Reaction-based bi-signaling chemodosimeter probe for selective detection of hydrogen sulfide and cellular studies

Article abstract of DOI:10.1039/c7nj04632d

A new quinoline-indolium-based chemical probe (DPQI) was synthesized and characterized for selective detection of hydrogen sulphide (H₂S). Probe DPQI displays highly selective and sensitive detection of hydrogen sulphide over other allied anions and thiol-containing amino acids in aqueous-DMSO at physiological pH. The probe DPQI acts as a bi-signaling fluorescent chemodosimeter for selective detection of hydrogen sulphide when excited at different wavelengths. The selectivity was guaranteed by the use of the unique nucleophilicity of HS^- for nucleophilic addition to the most electrophilic positively charged centre followed by thiolysis of the dinitrophenyl ether to yield DNB-SH and a fluorescent phenoxide moiety, which led to the fluorescence emission 'turn on' and characteristic visual fluorescent color change behavior of the sensing system. This is why the probe DPQI showed different optical properties at varying concentrations of hydrogen sulphide. The structural and electronic properties of the probe (DPQI) and its thiolysis product have been demonstrated using ab initio density functional theory (DFT) combined with time-dependent density functional theory (TDDFT) calculations. Utilizing this probe, we have successfully detected hydrogen sulphide in live cells.

Full text of DOI:10.1039/c7nj04632d

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Chem., 2018, DOI: 10.1039/C7NJ04632D.
Volume 40 Number
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January 2016 Pages 1–846
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PAPER
Jason B. Benedict et al.
The role of atropisomers on the photo-reactivity and fatigue of
diarylethene-based metal–organic frameworks
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Cite this: DOI: 10.1039/c0xx00000x
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ARTICLE TYPE
ReactionꢀBased Biꢀsignaling Chemodosimeter Probe for Selective Detection of
Hydrogen Sulfide and Cellular Studies
a
a
a
a
Uday Narayan Guria, Kalipada Maiti, Syed Samim Ali, Sandip Kumar Samanta, Debasish Mandal,
b
c
c
d
a
Ripon Sarkar, Pallab Datta, Asim Kumar Ghosh, Ajit Kumar Mahapatra*
5
Received (in XXX, XXX) Xth XXXXXXXXX 20XX, Accepted Xth XXXXXXXXX 20XX
DOI: 10.1039/b000000x
A new quinolineꢀindoliumꢀbased chemical probe (DPQI) was synthesized and characterized for selective detection of hydrogen sulphide
(
H S). Probe DPQI displays highly selective and sensitive detection of hydrogen sulphide over other allied anions and thiolꢀcontaining
2
1
0
amino acids in aqueousꢀDMSO at physiological pH. The probe DPQI acting as biꢀsignaling fluorescent chemodosimeter for selective
detection of hydrogen sulphide when excited at different wave lengths. The selectivity was guaranteed by use of the unique
−
nucleophilicity of HS for nucleophilic addition to the most electrophilic positively charged centre followed by thiolysis of the
dinitrophenyl ether to yield the DNBꢀSH and fluorescent phenoxide moiety, which led to fluorescence emission ‘turn on’ and
characteristic visual fluorescent color change behavior of the sensing system. This is why the probe DPQI showed different optical
properties at varying concentration of hydrogen sulphide. The structural and electronic properties of the probe (DPQI) and its thiolysis
product have been demonstrated using ab initio density functional theory (DFT) combined with timeꢀdependent density functional theory
1
5
(TDDFT) calculations. Utilizing this probe, we have successfully detected hydrogen sulphide in live cells.
.
Among these methods, fluorescent detection methods offered a
Introduction
Hydrogen sulfide (H S) is wellꢀknown as a volatile toxic gas with
the characteristic foulꢀsmell of rotten eggs. Now, it is also
considered the third most important gasotransmitter for regulating
cardiovascular, neuronal, immune, endocrine, and gastrointestinal
systems, along with nitric oxide and carbon monoxide. H S
contributes to a broad array of physiological responses to
maintain cellular health, including vasodilation, angiogenesis,
promising opportunity due to the unique advantages of rapidity,
sensitivity, convenience, simplicity, easy operation, and low cost.
Recently, the everꢀexpanding toolbox of reactionꢀbased probes
employ three primary strategies for H S detection, these probes
were primarily designed based on: (a) reduction reactions of
azide and nitro ; (b) nuclephilic reactions ; (c) reactions with
metal centered coordination complexes such as copper sulfide
50 precipitation. As the pKa of H S is 6.8 and it is calculated that
72% of the total H S exists as HS under physiological
conditions. Therefore, H S is a strong nucleophile in biological
2
2
2
3
3
4
0
5
0
5
0
4
5
2
1
2
19
20
2
3
21
4
5
6
oxygen
sensing,
apoptosis,
inflammation,
and
2
−
7
neuromodulation,
ischemia/reperfusion injury. On the other hand, studies have
established that unregulated, abnormal levels of H S may
contribute to diseases ranging from Alzheimer’s disease and
Down’s syndrome
Considering the challenges of measuring this volatile, reactive
small molecule to both healthy and disease states is the shortage
and it can also protect against
2
22
8
2
systems that should participate well in nucleophilic reactions.
2
9
Based on this property, the design of fluorescent probes for H2S
1
0
11
12
55 is mainly based on specific chemical reactions by taking
advantage of the reducing or nucleophilic properties of H S. For
example, LippertꢀChang and Wang et al. pioneered an
approach of using the reduction of aryl azide with H S to amine
to sense H S, which has been expanded to design azideꢀ
containing fluorescent probes by altering fluorophores.
al. used the nucleophilic attack of H S on the aldehyde
functionality to design a fluorescent probe to sense H S. Lin et
al. reported a nearꢀinfrared fluorescent probe for H S based on
to diabetes and liver cirrhosis.
2
23
24
of methods for selective tracking of H S molecule within living
2
2
biological specimens and biological samples. Therefore, easy and
convenient methods have been required for rapid assessment of
this biologically and environmentally important species. To date,
a large number of sensitive protocols have been reported for
sulfide detection. These methods include classical titrimetry
electrochemical
2
2
5ꢀ30
60
He et
2
3
1
2
1
3
2
32
1
4
15
thiolysis of dinitrophenyl ether.
techniques
chromatography
spectrophotometry
and fluorimetry
1
6
17
18
chemiluminescence
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OH
OH
Furthermore, a couple of elegant probes have been designed by
splitting the dinitrophenyl ether or dinitrobenzenesulfonyl group
N
CH
3
N
CHO
a
b
N
CHO
3
3
to restore their original fluorescence. The sulfide anion is a
very effective nucleophile and reacts readily with sulfonate
esters and thiolysis of aryl nitrate. Not only, being a good
nucleophile, it has a tendency to attack positively charged
electrophilic centre. This fact provided us with a clue for the
design of novel fluorescent probes for the sulfide anion having
both the reacting centre in one chromophore. In most of the
reported literatures the chemosensors are composed of either
ether group linked with the fluorophore moiety or positively
charged electrophilic centre. Even in few cases both the reactive
centre are present simultaneously. But in those cases only
thiolysis of ether occurred and hence fluorescence were
O
1
2
3
O
2
N
NO
2
5
0
5
0
5
5
c
N
1
1
2
O
N
O
2
N
NO
2
DPQI
Scheme 1. Synthesis of Probe DPQI: Reagents & Conditions: (a)
SeO , Dioxane, reflux, 12h. (b) 2,4ꢀdinitrofluorobenzene, Et N,
Dry AcCN, (c) 1,2,3,3ꢀTetramethylꢀ3Hꢀindolium iodide, ethanol,
rt, 6h.
