Chemical Engineering Journal 443 (2022) 136445

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Chemical Engineering Journal

journal homepage: www.elsevier.com/locate/cej

Precise probe design based ESIPT coupled AIE mechanism toward

endogenous cyanide in food detection and bioimaging
Cuibing Bai a, c, 1, Jie Zhang a, 1, Yuxin Qin a, Huanan Huang b, *, Zhenni Xia d, Qijun Zheng a,
Hongliang Dai a, Pengkai Lu d, Hui Miao a, Changqing Qu d, Rui Qiao a, c, *
a
School of Chemistry and Materials Engineering, Fuyang Normal University, Fuyang, Anhui Province 236037, PR China
b
College of Chemistry and Chemical Engineering, Jiangxi Province Engineering Research Center of Ecological Chemical Industry, Xinghuo Organosilicon Industry
Research Center, Jiujiang University, Jiujiang 332005, PR China
c
Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190,
PR China
d
Research Center of Anti-aging Chinese Herbal Medicine of Anhui Province, Fuyang, Anhui 236037, PR China

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

Keywords: Cyanide (CN–) is one of the strongest and fastest-acting toxic substances, and its excessive presence will cause
Cyanide ion great harm to the environment, food and human health. Although several fluorescent probes that recognize CN–
Fluorescent probe have been extensively reported, developing the fluorescent probe with aggregation-induced emission (AIE) and
ESIPT-AIE
large Stokes shift is still an extremely difficult problem due to lack of effective strategies. Thus, continuing efforts
Biological imaging
to seek novel, structurally distinct and functionally specific structures with large Stokes shift and AIE features are
highly anticipated. Herein, we design and synthesize a novel asymmetric bis-Schiff base block (named as HPBC)
based on the excited state intramolecular proton transfer (ESIPT) and AIE mechanism. Benefiting from restriction
of intramolecular rotation and –CN electron-withdraw nature promoting the ESIPT effect of HPBC in the
recognition system, HPBC shows large Stokes shift (243 nm), high selectivity, rapid identification of CN– and AIE
effects. Excitingly, the limit of detection (LOD) was calculated to be 1.32 × 10-7 M, which was much lower than
the 1.9 × 10-6 M specified by the World Health Organization and better than other previously reported probes.
More importantly, further investigations demonstrated that the molecule we designed was successfully applied to
sense endogenous CN– in food samples and the low cytotoxicity of HPBC was allowed to discover CN– in
C. elegans, mice and HeLa cells. Overall, this work provides a new viewpoint of the rational design and synthesis
of fluorescent probe toward CN– with AIE and large Stokes shift features, and triggers the discovery of new
functions and properties of biological materials, which would open a new frontier for probes.

1. Introduction
Cyanide (CN–) as one of the most toxic anions that is quite harmful to
mammals [1]. Even worse, it was widely used in many areas, including
electroplating, metallurgy, synthetic resins, preparation of paints, dyes
and so on [2-4]. Thus, the development of new materials that achieve
stabilized, simple and sensitive CN– detection is highly desirable [5]. In
recent years, fluorescent probes as a promising detection technology
have attracted more and more attention because of their prominent
advantages, such as visualization, high sensitivity and good selectivity,
real-time imaging [6,7]. Although several fluorescent probes that
recognize CN– have been reported [8-15], most of the reported CN–
probes still suffer from low Stokes shift (Δλ < 200 nm) and the
aggregation-caused quenching (ACQ) effect, which results in spectral
crosstalk, background fluorescence interference, and low detection
sensitivity [11-15]. These bottleneck greatly limit their application in
sophisticated biological systems [11-15]. Therefore, the development of
a new fluorescent probe with larger Stokes shift and AIE features that
can be used for accurate sensing and imaging of CN– is highly
anticipated.
In recent years, excited-state intramolecular proton transfer (ESIPT)
has been considered as an attractive fluorescence emission process for

* Corresponding authors at: School of Chemistry and Materials Engineering, Fuyang Normal University, Fuyang, Anhui Province 236037, PR China.
E-mail addresses: huanan200890@163.com (H. Huang), qiaorui@fynu.edu.cn (R. Qiao).
1
These authors contributes equally to this work.

