Bioluminescent Turn-On Probe for Sensing Hypochlorite in Vitro and in Tumors

Article abstract of DOI:10.1021/acs.analchem.7b01103

Hypochlorite (ClO^-) is one of the most important reactive oxygen species but using a BL probe for its selective detection (or imaging) still remains challenging. Herein, we report a latent BL probe benzoylhydrazine luciferin (1) for highly selective detection of ClO^- in vitro and imaging ClO^- in living cells and tumors. In vitro tests indicated that 1 could be applied for highly selective detection of ClO^- within the range of 0-62.5 μM with a limit of detection of 0.705 μM. Using these unique features of 1, we successfully applied it to image ClO^- in living cells and tumors. We envision that probe 1 might be applied to elucidate the biological roles of ClO^- in wider physiological and pathological processes in the near future.

Full text of DOI:10.1021/acs.analchem.7b01103

Subscriber access provided by CORNELL UNIVERSITY LIBRARY
Letter
A Bioluminescent Turn-On Probe for
Sensing Hypochlorite in Vitro and in Tumors
Peiyao Chen, Zhen Zheng, Yunxia Zhu, Yu Dong, Fuqiang Wang, and Gaolin Liang
Anal. Chem., Just Accepted Manuscript • Publication Date (Web): 09 May 2017
Downloaded from http://pubs.acs.org on May 9, 2017
Just Accepted
“Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted
online prior to technical editing, formatting for publication and author proofing. The American Chemical
Society provides “Just Accepted” as a free service to the research community to expedite the
dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts
appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been
fully peer reviewed, but should not be considered the official version of record. They are accessible to all
readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered
to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published
in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just
Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor
changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers
and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors
or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
Analytical Chemistry is published by the American Chemical Society. 1155 Sixteenth
Street N.W., Washington, DC 20036
Published by American Chemical Society. Copyright © American Chemical Society.
However, no copyright claim is made to original U.S. Government works, or works
produced by employees of any Commonwealth realm Crown government in the
course of their duties.

Page 1 of 5
Analytical Chemistry
1
2
3
4
5
6
7
8
9
A Bioluminescent Turn-On Probe for Sensing Hypochlorite in
Vitro and in Tumors
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
†
†
‡
†
,
†
,‡
,
†
Peiyao Chen, Zhen Zheng, Yunxia Zhu, Yu Dong, Fuqiang Wang,* and Gaolin Liang*
†
CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China,
Hefei, Anhui 230026, China.
‡
Analysis Center, Nanjing Medical University, Nanjing, Jiangsu 210093, China.
-
ABSTRACT: Hypochlorite (ClO ) is one of the most important reactive oxygen species but using a BL probe for its selective
detection (or imaging) still remains challenging. Herein, we report a latent BL probe benzoylhydrazine luciferin (1) for highly
-
-
selective detection of ClO in vitro and imaging ClO in living cells and tumors. In vitro tests indicated that 1 could be applied for
-
highly selective detection of ClO within the range of 0-62.5 μM with a limit of detection of 0.705 μM. Using these unique features
-
of 1, we successfully applied it to image ClO in living cells and tumors. We envision that probe 1 might be applied to elucidate the
-
biological roles of ClO in wider physiological and pathological processes in the near future.
-
Hypochlorite (ClO ) is one of the most important reactive
Herein, we report a BL turn-on probe for in vitro selective
- -
1-3
oxygen species (ROS),
sustaining human innate immunity during microbial invasion.
Endogenous ClO is generated by hydrogen peroxide (H
reacting with chloride ions (Cl ), catalyzed by heme enzyme
which plays an important role in
detection of ClO and in vivo imaging of ClO in tumors. As we
4
-
know, ClO is able to oxidize dibenzoylhydrazine into water-
-
38
2
2
O )
unstable dibenzoyl diimide. Thus, as outlined in Scheme 1, we
-
rationally designed a latent BL probe benzoylhydrazine luciferin
5,6
myeloperoxidase (MPO). Abnormal levels of hypochlorite are
reported to associate with certain diseases such as cardiovascular
diseases, neuron degeneration, lung injury, atherosclerosis,
(1) with the carboxyl acid group of whose -luciferin motif being
D
caged by benzoylhydrazine. Probe 1 itself is not the substrate for
-
fLuc. However, upon ClO oxidation and follow-up hydrolysis in
7-12
arthritis, and cancer. Thus, it is of great importance to detect or
physiological condition, probe 1 yields the active substrate -
luciferin for luciferase to generate a light readout (Scheme 1).
