Addis Ababa University

College of Health Sciences School of Pharmacy

Department of Pharmaceutical Chemistry and Pharmacognosy

Seminar II
Recent Advances in Electrochemiluminescence
By: Yegezu M.
1
July, 2022
Addis Ababa
CONTENTS

 Introduction
 Method
 Instrumentation
 Recent advancements
 Application
 Conclusion

2
INTRODUCTION
 The word luminescence comes from the Latin ‘lumen’,
which means light.
 It was first introduced as ‘luminescenz’ by the physicist E.
Wiedemann in 1888 to describe
 ‘All those phenomena of light which are not solely conditioned
by the rise in temperature’, as opposed to incandescence.
 Luminescence corresponds to cold light whereas incandescence is hot
light.
 Luminescence is a light closely related to spectroscopy
which study the absorption and emission of radiation by
matter
 Discovered by English physicist G.C. Stokes who identified the
luminescence phenomenon and formulated stokes law:
 Wavelength of emitted light is greater than that of the excited radiation 3
CONT---
 There are multiple luminescent methods:
 Photoluminescence (fluorescence, phosphorescence)
 Absorption of photons
 Chemiluminescence (CL)
 Chemical excitation of compounds
 Bioluminescence (BL)
 Biochemical reaction in a living organisms (Luminous organisms)

4
CONT---
 Electrochemiluminescence (ECL)
 Electrogenerated chemical excitation
 Crystalloluminescence
 Crystallization reaction
 Electroluminescence (EL)
 Radiative recombination of electrons and holes in a material (usually
a semiconductor) after an electrical current passes through the
material or a strong electric field is applied
 Radio chemiluminescence
 Radiation-induced chemical excitation

5
CONT
 Lyoluminescence
 Excitation induced by dissolution of an irradiated or other energy-
donating solid
 Sonoluminescence (SL)
 Excitation of compounds by ultra-sonication, either by energy
transfer from the intrinsic SL centers of water or by chemical
excitation by hydroxyl radicals and atomic hydrogen
 Mechanoluminescence
 Mechanical action on a solid

6
 ELECTROCHEMILUMINESCENCE
 Luminescence produced during electrochemical reactions in
solutions.
 The emission of light (luminescence) with limited emission of heat
 ECL is the combination of three words:
 Electro(electrical stimulation), chem (chemical reactions), and
luminescence (light).
 Initiated by an electrochemical triggering reaction ‘E’, continues with a
homogeneous chemical step ‘C’, which leads to the formation of the
excited state, and ends with a luminescent step, i.e. light emission, ‘L’.
 ECL is a way of changing electrical energy into radiative
energy

7
CONT---

8
CONT---

9
COMPONENTS OF ECL
 Luminophores: can be classified into three
 Inorganic ([Ru(bpy)3]2+ and its derivatives (Cyclometalated
iridium(III) complexes and metal complexes)
 Organic systems (polycyclic aromatic hydrocarbons (PAHs)),
Luminol ((5-amino-2, 3-dihydro1, 4-phthalazinedione) z
 Nanomaterials

 Electrode devices
 Where redox reaction occurs

 Optical devices
10
 Avalanche photodiodes (APD), PMTs and CCD
LUMINOPHORE
 [Ru(bpy)3]2+ has been the most used label because
a) Can undergo ECL reactions in an aqueous buffer medium,
which is perfect for biological matrices
b) It is not troubled by oxygen so degassing to remove the
dissolved oxygen is not necessary
c) Fully reversible one-electron transfer so the light emission
will be at its finest
d) Can be sufficiently connected to biological samples,
antibody, or antigen utilizing distinct bio-conjugates
strategies

11
PRINCIPLES

 the generation of reactive intermediates from stable

precursors at the surface of the electrode.
 Intermediates react to produce excited states that emit light.

 This intermediates can be produced from the same

precursor i.e annihilation
 Requires the luminophore, solvent and supporting electrolyte
to obtain ECL

 Or using coreactant 12
ANNIHILATION
 A - e- → A•+ (oxidation at electrode)
A + e- → A•- (reduction at electrode)
A•- + A•+ → A• + A (excited-state formation)
A• → A + hν (light emission)

13
COREACTANT

 Is a sacrificial molecule that is consumed irreversibly due to

the bond-breaking reaction during the process whereas
 The luminophore is regenerated and ready to start a new ECL
cycle.

