Electrochimica Acta 180 (2015) 196–201

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Electrochimica Acta
journal homepage: www.elsevier.com/locate/electacta

Electrogenerated Chemiluminescence (ECL) Quenching of the Ru

(bpy)32+/TPrA System by the Explosive TNT
Suman Parajuli, Xiaohui Jing1, Wujian Miao* ,2
Department of Chemistry and Biochemistry, The University of Southern Mississippi, Hattiesburg, MS 39406, United States

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

Article history: We report the quenching behavior of 2,4,6-trinitrotoluene (TNT) on the electrogenerated chemilumi-
Received 10 June 2015 nescence (ECL) of the Ru(bpy)32+/TPrA [bpy = tris(2,20 -bipyridine), TPrA = tri-n-propylamine] system at a
Received in revised form 30 July 2015 Pt electrode in acetonitrile containing tetra-n-butylammonium perchlorate supporting electrolyte.
Accepted 19 August 2015
Effective ECL quenching of the system upon the addition of TNT was observed, with a Stern-Volmer
Available online 22 August 2015
constant of 2  104 M1. The quenching constant calculated from the Stern-Volmer plot was found to be
2.1 1010 M1 s1 which suggests the efficient quenching of ECL by TNT. The competitive processes of
Keywords:
consuming TPrA free radicals and Ru(bpy)3+ species (produced as a result of reduction of Ru(bpy)32+ by
Electrogenerated chemiluminescence (ECL)
2,4,6-Trinitrotoluene (TNT)
TPrA) by TNT largely contributed to this quenching, in which the consumption of Ru(bpy)3+ by TNT was
Explosive more favorable than that of TPrA free radicals. TNT was also found to quench the fluorescence of Ru
Quenching (bpy)32+, with a Stern-Volmer and a quenching constant of 5.2  103 M1 and 5.5  109 M1 s1,
Stern-Volmer constant respectively. This collisional (dynamic) based fluorescence quenching process contributed approximately
one quarter of the overall ECL quenching efficiency. The present ECL quenching scheme could be used to
detect TNT at submicromolar levels.
ã 2015 Elsevier Ltd. All rights reserved.

1. Introduction
2,4,6-Trinitrotoluene (TNT) is a flammable odorless solid that
exists as colorless orthorhombic crystals or yellow monoclinic
needles but does not occur naturally in the environment [1]. The
groundwater or seawater contamination has occurred due to the
production, storage, testing, and disposal of explosives at military
installations, resulting in the environmental problems because
these toxic and persistent compounds can leach from soil [2–4]. At
the end of World War II, an estimated 300,000 tons of explosives
(mainly TNT) were disposed to the sea [1]. TNT is considered as
toxic as it produces toxic and mutagenic effects to life including
human being [5].
The methods with ability for rapid on-site detection of TNT are
highly desired. Currently, several methods of detection of TNT are
used, which include those based on fluorescence [6–17], amperom-
etry [18,19], surface plasmon resonance [5,20–28], mass spectrome-
try [29–34], chromatography [35–39], Raman spectroscopy [40–44],
and ion mobility spectroscopy [45,46]. In addition to amperometry,
* Corresponding author: Tel.: +1 601 266 4716; fax: +1 601 266 6075.
E-mail address: wujian.miao@usm.edu (W. Miao).
1
Visiting scholar from College of Chemistry and Chemical Engineering, Nantong
University, Natong 226007, PR China.
2
ISE member.
other electrochemical techniques have also been used to detect TNT
[47–51]. Each individual detection method described above has its
own advantages, but some drawbacks remain. For example,
fluorescence techniques generally suffer from the luminescent
impurities, the ion mobility spectroscopic technique suffers from the
matrix effects, and chromatographic separation-mass spectrometric
detection methods require sample preparation and are not
appropriate for field tests. These techniques use expensive instru-
ments that frequently require skilled manpower to operate. The
immunoassay-based electrogenerated chemiluminescence (ECL)
technique, which possesses very high sensitivity and selectivity
but requires extra effort in sample preparation, has been used to
detect TNT in soil and water samples [52–54]. Methods based on ECL
quenching have been recently employed in the detection and
quantification of e.g., phenols, quinones, and catechols [55,56], as
well as the explosives triacetone triperoxide and tetryl in acetonitrile
(via the Ru(bpy)32+ ion annihilation ECL scheme) [57,58].
Because each nitro group in TNT could undergo sequential 4e
to 6e reduction to form hydroxylamine and amine within a
potential range of 0 to -1.0 V vs SCE [59–64], TNT could be used to
quench or inhibit the ECL signals generated from a known
“oxidative-reduction” type standard such as the Ru(bpy)32+/TPrA
(TPrA = tri-n-propylamine) system as detailed below.

http://dx.doi.org/10.1016/j.electacta.2015.08.107
0013-4686/ ã 2015 Elsevier Ltd. All rights reserved.
 S. Parajuli et al. / Electrochimica Acta 180 (2015) 196–201 197

When the Ru(bpy)32+/TPrA system is used, both Ru(bpy)32+ and strategy that is simple, inexpensive, and sensitive for TNT
TPrA are oxidized upon the anodic potential scanning: detection and quantification.