https://doi.org/10.1016/j.cej.2022.136445
Received 14 December 2021; Received in revised form 20 February 2022; Accepted 14 April 2022
Available online 19 April 2022
1385-8947/© 2022 Elsevier B.V. All rights reserved.
C. Bai et al.
Scheme 1. Illustration of the probe design.
the design of fluorescent probes [16-18]. In the ESIPT process, the
intramolecular hydrogen bonds between the proton donor and acceptor
groups are close to each other [16-25]. Once molecules are excited by
light, the rapid proton transfer occurs, which tautomers is transformed
from the excited enol form (E*) to the excited ketone form (K*) [16-20].
The geometry of the excited ketone tautomer is quite different from the
enol form, and the Stokes shift generated by the absorption and emission
of the tautomers [16-20]. Therefore, the emission in the ESIPT system
exhibits the unusual large Stokes shift, which can effectively avoid self-
absorption and is very useful for accurate fluorescence detection and
analysis [16-20]. These advantages make ESIPT molecular as a prom­
ising candidate for fast and accurate sensing materials [19-25].
In addition, since the concept of aggregation-induced emission (AIE)
proposed by the Tang group in 2001 [26], the research on the properties
of AIE has attracted great attention. To date, many researchers are
involved in designing and discovering new AIE materials, which have
the excellent light stability, high signal-to-noise ratio, and investigating
their applicability in sensing systems [27-29]. Since AIE has shined in
many research fields, many researchers attempt to acquire more excel­
lent fluorescent materials with ESIPT and AIE because this may produce
“1 + 1 > 2” effect [30-37]. However, the biological materials with
ESIPT-AIE characteristics for CN– detection have not been fully inves­
tigated yet [38-42]. Therefore, regardless of fundamental research or
material application, it is fascinating to exploit the new ESIPT-AIE sys­
tems and probe their possible application in ion detection in living
organisms.
Herein, to understand and uncover the mechanism of CN– recogni­
tion with large Stokes shift at the molecule level, we designed and
synthesized a new probe HPBC base on the ESIPT and AIE strategy.
HPBC contained a hydroxyl group at the ortho position to form intra­
molecular hydrogen bonds. In addition, to enhance the conjugation and
electronic separation effects, the pyrene ring was introduced to the
Scheme 2. Synthetic route of HPBC.
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Chemical Engineering Journal 443 (2022) 136445
skeleton as the donor unit. On the other hand, cyano-substituted
biphenyl was introduced as the strong acceptor moiety. At the same
time, the introduction of double bonds can also play a role in restriction
of intramolecular rotation (Scheme 1). These main elements are crucial
issues in the design of current ESIPT and AIE materials. The systematic
analysis, spectral response and crystal structures indicate that HPBC
indeed possesses the ESIPT-AIE feature. More importantly, HPBC was
successfully applied to discriminate endogenous CN– in raw almond,
cassava root plant tissues, and to image CN– in C. elegans, mice and HeLa
cells with obvious “turn-on” fluorescence. To the best of our knowledge,
this is the first example that realized the detection of CN– with largest
Stokes shift (243 nm) and AIE in living cells. These meaningful findings
provide a new platform to construct ESIPT-AIE materials, which might
trigger the discovery of the new probes with amazing properties and
functions. It may provide a unique idea for the detection of CN–, and has
broad application prospects in food and biological detection.
2. Experimental
2.1. Materials and instruments
All reagents and solvents were purchased from commercial sources
and used without any further purification. 1H NMR and 13C NMR spectra
of the products were recorded on a Bruker 400 MHz NMR spectrometer
using tetramethylsilane (TMS) as the internal standard (chemical shifts
in ppm). FT-IR spectra were measured with a Horiba FT-720 infrared
spectrophotometer. High-resolution mass spectra (HR-MS) data were
obtained using a Thermo LTQ-Orbitrap mass spectrometer. UV–Vis
spectra were obtained using spectrophotometer (Shimadzu UV-2450).
Fluorescent spectra were acquired with fluorescence spectrophotom­
eter (Hitachi F-7000). X-ray crystallographic analysis was performed at
the X-ray crystallography facility, Shanghai Institute of Organic
C. Bai et al.
Fig. 1. (a) Fluorescence spectra of HPBC (1 × 10− 5 M) in various solvents, λex = 390 nm. (b) Normalized graph of HPBC in DMSO, λex = 390 nm. (c) Fluorescence
spectrum of HPBC (1 × 10− 5 M) in DMSO/H2O (fw: 0–95 %), λex = 390 nm. Inset: Images of HPBC in DMSO/H2O mixtures under the 365 nm UV lamp. (d)
