Article

Cite This: ACS Sens. 2018, 3, 1142−1148 pubs.acs.org/acssensors

Selective Detection of H2S by Copper Complex Embedded in Vesicles

through Metal Indicator Displacement Approach
Rahul Kaushik,† Rahul Sakla,† Amrita Ghosh,† G.Tamil Selvan,‡ P. Mosae Selvakumar,‡
and D. Amilan Jose*,†
†
Department of Chemistry, National Institute of Technology (NIT) Kurukshetra, Kurukshetra-136119, Haryana, India
‡
Department of Science & Humanities, Karunya Institute of Technology & Sciences, Coimbatore 641114, Tamil Nadu, India
*
S Supporting Information
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ABSTRACT: A new approach for the detection of hydrogen sulfide (H2S)

was constructed within vesicles comprising phospholipids and amphiphilic
copper complex as receptor. 1,2-Distearoyl-sn-glycero-3-phosphocholine
Downloaded via Amilan Jose Devadoss on July 2, 2018 at 06:01:28 (UTC).

(DSPC) vesicles with embedded metal complex receptor (1.Cu) sites have
been prepared. The vesicles selectively respond to H2S in a buffered
solution and show colorimetric as well as spectral transformation. Other
analytes such as reactive sulfur species, reactive nitrogen species, biological
phosphates, and other anions failed to induce changes. The H2S detection is
established through a metal indicator displacement (MIDA) process, where
Eosin-Y (EY) was employed as an indicator. Fluorescence, UV−vis
spectroscopy, and the naked eye as the signal readout studies confirm the
high selectivity, sensitivity, and lower detection limit of the vesicular
receptor. The application of vesicular receptors for real sample analysis was
also confirmed by fluorescence live cell imaging.
KEYWORDS: hydrogen sulfide, metal indicator displacement approach, phospholipid vesicles, colorimetric, fluorescent

H ydrogen sulfide (H2S) has been recognized as one of

three gasotransmitters, along with nitric oxide (NO) and
carbon monoxide (CO).1,2 The physiological and therapeutic
based on the formation of metal sulfide due to a high binding
constant of metal sulfides. Similar to MDA, the metal indicator
displacement approach (MIDA) is another approach, in which
importance of H2S is leading to quick progress in research both metal and indicator get displaced upon binding of H2S
activity involving H2S.3−6 Considering the complex biological
with metal center (Scheme 1). Nevertheless, MIDA based
functions,7,8 real-time and selective detection of endogenous
H2S is necessary to expand the knowledge about its exact sensor for the detection of H2S in vesicles not known in the
physiological and pathological role. But due to its volatile and literature until now.
reactive nature, accurate detection of H2S is heavily hampered Functionalized artificial membranes/bilayers with embedded
by sample preparation and detection methods. Classical chemical sensors represent a special class of chemosensors.41,42
methods such as gas chromatography, atomic absorption
spectroscopy (AAS), and electrochemical methods are available Scheme 1. Schematic Representation of (A) Metal
for the detection and estimation of H2S. However, due to harsh Displacement Approach (MDA) and (B) Metal Indicator
working conditions and tedious sample preparation, these Displacement Approach (MIDA)
methods are highly disadvantageous for biological samples. On
the other hand, UV−vis and emission spectroscopy methods
are highly reliable due to easy sample preparation, better
sensitivity, and quick sample analysis time. Therefore, several
chemical sensors have been recently reported for the detection
of H2S using change in fluorescence and UV/vis spectral
behavior.8−19 Most of the known H2S sensors work mainly
based on the H2S specific reactions such as reduction
reaction20−29 and nucleophilic attack.23,30−34 These strategies
have their own limitations such as low water solubility and
time-consuming response.35,36 As an alternative approach, we
and others have used the metal displacement approach Received: February 27, 2018
(MDA).18,37−40 It is promising and overcomes almost all the Accepted: June 1, 2018
disadvantages shown by reaction based H2S sensors. MDA is Published: June 1, 2018

© 2018 American Chemical Society 1142 DOI: 10.1021/acssensors.8b00174

ACS Sens. 2018, 3, 1142−1148
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The artificial bilayers mimic the functions of biomembranes
which are known to play a key role in different biological
processes associated with the transmission of signals across cell
membranes. The use of functionalized luminescent vesicles for
the detection of various biological analytes is known.43−45 But
in the literature, to the best of our knowledge, the vesicle-
embedded receptors for the real-time detection of H2S is not
known. Vesicles are the perfect platform for the detection of
endogenous analytes due to their monodisperse nature and
facile preparation. We envisaged that the design of vesicular
receptors with MIDA would allow us to replace the multistep
synthesis of signaling group attached receptors and also
improve biocompatibility that would certainly broaden their
applicability.