TPrA  e ! TPrAþ ð1Þ

2. Experimental

2.1. Chemicals
2þ 3þ
RuðbpyÞ3  e ! RuðbpyÞ3 ð2Þ

TPrA free radicals are produced after the deprotonation of the The chemicals used in this study were: acetonitrile (MeCN, 99.8%,
newly produced TPrA+: HPLC grade), tris-(2,2'-bypyridine) dichlororuthenium(II) hexahy-
drate (Ru(bpy)3Cl26H2O, 99.95%), and tri-n-propylamine (TPrA, 99+
Hþ 
%) (all from Sigma-Aldrich, St. Louis, MO, USA); 2,4,6-trinitrotoluene
TPrAþ ! TPrA ð3Þ
(TNT, 99.6%, 1000 mg/mL in MeCN, Supelco analytical, PA, USA);

The TPrA radical is a strong reducing agent with a redox sodium phosphate, monobasic monohydrate (J.T. Baker Chemicals
potential of -1.7 V vs SCE [65], and is the key species of ECL Co., Phillipsburg, NJ, USA); and silver nitrate (99.5 %) and tetra-n-
generation. In the absence of TNT, the TPrA radical reduces Ru butylammonium perchlorate (TBAP, 99+%, electrochemical grade)
(bpy)32+ to Ru(bpy)3+ that annihilates with Ru(bpy)33+ to form the (both from Fluka, Milwaukee, WI, USA). All chemicals were used as
excited state Ru(bpy)32+* that emits light [66]: received.
þ
þ TPrA ! RuðbpyÞ3 þ Product
2þ
RuðbpyÞ3 ð4Þ
2.2. Electrochemical and ECL studies

Cyclic voltammetry (CV) was performed with an electrochemical

3þ þ 2þ 2þ
Ru ðbpyÞ3 þ Ru ðbpyÞ3 ! RuðbpyÞ3 þ Ru ðbpyÞ3 ð5Þ workstation (Model 660A, CH Instruments, Inc., Austin, TX, USA). The
ECL and CV responses were simultaneously recorded with a
homemade ECL instrument [67,68], where the CHI electrochemical
2þ 2þ
Ru ðbpyÞ3 ! Ru ðbpyÞ3 þ hv ð6Þ workstation was combined with a photo-multiplier tube (PMT,
2+
Hamamatsu R928, Japan) installed under the electrochemical cell. A
Alternatively, the excited state Ru(bpy)3 * can be produced via voltage of -700 V was supplied to the PMT with a high voltage power
the following process: supply (Model 472A Brandenburg PMT power supply, England). The
RuðbpyÞ3
3þ
þ TPrA ! RuðbpyÞ3
2þ 
þ Product ð7Þ light signal (as photocurrent) was detected with a high sensitive
Keithley 6514 electrometer (Keithley, Cleveland, OH, USA) and
In the presence of TNT, however, part or the entire portion of the converted to a voltage (in 2 V) that was fed to the electrochemical
TPrA radicals could be consumed by TNT, resulting in significant workstation and displayed on the computer monitor. A conventional
decrease or complete quenching of the initial ECL signals of the Ru three-electrode electrochemical cell system was used with a Pt disk
(bpy)32+/TPrA system: (2 mm diameter) as the working electrode, a Pt gauge as the counter
 electrode, and a Ag/Ag+ (with 10 mM AgNO3 and 0.10 M TABP in
nTPrA þ TNT ! TNTn þ Product ð8Þ
MeCN) as the reference electrode (0.186 V vs NHE). The Pt working
Additionally, Ru(bpy)3+, another key species for ECL generation electrode was polished before every run with 0.3-0.05 mm alumina
(Eqs. (4)–(5))could be consumed by TNT: slurry, washed with water and dried with the Kim wipes facial tissue.
Note that the test solutionwas used without purging with nitrogen as
þ 2þ
nRuðbpyÞ3 þ TNT ! RuðbpyÞ3 þ TNTn ð9Þ the electrode potential was scanned in anodic direction and oxygen
which leads to further decrease in ECL intensity. from air had no effect on ECL production.
The aforementioned electron transfer processes could be better
understood with the redox potentials associated with TPrA, Ru 2.3. Fluorescence studies
(bpy)32+, Ru(bpy)32+*, TNT, and their derivatives as shown in
Scheme 1 [66]. Fluorescence experiments were carried out on a PTI Qunata
The present work focuses on the studies of the ECL quenching MasterTM 40 intensity based spectrofluorometer (Photon Technol-
behavior of the Ru(bpy)32+/TPrA system by TNT. Fluorescence ogy International, Inc., NJ, USA), with a slit width of 1.00 nm,
investigations will be conducted so that the respective contribu- excitation wavelength at 350 nm, and an emission wavelength
tions of the electron-transfer reactions and the collisional range from 500 to 690 nm.
(dynamic) process to the overall ECL quenching can be estimated. All experiments were conducted at the room temperature of
Data obtained from this study could be used to develop a new 20  1 C.