Normalized intensity of HPBC (1 × 10− 5 M) in DMSO/H2O, at 460 nm, λex = 390 nm. (e) SEM images of HPBC obtained in CH3CN. (f) SEM images of HPBC obtained
in CH3CN/H2O (fw = 70%).
Chemistry (SIOC), Chinese Academy of Sciences (CAS). Fluorescence
imaging was performed using fluorescence microscope (Nikon Eclipse
Ti-S). Density functional theory (DFT) calculations with B3LYP were
carried out by using the Gaussian 09 package.
All the spectroscopy was carried out in the DMSO/H2O solution. CN–
was used as the tetrabutyl ammonium salt (TBACN). All measurements
were carried out at ambient temperature. Moreover, the relevant details
were described in the supporting information.
2.2. Synthesis and characterization
3′ -formyl-4′ -hydroxy-[1, 1′ -biphenyl]-4-carbonitril and pyren-1-
ylmethylene hydrazinee were prepared according to the previous re­
ports, respectively [43]. 3′ -formyl-4′ -hydroxy-[1, 1′ -biphenyl]-4-car­
bonitril (1 mmol, 0.2232 g), pyren-1-ylmethylene hydrazinee (1 mmol,
0.2243 g), and ethanol were added into the flask. After refluxed for 6 h,
the mixture was cooled to ambient temperature. And the deposition was
gained. The crude product was filtered, rinsed by ethanol, filtered with
suction, and the solid was dried to obtain the pure sensor HPBC (Scheme
2). HPBC were characterized by 1H and 13C NMR spectroscopy, high-
resolution mass spectrometry (HRMS) (Figs. S1-S3). The single crystals
of HPBC suitable for X-ray diffraction were obtained after slow evapo­
ration from ethanol for several days at room temperature. Crystallo­
graphic data (CCDC 2035376) has been received by the Cambridge
Crystallographic Data Centre.
2.3. Analysis of real samples
The operation process of endogenous cyanide detection, cytotoxicity
and fluorescence bioimaging were shown in the supporting information.
3. Results and discussion
3.1. The ESIPT-AIE study
To verify our assumptions, the ESIPT process of HPBC was initial
explored. The dual fluorescence emission of the enol-form type (high
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Chemical Engineering Journal 443 (2022) 136445
energy, small Stokes shift) and the keto-form type (low energy, large
Stokes shift) were typical characteristics of ESIPT fluorophores [19-25].
Because the ESIPT process was vulnerable to be affected by the solvents
[44], thus, the optical properties of HPBC in different solvents were
investigated (Figs. 1 and S4). It’s clear that HPBC has obvious dual
emission of enol and keto tautomer under the excitation at 390 nm
(Fig. 1a), which unambiguously confirmed the ESIPT phenomenon. In
low-polar solvents (toluene, dichloromethane), strong ketone emitted at
about 600 nm. On the contrary, the emission intensity at 600 nm
decreased with raising the polarity of solvents (Fig. 1a) since hydrogen
bond could be developed between the solvent and –OH of HPBC in high-
polar solvents, which can stabilize the enol form to inhibit the occur­
rence of the ESIPT process [44,45]. And it’s evident that Stokes shift
reached the maximum value (207 nm) in DMSO (Fig. 1b). Due to its
compatibility with water and low biological toxicity, DMSO is often used
as a co-solvent for biological reagents compared to other organic sol­
vents. So, DMSO was chosen as the suitable solvent in our experiments.
To prove the AIE characteristics, we continued to study the fluores­
cence properties of HPBC in DMSO/H2O mixtures with different H2O
fractions (fw). When fw did not 30 %, HPBC presented the relatively
weak fluorescence emission band at 460 nm, which assigned to the
emission characteristics of enol tautomers (Fig. 1c and 1d). While fw
increased to 70% by degree, the fluorescent intensity reached the
maximum at 610 nm, indicating the AIE effect was triggered [46-49]. In
addition, the solution containing HPBC was dropped onto the filter
paper. And when the solvent evaporated, its fluorescence was found to
become obvious, which also confirmed that HPBC has AIE properties
(Figs. S5 and video). All results were also supported by SEM analysis
(Fig. 1e and 1f). It indicated that the probe HPBC still has a good fluo­
rescence emission effect in the aqueous system with AIE effect. For this
reason, DMSO/H2O (fw = 70%) solution as the test system for subse­
quent experiments.
Determination of the underlying mechanisms for the ESIPT and AIE
phenomenon is of great importance to a fundamental understanding of
photophysics, which will help deepen our understanding of lumines­
cence processes and guide our endeavors to design novel ESIPT and AIE
systems. It is worth noting that the crystal configurations play an
C. Bai et al. Chemical Engineering Journal 443 (2022) 136445