In this paper, we have reported a different approach by
embedding a copper complex functioning as an H2S reporter,
along with an indicator dye in the phospholipid vesicles (135 ±
10 nm) for the detection of H2S. The analyte binding at the
interface removes the metal in the form of metal sulfide. This
results in an expulsion of the indicator and metal from the
vesicular receptors, thereby triggering an optical response. For
the first time, we have introduced the MIDA in the vesicular
surface for the selective and sensitive detection of H2S in the
pure aqueous medium.
■
RESULTS AND DISCUSSION
To achieve the vesicular receptor synthesis, we have prepared
DPA (di-(2-picolyl)amine) based new amphiphilic copper
complex (1.Cu). 1.Cu was embedded into the phospholipids
1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) to form
vesicular receptor Ves-1.Cu (Chart 1). DPA based copper
Chart 1. Structures of 1.Cu, DSPC, and Indicator EY
complexes are known as prominent receptors to detect H2S due
to very low solubility product of CuS (Ksp = 6.36 × 10−36).18
Majority of the copper complex based H2S sensors are working
in semiaqueous system that may not be suitable for the
detection of H2S in biological system.12 Therefore, we have
prepared amphiphilic 1.Cu, that could be incorporated in
vesicles surface to work in pure aqueous environment.
Water-insoluble 1.Cu could be incorporated into the lipids
by a simple protocol to give a clear vesicular solution in water at
the physiological condition. Lipid bilayer vesicle embedded
copper complex (Ves-1.Cu) was prepared by using commer-
cially available synthetic lipid DSPC and amphiphilic complex
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1.Cu (25 mol % with respect to used DSPC) in HEPES buffer
(pH 7.4, 25 mM) by the well-known film-hydration method.46
The resulting heterogeneous liposomes Ves-1.Cu were
homogenized by extrusion method through polycarbonate
membranes to yield homogeneous small Ves-1.Cu. Homogen-
ized vesicular dispersions were assumed to be free from
impurities; therefore, the vesicle solution was not further
purified. The particle size distribution of the vesicles Ves-1.Cu
was examined by dynamic light scattering (DLS) method. The
average size of the receptor incorporated liposomes was found
to be 135 ± 10 nm (Figure S4). The dilution of the Ves-1.Cu
samples by water did not have much effect on the average size
of the Ves-1.Cu as determined by DLS (Figure S5). The
prepared vesicle dispersions were stored in the dark at 10 °C
and used within a week for the sensing study.
A convenient approach to detect analytes by using
nonfluorescent/colorimetric probes is the indicator displace-
ment approach, which is based on the principle of competitive
binding of an indicator and an analyte to the receptor.47,48
Here, we have selected Eosin-Y (EY) as a fluorescent and
colorimetric indicator. After screening numbers of indicators,
EY was selected due to the high water solubility, high quantum
yield, and ability to bind with copper ions. The characteristic
fluorescence emission of EY quenches after binding to copper
with the carboxylate group.49 Also, EY does not have any
binding effect with H2S even after addition of 1000 equiv
(Figure S6-A). Therefore, EY could be an ideal choice as an
indicator for the displacement assay in Ves-1.Cu solution.
Vesicular copper complex and indicator ensemble Ves-1.Cu-
EY was prepared by mixing Ves-1.Cu and EY in aqueous buffer
solution. The formation of sensing ensemble Ves-1.Cu-EY was
confirmed by UV−vis and fluorescence titration experiments.
EY (2.5 × 10−6 M) initially shows absorbance peak at 515 nm
(ε = 82,600 mol−1 cm−1) in water (phosphate buffer, 20 mM,
pH 7.4). After the successive addition of Ves-1.Cu (0−6.4 ×
10−6 M) to the EY solution, ratiometric change in absorption
spectrum was observed with the decrease in 515 nm peak and
increase in peak at 535 nm with about 20 nm red shift (Figure
1A). The color of the EY solution changed from pale pink to
dark pink, which could be easily detected by the naked eye. In
the case of the fluorescence spectrum, EY shows strong
emission intensity at 540 nm (λexc = 515 nm). However, the
emission intensity is decreased and quenched upon addition of
Ves-1.Cu (Figure 1B). The change in emission and UV−vis
absorption spectrum is due to the interaction between vesicle
embedded copper complex and carboxylic acid group of the EY.
The binding constant (KsV) for Ves-1.Cu to the EY calculated
as 9.09 × 102 M−1 in phosphate buffer (20 mM, pH 7.4). The
UV−vis and the fluorescent study confirm the formation of
theVes-1.Cu-EY ensemble in aqueous medium.
The use of copper complex in IDA (indicator displacement
approach) is familiar for the detection of anionic analytes such
as PPi and ATP.50,51 However, vesicle embedded copper
complex for the detection of H2S using IDA is not known.