3. Results and discussion

3.1. ECL generation from the Ru(bpy)32+/TPrA system

Fig. 1A shows the CV and ECL responses of 1.0 mM Ru(bpy)32

+
-25 mM TPrA in MeCN containing 0.10 M TBAP at a Pt electrode
with a scan rate of 50 mV/s. TPrA is oxidized at 0.6 V vs Ag/Ag+
and the generation of ECL light starts from 0.9 V vs Ag/Ag+ at which
Ru(bpy)32+ gets oxidized to Ru(bpy)33+. The optimum concentra-
tion of ECL coreactant was determined by running an ECL
experiment with 1.0 mM Ru(bpy)32+ and different concentrations
of TPrA with a Pt electrode. As shown in Fig. 1B, the maximum ECL
Scheme 1. Redox potentials of TPrA, Ru(bpy)32+, Ru(bpy)32+*, TNT, and their response occurs with a TPrA concentration of 25 mM, which is in
derivatives. good agreement with the data reported previously [69]. This
198 S. Parajuli et al. / Electrochimica Acta 180 (2015) 196–201

40

30 (a)

iECL (nA)
20

10 (b)

(c)
0
(d)
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8
+
E (V vs Ag/Ag )
Fig. 3. ECL quenching by TNT in the system containing 5 nM Ru(bpy)32+-25 mM
TPrA-0.10 M TBAP in MeCN with added TNT concentrations of (a) 0, (b) 44, (c) 88,
and (d) 132 mM. A 2-mm diameter of Pt electrode and a scan rate of 50 mV/s were
used for all tests.

To examine if the ECL could be completely quenched by TNT,

Fig. 1. (A) CV and ECL of 1.0 mM Ru(bpy)32+-25 mM TPrA in MeCN. (B) TPrA
concentration effect on the ECL generation of the Ru(bpy)32+/TPrA system.
Experiments were conducted at a 2-mm Pt electrode with a scan rate of 50 mV/s
using 1.0 mM Ru(bpy)32+ -0.10 M TBAP MeCN solutions. The ECL signal was the
photocurrent detected with the PMT (see Section 2.2 for details).
10 mM), the ECL intensity of the system became
concentration of TPrA, therefore, was used for the entire
experiment discussed below, unless otherwise indicated.
3.2. ECL quenching by TNT
The ECL quenching experiment by TNT was carried out initially
with 1.0 mM Ru(bpy)32+ and 25 mM TPrA in MeCN containing
0.10 M TBAP. As shown in Fig. 2, the ECL emission of the Ru(bpy)32
+
/TPrA system (Fig. 2a) is gradually decreased with successive
addition of TNT (stock solution: 1000 mg per mL in MeCN). The ECL
is reduced by 14% (Fig. 2b) and 68% (Fig. 2c) with a final TNT
concentration of 4.4 and 110 mM, respectively.
various concentrations of Ru(bpy)32+ and TPrA were tested. When
Ru(bpy)32+ concentration was down to nanomolar levels, complete
quenching was observed. For example, in a 5 nM Ru(bpy)32
+
-25 mM TPrA system (Fig. 3a), addition of 44, 88, and 132 mM
of TNT results in 76% (Fig. 3b), 90% (Fig. 3c), and 100% (Fig. 3d) of
the ECL quenching, respectively. Studies on the effect of lowing
TPrA concentration on the ECL quenching (with 1.0 mM Ru(bpy)32+)
revealed that with the decrease of TPrA concentration, TNT
quenching efficiency was slightly increased. However, at low TPrA
concentrations (
weak (Fig. 1B), which made the experimental observations
difficult. Furthermore, under low TPrA concentrations, no com-
plete quenching of ECL with a high TNT concentration of 132 mM
was observed. These data suggest that the quenching process
shown in Eq. (8) is probably a relatively slow electron transfer
process with respective to that of Eq. (9). In other words, ECL

Fig. 2. ECL quenching of the Ru(bpy)32+ (1.0 mM)/TPrA (25 mM) by TNT at a final Fig. 4. Ru(bpy)32+ concentration effect on ECL (a) without, and (b) with 4.4 mM TNT
concentration of (a) 0, (b) 4.4, and (c) 110 mM. Working electrode: 2-mm diameter addition to the MeCN solution containing 25 mM TPrA-0.10 M TBAP. A 2-mm
Pt, scan rate: 50 mV/s. For clarity, only forward scans of the ECL are plotted. diameter of Pt electrode and a scan rate of 50 mV/s were used for all tests.
 S. Parajuli et al. / Electrochimica Acta 180 (2015) 196–201 199

quenching by TNT via Ru(bpy)3+ consumption is more favorable

than direct TPrA free radical consumption. If that is the case, under
constant TPrA and TNT concentrations, ECL quenching efficiency
should increase with the increase in Ru(bpy)32+ concentration.
Fig. 4 illustrates the results of such an example, where the ECL is
quenched by 38% at 1.0 mM Ru(bpy)32+, and 45% at 10 mM Ru
(bpy)32+ after the system is added with 4.4 mM TNT (Fig. 4b).
Notably, increase in Ru(bpy)32+ concentration linearly increases
the ECL intensity with and without TNT, but the presence of a
constant amount of TNT shows quenching effect throughout the
experiment, which confirms that quenching effect is more related
to the Ru(bpy)32+ than to TPrA.
As shown in Fig. 5, a linear decrease in ECL intensity of the
standard Ru(bpy)32+ (1.0 mM)/TPrA (25 mM) system is evident with
the addition of TNT. Under the present experimental conditions,
this quenching method provides a TNT detection limit of 4.4 mM
(or 1.0 ppm).