Fig. 2. The crystal structures of HPBC: (a) showed the hydrogen bond, (b) viewing from the c axis, (c) crystal packing, (d) spacing data. (e) Frontier molecular
orbitals optimized and illustration of ESIPT process of HPBC. (f) Illustration of ESIPT process.

important role in bridging the natural properties of single molecules and
the macroscopic optoelectronic performance of organic materials, which
provides significant information for understanding the photophysical
processes of different kinds of luminescence behaviors. Therefore, to
gain insight into the cause of the ESIPT and AIE behaviors for HPBC, the
corresponding crystal structures were investigated in detail. As ex­
pected, HPBC revealed a highly planar conformation, the dihedral an­
gles between the –OH and –CH– – N– planes was almost the same plane,
which is very beneficial to form a strong intramolecular hydrogen bond.
Obviously, the strong intramolecular hydrogen bond distance was
1.916 Å between –OH and the adjacent N atom (Fig. 2a). The existing
intramolecular hydrogen bond and coplanar main functional groups
collectively promoted intramolecular proton transfer and the structural
transformation of HBPC under light induction to induce the occurrence
of ESIPT [44]. The intense intramolecular H-bonds not only ensure the
ESIPT behavior, but also help rigidize the molecular conformation,
which largely reduces the energy loss by suppressing the rotations of
-C–
– N and the biphenyl moieties. It is very conducive to the generation

Fig. 3. (a) The fluorescence spectrum of HPBC (6.0 × 10− 6 M) with the presence of different anions (3.0 × 10− 5 M) in DMSO/H2O (fw = 70%) solutions, λex = 390
nm. Inset: fluorescence changes of HPBC with addition of different anions (1–16: F-, Cl-, Br-, I-, SO2- 2- 2- - – – 3- – 2– -
4 , SO3 , S , AcO , CN , NO3, NO2, PO4 , HCO3, CO3 , H2PO4, and
–

blank) under 365 nm UV light. (b) Normalized absorption and emission spectra of HPBC + CN in DMSO/H2O (fw = 70%) solutions, λex = 390 nm. (c) Fluorescence
–

responses of HPBC + CN– at 623 nm in the present of various anions (1–15: F-, Cl-, Br-, I-, SO2- 2- 2- - – 3- – 2– -
4 , SO3 , S , AcO , NO3, NO2, PO4 , HCO3, CO3 , H2PO4, CN ). (d) The time
– –

course of spectra of HPBC before and after added with CN– in DMSO/H2O (fw = 70%) solutions, λex = 390 nm. Images of HPBC (e) and HPBC + CN– (f) in solid state
under 365 nm UV light.

