Full Papers
ChemistrySelect doi.org/10.1002/slct.202103098
z Analytical Chemistry
Pyrenyl-Based Aggregation-Induced Emission Luminogen
for Highly Sensitive and Selective Detection of 2,4,6-
Trinitrotoluene in Water
Hongmin Hao,[a] Zijian Ye,[a] Hui Dai,[a] Cong Liu,*[a] Aihua Yi,[b] Bingjia Xu,[a] Guang Shi,[a]
Shichen Su,*[c] Fahad Azad,[d] and Zhenguo Chi[e]
Trace detection of nitroaromatic explosives in aqueous solution
is important for environmental pollution evaluation and
terrorist prevention. Here, a new pyrenyl-based aggregation-
induced emission (AIE) fluorochrome (CPPyV) with desirable
fluorescent performance was synthesized and applied to detect
2,4,6-trinitrotoluene (TNT) in water. The fluorescence of CPPyV
can be selectivity and efficiently quenched by TNT. The Stern-
Volmer constant (KSV) of CPPyV for TNT was determined to be
6.2 × 105 M 1, with a low detection limit of 4 nM. Notably, CPPyV
could be used for TNT determinations in real water samples
Introduction
Nitroaromatic explosives (NAEs), such as 2,4,6-trinitrotoluene
(TNT) and 2,4,6-trinitrophenol (TNP), are mainly used in the
manufacture of explosive devices and are increasingly used in
the industrial field. When NAEs are impacted by high temper-
ature or strong impact, they will explode, which can seriously
threaten human safety and property.[1] In addition, due to the
carcinogenicity of NAEs and environmental pollution, they
have become an organic pollutant that can seriously affect
human health.[2] Selective and sensitive detection of NAEs in
water is thus important in national security, environmental
protection, and human health.[3]
[a] H. Hao, Z. Ye, H. Dai, Dr. C. Liu, B. Xu, Dr. G. Shi
Key Laboratory of Theoretical Chemistry of Environment, Ministry of
Education, School of Chemistry
South China Normal University, Guangzhou 510006, China
E-mail: liucong@m.scnu.edu.cn
[b] Dr. A. Yi
School of Materials Science and Engineering, Dongguan University of
Technology, Guangdong 523808, China
[c] Dr. S. Su
Institute of Semiconductor Science and Technology, South China Normal
University,Guangzhou 510631, China
E-mail: shichensu@126.com
[d] Dr. F. Azad
School of Natural Sciences (SNS), National University of Science and
Technology (NUST), H-12 Islamabad, Pakistan
[e] Dr. Z. Chi
State Key Laboratory of Optoelectronic Materials and Technologies,
School of Chemistry, Sun Yat-sen University
Key Laboratory for Polymeric Composite and Functional Materials of
Ministry of Education, School of Chemistry,Sun Yat-sen University
Guangzhou 510275, China
Supporting information for this article is available on the WWW under
https://doi.org/10.1002/slct.202103098
ChemistrySelect 2021, 6, 1 – 7
and also developed as a fluorescent paper sensor which could
efficiently detect TNT by naked-eye when the TNT concen-
tration was 1.0 × 10 8 M. The quenching process was dynamic
quenching mechanism and the theoretical calculations re-
vealed that photoin-duced electron transfer (PET) from excited
CPPyV to electron deficient TNT was the main reason for
fluorescence quenching. The excellent detection performance
of CPPyV yielded it a promising sensor material for actual field
applications.
Many techniques have been developed for detecting NAEs,
most analytical methods rely on complex instruments, which
are time-consuming and limit their application.[4] Fluorescence
measurement shows great simplicity and satisfactory sensitiv-
ity, so it has received extensive attention.[5] Although
fluorescence sensors has been well-known, most of lumino-
phores are insoluble and have aggregation-caused quenching
effects in water, which affect the detection of TNT.[6] Only a few
outstanding small fluorescent molecules reported for the
detection of TNT in water. In addition, rapid and selective assay
of TNT from other similar explosives that have strong electron
affinity, such as TNP, is still a challenge.[7]
Aggregation-induced emission (AIE) fluorescent materials,
because of its high luminous efficiency in the aggregated state,
are especially useful for the detection of explosives in water.[8]
However, many molecules have relatively high molecular
weight, high-cost precursors and problems of synthesis under
harsh reaction conditions.[9]
We aim to develop favorable electron-rich AIE luminogen
(AIEgen) for the efficient detection of TNT in a selective
manner. In this study, a small pyrenyl-based AIE fluorochrome
(CPPyV) was designed, which was constructed by linking two
electron-rich groups of carbazole to vinylbenzene and pyrene
moiety. Pyrene and their derivatives have been found to be
potential sensing fluorophores due to high quantum yield,
chemical stability, π-electron-rich property and electron trans-
fer ability.[10] The pyrene-based CPPyV was expected to have
twisted structures and high fluorescence efficiency in the
aggregated state. Herein, CPPyV was synthesized via a simple
route, and the precursors were considerably cost-effective
(Scheme 1). Then, the application of CPPyV for TNT detection in
water and real water samples were explored, the detection of
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ChemistrySelect doi.org/10.1002/slct.202103098