3.3. Fluorescence quenching of Ru(bpy)32+ by TNT

The fluorescence quenching study was carried out with 1.0 mM

Ru(bpy)32+ in MeCN with a quartz cuvette. The test solution was
excited at 350 nm and Ru(bpy)32+ produced the fluorescence
emission peak at 620 nm. The addition of TNT results in a decrease
in fluorescent emission of the test solution as shown in Fig. 6.
Upon the addition of 110 mM TNT, the fluorescence emission is
decreased by 42% with respect to the emission of Ru(bpy)32+
without TNT added (Fig. 6A). In order to verify the dilution effect by
the addition of TNT which was in MeCN, the control experiment
was carried out with the same concentration of Ru(bpy)32+ and
similar volumes of MeCN were added to the test solution and
fluorescence emissions were recorded. Results of the control
experiments (Fig. 6B) suggest that the effect of dilution on
fluorescence emission is negligible and within the range of
experimental errors.
Fig. 6. Fluorescence quenching of 3.0 mL of 1.0 mM Ru(bpy)32+ in MeCN by TNT with
3.4. Calculation of quenching constant [56,70,71] 1.0 cm quartz cuvette and an excitation at 350 nm. (A) TNT concentration
dependence, and (B) MeCN dilution effect.
The quantum yield of ECL (or fluorescence, “f”) of Ru(bpy)32+ in
MeCN in the absence of quenching agent (K ) is the rate at which mechanism as:
the excited state decays via ECL (or fluorescence) over sum of the 2þ
rates of its decay by ECL (or fluorescence) and non-radiative K ECLðfÞ ½RuðbpyÞ3  K ECLðfÞ
F0 ¼ ¼ ð10Þ
K ECLðfÞ ½RuðbpyÞ3
2þ
 þ K nr ½RuðbpyÞ3
2þ
 K ECLðfÞ þK nr

where KECL(f) and Knr stand for the rate constants for ECL (or
fluorescence) and non-radiative decay of the excited state Ru
(bpy)32+*, respectively. In the presence of a quenching agent like

Fig. 5. Relationship of ECL intensity with the TNT concentration in the system
containing 1.0 mM Ru(bpy)32+-25 mM TPrA-0.10 M TBAP in MeCN at a 2-mm Pt Fig. 7. Stern-Volmer plot of (a) ECL and (b) fluorescence quenching by TNT. See
electrode at a scan rate of 50 mV/s. Figs. 5 and 6 for experimental conditions.
200 S. Parajuli et al. / Electrochimica Acta 180 (2015) 196–201