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of AIE effect. In addition, the single crystal analyses reveal that HPBC is
constructed based on a monoclinic crystal system, crystallizing in the
space group P 1 21/c 1 with four molecules in one cell unit (Fig. 2b).
These four molecules present a unique spatial arrangement. For
instance, viewing from the c axis, the unique spatial arrangement
favored to construct an ordered antiparallel intersecting stacking modes
(Fig. 2b). It is worth mentioning that the antiparallel intersecting
stacking modes was enough to impede π-π stacking, which was another
favorable factor to generate the AIE behavior (Fig. 2c). More impor­
tantly, benefitting from the spatial arrangement, the adjacent three
molecules tend to associate with abundant intermolecular interactions.
As revealed in Fig. 2d, the OH⋅⋅⋅N, C–H (CH)⋅⋅⋅π, CH⋅⋅⋅N CH⋅⋅⋅CH in­ Fig. 4. The proposed sensing mechanism.
teractions have distances from 1.916 Å to 3.594 Å. These intense
intermolecular interactions help rigidize the molecular conformation
previously reported probes (Table S1). It’s concluded that HPBC could
and block the nonradiative pathway by restricting molecular motions.
discriminate CN– from other anions in aqueous solution with the large
Based on these data, we concluded that the increasingly strong in­
Stokes shift, high selectivity, ultra-sensitivity, and the low LOD.
teractions and spatial arrangement make HPBC pack in a more rigid
To grasp the interaction between HPBC and CN– intensively, 1H NMR
molecular conformation that leaded to the occurrence of AIE. Such
titrations, IR (Infrared Spectroscopy) and HR-MS were carried out at
studies not only expand the variety of ESIPT and AIEgens but also pro­
room temperature. According to the mole ratio method, the ratio was
vide a deep understanding of the structure-packing-performance rela­
1:1 between HPBC and CN– (Fig. S9). From the 1H NMR titration
tionship, guiding the design of organic molecules with AIE and ESIPT
(Fig. S10), the signal at 11.69 ppm, which represented the –OH (Ha,
properties.
Fig. 4) bond of HPBC disappeared in 1H NMR while CN– added gradu­
Besides, to investigate the distribution of electrons, the calculations
ally. Simultaneously, a new peak appeared at 3.63 ppm, which was
were carried out by density functional theory (DFT). The molecular
attributable to the proton of the –NH– (Hb, Fig. 4) group. And the result
orbital analysis of HPBC showed that HOMO and LUMO of the stable
was also approved by IR. After CN– were added, the new peaks of –CN
enol form delocalized mainly in the pyrene ring (Fig. 2e). And the
appears at 2156 cm− 1 and 2181 cm− 1 (Fig. S11). The peak appeared at
calculated HOMO and LUMO values were − 5.2675 eV and − 2.3429 eV.
1585 cm− 1, which was attributed to the newly formed carbonyl group.
However, HOMO of the ketone form was delocalized in the entire
The peak at 2958 cm− 1 belong to the methyl peak in the tetrabuty­
molecule, where HOMO and LUMO also became − 4.7479 eV and
lammonium cyanide (TBACN) used in the experiment. It’s pleased that
− 2.4973 eV. The calculation results and the change in distribution both
the mass data matched with the 1H NMR. From the ESI mass data, it
proved that the ESIPT occurred in HPBC (Fig. 2f) [45].
revealed the peak at 493.1641, which matched that of [HPBC +
CN–+H2O] + (calculated for C32H21N4O2, 493.1670) (Fig. S12). There­
3.2. The recognition properties of HPBC towards CN– fore, combined with the above data, we inferred the possible recognition
process (Fig. 4).
All the above experimental results confirm and explain that HPBC
has the characteristics of ESIPT and AIE, which is consistent with our
original design philosophy. In order to further study the properties of 3.3. Bioimaging of HPBC in HeLa cells