Scheme 1. Synthetic route for CPPyV compound.

TNT by fluorescent paper strip of CPPyV was also investigated.

The fluorescence quenching mechanism was confirmed by
fluorescence lifetime measurements and molecular orbital
energies calculation.

Results and Discussion

The designed CPPyV possessed a large conjugated skeleton of
pyrene moieties, which showed excellent fluorescence perform-
ance, and vinylbenzene that participated in forming a twisted
structure yielded the molecule to have AIE properties. The
linking of carbazole groups were aim to further improved the
fluorescence properties and develop favorable electron-rich
AIEgens for the efficient detection of TNT in a selective manner.
CPPyV was economically obtained using a simple Witting-
Horner method including two main reactants, diethyl(1,9-
dihydropyren-3-yl)methylphosphonate[11] and bis(4-(9H-carba-
zol-9-yl)phenyl)methanone,[12] which were prepared according
to published methods, respectively (Scheme 1). The structure
of CPPyV was characterized by 1H-NMR, MS (MALDI-TOF), and
FT-IR (Figures. S1-S3), which were consistent with the antici-
pated structure.
The fluorescence properties of CPPyV was explored. The
absorption spectrum of CPPyV in solution showed two peaks at
292 and 376 nm which were attributed to N-alkylcarbazole and
pyrene moieties, respectively (Figure 1A). The fluorescence
spectrum of CPPyV in water/THF mixtures and corresponding
fluorescence intensity change curves were collected (Figures 1B
and S4). The maximum fluorescence emission wavelength (λEm)
of CPPyV in solution was ∼ 505 nm. The fluorescence intensity
showed no clear change when the proportion of water in the
mixed solution was 0–60 %. When the proportion of water
exceeded 70 %, fluorescence intensity significantly increased
Figure 1. UV-Vis absorption spectra of CPPyV in THF (10 6 M, A) and
and, with the addition of 99 % water, exhibited fluorescence photoluminescence spectra of CPPyV in THF/water mixtures (λEx, 376 nm, B),
intensity ∼ 14 times that of pure THF solvent (Figure S4). After inset, optical photographs recorded under 365 nm UV light.
water addition, the materials gradually aggregated into nano-
particles, which was also confirmed by dynamic light scattering
analysis (Figure S5). These results demonstrated that CPPyV
possessed AIE performance and the excellent luminescent properties of this compound provided a theoretical basis for
TNT detection in aqueous solution.