(ECL) as the excited state collisional (dynamic) process as suggested by

Ru(bpy)32+* Collisional
the fluorescence studies. This quenching scheme as illustrated in
TPrA Electron-
Quenching Scheme 2 could be used to detect TNT at submicromolar levels.
Ru(bpy)33+ transf er1 ECL Quenched
-e
-H+ by consuming of Acknowledgments
TPrA TPrA
Pt TNT
O2N NO2
TPrA :):)
NO2
Ru(bpy)3+ :):):) The abstract was presented at PITTCON 2011, March 14-17,
(+) Ru(bpy)32+ Ru(bpy)3+
Ru(bpy)32+* :) Atlanta, GA. Financial support from the National Science Founda-
-e tion via an NSF CAREER award (CHE 0955878) is gratefully
Ru(bpy)33+
Electron-
Collisional
with TNT
transf er2 acknowledged.
Quenching
2+*
Ru(bpy)3 :):):) most ef ficient
(ECL) References
2+
Scheme 2. ECL quenching pathways of the Ru(bpy)3 /TPrA system by the [1] R. Wilson, C. Clavering, A. Hutchinson, Electrochemiluminescence enzyme
explosive TNT. immunoassay for TNT, Analyst 128 (2003) 480–485.
[2] J. Wang, R.K. Bhada, J. Lu, D. MacDonald, Remote electrochemical sensor for
TNT, the quantum yield could be expressed as: monitoring TNT in natural waters, Anal. Chim. Acta 361 (1998) 85–91.
[3] H. Sohn, R.M. Calhoun, M.J. Sailor, W.C. Trogler, Detection of TNT and picric acid
K ECLðfÞ ½RuðbpyÞ3 2þ  on surfaces and in seawater by using photoluminescent polysiloles, Angew.
F¼ Chem. Int. Ed. 40 (2001) 2104–2105.
K ECLðfÞ ½RuðbpyÞ3 2þ   þ K nr ½RuðbpyÞ3 2þ   þ K q ½TNT½RuðbpyÞ3 2þ  [4] A. Hilmi, J.H.T. Luong, A.-L. Nguyen, Determination of explosives in soil and
ground water by liquid chromatography-amperometric detection, J.
K ECLðfÞ Chromatogr. A 844 (1999) 97–110.
¼ ð11Þ
K ECLðfÞ þ K nr þ K q ½TNT [5] D.R. Shankaran, K.V. Gobi, T. Sakai, K. Matsumoto, K. Toko, N. Miura, Surface
plasmon resonance immunosensor for highly sensitive detection of 2,4,6-
where Kq is the quenching constant of ECL (or fluorescence). By trinitrotoluene, Biosens. Bioelectron. 20 (2005) 1750–1756.
[6] S. Content, W.C. Trogler, M.J. Sailor, Detection of nitrobenzene, DNT, and TNT
dividing Eq. (10) by Eq. (11) we get, vapors by quenching of porous silicon photoluminescence, Chem. -Eur. J. 6
(2000) 2205–2213.
F0 I0 Kq [7] G.M. Boudreaux, T.S. Miller, A.J. Kunefke, J.P. Singh, F.-Y. Yueh, D.L. Monts,
¼ ¼1þ ½TNT ¼ 1 þ K SV ½TNT ð12Þ
F I K ECLðfÞ þ K nr Development of a photofragmentation laser-induced-fluorescence laser
sensor for detection of 2,4,6-trinitrotoluene in soil and groundwater, Appl.
Opt. 38 (1999) 1411–1417.
This is the Stern-Volmer equation, in which the constant terms
[8] I.B. Bakaltcheva, L.C. Shriver-Lake, F.S. Ligler, A fiber optic biosensor for
give the slope of a plot and is called Stern-Volmer constant (KSV). I multianalyte detection: Importance of preventing fluorophore aggregation,
and I0 stand for the ECL (or fluorescence) intensities with and Sens. Actuators, B 51 (1998) 46–51.
without quenching agent, TNT. The reciprocal of (KECL(f) + Knr) gives [9] I.L. Medintz, E.R. Goldman, M.E. Lassman, A. Hayhurst, A.W. Kusterbeck, J.R.
Deschamps, Self-assembled TNT biosensor based on modular multifunctional
the lifetime (t ) of the excited state Ru(bpy)32+* species. surface-tethered components, Anal. Chem. 77 (2005) 365–372.
[10] L.C. Shriver-Lake, K.A. Breslin, P.T. Charles, D.W. Conrad, J.P. Golden, F.S. Ligler,
1
t¼ ð13Þ Detection of TNT in water using an evanescent wave fiber-optic biosensor,
K ECLðfÞ þ K nr Anal. Chem. 67 (1995) 2431–2435.
[11] G.P. Anderson, S.C. Moreira, P.T. Charles, I.L. Medintz, E.R. Goldman, M. Zeinali,
On the basis of data shown in Figs. 5 and 6A, the Stern-Volmer C.R. Taitt, TNT detection using multiplexed liquid array displacement
immunoassays, Anal. Chem. 78 (2006) 2279–2285.
plots of ECL and fluorescence quenching by TNT can be plotted as
[12] D. Wu, J.P. Singh, F.Y. Yueh, D.L. Monts, 2,4,6-trinitrotoluene detection by laser-
shown in Figs. 7a and 7b, respectively. The slopes of the Stern- photofragmentation-laser-induced fluorescence, Appl. Opt. 35 (1996)
Volmer plots for ECL and fluorescence quenching were calculated 3998–4003.
to be 2.0  104 M1 and 5.2  103 M1, respectively. The quenching [13] T. Naddo, Y. Che, W. Zhang, K. Balakrishnan, X. Yang, M. Yen, J. Zhao, J.S. Moore,
L. Zang, Detection of explosives with a fluorescent nanofibril film, J. Am. Chem.
constant (Kq) calculated with the Stern-Volmer constants and the Soc. 129 (2007) 6978–6979.