60
2
3
34
regenerated. Furtheremore, Lin et al. reported a typical
chemosensor which contain both the reactive centre and both of
35
them react with H S to show different optical properties. But,
2
Colorimetric and Fluorometric response towards
in that particular case the chromophore detecting Cys/GSH/Hcy
and H S simultaneously. But, there is no report where the probe,
composed of dual reacting centre and acting as a dual signaling
H S
2
2
65
At first, we tested the spectroscopic properties of DPQI (10 ꢁM)
by carrying out the fluorescence and absorption titrations of
DPQI with Na S (standard source of H S) in aqueous buffer
operator for selective detection of H S over the similar reactive
biothiols.
2
2
2
solution (20 mM HEPES buffer, pH 7.4) at 35 °C. The absorption
Keeping the above in perspective, herein, we have described a
dualꢀreactive groups with dualꢀfunctioning colorimetric and
spectra of DPQI in presence of H S shows some typical
2
7
0
character. DPQI in aqueous DMSO (1.0 µM, 2:1 v/v) HEPES
buffer (pH 7.4) displayed a maximum absorbance at 398 nm and
2
3
3
4
5
0
5
0
fluorometric probe for H S with easy structural manipulation by
protecting the hydroxyl group with 2,4ꢀdinitrofluorobenzene
2
5
20 nm responsible for DNB and rest of DPQI molecule
(DNFB). Here, fluorophore is attached to two reactive groups,
respectively.
DNFB and a highly activated iminium ion acceptor. We named
our probe DPQI (2ꢀ{2ꢀ[8ꢀ(2,4ꢀDinitroꢀphenoxy)ꢀquinolinꢀ2ꢀyl]ꢀ
vinyl}ꢀ1,3,3ꢀtrimethylꢀ3Hꢀindolium) and its quinoline skeleton is
used as a color reporting group and the DNB moiety can quench
the fluorescence via the photoinduced electron transfer (PET)
(a)
(b)
75
36
effect. Thus, the sensor has a very weak initial fluorescence
due to intramolecular charge transfer (ICT). Importantly, the
probe acting as a dual signaling moiety when excited at different
wave length and those signals are arises in different
concentration of H S. So the novelty of the probe is that two
2
reactive centre shows their reactivity in different concentration
Fig.1. (a) Absorption spectra of DPQI (1.0 ꢁM, 20 mM HEPES
of H S and exhibits different optical properties. This will finally 80 buffer PH 7.4, DMSOꢀH O; 1:2; v/v) upon gradual addition of
2
2
lead to the enhancement of the good selectivity and high
Na S (0–0.02 mM). (b) Absorption spectra of DPQI (1.0 ꢁM, 20
2
sensitivity of the probe.
mM HEPES buffer PH 7.4, DMSOꢀH O; 1:2; v/v) upon gradual
2
addition of Na S (0.02ꢀ0.1 mM); Inset: color change of DMSO
2
solution of DPQI before and after addition of Na S.
Results and discussions
2
8
9
9
5
0
5
With increasing concentration of H S (0ꢀ2 mM), we found that a
The probe molecule, DPQI, was synthesized in three steps
2
new peak arises at 468 nm up to 0ꢀ0.2 mM concentration of H2S
(Scheme 1). The reaction of 2ꢀmethylꢀ8ꢀhydroxyquinoline (1)
(20 equiv.) (Fig. 1a). From Fig. 1a it is observed that upon
4
5
with SeO /dioxane results in the formation of aldehyde
2
successive addition of H S, absorbance intensity at 520 nm is
derivative, 2, and its hydroxyl group protected by FꢀDNB under
basic conditions yielded the corresponding protected derivative 3,
which, upon coupling with 1,2,3,3ꢀtetramethylꢀ3Hꢀindolium
iodide resulted in DPQI. All of the compounds exhibited
satisfactory analytical and spectral data as given in the
Experimental Section (Fig. S1ꢀS6 †).
2
gradually decreased which is accompanied with a strong
isosbestic point at 500 nm indicated the formation of new
compound. Now, from the Fig. 1a we have noticed
a
hypsochromic shift occurred in absorbance spectra, which may
5
0
attribute to the nucleophilic addtion of H S to the Indolium
2
moiety thereby blocking the ICT process occurring from
quinoline to indolium moiety. Another vision from the graph is
that the peak at 398 nm remain undisturbed. This indicates that
2
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DNB part of the molecule is not cleaved. Thus, formation of
isosbestic point and hence formation of new compound and
vanished after addition of 20 equiv. of H S due to the blocking of
ICT process as DPQIꢀSH adduct was formed (Scheme 2).
2
hypsochromic shift in absorbance spectra leads to the formation 55 Importantly, we found no other emission band in visible region
of DPQIꢀSH adduct at the low concentration of H S.
even addition of higher concentration of H S. Then we excited
2
2
5
DPQI at 500 nm and found no fluorescence band in visible
Then, upon increasing the concentration of H S up to 0.1 mM
region. Upon incremental addition H S (˃ 20 equiv.) a new
2
2
(100 equiv) we noticed that the peak intensity at 398 nm
emission band at 593 nm was found up to 0.1 mM (Fig. 2b). A
gradually decreased and a new peak arises at 566 nm. An 60 deep orange fluorescent color was found in naked eye [Fig.
isosbestic point was found at 518 nm (Fig. 1b). This result
suggested that at high concentration of H S, H Sꢀmediated
cleavage of the electronꢀwithdrawing DNB group releases an
oxygen donor at pH 7.4, that increases the pushꢀpull character of
S12b(ii) †]. As the intensity of the newly generated emission
reaches the maximum values, we achieved more than 700 fold in
increase in fluorescence intensity. These results interpreted that
1
1
2
2
0
5
0
5
2
2
probe DPQI attacked by H S at a low concentration and
2
the probe and resulting in large bathochromic shifts (at 566 nm) 65 produces DPQIꢀSH adduct, while this intermediate product is
in absorption. So, the bathochromic shift is mainly due to strong
Intramolecular Charge Transfer (ICT) within quinolinol moiety.