ChemistrySelect 2021, 6, 1 – 7 2 © 2021 Wiley-VCH GmbH

 Full Papers
ChemistrySelect doi.org/10.1002/slct.202103098
Inspired by the excellent photoluminescence characteristics
and π-electron-richness of CPPyV, the potential applications of
CPPyV as a fluorescence probe for NAEs detection in water
were explored. Commonly used NAEs were chosen for this
investigation, including TNT, TNP, DNP, NP, 2-NT, and NB, the
fluorescence intensity of CPPyV monitored in their presence.
Notably, only TNT caused significant quenching of CPPyV
fluorescence emission (Figure 2). The fluorescence intensity of
CPPyV was almost completely quenched (96.18 %) by 0.01 mM
TNT, which indicated that CPPyV possessed high selectivity for
TNT. Interference from other NAEs was examined by testing
CPPyV fluorescence in the presence of different NAEs. The
CPPyV fluorescence intensity was quenched a little in the
presence of other NAEs mixture but significant quenched by
TNT (Figure 3). These results indicated that the superior
selectivity of CPPyV to TNT were not substantially interfered
with by commonly used NAEs.
Figure 2. Selectivity of CPPyV for sensing different nitro aromatic explosives
(0.01 mM) in aqueous solution (λEx = 376 nm).
Figure 3. Fluorescence quenching of CPPyV by TNT (0.01 mM) and commonly
used NAEs mixtures (TNP, DNP, NP, 2-NT, and NB) in aqueous solution
(λEx = 376 nm).
ChemistrySelect 2021, 6, 1 – 7
The detection effects of CPPyV toward TNT were further
explored by incrementally titrating TNT in CPPyV solutions. The
fluorescence intensity of CPPyV decreased with increased TNT
concentration (Figure 4A). Furthermore, the quenching effi-
ciency was quantitated by creating a Stern-Volmer(S V) plot,
according to equation (1):
K SV ½C� ¼ I0 =I-1 (1)
When the change of (I0/I-1) was plotted against the
concentration of quencher [C], a linear fitted line was obtained
(Figure 4B), which indicated that only one type of quenching
(dynamic or static) was predominant during quenching
processes.[13] The value of the fluorescence quenching constant
KSV for TNT, calculated from the S V plot was 6.2 × 105 M 1. For
comparison, relevant research data for TNT detection in
solution were summarized in Table 1, this calculated Ksv value
Figure 4. Fluorescence quenching diagram of CPPyV solutions with different
TNT concentrations (A) (λEx = 376 nm) and fitting diagram of relationship
between (I0/I-1) and concentration [C] (B).
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ChemistrySelect doi.org/10.1002/slct.202103098

Table 1. Comparison of TNT detection effects in CPPyV solution with

relevant literature.

Sample Detection status Ksv LOD Ref.

4 1 [14]
PCz-TPE THF/90 vol % 8.5 × 10 M ×
water.
TCMA acetone/hexane 1.399 × 104 M 1
× [15]

P2 poly- Water 1.5 × 105 M 1 6.4 × 10 8

M [16]

mers
Py-PAN Water 2.57 × 106 M 1
8.81 × 10 9
M [17]

NFM
AuNCs/ZIF- Water 6.64 × 104 M 1
5 × 10 9
M [18]

8
compound 70 % H2O/MeCN 4.3 × 107 M 1
3.2 × 10 9
M [9]