 The role of the coreactant is to provide energetic radicals

able to react with the luminophore in order to reach the
excited state.

14
CONT---
 Criteria’s for good coreactant:
 Must have good solubility in the reaction medium
 Electroactive

 The co-reactant and its redox must be free of any

electrochemical background in the swept potential as well as
any quenching effects which could lower the ECL intensity
 Good stability of the intermediate species of the co-reactant
 To allow enough time to react with luminophores to generate the
excited state
 The rapid rate of reaction between the reduced and oxidized
intermediates and luminophores 15
CONT---
 Coreactants have two mechanisms of reaction:
a. Oxidative-reduction co-reactants, where the electro-oxidation of
co-reactant yields intermediate species that are extremely
reducible.
 For example, oxalate ion (C2O42–), TPrA, and 2- (dibutylamino)
ethanol (DBAE)
 Oxalate ion (C O 2-) was the first coreactant discovered which produce
2 4
the strong reductant CO2•- up on oxidation in aqueous solution
 C2O42- - e- → [C2O4•-] → CO2•- + CO2
 The sequence of oxidative-reduction mechanism is:
a) The electrochemical oxidations at the electrode surface
b) Bond-breaking (or atom-transfer) reaction of the coreactant
giving a strong reducing radical and
c) The reduction of the oxidized luminophore by this coreactant 16
radical.
CONT---
b. Reductive-oxidation coreactants
 Electro-reduction of a coreactant yields intermediate species that are
extremely reducible such as peroxydisulfate and H2O2.
o The sequence of the reaction involves:
• Both luminophore and coreactant are first reduced.
• The reduced form of the coreactant undergoes a chemical step, such as,
for example, a bond cleavage, that generates a strong oxidizing radical.
• The electron-transfer reaction between the strong oxidizing coreactant
radical and the reduced luminophore is then exergonic enough to
populate the emissive state and to produce ECL

17
ADVANTAGES OF ECL
 Excellent sensitivity
 No radioisotopes are used

 Measurement is easy, inexpensive, simple and rapid

 The reagents are stable and relatively non-toxic

 Can achieve clinical quality data in a variety of samples

like
 Cell supernatant, serum, whole blood and plasma
 The entire reaction can be precisely controlled

18
ADVANCEMENTS IN ECL

 Time line of ECL

19
1. NANOMATERIALS
 Sensor probes made of materials like Au, Ag, Cu, Pt, Ni,
and alloy NCs have become more popular because of their
 High solubility, biocompatibility, size-dependent
luminescence, and catalysis.

 The ECL phenomenon of Au NCs/triethylamine (TEA)

system and Au NCs/potassium persulfate system was
found by Chen’s and Zhu’s group in 2011, respectively.
 They developed novel techniques for determining Pb2+,
dopamine, and hydrogen peroxide.
20
CONT---
 The metal NCs sensitivity was increased by the
following modifications:
 Valence state regulation
 Peroxidation
 Host-guest recognition
 Tuning ligand effects and
 Covalent bonding of co-reactants

 Potassium persulfate or hydrazine coreactants were used

for detection of
 Phenols, microRNAs, and dopamine

21
CONT---
 Carbon nanomaterials (CNMs) are among the most often
used nanomaterials due to their
 Eexcellent electrical conductivity
 Substantial surface area, and
 Variety of functionalisation options like electrochemical
aptasensors are being constructed
 By strong adsorption of aptamers to graphene and graphene oxide
(GO)

22
METAL-ORGANIC FRAMEWORK

 Are porous and crystalline materials built from metal

ions/clusters and organic linkers
 The features of MOFs such as
 Large surface area, tailorable structure and porosity, tunable
size and versatile functionality, make them preferable choice
for fields like
 Gas analysis, catalysis, separation, nano medical engineering
bioanalysis and biomedical research
23
APTASENSORS
 Aptamers (meaning “to fit”) is a synthetic single-stranded
DNA or RNA ligands with specificity similar to an
antibody
 Many unique properties of aptamers, such as
 Thecapability of structure switching, hybridization with css-
DNA, adsorption onto carbon materials, and nucleic acid
amplification, as well as susceptibility to the action of a specific
type of enzyme
 Have been utilized for the advancement of the different proof-of-
concepts in ECL aptasensors plat forms