The stoichiometry of the reaction between the probe DPQI and
further attacked by H S at a high concentration and thiolysis of
2
ether occurred to yield the orange fluorescent compound QIꢀSH
and a non fluorescent DNBꢀSH. Thus it is clear from above
phenomenon that the probe acting as a dual signaling moiety for
Na S has been derived to be 1:2 on the basis of a Jobs plot (Fig.
2
S11 †). The pattern of the Jobs plot (both by absorbance and 70 H S. Furthermore, H S has decent linear relationships with the
fluorescence method) indicate the stepwise reaction process. Thus
2
2
maximum intensity values of newly generated emission bands (at
593 nm) in a certain concentration range (Fig. S10 †), and
accordingly the limit of detection is calculated to be 3.5 µM.
the results indicate that H S did reaction with dual reaction
2
centers of the probe DPQI (Scheme 2). This results also
supported by the naked eye color change of the DMSO solution
of DPQI in presence of H S. We have noticed that in addition of
2
small amount of H S no significant color change was observed. 75 Selectivity of DPQI towards H2S
2
Whereas when concentration of H S was increased color of the
2
In order to check whether the DPQI is sensitive to only H S or
even to the other anions, similar fluorescence titrations were
solution changed drastically from yellow to deep red (Fig. 1b
inset).
2
carried out in the same medium with other different analytes, viz.
−
−
−
−
2−
−
−
−
−
2−
In receipt of decent outcome from UVꢀVis spectroscopy studies,
F , Cl , Br , I , SO
4
, HSO
3
, AcO , CN , H
2
PO
4
, S
2
O
3
as
we went for the fluorescence study. Initially, the probe DPQI , 80 well as thiol containing amino acids (Cys, Hcy and GSH) and
3
0
(10 µM in 2:1 water: DMSO mixture at pH 7.4) when excited at
98 nm, exhibit a low intense emission band at 461 nm (Fig. 2a)
responsible for weak blue fluorescence [Fig. S12b(i) †].
found no significant fluorescence enhancement in the presence of
any of these analytes (Fig. S8†). Therefore, DPQI is selective to
H S among other analytes studied. Visual fluorescent color
2
3
change experiments have been carried out to look at the behavior
of DPQI in the presence of various anions. Under UV light, the
solution of DPQI is low intense blue fluorescence whereas in the
(a)
(b)
85
presence of H S, it shows an intense orange fluorescent color
2
which is otherwise not present in case of the other analytes
3
5
studied (Fig. S13†). Therefore, H S can easily be differentiated
2
90
by visual color change among the other anionic species. In order
to show the practical utility of DPQI to detect H S selectively
2
even in the presence of other analytes, competitive anion
titrations were carried out. These studies reveal that the H Sꢀ
2
Fig.2. (a) Fluorescence spectra of DPQI (λex = 398 nm) (1.0 ꢁM,
2
induced fluorescence enhancement was unaffected in the
−
−
−
−
2−
−
0 mM HEPES buffer PH 7.4, DMSOꢀH O; 1:2; v/v) upon 95 presence of allied anions viz; F , Cl , Br , I , SO , HSO ,
2
4 3
−
−
−
2−
4
0
gradual addition of H S (0–0.02 mM). (b) Fluorescence spectra of
2
AcO , CN , H PO , S O . In addition, we also performed
2 4 2 3
DPQI (λex = 500 nm) (1.0 ꢁM, 20 mM HEPES buffer PH 7.4,
competitive selectivity studies with some representative thiol
containing amino acids (Cys, Hcy and GSH). These experimental
results implied that the DPQI probe is highly selective toward the
DMSOꢀH O; 1:2; v/v) upon gradual addition of H S (0.02ꢀ0.1
2
2
mM).
1
00 H S even though the concentration of other competitive analytes
4
5
Upon gradual addition of H S up to 0.02 mM, this low intense
2
2
is 100 equiv. higher than H S concentration (Fig. S9 †). Thus, it
fluorescence band intensity was decreased up to the saturation. It
is possible because intramolecular charge transfer (ICT) might
occur from the qunoline recognition unit to the indolium moiety,
resulting this low intense band, which is consistent with our
design strategy that the quinoline fluorescence in DPQI is
heavily quenched due to PET process occurring between DNB
2
is notable that DPQI can be used as a selective H S fluorescent
2
sensor in the presence of other relevant analytes.
To provide further support for nucleophilic addition followed by
5
0
1
05 the removal of DNB moiety from DPQI by H S, ESI MS
2
3
7
experiment was carried out for analysis of product.
and quinoline moiety. Thus the peak at 461 nm was completely
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nd
H S is the slow step of the reaction and high degree of 2 order
2
rate constant attribute to proceed the reaction.
(
a)
(b)
55
Scheme 2. Mechanism of reaction of H S with DPQI.
2
5
0
5
0
The reaction mechanism of DPQI to H S could be reasonably
explained by a nucleophilic reaction. To know if there happen
2
only nucleophilic addition of H S to the indolium moiety or only
2
thiolysis of ether pat or both, we have done mass spectroscopic
assay. In ESIꢀMS analysis the peak at m/z: 528.1517 responsible
for DPQIꢀSH adduct. Another peak at m/z: 362.1493 indicating
the formation of thiol adduct of quinonilolꢀindoline i.e. QIꢀSH.
Peak at 200.9918 confirm that thiolysis of ether was occurred and
DNBꢀSH was formed (Fig. S7 †). Importantly we did not have
any peak which might support that only thiolysis of ether was
occurred. These results strongly support the two step reaction of
Fig. 3. (a)Time dependent fluorescence change of DPQI with
1
1
2
6
0
H S (100 equiv.) as a function of time (at 593 nm). (b) Time
2
dependent fluorescence change of DPQI with H S (20 equiv) as a
2
0
function of time (at 461 nm, 35 C).
Density functional theory (DFT) calculations
the probe with H S and most importantly mechanism of the
2
To get insight into the optical properties of the new probe DPQI
reaction is first thiol addition then thiolysis of ether. To support
the stepwise reaction mechanism we have done MS (HRMS)
65
responding to H S, density functional theory (DFT) calculations
2
with the B3LYP exchange functional employing 6ꢀ31+G(d,p)
basis sets using a suite of Gaussian 09 programs were performed.
titration also. Upon reaction with 0.5 equiv. of H S, we found the
2
peak at m/z: 528.1518 (Fig. S19 †). This peak is the indication of
the formation of only DPQIꢀSH adduct. Addition of excess H2S
exhibited a peak at m/z: 362.1496 and at 200.9908 (Fig. S20 †).
Nevertheless, the former is for QIꢀSH and latter is responsible for
DNBꢀSH.
ICT
70
2
5
Reaction rate investigation
During the experiments, interestingly, we found that the reaction
rate between probe DPQI and H S was affected by the reaction
2
temperature. The reaction is slow at room temperature but quite
0
rapid at 60 C (the fluorescence intensity reached saturation
3
0
within 6 min). However, this temperature was too high to detect
ꢀ
0.12 a.u.
LUMO
H S in living cells. On the other hand, the probe exhibited almost
2
0
0
the same reaction rate under 35 C and 45 C conditions (Fig. 3).