D
-
image ClO in cells or in vivo. To date, numerous fluorescence
probes have been reported for the detection (or imaging) of ClO
-
1
3-17
-
.
However, most of the fluorescent probes suffer from low
Scheme 1. Schematic illustration of ClO oxidation of 1 to
photostability of photobleaching or autofluorescence interference
from the detecting samples.
yield -luciferin for bioluminescence generation.
D
Bioluminescent imaging (BLI) is a reliable, sensitive, non-
invasive imaging technique which has been widely used in
imaging a myriad of life processes such as cell proliferation and
18-24
migration, gene expression, and enzyme activities.
Among the
approximate 30 known bioluminescence (BL) systems, the firefly
25
luciferase-luciferin system is the most widely used one. In this
BL system, in the presence of adenosine triphosphate (ATP),
2
+
oxygen and magnesium ions (Mg ), firefly luciferase (fLuc)
catalyzes a two-step oxidation of luciferin to emit a visible
We began the study with the synthesis and characterization of
probe 1 (Scheme S1, Figure S1-S3). Then we used -luciferin to
26-29
(yellow to green) photon.
Notably, the generated
D
bioluminescent photons can even penetrate the tissues of intact
screen out an appropriate fLuc concentration to study the response
30,31
rodents, rendering BLI very suitable for in vivo imaging.
-
of probe 1 toward ClO . As shown in Figure S4, the BL intensity
Moreover, compared with fluorescence imaging, BLI does not
require external excitation and therefore possesses higher signal-
to-noise ratios. To date, a variety of BL probes have been
of -luciferin at fLuc concentration of 0.1 mg/mL was 5.3 folds of
D
that at fLuc concentration of 0.05 mg/mL. We thus choose 0.1
mg/mL fLuc to conduct the following in vitro experiment. As
developed2-36for the detection (or monitoring) of important
-
expected, in the absence of ClO , almost no BL signal was
3
analytes.
In 2015, Kojima et al. reported a sensitive
generated by 1 mM 1 after being incubated with 0.1 mg/mL fLuc
bioluminogenic probe for in vivo imaging the activity of highly
and 1 mM ATP in phosphate buffer (0.2 M, pH 7.4) (Figure 1A).
- 37
reactive oxygen species (hROS) including ClO . Nevertheless,
-
However, addition of 70-fold ClO to 1 mM 1 solution resulted in
-
this probe could not differentiate ClO from other hROS. Thus, it
a significant increase of BL intensity (a 58-fold increase at 560
nm, Figure 1A) under the same condition. This result suggested
still remains chanlleging to develop a bioluminescence probe for
-
the selective detection of ClO .
that free
D
-luciferin, which responds for the BL generation, was
-
yielded from ClO oxidation of 1, as we proposed in Scheme 1.
ACS Paragon Plus Environment

Analytical Chemistry
Page 2 of 5
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
-
Figure 1. (A) Bioluminescent wavelength scan for 1 mM 1 and 0.1 mg/mL fLuc in the presence (red) or absence (black) of 70 mM ClO in
-
phosphate buffer (0.2 M, pH 7.4). (B) Fitted calibration curve for BL intensity of 25 μM 1 at 560 nm with ClO concentration in the linear
-
region of 0-62.5 μM. (C) Selectivity studies of 1: Selectivity of 1 at 25 μM in the presence of 25 μM ClO or 100 μM NO, H
2
O
2
, hydroxyl
), glutathione (GSH), HCO ,
2 3
-
•-
1
-
radical (•OH), peroxynitrite (ONOO ), alkylperoxyl radical (ROO•), superoxide (O
2
), singlet oxygen ( O
-
-
-
2-
NO , NO , Cl , or SO respectively. The experiments were performed in triplicate. Error bars represent relative standard deviations (RSD)
of three independent experiments which are 1.1 × 10 , 2.3 × 10 , 6.9 × 10 , 4.0 × 10 , 1.3 × 10 , 3.8 × 10 , 4.2 × 10 , 5.2 × 10 , 0.018,
2
3
4
-3
-3
-3
-5
-3
-3
-3
-4
-3
-3
4.4 × 10 , 8.2 × 10 , respectively. Emission: 560 nm.