24
LABEL FREE ECL
 A novel label-free strategy was pursued to amplify ECL signal
and improve detection sensitivity via the novel combination of
Au NPs, CNOs and CS film.
 Was fabricated for the detection of ß-2-Microglobulin (ß2M)

 A single luminophor dual-potential ECL system based on

NGQDs was used
 For detection of Co2+ ion no extra coreactants or labeling with an
accurate and reliable sensing system.
 The dissolved oxygen used as co-reactant and participated in anodic and 25
cathodic reactions to generate NGQDs
APPLICATION
1. Analytical application
 Nanomaterials based electrochemical aptasensors for food
analysis
 A combination of [Ru(bpy)3]2+ and proline (as coreactant)
was used to identify paracetamol.
 ECL sensor based on electrode coated with a vertically
ordered mesoporous silica nanochannel film (VMSF)
 Was used for sensitive detection of clindamycin

26
CONT---
 VMSF/APTES/3DG Tris(2,2'-bipyridyl) ruthenium (Ru
(bpy)32+)
 To detect environmental contaminant 4-chlorophenol and effective
antihistamines chlorpheniramine
 Ru(bpy)32+/AZA sensor
 To detect Azamethiphos organophosphate

27
DIAGNOSIS OF CANCER CELLS
 CDs@ZrHf-MOF
 To detect human epidermal growth factor receptor-2 (HER2)
and living cancer cells (MCF-7) concurrently
 Useful tool for the early identification of breast cancer
 A novel Ru(bpy)2(dcbpy)-doped 2D MOF nanoplate
(Ru@Zr12-BPDC)
 Employed to construct an aptasensor for ultrasensitive assay
of mucin 1(MUC1) which is biomarker for many cancers
 Prostate cancer, pancreatic cancer and breast cancer
 ZnMOF(Ru)
 For detecting breast cancer 1 gene (BRCA1)

28
CONT---
 ZnCdS@ZnS quantum dots (QDs)–based label-free ECL
immunosensor
 Used for sensitive aflatoxin B1 detection (AFB1)
 Highly hepatotoxic, oncogenic, teratogenic, mutagenic, and immuno-
toxic properties in human and domestic animals
 Ru(bpy)32+@UiO-66-NH2 nanocomposite material
 For sensitive detection of a breast cancer biomarker CA15-3

29
IMMUNOASSAY
 Is a widespread technique in bioanalytical applications for
diagnosing diseases, drug monitoring, food testing, and
environmental monitoring.
 Antibodies are used to detect an analyte
 The use of radioisotope in RIA and the use of an enzyme
in ELISA that has limited stability due to simple
denaturation and treatment under particular conditions
 Can be overcomed by ECLIA

30
POINT-OF CARE TESTING

31
FORENSIC APPLICATION
 A new, robust and feasible ECL sensor was developed
for the detection of
 Amphetamine type stimulating drugs such as
Methamphetamine (MA) with high sensitivity
 Combining electrochemical properties of the glassy
carbon electrode improved with the composite film
containing [Ru(bpy)3]3+in Nafion with amperometric
response
 Paves great advantage for selective determination of MA

32
CONCLUSION

 The rapid development of ECL in both fundamentals and

applications over the past several years has clearly
demonstrated that ECL
 Is a powerful tool for ultrasensitive biomolecule detection
and quantification

 The nanomaterials allow for

 Miniaturization of biosensors, low production cost, ease of
use, the possibility of mass production, use of small sample
33
volumes and high analytical performance.
CONCLUSION
 The demands will drive the active study in:
 High-throughput analysis, aptasensors, nanomaterials, development
of new coreactants, MOF and ECL light-emitting molecules,
miniaturization of instruments, ECL screening techniques, new
analytical strategies, and new applications of ECL.

 Using the label free ECL was also another recent mechanism
which avoids
 Using different labels that may have different effect on the process
and increase sensitivity and reproducibility
34
Thank you!

35