In both cases the fluorescence intensity took 13 min to reach
saturation. Thus, 35 C was chosen as the optimal reaction
ꢀ0.17 a.u.
LUMO
75
0
3
5
temperature.
Encouraged by these preliminary results, we proceeded to study
the reaction kinetics under pseudoꢀfirstꢀorder conditions with a
ꢀ
0.28 a.u.
HOMO
DNBꢀOMe
large excess of H S (˃20 equiv.) over probe DPQI (10.0 µM) in
2
ꢀ0.32 a.u. HOMO
0
aqueous DMSO (2:1 v/v) pH 7.4 HEPES buffer at 35
temperature. The pseudoꢀfirstꢀorder rate constant
calculated as k = 0.23 min with the equation ln[(FmaxꢀF )/Fmax] 80 Fig. 4. Top: Energy optimized structure of DPQI and its emission
ꢀk t (Fig. S14a †), where F and F
C
/
QI
4
4
5
0
5
0
k
was
/
−1
38
t
/
=
are the fluorescence
max
response regulated by PET and ICT mechanism. Bottom: FMO
t
intensities at 593 nm at time t and the maximum value obtained
energy of DNBꢀOMe and QI at the excited state.
/
after the reaction was complete; k is the observed rate constant.
/
Emission of DPQI should be regulated by DNFB via
photoinduced electron transfer (PET) mechanism. To authenticate
the suggested fluorescence mechanism, the frontier orbital energy
of both separate DNBꢀOME and QI were also calculated. As
shown in Fig. 4, the energy of highest occupied molecular orbital
Furthermore, the plots of observe rate constant k vs. [H S] was a
2
straight line passing through the origin, suggesting that the
−
1 −1
85
reaction is overall second order with k = 775.44 M
s
(Fig.
2
S15 †). From Fig. 3b, it is obvious that at low concentration of
H S, nucleophilic reaction towards indolium cation of DPQI is
2
(HOMO) of DNB lies between HOMO and lowest unoccupied
much quicker (only 2 min required for saturation). So, the next
step reaction i.e. thiolysis of ether through addition elimination of
molecular orbital (LUMO) of DPQI molecule.
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This indicate that at excited state electron from HOMO of DNB
Therefore, the test strips could conveniently detect H S in
2
transfer to the HOMO of the DPQI and hence fluorescence is 55 solutions. The above result suggest that this type of solid system
quenched. Consequently, the πꢀelectrons cloud on both the
HOMO and LUMO efficiently distribute in the respective
quinoline and indole units in DPQI, suggesting that DPQI is a
typical intramolecular charge transfer (ICT)ꢀ based fluorophore.
These results suggesting the initial low intense blue fluorescence
at 461 nm. As, initially DPQIꢀSH adduct was formed the above
ICT process was turned off which is responsible for gradual
quenching in fluorescence at 461 nm. Whereas, thiolysis of ether
part diminish the PET process, producing the phenolate ion,
responsible for bathochromic shift both in absorption and
protocol may be used to perform as a sensitive and practical
‘‘dipꢀin’’ naked eye H S sensors in the near future.
2
5
(
a) (i)
(ii)
(iii)
(iv)
(v)
(v)
Ambient light
60
1
0
(i)
(ii)
(iii)
(iv)
Fluorescent light
(b)
emission spectra. Thus, the emission response of DPQI to H S is
2
delimited by the PET and ICT mechanisms together.
1
2
2
5
0
5
Calculations based on singleꢀexcitation timeꢀdependent DFT
(TDDFT) were performed to explain the electronic structural
Fig. 5. (a) Naked eye color change of TLC plates (i) DPQI (1.0
ꢀ
4
ꢀ4
ꢀ4
properties of the ground and excited states of DPQI. The vertical 65 x 10 M), (ii) 10.0 x 10 M Na S, (iii) 25.0 x 10 M Na S, (iv)
2
2
ꢀ
4
ꢀ4
transitions calculated by TDDFT (Table S1 †) were comparable
with the experimentally observed UV−vis spectra. The studies
suggest that the vertical transitions (HOMOꢀ1→LUMO, f =
40.0 x 10 M Na S, (v) 80.0 x 10 M Na S (b) Fluorescent color
2 2
change (i) DPQI (1.0 x 10 M), (ii) 10.0 x 10 M Na S, (iii) 25.0
x 10 M Na S, (iv) 40.0 x 10 M Na S, (v) 80.0 x 10 M Na S.
ꢀ
4
ꢀ4
2
ꢀ4
ꢀ
4
ꢀ4
2
2
2
0
.6002, the highest observed oscillator strength) observed at
415 nm are comparable to those of the experimentally observed
spectra at ∼398 nm for DPQI. Whereas, HOMO→LUMO (∆E =
.96 ev) observed at 495 nm is comparable with the experimental
Hydrogen sulphide coming as industrial waste materials and
∼
70
mixed with different water resources. Thus, detection of H S is
2
very much essential from natural surface water resources where
several interfering agents are present. We collected some surface
water samples from different areas of West Bengal, India (see
Supporting Information for complete details) and experienced the
2
peak at 520 nm. In the case of the phenolate ion, the observed
band at ∼542 nm is assigned to the transition of HOMO→LUMO
(∆E = 2.58 ev) (Fig. S17 †, Table S2 †) that results from a π →
75
effectiveness of our sensor in detecting H S in these various
aqueous media.
π* transition and corresponds to the experimental λmax ∼566 nm.
2
Practical application
3
3
4
0
5
0
Intense color change in naked eye as well as in fluorescent in
presence and absence of H S strongly recommends that the
2
sensors can be used in kits for the detection of H S. For the
2
complete study of the practical application of the probe DPQI,
test strips were made of by dipping TLC plates into acetonitrile
80
−3
solution of DPQI (c = 1.0× 10 M) and then drying them in air.
DPQI immersed TLC plates were then treated with different
concentration of H S. As shown in Fig. 5, when concentration of
2
H S was increased gradually, a noticeable change in color (from
2
pale yellow to deep brown) of the TLC plates were observed
under irradiation at 254 nm under a UV lamp.
Fig. 6. Sensing of H S from surface water sample by using
DPQI.
Similar phenomenon was also found in case of fluorescence
2
85
color. When H S concentration was increased, the fluorescent
2
color of the test strips changes from weak blue to orange (Fig. 5).
To do this, we prepared 10 ꢁM solutions of H S in different water
2
Most importantly, during solid state detection of H S we
2
samples and observed the change in emission of DPQI. In all
cases, substantial fluorescence (at 593nm) enhancement were
noticed (Fig. 6). These results suggest that irrespective of the
source of water, enhancement in fluorescence of DPQI is
4
5
observed that high concentration of H S was required for the
2
initial color change (both in ambient light and fluorescence).