High performance liquid chromatography (HPLC) analysis clearly
With its good in vitro property, probe 1 was subsequently
applied for BL imaging ClO in living cells. Before that,
cytotoxicity of 1 and ClO was evaluated on MDA-MB-231 cells
(non-luciferase transfected) with 3-(4,5-dimethylthiazol-2-yl) 2,5
diphenyltetrazplium bromide (MTT) assay. As shown in Figure S6,
90.0% of the cells survived after 24 h incubation with 100 µM 1,
suggesting that probe 1 under 100 µM is safe for live cell imaging.
For ClO , 95% of the cells survived after 4 h incubation with 20
µM ClO (Figure S7), suggesting that ClO concentration under 20
µM is safe for live cell imaging. In this work, we chose 1 at 50
µM and ClO at 5 µM to conduct the following cell imaging study.
FLuc–transfected MDA-MB-231 cells were pre-incubated with or
without 5 µM ClO in serum-free culture medium at 37 °C for 30
min, and washed with phosphate buffered saline (PBS) for three
times to remove extracellular ClO . Then the cells were placed in
the 96-well plate at 1 × 10 cells/well. BL signals generated by the
cells were recorded with a small animal imaging system in a real-
time manner. Generally, as shown in Figure 2, after the addition of
probe 1 to each well, the BL signals gradually increased to the
maximum at around 90 min, followed by a decrease, and then
maintained at comparable values over the observation window
Figure S8). To be specific, the brightest BL signal was observed
from the cells pre-incubated with ClO (middle row in Figure 2),
suggesting that probe 1 has good cell permeability, and can be
-luciferin inside cells. However, when
the cells were incubated with 25 mg/mL ClO scavenger taurine
for 5 min before the addition of 1 to the ClO pre-incubation cells,
their BL signal (bottom row in Figure 2) was significantly
-
indicated that, about 75.4% of 1 was converted to
D-luciferin
-
under this condition (Figure S5). After validation of the feasibility
-
of 1 for ClO detection, a linear relationship (Y = 0.599X + 0.694,
2
R = 0.998) between the BL enhancement (i.e., B - B
1
0
) of 25 µM
-
at 560 nm and ClO concentration was obtained over the range
of 0-62.5 µM (Figure 1B). The limit of detection (LOD) of ClO
of this assay was calculated to be 0.705 µM according to the
-
-
3
9
-
-
general 3σ method (3σ/s), comparable to those of reported
methods (Table S1).
-
Selectivity is one important parameter to evaluate the
performance of a new probe. Particularly, for a new probe which
is intentionally designed for biomedical applications, its highly
selective response to the target over other possible competitive
species is a must. Therefore, we screened a wide array of the
-
-
6
potential competitive species such as intracellular reactive oxygen
-
and nitrogen species (ROS/RNS) (e.g., NO, H
2
O
2
, •OH, ONOO ,
•-
1
ROO•, O
2 2
, and O ), intracellular abundant biomolecules (e.g.,
- - - -
GSH) and common anions (e.g., HCO
3
, NO
2 3
, NO , Cl , and
2
-
SO
4
) to study the selectivity of probe 1 by comparing its B - B
0
-
value induced by ClO over those of the selected species in the
(
presence of fLuc. As shown in Figure 1C, only 100 µM ROO• and
-
1
O showed weak bioluminescence after addition to 25 µM 1,
2
-
which was almost negligible compared to that induced by ClO
BL enhancement by ClO was 23-182 folds of those induced by
-
oxidized by ClO to yield
D
-
(
-
40
other species). Therefore, we concluded that probe 1 has excellent
selectivity toward ClO over other testing species.
-
-
-
decreased compared to that of ClO -treated cells. This
-
demonstrated that ClO could trigger 1 to release of
D
-luciferin for
BL generation in cells. For the cells without pre-incubation of
ClO , weak BL signal could be detected (top row in Figure 2).
This could be explained that the intracellular proteases hydrolyzed
the amide bond of 1 to yield -luciferin to generate the weak BL
because MDA-MB-231 cells themselves do not generate ClO .