Consequently, we have checked the ability of new ensemble
Ves-1.Cu-EY for the detection of H2S in aqueous phosphate
buffer solution. For all the experiments Na2S has been used as a
source for H2S gas. The change in UV−vis spectra of Ves-1.Cu-
EY was monitored in the presence excess amount (10 equiv) of
biologically important analytes such as reactive sulfur species
(glutathione, cysteine, methionine, S2O32−), reactive nitrogen
species (NO•, NO2− and N3−), biological phosphates (ATP,
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Figure 1. (A) UV−vis titration of EY (2.5 × 10−6 M) with Ves-1.Cu
(0−6.4 × 10−6 M) in aqueous medium (phosphate buffer, 20 mM, pH
7.4). (B) Emission titration of EY (2.5 × 10−6 M) with Ves-1.Cu (0−
6.4 × 10−6 M) in aqueous medium (phosphate buffer, 20 mM, pH
7.4).
ADP, AMP, PPi, H2PO4−, and HPO42−), and other anions (F−,
Cl−, Br−, I−, and SO42−).
As shown in Figure 2A the spectrum drastically changes only
with the H2S. Other analytes exhibited an insignificant change
in the absorbance spectrum. The UV−vis absorption spectrum
(λmax = 515 nm) of ensemble Ves-1.Cu-EY with H2S matches
perfectly with the absorption spectrum of EY.
Copper ions are known to react with sulfide to form stable
CuS species,52 which has a low solubility product constant
(Ksp) = 1.6 × 10−24. So, it was expected that, in the presence of
H2S, the copper ion was removed from the Ves-1.Cu-EY
ensemble as CuS; consequently, the indicator dye EY is
displaced from the system and the original absorption spectrum
is recovered (Scheme 2).
The sensitivity of H2S binding was determined via systematic
titration by successive addition of H2S (0−2.5 × 10−5 M) to the
Ves-1.Cu-EY ensemble (Figure 2B). Subsequently, the
reversibility in the absorption spectrum was observed by
restoration of the absorbance peak at 515 nm. As shown in
Figure 3, H2S detection could also be monitored by the naked
eye, through a prominent color change from dark pink to pale
pink. Other analytes failed to induce any color change (Figure
3B). Time dependent study shows that H2S detection was fast
and instantaneous (Figure S6-B). Real-time detection is very
important for the practical application; consequently, the quick
response time of the present sensing ensemble has the
advantage over other probes working based on the chemical
reactivity of H2S.
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Figure 2. (A) Selectivity studies of Ves-1.Cu-EY with different anions
such as reactive sulfur species (glutathione, cysteine, methionine,
S2O32−), reactive nitrogen species (NO•, NO2−, and N3−), biological
phosphates (ATP, ADP, AMP, PPi, H2PO4−, and HPO42−), and other
anions (F−, Cl−, Br−, I−, and SO42−) using UV−vis spectroscopy in
aqueous buffer medium (phosphate buffer, 20 mM, pH 7.4). (B)
Absorbance titration of Ves-1.Cu-EY ensemble with H2S (0−2.5 ×
10−5 M) in aqueous medium.
Scheme 2. Schematic Representation of H2S Sensing Using
Ves-1.Cu-EY through MIDA Approach
Selectivity of the Ves-1.Cu-EY was also tested in the
presence of interfering species such as relevant biothiols and
biological phosphates. The competitive experiments revealed
that negligible interference or no interference was observed in
the coexistence of various species and H2S (Figure S7).
Accordingly, the probe Ves-1.Cu-EY could be applicable for
the selective determination of H2S even in the presence of
other biological species like ATP, ADP, AMP, PPi, GSH, and
cysteine. This result predicts the possible use of the probe even
when H2S coexists with other biologically relevant biothiols like
glutathione, cysteine, methionine, and phosphates.
EY dye is well-known for its characteristic fluorescent
behavior; therefore, we have recorded the emission spectrum
of Ves-1.Cu-EY in the presence and absence of H2S and other
DOI: 10.1021/acssensors.8b00174
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Figure 3. (A) Fluorescence change under UV light and (B) naked eye
color change; of Ves-1.Cu-EY with different anions (10 equiv) in
phosphate buffer (pH 7.4).
analytes. EY has strong emission at 540 nm (λex. = 515 nm) but
after binding with Ves-1.Cu it shows very weak emission
(Figure 3A). The weak emission peak observed at 540 nm for
Ves-1.Cu-EY was increased only in the presence of H2S. Other
analytes did not show any change in emission intensity (Figure
S8). This is due to the displacement of Cu as CuS and thereby
removal of EY from the ensemble. The limit of detection
(LOD) for H2S was calculated as 4.06 μM from systematic
fluorescent titration (Figure 4A). The observed LOD (Figure
4B) is lower as compared to polymer-based/nanomaterial-
Figure 4. (A) Selectivity studies of Ves-1.Cu-EY with different anions
such as reactive sulfur species (glutathione, cysteine, methionine,
S2O32−), reactive nitrogen species (NO•, NO2−, and N3−), biological
phosphates (ATP, ADP, AMP, PPi, H2PO4−, and HPO42−), and other
anions (F−, Cl−, Br−, I−, and SO42−) using fluorescence spectroscopy
in aqueous buffer medium (phosphate buffer, 20 mM, pH 7.4; λexc =
515 nm). (B) Emission titration of Ves-1.Cu-EY ensemble with H2S
(0−2.5 × 10−5 M) in aqueous medium (phosphate buffer, 20 mM, pH
7.4; λexc = 515 nm).