This observation again suggests that two step reactions was
90
occurred. Primarily, some H S was consumed for nucleophilic
2
manifested in the presence of H S. These results undeniably set
2
addition to the indolium moiety which has a minute effect to the
color and fluorescence properties (pale yellow color and weak
blue fluorescence). Secondly, addition of excess H2S is
responsible for thiolysis of dinitrophenyl ether which is
accompanied with the color as well as fluorescence change.
up the capability of DPQI as a prevalent fluorescent sensor for
H S.
2
5
0
This journal is © The Royal Society of Chemistry [year]
Journal Name, [year], [vol], 00–00 | 5

New Journal of Chemistry
Page 6 of 10
View Article Online
DOI: 10.1039/C7NJ04632D
treated with DPQI (0.1 µM) (red channel λ =561 nm, λ = 585ꢀ
630 nm.). (C2) Red channel Fluorescence image of DPQI (red
Detection of hydrogen sulphide with DPQI in
living cells
ex
em
50
channel λ =561 nm, λ = 585ꢀ630 nm.) treated with 20.0 µM
ex
em
Na S.. (C3) fluorescence image of DPQI inside cells treated with
2
Next, we evaluate the abilities of DPQI to visualize changes in
1
00 µM Na S ( red channel λ =561 nm, λ = 585ꢀ630 nm.).
2 ex em
H S (here we use Na S for standard of H S) levels in liveꢀcell
2
2
2
5
0
5
imaging mode using fluorescence microscopy (Fig. 7) (detailed
of cell study is described in Experimental section). Hence, to
know the effectiveness of DPQI as a probe for in vitro detection
It is quite obvious that as bright field images in different cases did
not revealed any extensive morphological change the components
are cell viable. These findings open up the avenue for future in
vivo biomedical applications of this probe.
5
5
of H S by fluorescence microscopy, Vero cells were used to
2
detect endogenous H S in live cells. To do this, first we did MTT
2
1
assay of all the components to see the cell viability (Fig. S18 †).
From Fig. S17, it is found that probe DPQI hardly has an adverse
Conclusion
effect towards the cell; however the H S had concentration
2
In conclusion, we have synthesized a new quinolineꢀindoliumꢀ
dependent adverse effect when cells were treated with varying
6
6
7
7
0
5
0
5
based an improved fluorescent probe DPQI for H S detection.
2
−
concentrations of H S. As shown in the bright field and
2
The design was based nucleophilic addition of HS to iminium
1
fluorescence images of the cells incubated only with Na S (0.1
2
ion followed by thiolysis of dinitrophenyl ether to release the
fluorophore. This twoꢀstep reaction was observed by slowly
mM) for 30 min no fluorescence can be observed (Fig 7a and 7b).
After incubating Vero cells with 0.1 µM of DPQI for 30 min at
increasing concentration of H S. Concomitantly, this probe
0
2
3
7 C in presence of Na S we found different color inside cells at
2
showed fluorescent emission colour changes from colorless to
orange and visible colour changes from pale yellow to bright
different concentration of H S.
2
brown at higher concentration in response to H S. The probe has
2
2
0
Initially, blue color inside cells was found when the cells were
a high selectivity for H S over competitive analytes. The H S
2
2
only treated with DPQI (Fig. 7d) (blue channel, λ =350nm, λ =
ex
em
sensing mechanism, namely, H S triggered addition and then
2
3
80ꢀ480 nm). Then we observed cells in fluorescence microscope
thiolysis reaction, was proved by LC–MS. TDDFT calculations
were carried out to demonstrate the electronic properties of DPQI
and its reaction products. The practical phenomena are fully
supported by DFT calculations. Test kit experiment implies that
with increasing concentration of H S. Fluorescence inside cells
2
was tend to vanish when treated with 0.02 mM of H S (20 equiv)
2
2
5
(Fig. 7e). Upon increasing the concentration (up to 0.1 mM) (red
channel λ =561 nm, λ = 585ꢀ630 nm) we found that intensity of
ex
em
the probe is able to detect H S even in solid state. This probe is
2
orange fluorescence inside the cells was gradually increased (Fig.
fꢀ7h). This result also prove the two step reaction of the probe
also applicable to detect H S in live cell and has the ability to
2
6
detect intracellular H S.
2
and H S.
2
3
0
The fluorescence microscopic study strongly recommended that
Experimental Section
probe DPQI could readily cross the membrane barrier of the
Vero cells, and detect intracellular H S.
2
Materials and Methods. Unless otherwise mentioned, materials
B r igh t F ield
A 1)
B lu e ch a n n e l R e d ch a n n e l
8
0
were obtained from commercial suppliers and were used without
(
(B 1 )
(B 2 )
(B 3 )
(C 1)
(C 2)
(C 3)
1
13
further purification. H and C NMR spectra were recorded on a
Brucker 400 and 300 MHz instrument. For NMR spectra,
DMSOꢀd and CDCl were used as solvent using TMS as an
D P Q I
6
3
(
A 2)
3
5
internal standard. Chemical shifts are expressed in δ ppm units
D P Q I
1
1
1
85
and H– H and H–C coupling constants in Hz. Mass spectra
were carried out using a Waters QTOF Micro YA 263 mass
spectrometer. Fluorescence spectra were recorded on a Perkin
Elmer Model LS 55 spectrophotometer. UV spectra were
recorded on a JASCO Vꢀ530 spectrophotometer.
+
+
2
H S (2 0 µM )
(
A 3)
D P Q I
S (10 0µ M )
H
2
9
0
Cell Imaging Study. For fluorescence imaging studies Vero cell
Fig. 7. Fluorescence images of probe in vero cells (40× objective
lens): (A1) Bright field image of only cells treated with DPQI
(Vero 76, ATCC No CRLꢀ1587) lines, were seeded on sterile 35
4
0
mm µꢀDish, glass bottom culture dish (ibidi GmbH, Germany),
(
0.1 µM) (A2) Bright field image of DPQI treated with 20.0 µM
and incubated at 37 °C in a CO incubator for 24ꢀ30 hours. The
2
Na S (A3) Bright field cell image of DPQI with 100 µM Na S.
