A highly selective fluorescence turn-on detection of hydrogen peroxide and d-glucose based on the aggregation/deaggregation of a modified tetraphenylethylene

Article abstract of DOI:10.1016/j.tetlet.2014.01.056

A new selective fluorescence turn-on detection of hydrogen peroxide was established by taking advantage of the aggregation induced-emission (AIE) behavior of tetraphenylethylene unit and the reaction of hydrogen peroxide toward the arylboronic ester group in compound 1. Moreover, compound 1 was successfully utilized for the selective detection of d-glucose in aqueous solution.

Full text of DOI:10.1016/j.tetlet.2014.01.056

Tetrahedron Letters 55 (2014) 1471–1474
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Tetrahedron Letters
journal homepage: www.elsevier.com/locate/tetlet
A highly selective fluorescence turn-on detection of hydrogen
peroxide and D-glucose based on the aggregation/deaggregation
of a modified tetraphenylethylene
⇑
⇑
Fang Hu, Yanyan Huang, Guanxin Zhang , Rui Zhao, Deqing Zhang
Beijing National Laboratory for Molecular Sciences, Key Laboratories of Organic Solids and Analytical Chemistry for Living Systems, Chinese Academy of Sciences, Beijing 100190, China
a r t i c l e i n f o
a b s t r a c t
Article history:
A new selective fluorescence turn-on detection of hydrogen peroxide was established by taking advan-
tage of the aggregation induced-emission (AIE) behavior of tetraphenylethylene unit and the reaction
of hydrogen peroxide toward the arylboronic ester group in compound 1. Moreover, compound 1 was
Received 22 November 2013
Revised 31 December 2013
Accepted 14 January 2014
Available online 18 January 2014
successfully utilized for the selective detection of
D-glucose in aqueous solution.
Ó 2014 Elsevier Ltd. All rights reserved.
Keywords:
Aggregation induced-emission
Fluorescent probes
Hydrogen peroxide
D-Glucose
Tetraphenylethylene
Reactive oxygen species (ROS) as a class of radical or nonradical
oxygen-containing species include hydrogen peroxide (H2O2),
singlet oxygen (1O2), hydroxy radicals (ÅOH), superoxide anions
ðOꢀ2ꢁÞ, and nitric oxide.1 In general, ROS are toxic to cells if the levels
of ROS exceed the tolerable physiological range.2 Among ROS spe-
cies, hydrogen peroxide is the least reactive and mild oxidant.
However, it can be transformed into strong oxidants. For example,
H2O2 is readily converted to the highly reactive hydroxyl radical
via the Fenton reaction.3 Despite its hazardness to organisms,
hydrogen peroxide is ubiquitous as it is a by-product of many
metabolic reactions.1d Also, hydrogen peroxide is widely used for
environmental and industrial applications such as drinking water
purification.4 Therefore, it is highly desirable to develop efficient
and simple sensing systems to monitor H2O2 concentration level
in not only living cells but also environment.
selectivity and sensitivity of these probes are excellent, most
syntheses are usually time-consuming and laborious.
Herein we report a new fluorescence turn-on detection of H2O2
in aqueous solution with a new tetraphenylethylene (TPE) mole-
cule 1 (Scheme 1) with N-4-(benzyl boronic pinacol ester) pyridini-
um bromide moiety. The sensing mechanism of 1 toward H2O2 in
aqueous solution is illustrated in Scheme 1 and is explained as
follows: (i) the pyridinium moiety may render 1 water-soluble.
As a result it is anticipated that 1 is weakly emissive in aqueous
Up to now, a large number of sensors for H2O2 have been
invented based on either electrochemical or optical methods.5–7
Among them, fluorometry has caught more attention due to its
higher sensitivity and can be applied in cells and tissue. For
instance, pentafluorobenzenesulfonyl fluoresceins were utilized
for
sensing
H2O2
based
on
the
fact
that
the
pentafluorobenzenesulfonyl group can be cleaved off by H2O2 via
perhydrolysis and thus the fluorescence is enhanced.6a While the
⇑
Corresponding authors. Tel.: +86 10 62639355; fax: +86 10 62559373 (D.Z.).
E-mail address: dqzhang@iccas.ac.cn (D. Zhang).
Scheme 1. The chemical structure of 1 and 2, and the design rationale for the
fluorescence turn-on detection of H2O2.
0040-4039/$ - see front matter Ó 2014 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.tetlet.2014.01.056