lifetime of the excited state Ru(bpy)32+* species in MeCN (0.943 ms [14] M. Altamirano, L. Garcia-Villada, M. Agrelo, L. Sanchez-Martin, L. Martin-Otero,
[72]) came out to be 2.1 1010 M1 s1 for ECL quenching and A. Flores-Moya, M. Rico, V. Lopez-Rodas, E. Costas, A novel approach to
improve specificity of algal biosensors using wild-type and resistant mutants:
5.5  109 M1 s1 for fluorescent quenching. The Stern-Volmer An application to detect TNT, Biosens. Bioelectron. 19 (2004) 1319–1323.
plots suggest that the ECL quenching efficiency is about four times [15] S. Tao, G. Li, J. Yin, Fluorescent nanofibrous membranes for trace detection of
of the fluorescence quenching efficiency. This is not surprising, TNT vapor, J. Mater. Chem. 17 (2007) 2730–2736.
[16] U. Narang, P.R. Gauger, A.W. Kusterbeck, F.S. Ligler, Multianalyte detection
because the ECL quenching processes involve pathways that are using a capillary-based flow immunosensor, Anal. Biochem. 255 (1998) 13–19.
associated with both electron-transfer reactions (Eqs. (8)-(9)) and [17] A. Narayanan, O.P. Varnavski, T.M. Swager, T. Goodson, III, Multiphoton
a fluorescence collisional (dynamic) [71] quenching step as shown fluorescence quenching of conjugated polymers for TNT detection, J. Phys.
Chem. C 112 (2008) 881–884.
in Eq. (14). Quantitatively, the first two pathways contribute about [18] W.J. Buttner, M. Findlay, W. Vickers, W.M. Davis, E.R. Cespedes, S. Cooper, J.W.
three quarters of the overall ECL quenching efficiency. As a result, Adams, In situ detection of trinitrotoluene and other nitrated explosives in
ECL quenching is more efficient than fluorescence quenching for soils, Anal. Chim. Acta 341 (1997) 63–71.
[19] A. Hilmi, J.H.T. Luong, A.-L. Nguyen, Development of electrokinetic capillary
the system reported in this paper.
electrophoresis equipped with amperometric detection for analysis of
þ TNT explosive compounds, Anal. Chem. 71 (1999) 873–878.
RuðbpyÞ3 2þ ! RuðbpyÞ3 2þ ð14Þ [20] M. Riskin, R. Tel-Vered, O. Lioubashevski, I. Willner, Ultrasensitive surface
plasmon resonance detection of trinitrotoluene by a bis-aniline-cross-linked
au nanoparticles composite, J. Am. Chem. Soc. 131 (2009) 7368–7378.
4. Conclusions [21] P. Singh, T. Onodera, K. Matsumoto, N. Miura, K. Toko, Highly sensitive surface
plasmon resonance sensor on nanoscale bioactive surfaces for specific
detection of tri-nitro toluene, Mater. Res. Soc. Symp. Proc 951E (2007) Paper #:
The high explosive compound 2,4,6-trinitrotoluene (TNT) was 0-E0907-0912.
used as an ECL quenching agent in the Ru(bpy)32+/TPrA system. The [22] P. Singh, T. Onodera, Y. Mizuta, K. Matsumoto, N. Miura, K. Toko, Novel dnp-klh
protein conjugate surface for sensitive detection of TNT on spr immunosensor,
TNT was found to quench both ECL and fluorescence emission of Ru
Sens. Mater. 19 (2007) 261–273.
(bpy)32+* in MeCN. It was realized that the Ru(bpy)32+/TPrA ECL [23] J. Bowen, L.J. Noe, B.P. Sullivan, K. Morris, V. Martin, G. Donnelly, Gas-phase
was quenched by depleting Ru(bpy)3+ and TPrA free radical from detection of trinitrotoluene utilizing a solid-phase antibody immobilized on a
gold film by means of surface plasmon resonance spectroscopy, Appl.
the system which otherwise would have produced ECL light as well
Spectrosc. 57 (2003) 906–914.
 S. Parajuli et al. / Electrochimica Acta 180 (2015) 196–201
[24] A.A. Strong, D.I. Stimpson, D.U. Bartholomew, T.F. Jenkins, J.L. Elkind, Detection
of trinitrotoluene (TNT) extracted from soil using a surface plasmon resonance
(SPR)-based sensor platform, Proc. SPIE-Int. Soc. Opt. Eng. 3710 (1999)
362–372.
[25] D.R. Shankaran, K.V. Gobi, T. Sakai, K. Matsumoto, T. Imato, K. Toko, N. Miura, A
novel surface plasmon resonance immunosensor for 2,4,6-trinitrotoluene
(TNT) based on indirect competitive immunoreaction: A promising approach
for on-site landmine detection, IEEE Sens. J. 5 (2005) 616–621.
[26] Y. Mizuta, T. Onodera, P. Singh, K. Matsumoto, N. Miura, K. Toko, Highly
sensitive detection of TNT using a poly(amidoamine) dendron-based SPR
immunosensor, Sens. Mater. 22 (2010) 193–200.
[27] T. Kawaguchi, D.R. Shankaran, S.J. Kim, K. Matsumoto, K. Toko, N. Miura,
Surface plasmon resonance immunosensor using au nanoparticle for detection
of TNT, Sens. Actuators, B 133 (2008) 467–472.
[28] K. Matsumoto, A. Torimaru, S. Ishitobi, T. Sakai, H. Ishikawa, K. Toko, N. Miura,