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based probes reported for the detection of H2S.53−59 The LOD
is also comparable to or better than other small molecule-based
probes reported for the H2S detection (Table S1).
Time-resolved fluorescence studies were carried out to
determine the emission decay parameters for EY, Ves-1.Cu-
EY, and Ves-1.Cu-EY with H2S (Figure S9). EY exhibited a
lifetime τ1 = 1.17 ns (quantum yield = 0.236) with single
exponential decay. The ensemble probe displayed a lifetime of
τ1 = 1.37 ns and τ2 = 2.74 ns (quantum yield = 0.087) with
biexponential decay. However, in the presence of H2S (2.5 ×
10−5 M), the lifetime was measured as τ1 = 1.04 ns and τ2 =
2.06 ns (quantum yield = 0.213). The recovery of EY lifetime
(τ1) and quantum yield further confirms the displacement of
dye from the vesicular surface due to the formation of CuS via
MIDA. Detailed UV−vis and fluorescence studies ensured that
the detection of H2S could be done with high sensitivity using
Ves-1.Cu-EY sensing ensemble.
Further, we have also used Ves-1.Cu-EY to monitor the
release of H2S from the donor molecule benzoic (methyl
carbonic) dithioperoxyanhydride.39 Time dependent fluores-
cence measurement was performed at 530/645 nm as
excitation/emission wavelength for 30 min at RT. The kinetic
study showed that the spontaneous release of H2S by donor
molecules easily monitored by the probe Ves-1.Cu-EY based
on fluorescent change (Figure S20). This experiment
demonstrated that the use of Ves-1.Cu-EY for the detection
and monitoring of released H2S from donor molecules.
The morphology of vesicles was investigated under the light
microscope. The Ves-1.Cu are spherical in shape and well
separated from each other (Figure S10). The ensemble Ves-
1.Cu-EY alone did not show any fluorescence property in light
microscope, but upon binding with H2S, the vesicle solution
shows fluorescent behavior due to the displacement of copper
and the existence of free EY dye unit (Figure S11). DLS
measurement of Ves-1.Cu-EY was also recorded in the absence
and presence of H2S. In the presence of H2S, the particle size
distribution shows two different-sized particles in the solution
such as 200 nm and about 1050 nm. The higher-sized particle
may be attributed to the CuS precipitation. These results also
confirm the MIDA mechanism on the vesicular surface (Figure
S4).
H2S is synthesized both enzymatically and nonenzymatically
in the human body; therefore, it is highly important to detect
H2S in the real biological samples.15,60,61 To explore the
efficiency of the Ves-1.Cu-EY ensemble in real biological
samples, a live cell imaging study was performed. The cell
images were captured using mouse skeletal muscle cells.
Fluorescence images of cells show that the cells were highly
fluorescent in the presence of EY (2.5 × 10 −6 M).
Consequently, when the cells were treated with vesicles Ves-
1.Cu the decrease in fluorescence intensity was observed due to
the formation of nonfluorescent ensemble Ves-1.Cu-EY.
Further, when the cells were incubated with H2S, the
fluorescence intensity of the cell lines was recovered. This
showed that the MIDA mechanism is well supported by the live
cell imaging experiment and the Ves-1.Cu-EY ensemble could
be used for the detection of H2S in a biological sample at the
cellular level (Figure 5).
As of now, we have performed the entire H2S binding studies
in phosphate buffer. However, inorganic and biological
phosphates are known for binding with copper complex.45,62
Therefore, we have investigated the effect of the buffer by
replacing phosphate buffer with HEPES buffer. UV−vis and
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Figure 5. (A) Fluorescence microscopy images of C2C12 mouse
skeletal muscle fixed cell lines with EY (2.5 μM) only (B) With Ves-
1.Cu-EY. (C) After treatment with H2S (37 μM). C2C12 mouse
skeletal muscle fixed cell lines with 50−60% confluency using FITC
UV filter with a total magnification of 100 X.
fluorescent results suggest that the probe Ves-1.Cu-EY was not
very selective for H2S in HEPES buffer, as it also displayed
some changes with PPi, GSH, and Cys. The emission spectra
showed 15-, 8-, 4-, and 2-fold decrement in emission intensity
with H2S, Cys, GSH, and PPi, respectively (Figure S12).