1472
F. Hu et al. / Tetrahedron Letters 55 (2014) 1471–1474
solutions according to previous studies;8–10 (ii) it is well known
that H2O2 can stoichiometrically convert phenylboronic ester
moiety into the phenol group.5c,11 Thus, incubation of hydrogen
peroxide with 1 should result in oxidation of the phenylboronic
pinacolester, followed by hydrolysis and 1,6-elimination of p-qui-
none-methide to yield the p-pyridine substituted TPE (2,
Scheme 1). Compound 2 is expected to show low solubility in
aqueous solutions, thus aggregation will occur and turn on the
fluorescence of TPE moiety based on the aggregation-induced
emission (AIE) feature of TPE compounds.8–10 In this way, com-
pound 1 can be employed for the fluorescence turn-on detection
of H2O2 in aqueous solutions. The results reveal that 1 exhibits high
selectivity toward H2O2. Moreover, by combining the cascade
enzymatic (oxidation of D
-glucose with GOx to produce H2O2)12
and chemical (oxidation of phenylboronic ester by H2O2) reactions,
compound 1 is successfully utilized for the selective detection of
D-glucose in aqueous solution.
The synthesis of started from monobromo-substituted
1
Figure 1. Fluorescence spectra of 1 (5.0 lM) after incubation with different
amounts of H2O2; the reaction was performed at 37 °C for 60 min in 10 mM HEPES
buffer at pH 7.4; the excitation wavelength was 390 nm; inset shows photos of the
tetraphenylethylene derivative (3) as shown in Scheme 2. Palla-
dium-catalyzed Suzuki coupling reactions between compound 3
and 4-pyridinylboronic acid yielded compound 2, which was
allowed to react with 4-(bromomethyl) benzene boronic pinacol
ester to afford 1 in 95.6% yield. The chemical structure of 1 was
characterized with 1H NMR, 13C NMR, and mass spectra (detailed
procedures are given in Supplementary data).13
solution of 1 (5.0
lM) before (A) and after (B) incubation with 150.0 lM of H2O2
under UV light (365 nm).
As expected, compound 1 is soluble in water, and the HEPES
buffer solution (10 mM, pH = 7.4) of 1 (5.0 lM) can be easily pre-
pared. Figure 1 shows the fluorescence spectra of 1 before and after
addition of different amounts of H2O2. As anticipated, the aqueous
solution of 1 (5.0 lM) was almost non-emissive in the absence of
H2O2 (see Fig. 1, curve a). However, after incubation with H2O2 at
37 °C the fluorescence intensity of 1 increased gradually concomi-
tant with the emission peak progressively blue-shifting from 610
to 510 nm.14 For instance, the fluorescence intensity at 510 nm
(see Fig. 1, curve b) increased by more than 260 times after the
solution of 1 was incubated with 150.0
fact, the fluorescence quantum yield of the HEPES buffer solution
(10 mM, pH = 7.4) of 1 (5.0 M) increased from 0.006 to 0.121
(by reference to quinine hemisulfate monohydrate) after H2O2
(150.0 M) was introduced. Such fluorescence enhancement can
lM of H2O2 for 30 min. In
l
l
be distinguished with naked-eye as shown in the inset of Figure 1,
where photos of solutions of 1 in the absence and presence of H2O2
under UV light (365 nm) illumination are displayed. Figure 2 de-
picts the plot of the relative fluorescence intensity (I/I0 ꢁ 1) at
510 nm versus concentration of H2O2. Interestingly, the fluores-
cence intensity of 1 increases almost linearly with the concentra-
Figure 2. The plot of (I/I0 ꢁ 1) at 510 nm versus the concentration of hydrogen
peroxide; inset shows the linear relation for concentration of hydrogen peroxide in
the range of 10.0–110.0 lM.
inset of Figure 2. Accordingly, the detection limit of H2O2 was
estimated to be 180.0 nM (n = 11 and S/N = 3).
tion of H2O2 in the range of 10.0–110.0 lM as displayed in the
Moreover, fluorescence responses of 1 to other reactive oxygen
species were examined. As depicted in Figure 3, significant fluores-
cence enhancement was observed only after incubation with H2O2.
Other ROS, such as singlet oxygen, hydroxy radical, superoxide
anion, and nitric oxide etc., induced only negligible fluorescence
enhancement for 1 under the same conditions. Thus, compound
1 shows high selectivity toward H2O2.
As illustrated in Scheme 1, such fluorescence enhancement is
attributed to the aggregation of compound 2 which is generated
from the oxidation reaction of 1 with H2O2. The formation of fluo-
rescent aggregates was confirmed by both confocal laser scanning
microscopic (CLSM) and dynamic light scattering (DLS) studies. As
displayed in Figure 4, there were almost no fluorescent aggregates
for the solution of 1 (10.0
ever, fluorescent aggregates emerged after incubation with H2O2
(200.0 M) based on the CLSM images. DLS data (see Fig. 5) also
indicated the formation of aggregates of 200–600 nm for the
solution of 1 (5.0 M) after incubation with H2O2 (200.0 M).
lM) before incubation with H2O2; how-
l
l
l
Scheme 2. Synthetic approach to compound 1.