HPLC analysis indicated that, after the MDA-MB-231 cells (non-
luciferase transfected) were incubated with 200 μM 1 for 30 min,
-luciferin inside
cells (Figure S9). For the ClO -pretreated cells, incubation with
probe 1 resulted in a three-time increased amount of -luciferin
-
D
-
Figure 2. Time-course bioluminescence images of fLuc-
transfected MDA-MB-231 cells incubated with 50 μM 1 (top row),
15.4% of uptaken 1 was proteolysed to yield
D
-
-
pre-incubated with 5 μM exogenous ClO and then with 50 μM 1
-
(
middle row), pre-incubated with 5 μM exogenous ClO followed
D
-
-
by 25 mg/mL ClO scavenger taurine and then with 50 μM 1
bottom row) acquired at 0, 15, 30, 50, 70, 90, 110, 130 and 150
min in serum-free culture medium at 37 °C.
inside cells, compared with those cells without ClO treatment
(Figure S9). This further comfirmed that most of the intracellular
-luciferin was generated by 1 that reacted with ClO for the BL
(
-
D
ACS Paragon Plus Environment

Page 3 of 5
Analytical Chemistry
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
augmentation. We also conducted cell imaging experiment with
the cells pre-incubated with 1 before ClO addition. As shown in
taurine at 50 mg/mL 5 min later and 100 μL 1 at 2.5 mM 5 min
later (bottom row) in PBS at 20, 70, 120, 170, and 220 min.
-
Figure S10, the brightest BL signal was also observed from the
-
In summary, we have successfully developed
a latent
-
cells with ClO addition. However, the BL signal increased to the
-
bioluminescent probe 1 for highly selective detection of ClO in
maximum much quicker (15 min after ClO addition) and then
-
-
vitro and imaging ClO in tumors of living mice. Upon ClO
oxidation and the follow-up hydrolysis, 1 was converted to -
decreased gradually (Figure S11). This could be explained that,
-
D
upon the addition of ClO , probe 1 was turned into
D
-luciferin in
luciferin for BL generation in the presence of fLuc. This property
the medium, resulting in similar cell imaging pattern to that of
3
2
of 1 was successfully applied for highly selective detection of
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
cells directly incubated with
D
-luciferin. The above results
-
-
ClO within 0-62.5 μM and a LOD of 0.705 μM in vitro. The
suggest that probe 1 is suitable for BLI of ClO in living cells.
-
feasibility of this assay for imaging ClO in living cells and in
-
We then applied 1 for BLI of ClO in living animals. The nude
tumors was also validated. With these unique features of 1, we
mice, xenografted with fLuc-transfected MDA-MB-231 tumor in
envision that it might be applied to elucidate the biological roles
the right thigh for each, were randomly divided into three groups
-
of ClO in wider physiological and pathological processes in the
-
(
one group for the injection of ClO followed by injection of 1 as
near future.
the experimental group, one group for the injection of 1 only and
-
one group for the successive injections of ClO , taurine, and 1 as
the two control groups). BL signal generated from the tumors in
the nude mice was then recorded in real-time by a small animal
ASSOCIATED CONTENT
Supporting Information
-
imaging system. As shown in Figure 3, even without ClO
injection, tumors of 1-injected mice also exhibited weak BL signal
(
top row in Figure 3). This could be explained that small amount
of 1 was hydrolyzed by intracellular proteases of tumor cells to
yield -luciferin for BL generation. However, within the
General methods; Syntheses and characterizations of 1;
Supporting scheme, figures, and tables (Scheme S1, Figure S1-
S12, and Table S1-S3). This material is available free of charge
via the Internet at http://pubs.acs.org.
D
observation time (i.e., 20-220 min), the BL intensity of the
experimental group remained the highest (middle row in Figure 3).
It increased to the peak at 70 min, and then decreased slowly
-
thereafter (Figure S12), suggesting that, under ClO oxidation,
AUTHOR INFORMATION
Corresponding Authors
-
most of 1 yielded
D
-luciferin for the strong BL. ClO -scavenging
control experiment indicated that the taurine-treated mice had
obviously lower BL intensity than that of the experimental group
*
*
E-mail: wangfq@njmu.edu.cn
E-mail: gliang@ustc.edu.cn
(
bottom row in Figure 3). This further affirmed that above strong
-
BL from experimental group was resulted from ClO oxidation of
1 to yield -luciferin. These in vivo results demonstrated that
probe 1 could be applied for imaging ClO in tumors of living
D
-
Notes
animals.