Therefore, it is evident from UV−vis and emission studies that
the Ves-1.Cu-EY ensemble shows less selectivity (Figures S13−
S14). However, the calculated LOD (0.59 μM) suggested that
HEPES buffer is useful for sensitive detection of H2S (Figures
S15−S18). The LOD calculated by using HEPES buffer is 7
times less than the phosphate buffer. The study of the effect of
different buffer, lipid, and complex loading on H2S binding is in
progress; these results will be published in due time.
■ CONCLUSION
In summary, a new strategy for H2S detection was developed
through an MIDA process on the nanovesicular surface, where
Cu(II) acts as a binding site and EY acts as an indicator for the
detection mechanism. The vesicular ensemble developed
showed high selectivity and sensitivity for H2S. The practical
application of this vesicular probe was proven by the biological
cell imaging study. The Ves-1.Cu- EY ensemble showed a
number of advantages over other reported probes. Most of the
reported H2S sensors work in a semiaqueous environment and
lack applications in biological samples. On the other hand,
vesicles as receptors provide a pure aqueous environment at
physiological pH with better sensitivity. The vesicular platform
also provides the liberty to modulate the lipid membrane
depending upon the cellular environment in the biological
sample. However, no such advantage was provided by small
molecular probes. Vesicles embedded with copper complex
based on MIDA approach open up the platform to modulate
the emission and excitation wavelengths with suitable indicator
dyes.
■
ASSOCIATED CONTENT
*
S Supporting Information
The Supporting Information is available free of charge on the
ACS Publications website at DOI: 10.1021/acssen-
sors.8b00174.
Experimental sections, general procedure for spectro-
scopic measurements, UV−vis absorption, lifetime and
fluorescence studies; DLS analyses and color change
images (PDF)
■ AUTHOR INFORMATION
Corresponding Author
*E-mail: amilanjosenit@nitkkr.ac.in.
ORCID
Rahul Sakla: 0000-0001-5538-1830
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Article
P. Mosae Selvakumar: 0000-0002-8712-1168
D. Amilan Jose: 0000-0003-2978-3400
Author Contributions
The manuscript was written through contributions of all
authors.
Notes
The authors declare no competing financial interest.
■ ACKNOWLEDGMENTS
The authors acknowledge Department of Chemistry, NIT-
Kurukshetra for the lab facilities. We are thankful to Dr.
Ashwani Mittal, Biochemistry Department, University College,
Kurukshetra University for fluorescence microscopic images
and cell imaging experiment. D.A.J. acknowledges the financial
support for the SERB-DST, New Delhi project grant SB/FT/
CS-195/2013 and CSIR grant No. 01 (2855)/16/EMR-II. A.G.
is thankful to the financial support of DST, India for the SERB-
DST young scientist research project grants (SB/FT/CS-193/
2013). R.S. acknowledges DST-Haryana for a HSCST research
fellowship.
■ REFERENCES
(1) Abe, K.; Kimura, H. The possible role of hydrogen sulfide as an
endogenous neuromodulator. J. Neurosci. 1996, 16, 1066.
(2) Reiffenstein, R. J.; Hulbert, W. C.; Roth, S. H. Toxicology of
hydrogen sulfide. Annu. Rev. Pharmacol. Toxicol. 1992, 32, 109−34.
(3) Szabo, C. Hydrogen sulphide and its therapeutic potential. Nat.
Rev. Drug Discovery 2007, 6, 917−935.
(4) Yang, G.; Wu, L.; Jiang, B.; Yang, W.; Qi, J.; Cao, K.; Meng, Q.;
Mustafa, A. K.; Mu, W.; Zhang, S.; Snyder, S. H.; Wang, R. H2S as a
Physiologic Vasorelaxant: Hypertension in Mice with Deletion of
Cystathionine γ-Lyase. Science 2008, 322, 587−590.
(5) Drew, K. L.; Rice, M. E.; Kuhn, T. B.; Smith, M. A.
Neuroprotective adaptations in hibernation: therapeutic implications
for ischemia-reperfusion, traumatic brain injury and neurodegenerative
diseases. Free Radical Biol. Med. 2001, 31, 563−73.
(6) Boehning, D.; Snyder, S. H. Novel neural modulators. Annu. Rev.
Neurosci. 2003, 26, 105−131.
(7) Eto, K.; Asada, T.; Arima, K.; Makifuchi, T.; Kimura, H. Brain
hydrogen sulfide is severely decreased in Alzheimer’s disease. Biochem.
Biophys. Res. Commun. 2002, 293, 1485−1488.
(8) Kimura, H. Hydrogen sulfide: its production, release and
functions. Amino Acids 2011, 41, 113−121.