2
2
next day, cells were washed three times with phosphate buffered
saline (PBS) and fixed using 4% paraformaldehyde in PBS (pH
(B1) Blue channel fluorescence images of cells treated with
95
DPQI (0.1 µM) ( blue channel, λ =350nm, λ = 380ꢀ480 nm).
ex
em
7
.4) for 10 minutes at room temperature. Thereafter the cells were
4
5
(B2) Fluorescence image of DPQI ( blue channel, λ =350nm,
ex
washed with PBS followed by permeabilization using 0.1%
λ = 380ꢀ480 nm) treated with 20.0 µM Na S. (B3) Fluorescence
em
2
−7
saponin for 10 min followed by incubation with 1.0 × 10 M of
image of DPQI ( blue channel, λ =350nm, λ = 380ꢀ480 nm)
ex
em
probe DPQI dissolved in 100 ꢁL DMEM at 37 °C for 1 h in a
treated with 100 µM Na S (C1) Red channel image of cells
2
6
|
Journal Name, [year], [vol], 00–00
This journal is © The Royal Society of Chemistry [year]

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New Journal of Chemistry
View Article Online
DOI: 10.1039/C7NJ04632D
CO2 incubator. Before microscopic imaging, all the solutions
55
Synthesis and Structure Characterization.
were aspirated and mounted on slides in a mounting medium
containing DAPI (1 ꢁg/mL) and stored in dark before
microscopic images were acquired. The cells were observed
under Andor spinning disk confocal microscope (SDꢀCM) with
excitation at 350 nm monochromatic laser beam and the collected
Preparation of 2ꢀFormylꢀ8ꢀHydroxyꢀQuinoline (2): A mixture
of Selenium dioxide (1041.92mg, 9.378mmol), (dissolved in a
minimum amount of water), and 2ꢀmethylꢀ8ꢀhydroxyquinolinol
(1) (1500mg, 9.423mmol) were dissolved in 1, 4ꢀdioxane.The
5
range of emission wavelength was between 420 and 480 nm (blue 60 resultant reactant mixture was refluxed for 2ꢀ3hours. After the
channel) and excited at 561 nm and the collected range of
emission wavelength was between 585 and 630 nm (red channel).
Cells were imaged live by SDꢀCM 63× oilꢀimmersion objective.
Images were acquired in zꢀstacks of 28 planes at 0.3ꢀꢁm intervals
with 400ꢀms exposure times every 20 seconds over a period of 30 65 yellow powder obtained. The yield of the crude product is
minutes. In another dish new cells were again cultured in the
completion of the reaction (TLC monitored) the crude product
was subjected to filtration, while in hot condition, followed by
purification of the filtrate obtained, by the column
chromatography on silica gel (Hexane: EtOAc; 8:2(v/v)).A light
1
1
2
0
5
0
0
ꢀ1
1065mg. (65.33%), M.P. =102 C: IR: KBr (Cm ): 3466, 2815,
+
same manner followed by treatment with Na S (0.1 mM).
1711.MS (HR MS): (m/z, %): 174.0547 [(M+H ), 100 %]
2
Thereafter the cells were washed thrice with PBS (pH 7.4) to
remove any excess Na S and incubated in DMEM containing
Preparation of O -protected 2-Formyl-8-Hydroxy-Quinoline
(3): A mixture of compound 2 (150 mg, 0.86 mmol), 2,4ꢀ
2
probe DPQI. Finally the cells were washed again with PBS (pH
7
.4) three times to remove excess probe outside the cells. The 70 dinitrofluorobenzene(165mg, 0.9mmol) and triethylaminewere
emissions were obtained at above recommended wave length for
different concentration of Na2S (blue channel for low
concentration and red channel for high concentration).
dissolved in dry acetonitrile. The reaction mixture was refluxed.
After the completion of the reaction (TLC monitored), solvent
was evaporated in vacuum. Water was added to the reaction
mixture and the organic part of the mixture was separated by
Cytotoxic effect on Cells. The cytotoxic effects of DPQI and 75 using dichloromethane (2×30 ml). The combine organic layer
Na2S were determined by MTT assay following the
manufacturer’s instruction (MTT 2003, SigmaꢀAldrich, MO).
Vero cells were seeded onto 96ꢀwell plates (approximately 104
cells per well) for 24 h. Next day media was removed and various
was then purified by column chromatography. A yellow
crystalline compound of 3 was obtained as a pure product. The
yield of the crude product is 51.2mg (80.15%), MS (HR MS):
2
3
3
5
0
5
+
1
(m/z, %): 340.0509 [(M+H ), 100 %];. HꢀNMR (DMSOꢀd6, 300
concentrations of probe DPQI, Na S (0, 15, 25, 50, 75, and 100 80 MHz): δ (ppm) 7.06 (d, 1H, J=9.0Hz), 7.88 (m, 2H), 8.01
2
ꢁM) made in DMEM were added to the cells and incubated for
(d,1H,J= 9.0Hz), 8.10 (m, 1H), 8.26 (m, 1H), 8.70
(d,1H,J=9.0Hz), 8.92 (d,1H, J=3.4Hz), 9.94 (1H,s).
2
4 h. Solvent control samples (cells treated with DMSO in
DMEM), no cells and cells in DMEM without any treatment were
also included in the study. Following incubation, the growth
media was removed, and fresh DMEM containing MTT solution
Preparation of DPQI: Ethanolic solution compound 3 (95mg,
0.280mmol) was added with 1,2,3,3ꢀTetramethylꢀ3Hꢀindolium
was added. The plate was incubated for 3−4 h at 37°C. 85 iodide (48mg, 0.28mmol) and reflux for overnight, a redcolor
Subsequently, the supernatant was removed, the insoluble colored
formazan product was solubilized in DMSO, and its absorbance
was measured in a micro titer plate reader (PerkinꢀElmer) at 570
nm.
precipitate was appeared during stirring at room temperature,
washed with EtOH. Then the solid was reꢀcrystallized from
EtOHꢀether (3:2; v/v) mixture to afford pure DPQI as a red soil.
Yield: 78 mg (56 %), M.P.> 250ºC. MS (HR MS): (m/z, %):
+
1
90
495.1451 [(M ), 100 %];. HꢀNMR (CDCl , 400 MHz): δ
3
Computational Methods. Geometries have been optimized
using the B3LYP/6ꢀ31G (d, p) level of theory. The geometries are
verified as proper minima by frequency calculations. Timeꢀ
dependent density functional theory calculation has also been
(ppm)1.24 (s, 6H), 1.81 (s, 3H), 6.92 (d, 2H,J=9.2Hz), 7.61 (t,
4H, J=9.0Hz), 7.75 (t, 2H,J=6.0Hz), 7.88 (d, 1H,J=9.0Hz),8.01
(d, 1H,J=12.0Hz), 8.24(m,1H,J=5.4Hz), 8.93(d,1H,J=6.0Hz),
4
0
13
8.57(d,1H,J=12.0Hz), 8.86(d,1H, J=8.1Hz). CꢀNMR (DMSOꢀ
performed at the same level of theory. All calculations have been 95 d6, 100 MHz): δ (ppm) 24.96, 35.02, 49.23, 52.62, 56.94, 110.12,
carried out using Gaussian 09 program.
115.73, 122.24, 125,17, 127.37, 128.79, 129.32, 129.78, 130.18,
1
1
34.05, 138.82, 142.07, 143.87, 143.92, 147.82, 149.42, 150.85,
52.56, 153.72, 162.53, 166.03, 181.73.
General method of UVꢀvis and fluorescence titration. For
UVꢀvis and fluorescence titrations, stock solution of Probe DPQI
4
5
was prepared (c = 10.0 µM) in H O–DMSO (2:1, v/v) solution
2
Acknowledgements
(
20.0 mM HEPES buffer, pH 7.4).The solution of the guest
ꢀ
3
ꢀ1
analytes in the order of 1.0 x 10 ML was also prepared in 100 We gratefully acknowledge DAEꢀBRNS [Project No.