T. Imato, Preparation and characterization of a polyclonal antibody from rabbit
for detection of trinitrotoluene by a surface plasmon resonance biosensor,
Talanta 68 (2005) 305–311.
[29] J. Yinon, H.G. Boettger, W.P. Weber, Negative ion mass spectrometry. New
analytical method for detection of trinitrotoluene, Anal. Chem. 44 (1972)
2235–2237.
[30] N.L. Sanders, S. Kothari, G. Huang, G. Salazar, R.G. Cooks, Detection of
explosives as negative ions directly from surfaces using a miniature mass
spectrometer, Anal. Chem. 82 (2010) 5313–5316.
[31] Y. Zhang, X. Ma, S. Zhang, C. Yang, Z. Ouyang, X. Zhang, Direct detection of
explosives on solid surfaces by low temperature plasma desorption mass
spectrometry, Analyst 134 (2009) 176–181.
[32] Y.-M. Zhou, W. Jin, H.-W. Chen, R.-F. Xu, Rapid detection of trace trinitrotoluene
on various surfaces using desorption electrospray ionization mass
spectrometry, Fenxi Huaxue 35 (2007) 621.
[33] Y. Song, H. Chen, R.G. Cooks, Reactivity of acetonyl anion with nitroaromatics:
An atmospheric pressure chemical ionization study, Rapid Commun. Mass
Spectrom. 19 (2005) 3493–3499.
[34] J.F. Garcia-Reyes, J.D. Harper, G.A. Salazar, N.A. Charipar, Z. Ouyang, R.G. Cooks,
Detection of explosives and related compounds by low-temperature plasma
ambient ionization mass spectrometry, Anal. Chem. 83 (2011) 1084–1092.
[35] J. Yinon, Field detection and monitoring of explosives, TrAC, Trends Anal. Chem
21 (2002) 292–301.
[36] R. Batlle, H. Carlsson, E. Holmgren, A. Colmsjo, C. Crescenzi, On-line coupling of
supercritical fluid extraction with high-performance liquid chromatography
for the determination of explosives in vapor phases, J. Chromatogr. A 963
(2002) 73–82.
[37] R. Batlle, H. Carlsson, P. Tollbaeck, A. Colmsjoe, C. Crescenzi, Enhanced
detection of nitroaromatic explosive vapors combining solid-phase
extraction-air sampling, supercritical fluid extraction, and large-volume
injection-GC, Anal. Chem. 75 (2003) 3137–3144.
[38] M.E. Walsh, Determination of nitroaromatic, nitramine, and nitrate ester
explosives in soil by gas chromatography and an electron capture detector,
Talanta 54 (2001) 427–438.
[39] A. Bromberg, R.A. Mathies, Homogeneous immunoassay for detection of TNT
and its analogues on a microfabricated capillary electrophoresis chip, Anal.
Chem. 75 (2003) 1188–1195.
[40] R.J.I. Jerez, M.d.R. Balaguera, A. Cabanzo, l.C.M.E. de, S.P. Hernandez-Rivera,
Enhanced raman scattering of nitro-explosives on nanoparticles substrates:
Au-ag alloy, tin oxide, and scandium oxide, Proc. SPIE-Int. Soc. Opt. Eng. 6201
(2006) 62.012G/62011-62012G/62018.
[41] A.M. Chamoun-Emanuelli, O.M. Primera-Pedrozo, M.A. Barreto-Caban, J.I.
Jerez-Rozo, S.P. Hernandez-Rivera, Enhanced raman scattering of TNT on
nanoparticles substrates: Ag, Au and bimetallic Au/Ag colloidal suspensions,
ACS Symp. Ser. 1016 (2009) 217–232.
[42] J.I. Jerez-Rozo, O.M. Primera-Pedrozo, M.A. Barreto-Caban, S.P. Hernandez-
Rivera, Enhanced raman scattering of 2,4,6-TNT using metallic colloids, IEEE
Sens. J. 8 (2008) 974–982.
[43] X. Wang, S. Chang, J. Yang, J. Tan, H. Jia, H. Yin, X. Li, G. Peng, Detection of TNT in
acetone using raman spectroscopic signature, Proc. SPIE 6622 (2008) 662.219/
662211-662219/662218.
[44] K. Kneipp, Y. Wang, R.R. Dasari, M.S. Feld, B.D. Gilbert, J. Janni, J.I. Steinfield,
Near-infrared surface-enhanced raman scattering of trinitrotoluene on
colloidal gold and silver, Spectrochim. Acta, Part A 51 A (1995) 2171–2175.
[45] J.K. Lokhnauth, N.H. Snow, Stir-bar sorptive extraction and thermal
desorption-ion mobility spectrometry for the determination of trinitrotoluene
and 1,3,5-trinitro-l,3,5-triazine in water samples, J. Chromatogr., A 1105 (2006)
33–38.
[46] S.E. Klassen, P. Rodacy, R. Silva, Reactant ion chemistry for detection of TNT,
RDX, and PETN using an ion mobility spectrometer, Sandia National
Laboratories, 1997, 2015 pp. 1–26.
201
[47] S.A. Trammell, F. Velez, P.T. Charles, A. Kusterbeck, Electrochemical detection
of 2,4,6-trinitrotoluene using interdigitated array electrodes, Anal. Lett. 41
(2008) 2634–2645.
[48] F. Wang, W. Wang, B. Liu, Z. Wang, Z. Zhang, Copolypeptide-doped polyaniline
nanofibers for electrochemical detection of ultratrace trinitrotoluene, Talanta
79 (2009) 376–382.
[49] R.G. Bozic, A.C. West, R. Levicky, Square wave voltammetric detection of 2,4,6-
trinitrotoluene and 2,4-dinitrotoluene on a gold electrode modified with self-