(9) Lin, V. S.; Chang, C. J. Fluorescent probes for sensing and
imaging biological hydrogen sulfide. Curr. Opin. Chem. Biol. 2012, 16,
595−601.
(10) Xuan, W.; Sheng, C.; Cao, Y.; He, W.; Wang, W. Fluorescent
Probes for the Detection of Hydrogen Sulfide in Biological Systems.
Angew. Chem., Int. Ed. 2012, 51, 2282−2284.
(11) Duan, C.-X.; Liu, Y.-G. Recent Advances in Fluorescent Probes
for Monitoring of Hydrogen Sulfide. Curr. Med. Chem. 2013, 20,
2929−2937.
(12) Kumar, N.; Bhalla, V.; Kumar, M. Recent developments of
fluorescent probes for the detection of gasotransmitters (NO, CO and
H2S). Coord. Chem. Rev. 2013, 257, 2335−2347.
(13) Peng, H.; Chen, W.; Burroughs, S.; Wang, B. Recent advances in
fluorescent probes for the detection of hydrogen sulfide. Curr. Org.
Chem. 2013, 17, 641−653.
(14) Peng, B.; Xian, M. Fluorescent Probes for Hydrogen Sulfide
Detection. Asian J. Org. Chem. 2014, 3, 914−924.
(15) Yu, F.; Han, X.; Chen, L. Fluorescent probes for hydrogen
sulfide detection and bioimaging. Chem. Commun. 2014, 50, 12234−
12249.
(16) Guo, Z.; Chen, G.; Zeng, G.; Li, Z.; Chen, A.; Wang, J.; Jiang, L.
Fluorescence chemosensors for hydrogen sulfide detection in
biological systems. Analyst 2015, 140, 1772−1786.
DOI: 10.1021/acssensors.8b00174
ACS Sens. 2018, 3, 1142−1148
ACS Sensors
(17) Lin, V. S.; Chen, W.; Xian, M.; Chang, C. J. Chemical probes for
molecular imaging and detection of hydrogen sulfide and reactive
sulfur species in biological systems. Chem. Soc. Rev. 2015, 44, 4596−
4618.
(18) Kaushik, R.; Ghosh, A.; Jose, D. A. Recent progress in hydrogen
sulphide (H2S) sensors by metal displacement approach. Coord. Chem.
Rev. 2017, 347, 141−157.
(19) Xin, X.; Wang, J.; Gong, C.; Xu, H.; Wang, R.; Ji, S.; Dong, H.;
Meng, Q.; Zhang, L.; Dai, F.; Sun, D. Cyclodextrin-Based Metal-
Organic Nanotube as Fluorescent Probe for Selective Turn-On
Detection of Hydrogen Sulfide in Living Cells Based on H2S-Involved
Coordination Mechanism. Sci. Rep. 2016, 6, 21951.
(20) Wu, Z.; Feng, Y.; Geng, B.; Liu, J.; Tang, X. Fluorogenic sensing
of H2S in blood and living cells via reduction of aromatic dialkylamino
N-oxide. RSC Adv. 2014, 4, 30398−30401.
(21) Bamesberger, A.; Kim, G.; Woo, J.; Cao, H. Reduction of Nitro
Group on Derivative of 1,8-Naphthalimide for Quantitative Detection
of Hydrogen Sulfide. J. Fluoresc. 2015, 25, 25−29.
(22) Li, J.; Yin, C.; Huo, F. Chromogenic and fluorogenic
chemosensors for hydrogen sulfide: review of detection mechanisms
since the year 2009. RSC Adv. 2015, 5, 2191−2206.
(23) Das, A. K.; Goswami, S.; Dutta, G.; Maity, S.; Mandal, T.k.;
Khanra, K.; Bhattacharyya, N. A concentration dependent auto-relay-
recognition by the same analyte: a dual fluorescence switch-on by
hydrogen sulfide via Michael addition followed by reduction and
staining for bio-activity. Org. Biomol. Chem. 2016, 14, 570−576.
(24) Zhou, G.; Wang, H.; Ma, Y.; Chen, X. An NBD fluorophore-
based colorimetric and fluorescent chemosensor for hydrogen sulfide
and its application for bioimaging. Tetrahedron 2013, 69, 867−870.
(25) Santos-Figueroa, L. E.; de laTorre, C.; El Sayed, S.; Sancenon,
F.; Martinez-Manez, R.; Costero, A. M.; Gil, S.; Parra, M. A
Chemosensor Bearing Sulfonyl Azide Moieties for Selective
Chromo-Fluorogenic Hydrogen Sulfide Recognition in Aqueous
Media and in Living Cells. Eur. J. Org. Chem. 2014, 2014, 1848−1854.
(26) Elsayed, S.; de la Torre, C.; Santos-Figueroa, L. E.; Marin-
Hernandez, C.; Martinez-Manez, R.; Sancenon, F.; Costero, A. M.; Gil,
S.; Parra, M. Azide and sulfonylazide functionalized fluorophores for
the selective and sensitive detection of hydrogen sulfide. Sens.