H O–DMSO (2:1, v/v) solution (20.0 mM HEPES buffer, pH
36(1)/14/12/2016ꢀBRNS/36012] for financial support. SSA
thanks to UGCꢀMANF (Student ID: 2015ꢀ17ꢀWESꢀ67786), New
Delhi for the financial and research support.
2
5
0
7.4). The test solution of sensor Probe 1 was prepared by proper
dilution method. The spectra of these solutions were recorded by
means of UVꢀvis and the fluorescence methods. All the solvents
were purchased from local suppliers and were distilled by
standard procedure before use.
This journal is © The Royal Society of Chemistry [year]
Journal Name, [year], [vol], 00–00 | 7

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DOI: 10.1039/C7NJ04632D
5
5
310–311. (b) L. F. Hu, M. Lu, C. X. Tiong, G. S. Dawe, G. Hu
and J. S. Bian, Aging Cell, 2010, 9, 135.
Conflicts of interest
There are no conflicts to declare
Author information
1
2
1. W.Yang, G. Yang, X. Jia, L. Wu and R. Wang, J. Physiol.
005, 569, 519–531.
60
1
2. S. Fiorucci, E. Antonelli, A. Mencarelli, S. Orlandi, B. Renga,
a
Department of Chemistry, Indian Institute of Engineering
G. Rizzo, E. Distrutti, V. Shah and A. Morelli, Hepatology.,
2005, 42, 539–548.
5
0
5
Science and Technology, Shibpur, Howrahꢀ711103, West Bengal,
India,
Email:
akmahapatra@rediffmail.com,
Fax:
+
913326684564
65 13. C. Liu and S. Shen, Anal. Chem., 1964, 36, 1652ꢀ1654.
b
School of Chemistry and Biochemistry, Thapar Institute of
1
4. (a) N. S. Lawrence, J. Davis, L. Jiang, T. G. J. Jones, S. N.
Davies and R. G. Compton, Electroanalysis 2000, 18, 1453–
460; (b) I. S. P. Savizi, H.ꢀR. Kariminia, M. Ghadiri and R.
RoostaꢀAzad, Biosens. Bioelectron., 2012, 35, 297ꢀ301.
Engineering and Technology, Patiala 147 004 Punjab, 486 India.
1
c
1
Centre for Healthcare Science and Technology, Indian Institute
70
of Engineering Science and Technology, Shibpur, India
1
2
5.(a) J. Čmelik, J. Machat, V. Otruba and V. Kanicky, Talanta,
010, 80, 1777ꢀ1781; ( b) J. M. Davidson, Z. Pikramenou, A.
d
Bhabha Atomic Research Centre,Trombay.Mumbaiꢀ400085,
India.
Ponce and R. E. P. Winpenny, Anal. Chem., 2009, 81, 3669ꢀ3675.
75
†
Electronic Supplementary Information (ESI) available: [details
1
1
6. A. Safavi and M. A. Karimi, Talanta, 2002, 57, 491ꢀ500.
7. (a) J. Furne, A. Saeed and M. D. Levitt, Am. J. Physiol. 2008,
1
of any supplementary information available should be included
here]. See DOI: 10.1039/b000000x/
2
95, R1479–R1485; (b) S. E. Mylon and G. Benoit, Environ.
Notes and References
80 Sci. Technol., 2001, 35, 4544ꢀ4548; (c) J. Radfordknoery and G.
A. Cutter, Anal. Chem., 1993, 65, 976ꢀ982.
1
. (a) H. Kimura, Amino Acids 2011, 41, 113. (b) D. R. Linden,
Antioxid. Redox Signaling, 2014, 20, 818. (c) S. Fiorucci, E.
Distrutti, G. Cirino and J. L. Wallace, Gastroenterology, 2006,
18.(a) V.S. Lin, W. Chen, M. Xian and C.J. Chang Chem. Soc.
Rev., 2015, 44, 4596—4618; (b) B. Peng and M. Xian Asian J.
2
2
3
3
4
4
5
0
5
0
5
0
5
0
1
9
31, 259. (d) C. Szabó, Nature Reviews Drug Discovery 2007, 6, 85 Org. Chem. 2014, 3, 914 – 924.
17. (e) O. Kabil and R. Banerjee, Journal of Biological
Chemistry 2010, 285, 21903.
19. (a) Y. Yan, H. Yu, Y. Zhang, K. Zhang, H. Zhu, T. Yu, H.
Jiang and Wang, S. ACS Appl. Mater. Interfaces 2015, 7, 3547;
(b) T. Liu, J. Lin, Z. Li, L. Lin, Y. Shen, H. Zhu and Y. Qian,
2. G.Yang, L. Wu, B. Jiang, B.Yang, J. Qi, Cao. K, Q. Meng,
A.K.Mustafa, W. Mu, S.Zhang, S. H. Snyder and R.Wang, 90 Analyst 2015, 140, 7165; (c) L. Yang, X. Liu, L. Gao, F. Qi, H.
Science 2008, 322, 587–590.
Tian and X. Song, RSC Adv. 2015, 5, 98154; (d) Y. Cai, L. Li, Z.
Wang, Sun, Z. J. A. Qin and B. Z. Tang, Chem. Commun.
2014,52, 8892; (e) L. He, W. Lin, Q. Xu and H. Wei, Chem.
Commun. 2015, 51, 1510; (f) X.L. Liu, X.J. Du, C.ꢀG. Dai and
3
. A. Papapetropoulos, A. Pyriochou, Z. Altaany, G. Yang, A.
Marazioti, Z. Zhou, M. G. Jeschke, L. K. Branski, D. N. Herndon
and R.Wang, Szabo, C. Proc. Natl. Acad. Sci. U.S.A. 2009, 106, 95 Q.ꢀH. Song, J. Org. Chem. 2014, 79, 9481. (g) K. Maiti,
2
1972–21977.
A.K.Mahapatra, R.Maji, S.Mondal, S.S.Ali, A.Gangopadhyay,
S.K.Manna and S.Mandal, Chemistry Select, 2016, 1, 5066ꢀ5073.
4
. Y.J. Peng, J. Nanduri, G. Raghuraman, D. Souvannakitti, M.
M. Gadalla and G. K. Kumar, Proc. Natl. Acad. Sci. U.S.A. 2010,
07, 10719–10724.
20.(a) H. Li, Q. Yao, J. Fan, N. Jiang, J. Wang, J. Xia and X.
100 Peng, Chem. Commun. 2015, 51, 16225; (b) Y. Qian, J. Karpus,
O. Kabil, S.ꢀY. Zhang, H.ꢀL. Zhu, R. Banerjee, J. Zhao and C.
He, Nat. Commun. 2011, 2, 495; (c) J. Liu, Y.ꢀQ. Sun, J. Zhang,
T. Yang, J. Cao, L. Zhang and W. Guo, Chem. Eur. J. 2013, 19,
4717; (d) X. Wang, J. Sun, W. Zhang, X. Ma, J. Lv and B. Tang,
105 Chem. Sci. 2013, 4, 2551; (e) Y. Chen, C. Zhu, Z. Yang, J. Chen,
Y. He, Y. Jiao, W. He, L. Qiu, J. Cen and Z. Guo, Angew.
Chem., Int. Ed. 2013, 52, 1688.