assembled monolayers, Sens. Actuators, B 133 (2008) 509–515.
[50] E.R. Cespedes, S.S. Cooper, W.M. Davis, W.J. Buttner, W.C. Vickers, In situ
detection of TNT contamination using electrochemical sensors in cone
penetrometer system, Proc. SPIE-Int. Soc. Opt. Eng. 2367 (1995) 33–42.
[51] A.M. O'Mahony, J. Wang, Nanomaterial-based electrochemical detection of
explosives: A review of recent developments, Analytical Methods (2013) .
[52] T.L. Pittman, B. Thomson, W. Miao, Ultrasensitive detection of TNT in soil,
water, using enhanced electrogenerated chemiluminescence, Anal. Chim. Acta
632 (2009) 197–202.
[53] R. Wilson, C. Clavering, A. Hutchinson, Paramagnetic bead based enzyme
electrochemiluminescence immunoassay for TNT, J. Electroanal. Chem 557
(2003) 109–118.
[54] R. Wilson, C. Clavering, A. Hutchinson, Electrochemiluminescence enzyme
immunoassays for TNT and pentaerthyritol tetranitrate, Anal. Chem. 75 (2003)
4244–4249.
[55] J. McCall, C. Alexander, M.M. Richter, Quenching of electrogenerated
chemiluminescence by phenols, hydroquinones, catechols, and
benzoquinones, Anal. Chem. 71 (1999) 2523–2527.
[56] H. Zheng, Y. Zu, Highly efficient quenching of coreactant electrogenerated
chemiluminescence by phenolic compounds, J. Phys. Chem. B 109 (2005)
16047–16051.
[57] A. Shaw, P. Lindhome, R.L. Calhoun, Electrogenerated chemiluminescence
(ECL) quenching of Ru(bpy)32+ by the explosives TATP and tetryl, J.
Electrochem. Soc. 160 (2013) H782–H786.
[58] P. Lindhome, R.L. Calhoun, Detection of tetryl by electrogenerated
chemiluminescence (ECL) quenching of Ru(bpy)32+, ECS Trans 50 (2013) 1–7.
[59] J. Wang, Electrochemical sensing of explosives, in: J. Yinon (Ed.),
Counterterrorist detection techniques of explosives, Elsevier B. V., Amsterdam,
2007, pp. 91–108 Chapter 4.
[60] H.-X. Zhang, A.-M. Cao, J.-S. Hu, L.-J. Wan, S.-T. Lee, Electrochemical sensor for
detecting ultratrace nitroaromatic compounds using mesoporous SiO2-
modified electrode, Anal. Chem. 78 (2006) 1967–1971.
[61] N.P. Saravanan, S. Venugopalan, N. Senthilkumar, P. Santhosh, B. Kavita, H.G.
Prabu, Voltammetric determination of nitroaromatic and nitramine explosives
contamination in soil, Talanta 69 (2006) 656–662.
[62] J. Wang, R.K. Bhada, J. Lu, D. MacDonald, Remote electrochemical sensor for
monitoring TNT in natural waters, Anal. Chim. Acta 361 (1998) 85–91.
[63] A. Hilmi, J.H.T. Luong, A.-L. Nguyen, Determination of explosives in soil and
ground water by liquid chromatography-amperometric detection, J.
Chromatogr. A 844 (1999) 97–110.
[64] H. Sohn, R.M. Calhoun, M.J. Sailor, W.C. Trogler, Detection of TNT and picric acid
on surfaces and in seawater by using photoluminescent polysiloles, Angew.
Chem. Int. Ed. 40 (2001) 2104–2105.
[65] R.Y. Lai, A.J. Bard, Electrogenerated chemiluminescence. 70. The application of
ECL to determine electrode potentials of tri-n-propylamine, its radical cation,
and intermediate free radical in MeCN/benzene solutions, J. Phys. Chem. A 107
(2003) 3335–3340.
[66] W. Miao, J.-P. Choi, A.J. Bard, Electrogenerated chemiluminescence 69: The tris
(2,20 -bipyridine) ruthenium(II), (Ru(bpy)32+)/Tri-n-propylamine (TPrA)
system revisited - a new route involving TPrA+. Cation radicals, J. Am. Chem.
Soc. 124 (2002) 14478–14485.
[67] W. Miao, Electrogenerated chemiluminescence and its biorelated applications,
Chem. Rev. 108 (2008) 2506–2553.
[68] D.J. Rosado Jr., W. Miao, Q. Sun, Y. Deng, Electrochemistry and electrogenerated
chemiluminescence of all-trans conjugated polymer poly[distyrylbenzene-b-
(ethylene oxide)]s, J. Phys. Chem. B 110 (2006) 15719–15723.
[69] W. Miao, A.J. Bard, Electrogenerated chemiluminescence. 72. Determination of
immobilized DNA and C-reactive protein on Au(111) electrodes using tris(2,2'-
bipyridyl) ruthenium(II) labels, Anal. Chem. 75 (2003) 5825–5834.
[70] M.W. Legenza, C.J. Marzzacco, Rate constant for fluorescence quenching. An
undergraduate experiment using the spectronic 20, J. Chem. Edu 54 (1977)
183.
[71] P.P.H. Cheng, D. Silvester, G. Wang, G. Kalyuzhny, A. Douglas, R.W. Murray,
Dynamic and static quenching of fluorescence by 1-4 nm diameter gold
monolayer-protected clusters, J. Phys. Chem. B 110 (2006) 4637–4644.
[72] J.V. Caspar, T.J. Meyer, Photochemistry of tris(2,20 -bipyridine) ruthenium(2+)
ion (Ru(bpy)32+). Solvent effects, J. Am. Chem. Soc. 105 (1983) 5583–5590.