F. Hu et al. / Tetrahedron Letters 55 (2014) 1471–1474
1473
Figure 3. Variation of the relative fluorescence intensity at 510 nm of 1 (5.0
lM)
after incubation with 100.0 M of hydrogen peroxide and 200 M of other reactive
l
l
oxygen species; the reaction was performed at 37 °C for 60 min in HEPES buffer
(10 mM, pH 7.4); the excitation wavelength was 390 nm.
Figure 5. The DLS data of 1 (5.0
(200.0
10 mM HEPES buffer at pH 7.4 before the collection of DLS data.
l
M) before and after incubation with H2O2
l
M). The reaction between 1 and H2O2 was performed at 37 °C for 30 min in
Figure 6. Fluorescence spectra of 1 (5.0
lM) after incubation with GOx (1.1 U/mL)
and different concentrations of -glucose; the reaction was performed at 37 °C for
90 min in 10 mM HEPES buffer at pH 7.4; the excitation wavelength was 390 nm.
D
Figure 4. Fluorescence confocal laser scanning images of 1 (10.0
after (b) incubation with H2O2 (200.0 M); the reaction was performed at 37 °C for
60 min in 10 mM HEPES buffer at pH 7.4. The scale bar represents 20 m.
lM) before (a) and
l
l
ensemble solution. The fluorescence intensity ratio (I/I0 ꢁ 1) at
510 nm as the function of concentration of D-glucose is shown in
The formation of 2 after incubation of 1 with H2O2 (see
Scheme 1) is confirmed by the mass spectral (Fig. S2) and HPLC
analysis (Fig. S3). The mass signal at m/z = 470.3, corresponding
to the molecular weight of 2+H+, was detected after the solution
of 1 was incubated with H2O2. Moreover, HPLC analysis also indi-
cated the formation of 2 by comparing with the authentic sample
of 2 (see Fig. S3).
Figure S6, where I and I0 represent the fluorescence intensities of
the ensemble solutions after and before incubation with -glucose
D
and GOx, respectively. In the concentration range of 0.05–0.25 mM,
the fluorescence intensity ratio (I/I0 ꢁ 1) increases almost linearly
with the concentration of D-glucose (see inset of Fig. S6). D-Glucose
with concentration as low as 3.0
fluorescent assay.
lM can be detected with this new
In the following, we demonstrate the application of 1 for
the selective detection of
-glucose can be oxidized in the presence of GOx (glucose oxidase)
D
-glucose. As reported previously,12
In order to demonstrate the selectivity of this fluorescence
detection of -glucose, the fluorescence spectra of compound
1 were also recorded after separate incubation with -mannose,
-fructose, and -galactose in the same way as for -glucose. As
depicted in Figure 7, a remarkable fluorescence enhancement
was only observed for 1 after incubation with GOx and -glucose.
This is understandable by considering the fact that GOx can selec-
tively oxidize -glucose to generate H2O2.
D
D
D
to generate H2O2 which can further convert 1 into 2 (see Scheme 1).
Therefore, it is anticipated the compound 1 can be utilized for
D
D
D
the detection of
enzymatic and chemical reactions. The ensemble HEPES buffer
(10 mM, pH = 7.4) solutions containing 1 (5.0 M), GOx (1.1 U/mL),
and different amounts of -glucose (0–0.5 mM) were incubated
D-glucose by taking advantage of the cascade
D
l
D
D
In summary, we have successfully established a selective fluo-
rescence turn-on detection of H2O2 by making use of the abnormal
fluorescent behavior of TPE compounds and the oxidation of the
phenylboronic pinacol ester group by H2O2. This fluorometric
turn-on detection of H2O2 possesses the following features: (1) 1
at 37 °C for 90 min. Then, the fluorescent spectrum of each solution
was measured as shown in Figure 6. Obviously, the fluorescence
intensity of the ensemble increased gradually after the addition
of
D
-glucose; moreover, more fluorescence enhancement was
-glucose in the
observed by increasing the concentration of
D