The authors declare no competing financial interests.
ACKNOWLEDGMENT
This work was supported by Collaborative Innovation Center of
Suzhou Nano Science and Technology, Hefei Science Center
CAS (2016HSC-IU010), Ministry of Science and Technology of
China (2016YFA0400904), and the National Natural Science
Foundation of China (Grants 21675145 and U1532144).
REFERENCES
(1) McCord, J. M. Science 1974, 185, 529-531.
(2) D'Autréaux, B.; Toledano, M. B. Nat. Rev. Mol. Cell Biol. 2007,
8
, 813-824.
3) Mittal, M.; Siddiqui, M. R.; Tran, K.; Reddy, S. P.; Malik, A. B.
Antioxid. Redox Signal. 2014, 20, 1126-1167.
4) Hidalgo, E.; Bartolome, R.; Dominguez, C. Chem.-Biol.
Interact. 2002, 139, 265-282.
5) Goiffon, R. J.; Martinez, S. C.; Piwnica-Worms, D. Nat.
Commun. 2015, 6, 6271.
(
(
(
(
(
6) Harrison, J.; Schultz, J. J. Biol. Chem. 1976, 251, 1371-1374.
7) Sugiyama, S.; Okada, Y.; Sukhova, G. K.; Virmani, R.;
Heinecke, J. W.; Libby, P. J. Pathol. 2001, 158, 879-891.
(8) Yap, Y. W.; Whiteman, M.; Cheung, N. S. Cell. Signal. 2007,
Figure 3. Time-course bioluminescence imaging of nude mice
xenografted with fLuc-transfected MDA-MB-231 tumors after
intraperitoneal injection of 100 μL 1 at 2.5 mM (top row), 100 μL
1
9, 219-228.
9) Hammerschmidt, S.; Buchler, N.; Wahn, H. Chest 2002, 121,
573-581.
(10) Daugherty, A.; Dunn, J. L.; Rateri, D. L.; Heinecke, J. W. J.
Clin. Invest. 1994, 94, 437-444.
(
-
ClO at 1 mM followed by 100 μL 1 at 2.5 mM 5 min later
-
(
middle row), or 100 μL ClO at 1 mM followed by 100 μL
ACS Paragon Plus Environment

Analytical Chemistry
Page 4 of 5
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
(11) Wu, S. M.; Pizzo, S. V. Arch. Biochem. Biophys. 2001, 391,
119-126.
(26) Li, J.; Chen, L.; Du, L.; Li, M. Chem. Soc. Rev. 2013, 42, 662-
676.
(27) Marques, S. M.; Esteves da Silva, J. C. IUBMB Life 2009, 61,
6-17.
(28) McCutcheon, D. C.; Paley, M. A.; Steinhardt, R. C.; Prescher,
J. A. J. Am. Chem. Soc. 2012, 134, 7604-7607.
(29) Conley, N. R.; Dragulescu‐Andrasi, A.; Rao, J.; Moerner, W.
Angew. Chem. Int. Ed. 2012, 51, 3350-3353.
(30) Van de Bittner, G. C.; Bertozzi, C. R.; Chang, C. J. J. Am.
Chem. Soc. 2013, 135, 1783-1795.
(31) Heffern, M. C.; Park, H. M.; Au-Yeung, H. Y.; Van de Bittner,
G. C.; Ackerman, C. M.; Stahl, A.; Chang, C. J. Proc. Natl. Acad. Sci.
U. S. A. 2016, 113, 14219-14224.
(32) Zheng, Z.; Wang, L.; Tang, W.; Chen, P.; Zhu, H.; Yuan, Y.;
Li, G.; Zhang, H.; Liang, G. Biosens. Bioelectron. 2016, 83, 200-204.
(33) Takakura, H.; Kojima, R.; Kamiya, M.; Kobayashi, E.;
Komatsu, T.; Ueno, T.; Terai, T.; Hanaoka, K.; Nagano, T.; Urano, Y.
