian, S. Friedbauer, D. Kabashi, K.M. Ferencz, J.C. Barajas, K. O’Brien, lore Institute of Technology, Vellore. Her research areas are
A simplified digestion protocol for the analysis of hg in fish by cold vapor atomic focused on the synthesis of acridinedione and its chemosensor
absorption spectroscopy, J. Chem. Educ. 92 (4) (2015) 698–702, [Link] applications.
10.1021/ed500687b.
[17] A. Caballero, V. Lloveras, D. Curiel, A. Tárraga, A. Espinosa, R. García, J. Veciana,
Electroactive thiazole derivatives capped with ferrocenyl units showing charge-
transfer transition and selective ion-sensing properties: a combined experimental
and theoretical study, Inorg. Chem. 46 (3) (2007) 825–838, [Link]
10.1021/ic061803b.
[18] A.N. Anthemidis, G.A. Zachariadis, C.E. Michos, J.A. Stratis, Time-based on-line
preconcentration cold vapour generation procedure for ultra-trace mercury
determination with inductively coupled plasma atomic emission spectrometry,
Anal. Bioanal. Chem. 379 (2004) 764–769, [Link] D. Thirumalai is an Associate Professor in the Department of
2593-2. Chemistry, Thiruvalluvar University, Vellore, India. Received
[19] C.M. Tseng, A. De Diego, F.M. Martin, D. Amouroux, O.F. Donard, Rapid Ph.D degree from the Department of Organic Chemistry, Uni­
determination of inorganic mercury and methylmercury in biological reference versity of Madras, in 2004 and worked as a Post-Doctoral
materials by hydride generation, cryofocusing, atomic absorption spectrometry Researcher at National Tsing Hua University and at ITRI,
after open focused microwave-assisted alkaline digestion, J. Anal. At. Spectrom. 12 Taiwan (2004-2009). He has also worked as an Assistant Pro­
(7) (1997) 743–750, [Link] fessor (Senior) at VIT, Vellore, India (2009-2013). His current
[20] W. Yantasee, Y. Lin, T.S. Zemanian, G.E. Fryxell, Voltammetric detection of lead research interests include the synthesis of dendrimers and small
(II) and mercury (II) using a carbon paste electrode modified with thiol self- organic molecules and studying the anti-cancer and anti-
assembled monolayer on mesoporous silica (SAMMS), Analyst 128 (5) (2003) inflammatory activities. He has published 52 research articles
467–472, [Link] in peer-reviewed journals, 6 book chapters and holds 3 US
[21] M.L. Ho, K.Y. Chen, G.H. Lee, Y.C. Chen, C.C. Wang, J.F. Lee, P.T. Chou, Mercury patents.
(II) recognition and fluorescence imaging in vitro through a 3D-complexation
structure, Inorg. Chem. 48 (21) (2009) 10304–10311, [Link]
ic901613s.
[22] X. Zhang, Y. Li, H. Su, S. Zhang, Highly sensitive and selective detection of Hg2+
using mismatched DNA and a molecular light switch complex in aqueous solution, Jebiti Haribabu received his Ph.D. in Bioinorganic and Me­
Biosens. Bioelectron. 25 (6) (2010) 1338–1343, [Link] dicinal Chemistry from National Institute of Technology Tir­
bios.2009.10.023. uchirappalli, India, in 2018. He subsequently completed a JSPS
[23] S.P. Ragavi, D. Thirumalai, I.V. Asharani, V. Radhakrishnan, P. Jerome, Dual- Postdoctoral Fellowship at Tokyo University of Science, Japan,
channel fluorescent sensor for rapid Cu2+ and Fe3+ detection: enhanced sensitivity followed by a FONDECYT Postdoctoral Fellowship at Uni­
and selectivity with triazole-substituted acridinedione derivative, Heliyon 10 (19) versidad de Atacama (UDA), Chile. He is currently serving as an
(2024) e38318, [Link] Assistant Professor in the Faculty of Medicine at UDA. Dr.