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F. Hu et al. / Tetrahedron Letters 55 (2014) 1471–1474
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Figure 7. Variation of the relative fluorescence intensity at 510 nm of 1 (5.0
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is easily synthesized; (2) the determination procedure can be car-
ried out in aqueous solutions; (3) H2O2 with concentration as low
as 180 nM can be detected and interferences from other ROS are
negligible. Moreover, the ensemble of compound 1 and GOx was
successfully employed for the detection of D-glucose with good
sensitivity and selectivity by utilizing the cascade enzymatic and
chemical reaction.
Acknowledgments
The present research was financially supported by the Chinese
Academy of Sciences, NSFC and State Key Basic Research Program.
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Supplementary data
13. Characterization data for compound 1: 1H NMR (400 MHz, DMSO-d6): d 9.15 (d,
J = 4.0 Hz, 2H), 8.48 (d, J = 4.0 Hz, 2H), 7.91 (d, J = 8.0 Hz, 2H), 7.73 (d, J = 8.0 Hz,
2H), 7.52 (d, J = 8.0 Hz, 2H), 7.26–7.11 (m, 5H), 7.00 (d, J = 4.0 Hz, 2H), 6.93 (d,
J = 8.0 Hz, 2H), 6.88 (d, J = 8.0 Hz, 2H), 6.74–6.69 (t, J = 8.0 Hz, 4H), 5.85 (s, 2H),
3.68 (s, 6H), 1.28 (s, 12H); 13C NMR (100 MHz, DMSO-d6): d 158.1, 158.0, 154.3,
148.4, 144.7, 143.2, 141.7, 137.6, 137.5, 135.2, 135.1, 132.2, 132.1, 132.0, 130.8,
128.1, 128.0, 127.7, 126.6, 124.3, 113.4, 113.2, 83.9, 62.1, 24.6; HR-MS (ESI,
positive): calcd for C46H45BNO4 [MꢁBrꢁ]+: 686.3454; found: 686.3441.
14. Our previous studies indicated that the absorption and fluorescence of
tetraphenylethylene derivatives could be tuned by intramolecular electron-
donor (D) and acceptor (A) interactions; strong intramolecular D–A interaction
could yield red-shifts for both absorption and fluorescence spectra (see, Gu, X.;
Yao, J.; Zhang, G.; Zhang, C.; Yan, Y.; Zhao, Y.; Zhang, D. Chem. Asian J. 2013, 8,
2362). Compared with pyridium unit, pyridine unit possesses weaker electron-
accepting ability, leading to weak intramolecular D–A interaction within 2 and
the fluorescence blue-shift after pyridium moiety in 1 is transformed into
pyridine in 2.
Supplementary data (synthesis and characterization; UV spec-
tra, DLS data and relevant data) associated with this article can
be found, in the online version, at http://dx.doi.org/10.1016/
j.tetlet.2014.01.056.
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