HPBC, the effects of different anionic recognition and detection were
tested. The change of UV absorption and fluorescence emission spectra Firstly, we determined the cytotoxicity and bioimaging of HPBC and
caused by various ions were researched in DMSO/H2O (fw = 70%) so­ HPBC + CN– in HeLa cells (Fig. S13). Even HPBC and HPBC + CN–
lution separately. It was found that the absorption and emission of the concentration was as high as 400 μM, the cell viability of HeLa cells still
HPBC showed manifest changes only in the presence of CN– (Fig. 3a and maintained the high level, which indicated that HPBC had good
S6a). Upon addition of CN– to the HPBC solution, the absorption in­ biocompatibility. After HeLa cells incubated with HPBC, there was no
tensity decreased. And it also exhibited dual emitted peaks at 420 nm fluorescence (Fig. 5). However, HPBC-stained HeLa cells exhibited vivid
and 623 nm (Fig. 3a), which indicated CN– might promote the progress fluorescence signals in living cells after being cultured with CN–. And the
of ESIPT when HPBC interacted with CN–. As the result, the maximum fluorescence intensity of the dual channels also increased with the CN–
Stokes shift reached 243 nm (Fig. 3b). From the anti-interference concentration increase. The results indicated that HPBC and HPBC þ
experiment (Fig. 3c, S6b and S6c), the results showed that other ions CN– not only had the potential to be internalized by cells, but also
didn’t have any effect on the interaction between HPBC and CN–, which showed clear self-indicating behavior, which could reveal their location
indicated that HPBC recognized CN– selectively. through the AIE effect. Moreover, the dual-channel fluorescence images
To evaluate the sensitivity of HPBC toward CN–, the fluorescence were obtained based on ESIPT. It’s meaningful that the large Stokes shift
intensity of HPBC at 623 nm changing with time were recorded in the greatly reduced the background fluorescence interference. And high-
absence and presence of CN–. According to the results (Fig. 3d), the quality biological imaging images were received at the same time. The
emission intensity of HPBC in solution was not change obviously with above data illustrated that HPBC easily penetrated cells and manifested
time. However, the emission intensity increased immediately after the clear changes in fluorescence signal via the ESIPT-AIE effect, which were
CN– added into the solution system. And the fluorescence intensity suitable to image CN– in living cells. It’s conducive to use HPBC as the
remained unchanged after 8 min. To our surprises, the fluorescence also advanced material to identify CN– and their influence on cells.
changed significantly in the solid state when HPBC encountered CN–
(Fig. 3e and 3f). Then, we recorded the fluorescence spectra of HPBC + 3.4. Endogenous cyanide detection
CN– change in DMSO/H2O mixed solutions with different fw (Fig. S7).
While fw improved gradually, the emission intensity enhanced. It hinted Then, in order to verify the practical application of HPBC, the
that AIE occurred once H2O added [46-49]. All facts showed that HPBC recognition of endogenous CN– was fulfilled. As we all know, raw bitter
could be applied in real-time detection of CN– potentially on the basis of almonds and cassava contained a lot of CN– [50]. After raw almonds and
ESIPT-AIE. Moreover, the limit of detection (LOD) was calculated to be cassava were soaked in solutions for 2 h with or without HPBC, the cell
1.32 × 10-7 M (Fig. S8) [49,50], which was lower than the 1.9 × 10-6 M imaging were collected. As the results, normal raw almond cells did not
specified by the World Health Organization [51] and better than other produce fluorescence in the green and red channels (Fig. 6). However,