J. Am. Chem. Soc. 2015, 137, 4010-4013.
(34) Wu, W.; Li, J.; Chen, L.; Ma, Z.; Zhang, W.; Liu, Z.; Cheng,
Y.; Du, L.; Li, M. Anal. Chem. 2014, 86, 9800-9806.
(35) Cohen, A. S.; Dubikovskaya, E. A.; Rush, J. S.; Bertozzi, C. R.
J. Am. Chem. Soc. 2010, 132, 8563-8565.
(36) Jiang, T.; Ke, B.; Chen, H.; Wang, W.; Du, L.; Yang, K.; Li,
M. Anal. Chem. 2016, 88, 7462-7465.
(
(
12) Weitzman, S. A.; Gordon, L. Blood 1990, 76, 655-663.
13) Shepherd, J.; Hilderbrand, S. A.; Waterman, P.; Heinecke, J.
W.; Weissleder, R.; Libby, P. Chem. Biol. 2007, 14, 1221-1231.
14) Chen, T.; Hu, Y.; Cen, Y.; Chu, X.; Lu, Y. J. Am. Chem. Soc.
2013, 135, 11595-11602.
15) Goswami, S.; Das, A. K.; Manna, A.; Maity, A. K.; Saha, P.;
Quah, C. K.; Fun, H.-K.; Abdel-Aziz, H. A. Anal. Chem. 2014, 86,
315-6322.
16) Hou, J.; Wu, M.; Li, K.; Yang, J.; Yu, K.; Xie, Y.; Yu, X.
Chem. Commun. 2014, 50, 8640-8643.
17) Wu, Y.; Wang, J.; Zeng, F.; Huang, S.; Huang, J.; Xie, H.; Yu,
C.; Wu, S. ACS Appl. Mater. Interfaces 2016, 8, 1511-1519.
18) Dragulescu-Andrasi, A.; Liang, G.; Rao, J. Bioconjugate
(
(
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
6
(
(
(
Chem. 2009, 20, 1660-1666.
(19) Jacobson, G. B.; Shinde, R.; Contag, C. H.; Zare, R. N. Angew.
Chem. Int. Ed. 2008, 47, 7880-7882.
(
20) Yuan, Y.; Wang, F.; Tang, W.; Ding, Z.; Wang, L.; Liang, L.;
Zheng, Z.; Zhang, H.; Liang, G. ACS Nano 2016, 10, 7147-7153.
21) Yang, Y.; Shao, Q.; Deng, R.; Wang, C.; Teng, X.; Cheng, K.;
(
Cheng, Z.; Huang, L.; Liu, Z.; Liu, X. Xing, B. Angew. Chem. Int. Ed.
2012, 51, 3125-3129.
(
22) Zheng, Z.; Li, G.; Wu, C.; Zhang, M.; Zhao, Y.; Liang, G.
Chem. Commun. 2017, 53, 3567-3570.
23) Evans, M. S.; Chaurette, J. P.; Adams Jr, S. T.; Reddy, G. R.;
(37) Kojima, R.; Takakura, H.; Kamiya, M.; Kobayashi, E.;
Komatsu, T.; Ueno, T.; Terai, T.; Hanaoka, K.; Nagano, T.; Urano, Y.
Angew. Chem. Int. Ed. 2015, 54, 14768-14771.
(38) Chen, X.; Wang, X.; Wang, S.; Shi, W.; Wang, K.; Ma, H.
Chem.-Eur. J. 2008, 14, 4719-4724.
(39) Ogren, P. J.; Meetze, A.; Duer, W. C. J. Anal. Toxicol. 2009,
33, 129-142.
(40) Weiss, S. J.; Klein, R.; Slivka, A.; Wei, M. J. Clin. Invest.
(
Paley, M. A.; Aronin, N.; Prescher, J. A.; Miller, S. C. Nat. Methods
2014, 11, 393-395.
(
24) Adams, S. T.; Mofford, D. M.; Reddy, G.; Miller, S. C. Angew.
Chem. Int. Ed. 2016, 55, 4943-4946.
25) Kaskova, Z. M.; Tsarkova, A. S.; Yampolsky, I. V. Chem. Soc.
Rev. 2016, 45, 6048-6077.
(
1982, 70, 598-607.
ACS Paragon Plus Environment

Page 5 of 5
Analytical Chemistry
1
2
3
For TOC only
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
ACS Paragon Plus Environment