Actuators, B 2015, 207, 987−994.
(27) Buragohain, A.; Biswas, S. Cerium-based azide- and nitro-
functionalized UiO-66 frameworks as turn-on fluorescent probes for
the sensing of hydrogen sulphide. CrystEngComm 2016, 18, 4374−
4381.
(28) Thirumalaivasan, N.; Venkatesan, P.; Wu, S.-P. Highly selective
turn-on probe for H2S with imaging applications in vitro and in vivo.
New J. Chem. 2017, 41, 13510−13515.
(29) Zhang, J.; Gao, Y.; Kang, X.; Zhu, Z.; Wang, Z.; Xi, Z.; Yi, L. o,o-
Difluorination of aromatic azide yields a fast-response fluorescent
probe for H2S detection and for improved bioorthogonal reactions.
Org. Biomol. Chem. 2017, 15, 4212−4217.
(30) Peng, B.; Xian, M. Hydrogen Sulfide Detection Using
Nucleophilic Substitution−Cyclization-Based Fluorescent Probes. In
Methods in Enzymology; Academic Press, 2015; pp 47−62.
(31) Liu, J.; Sun, Y.-Q.; Zhang, J.; Yang, T.; Cao, J.; Zhang, L.; Guo,
W. A Ratiometric Fluorescent Probe for Biological Signaling Molecule
H2S: Fast Response and High Selectivity. Chem. - Eur. J. 2013, 19,
4717−4722.
(32) Wu, X.; Shi, J.; Yang, L.; Han, J.; Han, S. A near-infrared
fluorescence dye for sensitive detection of hydrogen sulfide in serum.
Bioorg. Med. Chem. Lett. 2014, 24, 314−316.
(33) Saha, T.; Kand, D.; Talukdar, P. Performance comparison of two
cascade reaction models in fluorescence off-on detection of hydrogen
sulfide. RSC Adv. 2015, 5, 1438−1446.
(34) Kawagoe, R.; Takashima, I.; Uchinomiya, S.; Ojida, A.
Reversible ratiometric detection of highly reactive hydropersulfides
using a FRET-based dual emission fluorescent probe. Chem. Sci. 2017,
8, 1134−1140.
1147
Article
(35) Lippert, A. R. Designing reaction-based fluorescent probes for
selective hydrogen sulfide detection. J. Inorg. Biochem. 2014, 133, 136−
142.
(36) Sathyadevi, P.; Chen, Y.-J.; Wu, S.-C.; Chen, Y.-H.; Wang, Y.-M.
Reaction-based epoxide fluorescent probe for in vivo visualization of
hydrogen sulfide. Biosens. Bioelectron. 2015, 68, 681−687.
(37) Kaushik, R.; Singh, A.; Ghosh, A.; Jose, D. A. Selective
Colorimetric Sensor for the Detection of Hg2+ and H2S in Aqueous
Medium and Waste Water Samples. ChemistrySelect 2016, 1, 1533−
1540.
(38) Kaushik, R.; Ghosh, A.; Jose, D. A. Simple terpyridine based
Cu(II)/Zn(II) complexes for the selective fluorescent detection of
H2S in aqueous medium. J. Lumin. 2016, 171, 112−117.
(39) Kaushik, R.; Kumar, P.; Ghosh, A.; Gupta, N.; Kaur, D.; Arora,
S.; Jose, D. A. Alizarin red S-zinc(II) fluorescent ensemble for selective
detection of hydrogen sulphide and assay with an H2S donor. RSC
Adv. 2015, 5, 79309−79316.
(40) Xin, X.; Dai, F.; Li, F.; Jin, X.; Wang, R.; Sun, D. A visual test
paper based on Pb(ii) metal-organic nanotubes utilized as a H2S
sensor with high selectivity and sensitivity. Anal. Methods 2017, 9,
3094−3098.
(41) Jose, D. A.; Stadlbauer, S.; Koenig, B. Polydiacetylene-Based
Colorimetric Self-Assembled Vesicular Receptors for Biological
Phosphate Ion Recognition. Chem. - Eur. J. 2009, 15, 7404−7412.
(42) Jose, D. A.; Koenig, B. Polydiacetylene vesicles functionalized
with N-heterocyclic ligands for metal cation binding. Org. Biomol.
Chem. 2010, 8, 655−662.
(43) Huang, T.; Hou, Z.; Xu, Q.; Huang, L.; Li, C.; Zhou, Y. Polymer
Vesicle Sensor for Visual and Sensitive Detection of SO2 in Water.
Langmuir 2017, 33, 340−346.