1
5
6
. G. Yang, L.Wu and R. Wang, FASEB J. 2006, 20, 553–555.
. L. Li, M. Bhatia, Z. Zhu, Y. Zhu, C. Y. R. D. Ramnath, Z. J.
Wang, F. B. Anuar, M. Whiteman, M. SaltoꢀTellez and P. K.
Moore, FASEB J. 2005, 19, 1196–1198.
7
. K. Abe and H. Kimura, J. Neurosci. 1996, 16, 1066–1071.
8. C.K. Nicholson and J.W. Calvert, Pharmacol. Res. 2010, 62,
89–297.
21. (a) V. S. Lin and C. J. Chang, Curr. Opin. Chem. Biol., 2012,
110 16, 595ꢀ601; (b) A. R. Lippert, J. Inorg. Biochem., 2014, 133,
2
9
1
36ꢀ142; (c) R. Wang, F. Yu, L. Chen, H. Chen, L. Wang and
. (a) D. Giuliani, A. Ottani, D. Zaffe, M. Galantucci, F. Strinati,
W. Zhang, Chem. Commun., 2012, 48, 11757ꢀ11759.
R. Lodi and S. Guarini, Neurobiol. Learn. Mem, 2013, 104, 82.
(b) K. Eto, T. Asada, K. Arima, T. Makifuchi and H. Kimura,
Biochem. Biophys. Res. Commun. 2002, 293, 1485–1488.
22. Q. Li and J. R. Lancaster, Jr., Nitric Oxide 2013, 35, 21 –34.
115
2
3. A. R. Lippert, E. J. New, C. J. Chang, J. Am. Chem. Soc.
1
0. (a) P. Kamoun, M.C. Belardinelli, A. Chabli, K. Lallouchi
2011, 133, 10078.
and B. ChadefauxꢀVekemans, Am. J. Med. Genet. 2003, 116A,
8
|
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This journal is © The Royal Society of Chemistry [year]

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2
4. H. Peng, Y. Cheng, C. Dai, A. L. King, B. L. Predmore, D. J.
60
Lefer and B. Wang, Angew. Chem., Int. Ed. 2011, 50, 9672.
2
5. S. K. Das, C. S. Lim, S. Y. Yang, J. H. Han and B. R. Cho,
Chem. Commun. 2012, 48, 8395.
5
2
4
6. L. A. Montoya and M. D. Pluth, Chem. Commun. 2012, 48,
767.
2
7. Q. Wan, Y. Song, Z. Li, X. Gao and H. Ma, Chem. Commun.
1
1
2
2
3
3
4
4
5
0
5
0
5
0
5
0
5
0
2013, 49, 502.
2
2
8. Z. Wu, Z. Li, L. Yang, J. Han and S. Han, Chem. Commun.
012, 48, 10120.
29. F. Yu, P. Li, P. Song, B. Wang, J. Zhao and K. Han, Chem.
Commun. 2012, 48, 2852.
3
2
0. S. Chen, Z.ꢀj. Chen, W. Ren and H.ꢀw.Ai, J. Am. Chem. Soc.
012, 134, 9589.
3
1. Y. Qian, J. Karpus, O. Kabil, S.ꢀY. Zhang, H.ꢀL. Zhu, R.
Banerjee, J. Zhao and C. He, Nat. Commun. 2011, 2, 495.
3
2
2.(a) X. Cao, W. Lin, K. Zheng and L. He, Chem. Commun.
012, 48, 10529.
3
3. A. K. Das, S. Goswami , C. K. Quah and H. K. Fun, New J.
Chem., 2015, 39, 5669ꢀ5675.
3
4. P. Qi, D. Zhang, Y. Suna and Y. Wana, Anal. Methods,
2016, 8, 3339ꢀ3344.
3
2
5. L. He, X. Yang, K. Xu, X. Kong and W. Lin, Chem. Sci.,
017, 8, 6257.
36. (a) S. Chen, P. Hou and X. Z. Song, Sens. Actuators, B,
015, 221, 951–955. (b) J. M. Wang, H. Yu, Q. Li and S. J.
2
Shao, Talanta, 2015, 144, 763–768. X. J. Yang, L. Q. Shen, H. B.
Bao, X. X. Fang, J. W. Xu, Y. X. Zhao and W. Yang, Sens.
Actuators, B, 2015, 220, 1361–1367.
3
8
7. Q. Huang, X. F. Yang and H. Li, Dyes Pigm., 2013, 99, 871–
77. C. H. Zhang, B. Peng, W. Chen, S. M. Shuang, M. Xian and
C. Dong, Dyes Pigm., 2015, 121, 299–304. X. Li, J. Cheng, Y. L.
Gon, B. Yang and Y. Z. Hu, Biosens. Bioelectron., 2015, 65,
302–306.
3
8. (a) H. Zhang, C. Zhang, R. Liu, L. Yi and H. Sun Chem.
Commun., 2015, 51, 2029ꢀ2032; (b) H. Zhang, C. Zhang, R. Liu,
L. Yi and H. Sun Chem. Commun., 2015, 51, 2029; (c) H. Zhang,
P. Wang, Y. Yang and H. Sun, Chem. Commun., 2012, 48,
1
0672; (d) J. Jo and D. Lee, J. Am. Chem. Soc., 2009, 131, 16283.
5
5
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Journal Name, [year], [vol], 00–00 | 9

New Journal of Chemistry
Page 10 of 10
View Article Online
DOI: 10.1039/C7NJ04632D
Graphical abstract
Reaction-Based Bi-signaling Chemodosimeter Probe for Selective Detection
of Hydrogen Sulfide and Cellular Studies
a
Uday Narayan Guria, Kalipada Maiti, Syed Samim Ali, Sandip Kumar Samanta, Debasish
a
a
a
b
Mandal, Ripon Sarkar, Pallab Datta, Asim Kumar Ghosh, Ajit Kumar Mahapatra*
c
c
d
a
a
Department of Chemistry, Indian Institute of Engineering Science and Technology, Shibpur, Howrah-711103, West
Bengal, India, Email: akmahapatra@rediffmail.com, Fax: +913326684564
b
School of Chemistry and Biochemistry, Thapar Institute of Engineering and Technology, Patiala 147 004 Punjab, 486
India.
c
Centre for Healthcare Science and Technology, Indian Institute of Engineering Science and Technology, Shibpur, India
d
Bhabha Atomic Research Centre,Trombay.Mumbai-400085, India.
HS
HS
N
H2S(20-100 equiv)
N
N
2
H S(0-20equiv)
O
N
OH
N
O
N
O2N
NO2
O
N
NO
2
2