1 (v/v)
CPPyV Water 6.15 × 105 M 1
4.25 × 10 9
M This
work

Figure 5. Quenching of CPPyV by different TNT concentrations and

for TNT was comparable to those reported in the
literature.[9,14–18]
The LOD value was obtained by monitoring the
fluorescence quenching after gradually adding low concen-
tration TNT analyte (0-3 μM). The change in fluorescence
intensity was plotted against analyte concentration (Figure S6)
and the LOD was calculated using the equation
LOD ¼ 3s=m,
The LOD of CPPyV for TNT detection was found to be
4.24 nM (0.424 ppb). Compared with documented sensors in
the literature, CPPyV had comparable fluorescence quenching
coefficient and lower detection limit for TNT.[9,14–18] However,
most sensors documented in the literature can only used in
organic or mixed aqueous media, while CPPyV was purposely
designed for detection of TNT in aqueous media, which was
more suitable for far-ranging applications.
At the same time, CPPyV sensitivity to TNP was examined.
The Ksv constant of TNP (5.24 × 104 M 1) was ∼ 1 order lower
than that of TNT (6.2 × 105 M 1) and the LOD of TNP (56.3 nM)
was also much higher than that of TNT (4.24 nM, Figure S7).
Clearly, the sensitivity of CPPyV for TNT was much higher than
for TNP. As expected, CPPyV can selectively detect TNT in the
presence of TNP in aqueous solution.
The mechanism of the fluorescence quenching processes of
CPPyV was confirmed by performing time-resolved
fluorescence measurements of CPPyV in fluorescence titrations
(Figure 5). The Photoluminescence decays showed that the
lifetime value of CPPyV was 2.15 ns. With incremental TNT
addition, lifetime values were reduced to 1.86, 1.62, 1.48, 1.37,
and 1.26 ns in the presence of 2, 4, 6, 8, and 10 μM TNT,
respectively. The shorter lifetime values of CPPyV upon TNT
addition suggested a dynamic quenching process. In fact, the
UV-Vis spectrum also revealed that there was almost no change
of CPPyV UV-Vis spectrum after the addition of TNT, which
indicating that there was no significant interaction of CPPyV
with TNT at the ground state, and hence no complex was
formed (Figure S8). According to linearity of the S V plot,
dynamic quenching may be dominant mechanism (Figure 3B).
fluorescence lifetime changes with TNT concentration.
In the dynamic quenching process, the excited state of
fluorophore is quenched by a collision with quencher instead
of forming a ground state complex.
The emission spectrum of CPPyV and absorption spectrum
of TNT were also examined (Figure S9). Generally, if there was
an effective spectral overlap between the emission spectrum of
a fluorophore and absorption spectrum of the analyte,
fluorescence resonance energy transfer (FRET) might occur.[19]
Here, there almost no overlap between the emission spectrum
of CPPyV and absorption spectrum of TNT, which indicated
that FRET might not occur. The DLS studies after addition of
TNT were explored and showed that the aggregates of CPPyV
remain stable (Figure S10).
Based on the above analysis, photoinduced electron trans-
fer (PET) from the excited CPPyV to TNT may be the
mechanisms for the fluorescence quenching.[18] In PET process,
an excited state donor-acceptor complex was formed. To prove
the quenching process, the geometry for CPPyV, TNT and the
complex of CPPyV and TNT were optimized by density
functional theory and the electron densities of highest
occupied molecular orbitals (HOMO) and lowest occupied
molecular orbitals (LUMO) were determined (Figure 6). For
CPPyV, the LUMO was 2.24 eV. In comparison, TNT had a
much lower LUMO than that of CPPyV, at only 4.52 eV. The
orbital energy gap (2.28 eV) between the LUMOs of CPPyV and
TNT provided thermodynamic driving force for the electrons
transfer from electron-rich sensor (CPPyV) to electron-deficient
analytes (TNT). On the other hand, after adding TNT, the LUMO
energy of CPPyV (-2.24 eV) decreased to 4.24 eV in [CPPyV +
TNT] complex, while the newly formed HOMO energy did not
change (-5.54 eV). The initial HOMO and LUMO energy differ-
ence of CPPyV (3.21 eV) is reduced to 1.12 eV in [CPPyV + TNT]
complex. In addition, the LUMO mainly localized on TNT in
[CPPyV + TNT] complex, which also proved the transfer of
electrons from the excited state of CPPyV to TNT.
In conclusion, the PET process from excited CPPyV to
electron deficient TNT was concluded to be the main reason
for fluorescence quenching. The superior selectivity was