(44) Kim, H. M.; An, M. J.; Hong, J. H.; Jeong, B. H.; Kwon, O.;
Hyon, J.-Y.; Hong, S.-C.; Lee, K. J.; Cho, B. R. Two-Photon
Fluorescent Probes for Acidic Vesicles in Live Cells and Tissue. Angew.
Chem., Int. Ed. 2008, 47, 2231−2234.
(45) Banerjee, S.; König, B. Molecular Imprinting of Luminescent
Vesicles. J. Am. Chem. Soc. 2013, 135, 2967−2970.
(46) Zhang, H. Thin-Film Hydration Followed by Extrusion Method
for Liposome Preparation. In Liposomes: Methods and Protocols,
D’Souza, G. G. M., Ed.; Springer New York: New York, NY, 2017; pp
17−22.
(47) You, L.; Anslyn, E. V. Competition Experiments. In
Competition Experiments. In Supramolecular Chemistry: From
Molecules to Nanomaterials; John Wiley & Sons, Ltd, 2012.
(48) Nguyen, B. T.; Anslyn, E. V. Indicator−displacement assays.
Coord. Chem. Rev. 2006, 250, 3118−3127.
(49) Hu, M.; Feng, G. Highly selective and sensitive fluorescent
sensing of oxalate in water. Chem. Commun. 2012, 48, 6951−6953.
(50) Kim, S. K.; Lee, D. H.; Hong, J.-I.; Yoon, J. Chemosensors for
Pyrophosphate. Acc. Chem. Res. 2009, 42, 23−31.
(51) Umali, A. P.; Anslyn, E. V.; Wright, A. T.; Blieden, C. R.; Smith,
C. K.; Tian, T.; Truong, J. A.; Crumm, C. E.; Garcia, J. E.; Lee, S.;
Mosier, M.; Nguyen, C. P. Analysis of Citric Acid in Beverages: Use of
an Indicator Displacement Assay. J. Chem. Educ. 2010, 87, 832−835.
(52) Wu, P.; Zhang, J.; Wang, S.; Zhu, A.; Hou, X. Sensing during In
Situ Growth of Mn-Doped ZnS QDs: A Phosphorescent Sensor for
Detection of H2S in Biological Samples. Chem. - Eur. J. 2014, 20, 952−
956.
(53) Gao, J.; Li, Q.; Wang, C.; Tan, H. Copper (II)-mediated
fluorescence of lanthanide coordination polymers doped with carbon
dots for ratiometric detection of hydrogen sulfide. Sens. Actuators, B
2017, 253, 27−33.
(54) Sun, K.; Liu, X.; Wang, Y.; Wu, Z. A polymer-based turn-on
fluorescent sensor for specific detection of hydrogen sulfide. RSC Adv.
2013, 3, 14543−14548.
(55) Liu, B.; Chen, Y. Responsive Lanthanide Coordination Polymer
for Hydrogen Sulfide. Anal. Chem. 2013, 85, 11020−11025.
(56) Xu, H.; Wu, J. e.; Chen, C.-H.; Zhang, L.; Yang, K.-L. Detecting
hydrogen sulfide by using transparent polymer with embedded CdSe/
CdS quantum dots. Sens. Actuators, B 2010, 143, 535−538.
DOI: 10.1021/acssensors.8b00174
ACS Sens. 2018, 3, 1142−1148
ACS Sensors Article

(57) Zhang, L.; Lou, X.; Yu, Y.; Qin, J.; Li, Z. A New Disubstituted
Polyacetylene Bearing Pyridine Moieties: Convenient Synthesis and
Sensitive Chemosensor toward Sulfide Anion with High Selectivity.
Macromolecules 2011, 44, 5186−5193.
(58) Yan, Q.; Sang, W. H2S gasotransmitter-responsive polymer
vesicles. Chem. Sci. 2016, 7, 2100−2105.
(59) Zhang, J.; Hao, X.; Sang, W.; Yan, Q. Hydrogen Polysulfide
Biosignal-Responsive Polymersomes as a Nanoplatform for Distin-
guishing Intracellular Reactive Sulfur Species (RSS). Small 2017, 13,
1701601.
(60) Qi, P.; Zhang, D.; Sun, Y.; Wan, Y. A selective near-infrared
fluorescent probe for hydrogen sulfide and its application in sulfate-
reducing bacteria detection. Anal. Methods 2016, 8, 3339−3344.
(61) Hartle, M. D.; Pluth, M. D. A practical guide to working with
H2S at the interface of chemistry and biology. Chem. Soc. Rev. 2016,
45, 6108−6117.
(62) Jose, D. A.; Ghosh, A.; Schiller, A. Discovering the Future of
Molecular Sciences: Supramolecular receptors for the recognition of
bioanalytes; Wiley-VCH Verlag GmbH & Co. KGaA, 2014; pp 3−30.

1148 DOI: 10.1021/acssensors.8b00174

ACS Sens. 2018, 3, 1142−1148