[24] X. Ke, Y. Fan, J.Z. Zhou, A. Huang, A novel coumarin-derived dithioacetal Haribabu’s research focuses on organometallic and lumines­
chemosensor for trace detection of Hg2+ in real water samples, J. Chem. Res. 44 cent complexes for biological and sensing applications. He has
(3–4) (2020) 142–147, [Link] published over 150 peer-reviewed articles, one book and one
[25] S.P. Ragavi, I.V. Asharani, Acridinedione-based fluorescence ‘turn-off’ sensor for book chapter. He has been recognised among the world’s top
real-time detection of carcinogenic Cr (VI) ions, Inorg. Chem. Commun. 164 (2024) 2% of scientists (2023–2025).
112443, [Link]
[26] S.P. Ragavi, D. Thirumalai, V. Radhakrishnan, I.V. Asharani, A ‘turn-on’ hydrazine
carbothioamide-functionalized acridinedione fluorescent probe for nanomolar
detection of Al3+: applications in water, pharmaceuticals, and live-cell imaging,
Spectrochim. Acta A Mol. Biomol. Spectrosc. (2025) 127197, [Link]
Tanay Debnath is an Assistant Professor in the Department of
10.1016/[Link].2025.127197.
Chemistry, School of Advanced Sciences, Vellore Institute of
[27] G. Yang, X. Meng, S. Fang, H. Duan, L. Wang, Z. Wang, A highly selective
Technology (VIT), India. He earned his Ph.D. in 2019 from the
colorimetric fluorescent probe for detection of Hg2+ and its application on test
Indian Association for the Cultivation of Science, India. He was
strips, RSC Adv. 9 (15) (2019) 8529–8536, [Link]
a Fulbright-Nehru Postdoctoral Fellow at the University of
[28] M. Ilakiyalakshmi, S. Mohandoss, S.M. Roopan, A.A. Napoleon, Nanomolar Hg2+
Texas at Dallas, USA, working on unnatural base pair systems
detection via phenothiazine fluorogenic chemosensor: applications in water
using Density Functional Theory (DFT) and classical Molecular
analysis and intracellular imaging, Spectrochim. Acta A Mol. Biomol. Spectrosc.
Dynamics (MD). He later served as a postdoctoral fellow at
326 (2025) 125157, [Link]
Queen’s University, Canada, studying histone proteins using
[29] A. Pal, K.M. Das, B. Goswami, A. Thakur, Microwave-assisted neat synthesis of a
computational approaches. His expertise lies in computational
ferrocene appended phenolphthalein diyne: a designed synthetic scaffold for Hg2+
chemistry, particularly DFT and MD.
ion, Inorg. Chem. 59 (14) (2020) 10099–10112, [Link]
inorgchem.0c01236.
[30] J. Haribabu, S. Srividya, D. Mahendiran, D. Gayathri, V. Venkatramu,
N. Bhuvanesh, R. Karvembu, Synthesis of palladium (II) complexes via Michael
addition: antiproliferative effects through ROS-mediated mitochondrial apoptosis
and docking with SARS-CoV-2, Inorg. Chem. 59 (23) (2020) 17109–17122,
[Link] Dr. I. V. Asharani is currently a Professor in the Department of
[31] J. Haribabu, Y. Tamura, K. Yokoi, C. Balachandran, M. Umezawa, K. Tsuchiya, Chemistry at Vellore Institute of Technology, Vellore, India.
S. Aoki, Synthesis and anticancer properties of Bis- and mono (cationic peptide) She obtained her Ph.D. in Chemistry from the University of
hybrids of Cyclometalated iridium (III) complexes: effect of the number of peptide Madras, Tamil Nadu, India. She has 17 years of teaching
units on anticancer activity, Eur. J. Inorg. Chem. 2021 (18) (2021) 1796–1814, experience and has published 60 research articles in reputed
[Link] journals. Six students have completed their Ph.D. under her
guidance, and six research scholars are currently pursuing their
doctoral studies under her supervision. Her research interests
include green synthesis of nanomaterials and their environ­
mental applications, as well as the design and synthesis of ac­
ridine derivatives for chemosensing.

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