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Fig. 5. HeLa cells incubated with HPBC and HPBC + CN–, respectively. Left, middle, and right column were bright field images, blue, red, and merge channel (blue
and red field) from fluorescence images, respectively.

Fig. 6. (a) Image of raw almonds, cassava and edible almonds. (b-d) Fluorescence images of raw almonds cassava and edible almonds cells. Left and right column
were bright field images, green and red channel from in vitro fluorescence images, respectively.

the cells with HPBC produced obvious fluorescence. And the same re­
sults were obtained on cassava. To consolidate this result, the cooked
almonds were observed under the same conditions. It’s determined that
there was no fluorescence in cell imaging. Because raw almond and
cassava slice cells contain high levels of CN–, where the content of CN– in
cooked almonds was decreased. So, HPBC could distinguish endogenous
CN– availably.
3.5. C. Elegans and mice imaging
At last, to examine its detection effect in vivo, animal cell imaging
was performed using C. elegans and mice. Due to the ESIPT-AIE
characteristics of HPBC, the dual-channel mode with green channel
and red channel was used in the bioimaging. From Fig. 7a and 7b, the
elegans cultivated with HPBC displayed very dim fluorescence. How­
ever, the fluorescence was enhanced and covered the entire elegans after
the elegans were exposed to CN– (Fig. 7c). It demonstrated that HPBC
could be applied to recognize CN– in vivo. Then, HPBC was squirted into
the two mice in the muscles. And CN– was injected into one of the mice
at the same location by the same way. After 6 h, the liver, kidney and
heart cells of the mice were subjected to fluorescence imaging obser­
vation (Fig. 8). When HPBC interacted with CN–, the tissue cells emitted
significant fluorescence by dual-channel because of the ESIPT-AIE ef­
fect. Moreover, the fluorescence observed in the liver and kidney tissue

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C. Bai et al.
Fig. 7. In vivo fluorescence imaging of Caenorhabditis elegans:(a) images of
live C. elegans, (b) with HPBC (1.0 × 10− 4 M), (c) HPBC (1.0 × 10− 4 M) upon
fluorogenic response with CN– (1.0 × 10− 4 M). Left and right column were
bright field images, green and red channel from fluorescence images,
respectively.
cells of mice was brighter than that in the heart cells because the liver
and kidney were both detoxification organs, where the HPBC and HPBC
+ CN– concentration were higher.
Fig. 8. The fluorescence image of liver, kidney, and heart cells of mice after thigh intramuscular injection of HPBC (1.0 × 10− 4 M) and HPBC (1.0 × 10−
(1.0 × 10− 4 M) 6 h later, respectively. Left and middle were bright field images, green and red channel from fluorescence images, respectively.
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Chemical Engineering Journal 443 (2022) 136445
4. Conclusions
In conclusion, we firstly designed and synthesized a probe based on
the ESIPT-AIE mechanism, which could recognize CN– effectively with
large Stokes shift, high selectivity, and rapid off–on response and lower
LOD. The mechanism was verified by some methods. More importantly,
it was established that HPBC could detect endogenous CN– effectively
through fluorescence imaging in plants. And it also revealed that HPBC
could be suitable for the effective fluorescent sensor for detecting CN– in
living systems. This work might contribute to research about the
detection of CN– based on ESIPT-AIE and it might provide the new di­
rection to get some fluorescent probes with large Stokes shift. In addi­
tion, it also confirmed that the properties of hydrogen bonding,
electronic effects and restriction of intramolecular rotation are com­
bined to realize the joint function of ESIPT and AIE characteristics by
designing and synthesizing specific structures. This strategy has a good
enlightenment for the design of multifunctional probes and it also pro­
vides ideas for obtaining advanced luminescent materials with multiple
properties for detecting specific substances in the environment, food and
organisms.
CRediT authorship contribution statement
Cuibing Bai: Writing – review & editing. Jie Zhang: Writing – re­
view & editing. Yuxin Qin: . Huanan Huang: Supervision, Writing –
review & editing. Zhenni Xia: . Qijun Zheng: . Hongliang Dai: .
Pengkai Lu: . Hui Miao: Supervision. Changqing Qu: Supervision. Rui
Qiao: Investigation, Supervision, Writing – review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
4
M) + CN–
C. Bai et al.
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgments
Many thanks to the financial assistance from the National Natural
Science Foundation of China (21302019 and 21861022), the Key Project
of Natural Science Foundation of Jiangxi Province
(20202ACBL213003), and the Open Project Grant of Key Laboratory of
Photochemical Conversion and Optoelectronic Materials
(PCOM202002), TIPC, Chinese Academy of Sciences. This work was also
supported by Anhui Provincial Natural Science Foundation
(1908085QB78), Undergraduate Training Programs for Innovation and
Entrepreneurship (S202010371050 and 202010371029) and the
Cooperation Project of Fuyang Municipal Government and Fuyang
Normal University (SXHZ202002).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.cej.2022.136445.
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