ChemistrySelect 2021, 6, 1 – 7 4 © 2021 Wiley-VCH GmbH

 Full Papers
ChemistrySelect doi.org/10.1002/slct.202103098
Figure 6. Orbital energy levels of CPPyV and nitroaromatic explosives.
considered to be related to the electron-withdrawing ability of
the analyte. TNT and TNP had the most nitro groups on their
aromatic rings and thus had strong electron-withdrawing
ability, which led to relatively high quenching efficiency. The
LUMO levels of TNT and TNP were comparable.[4] However, TNT
exhibited relatively higher quenching efficiency than TNP, as
TNP had a hydroxyl group which had lone electron pairs. In
addition, TNP formed a negatively charged anion in aqueous
solution and the resulting electrostatic repulsion hindered the
PET process, which lead to a lower quenching effects.[20]
Considering the observed excellent monitoring perform-
ance of CPPyV for TNT, the practical application of CPPyV was
performed. The real water asmples, including deionized water,
purified water, tap water, lake water (a lake at South China
Normal University), and Zhujiang River water (a river in
Guangzhou, China) were collected, and then filtered with
disposable filter membrane. Photographs of CPPyV in these
real water samples before and after adding TNT solution
(0.01 mM) were investigated. It was found that the fluorescence
intensity of these samples showed clear quenching in the
present of TNT (Figure S11 and 12), which suggested the great
potential for sensing TNT in real water samples.
Developing portable sensors for on-site detection is
significance.[21] Filter paper can be used for preparing visual
detection materials of TNT because of its portability, flexibility,
and low cost. A simple CPPyV filter paper sensor was produced
by soaking filter paper in CPPyV solution (10 4 M) and then the
treated paper dried naturally. It was observed that CPPyV filter
paper exhibited strong emission under 365 nm excitation
(Figure 7). After different concentrations of TNT solution (0.01-
10 μM) were applied by fingertip to the treated filter paper, the
noticeable fluorescence quenching of CPPyV paper-based
sensor was observed when the concentration of TNT was more
than 1.0 × 10 8 M. These results illustrated that simple CPPyV
ChemistrySelect 2021, 6, 1 – 7
Figure 7. Photographs of filter paper after immersion in CPPyV solution
(10 4 M) and contact with different fingertip-applied TNT concentrations
under 365 nm irradiation. From left to right, TNT concentrations 1.0 × 10 5,
1.0 × 10 6, 1.0 × 10 7, and 1.0 × 10 8 M (A–D, respectively).
paper sensors could be also used for real-time field and visual
sensing of TNT.
Conclusion
A new pyrenyl-based AIE fluorochrome (CPPyV) with desirable
fluorescent performance was successfully synthesized for the
detection of a specific nitroaromatic explosive, TNT, in water.
This fluorochrome had good luminescent properties and
displayed high selectivity and sensitivity to TNT in aqueous
solution. The fluorescence emission quenching of CPPyV
reached 96.18 % with 0.01 mM TNT solution, while there were
no significant changes when other nitroaromatic explosives
were added. CPPyV can selectively detect TNT in the presence
of other commonly used NAEs in aqueous solution. For TNT
detection, CPPyV showed a high Ksv constant of 6.2 × 105 M 1
and low detection limit of 4 nM. The PET process from excited
CPPyV to electron deficient TNT was the main reason for the
fluorescence quenching. This molecule was simple but unique,
showed great potential for TNT determination in real water
samples and could be processed as a fluorescent paper for
naked eye detection of trace TNT.
Supporting Information Summary
The detailed experimental procedures are given in the
supporting information.
Acknowledgements
This work was supported by National Natural Science Foundation
of China (51603233), Natural Science Foundation of Guangdong
Province of China (2020 A1515010439 and 2019 A1515010550),
Pearl River S&T Nova Program of Guangzhou (201806010125),
Opening Foundation of Key Laboratory for Polymeric Composite
and Functional Materials of Ministry of Education(Sun Yat-sen
University, PCFM-2019-05), Opening Foundation of StateKey
Laboratory of Optoelectronic Materials and Technologies(Sun Yat-
sen University, OEMT-2018-KF-11 and OEMT-2019-KF-08).
Conflict of Interest
The authors declare no conflict of interest.
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ChemistrySelect doi.org/10.1002/slct.202103098

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FULL PAPERS
A small pyrenyl-based < url
href = “http://www.baidu.com/link?
url = FqaZPO-
K6eKSTs5xsNj4LjmMKrtjRCCKqOclKq-
7KNayZi6bj54CNKHUK88kAV-
34 h8IlDtN46_OLYWWIx1ILlK” > AIE
fluorochrome (CPPyV) was reported,
which was synthesized via a simple
route. CPPyV had the advantages of
H. Hao, Z. Ye, H. Dai, Dr. C. Liu*,
Dr. A. Yi, B. Xu, Dr. G. Shi, Dr. S. Su*,
Dr. F. Azad, Dr. Z. Chi
1–7
Pyrenyl-Based Aggregation-
Induced Emission Luminogen for
Highly Sensitive and Selective
Detection of 2,4,6-Trinitrotoluene
in Water
high selectivity and sensitivity in the
detection of TNT, with a low detection
limit of 4 nM. Notably, CPPyV could
be used for TNT determinations in
real water samples and also
developed as a fluorescent paper
sensor which could efficiently detect
TNT by naked-eye when the TNT con-
centration was 1.0 × 10 8 M.