Chemiluminescence in Analytical Chemistry
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[93]. The sensor was stable up to 2 weeks and could be easily regenerated in water making it suitable for use in fermentation industries.
Table 2 lists some typical examples of non-enzyme-based CL sensors.
5. CL IMMUNOSENSORS
In immunoassays the main advantages of CL as a detection principle are the high sensitivity and rapidity of measurement without lengthy incubations. These advantages are more than potential so as to design a CL immunosensor. Aizawa [94] reported on a fiberoptic CL immunosensor by using an aromatic hydrocarbon and enzyme as labeling agents. The antibody was not directly immobilized on an optical fiber. Hara et al. [95] developed a fiberoptic immunosensor by immobilizing the antibody on an optical fiber. A metal-complex compound iron(III)- 2,9,16,23-tetrakis(chlorocarbonyl) phthalocyanine [TCCP-Fe(III)] was used as a CL catalyst. Competitive immunoassay was carried out by immersing the end of the optical fiber in a solution of human serum albumin (HSA) complexed with the metal complex. The flow rates for phosphate buffer, H2O2, and luminol were 1.50, 0.25, and, 0.25 mL/min, respectively. The HSA was determined in the 0.1– 100-mg/L concentration range, with a detection limit of 40 pg. The selectivity can be improved by utilization of a monoclonal antibody. A schematic flow diagram of a fiberoptic immunosensor designed by Hara et al. [95] is illustrated in Figure 6.
Zhang et al. [96] developed a FIA-CL immunosensor by immobilizing antibody on controlled pore glass filled into a column (0.2 5 cm). With this method, the time required per assay was 20 min compared to 20 h with conventional enzyme-linked immunosorbent assay (ELISA). Moreover, the whole assay could be proceeded automatically by control through a microprocessor. The technique was used for determination of hepatitis B surface antigen (HBsAg) in serum. Gatto-Menking et al. [97] developed an immunomagnetic ECL sensor for sensitive detection of biotoxoid and bacterial spores based on the CL from the ruthenium(III) trisbipyridal chelate-labeled reporter antibodies induced by a potential of 650–800 V. Detection limits of the order of fg were achieved for botulinus A, cholera β subunit, ricin, and staphylococcal enterotoxoid B.
Starodub et al. [98] studied different constructions and biomedical applications of immunosensors based on fiberoptic and enhanced CL. They discussed three different approaches of immobilization of one of the immunocomponents on the fiberoptic surface. Good results could be achieved by the use of a special membrane closely connected to the fiberoptic, with sensitivities compared to that obtained by the ELISA method but with a faster rate of analysis. The sensor was
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Figure 6 (A) Schematic flow diagram of a fiberoptic immunosensor. (a) Buffer solution;
(b) H2O2 solution; (c) luminol solution; (d) four-way cock; (e) cell; (f) optical fiber; (g) PMT; (h) photon-counter; (I) integrator; (P1, P2, P3), pumps. (B) The cell set with an optical fiber. (a) optical fiber; (b) silicone tube (3 mm od 1 mm id); (c) Teflon-made three-way joint (3 mm id); (d) immobilized antibody.
applied to determination of several antigens such as estradiol-17, α-2-interferon, chorionic gonadotropine, antibodies, and cells of Salmonella [99].
6. DNA SENSORS
A DNA optical sensor system was proposed based on the combination of sandwich solution hybridization, magnetic bead capture, FI, and CL for rapid detection of DNA hybridization [100]. Bacterial alkaline phosphatase (phoA) gene and hepatitis B virus (HBV) DNA were used as target DNA. A biotinylated DNA probe was used to capture the target gene onto the streptavidin-coated magnetic beads and a calf intestine alkaline phosphatase (CAP)-labeled DNA probe was used for subsequent enzymatic CL detection. The detection cycle was less than 30 min, excluding the DNA hybridization time, which was about 100 min. Both the phoA gene and HBV DNA could be detected at picogram or femtomole levels. Successive sample detection could be made by removing the magnetic field and using a washing step.
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DNA probes have been covalently immobilized onto the distal end of the optical fiber bundle in the construction of a novel CL DNA bisensor [101]. Hybridization of HRP-labeled complementary nucleotides to the immobilized probes was detected by enhanced CL. This approach was specific, sensitive, and available for diagnostic application with a detection limit of 0.1 ppm.
7. CONCLUSIONS
Analytical techniques using CL reactions in air or in solution as detection methods have received much attention in various fields owing to their extremely high sensitivity along with extra advantages such as the simple instrumentation required, fast dynamic response properties, wide calibration ranges, the easy coupling to various separation techniques such as HPLC and capillary electrophoresis (CE), and suitability for miniaturization in analytical chemistry. Most of these techniques, however, require several types of reagents, which are continuously delivered and thus wasted in large quantities. This is undesirable not only in view of the simplification of the detection device, but also because of cost, apart from environmental and resource considerations. Some efforts so far have been devoted to reduce the consumption of reagents, including controlled release, cyclic use, immobilization, and regeneration. These proceedings may be directed toward the construction of CL sensors. Compared with chemical sensors based on other sensing principles, very few CL sensors have been reported so far. Although one reason might be the fact that the CL technique is unfamiliar to most scientists, the consumption of reagents accompanying CL reactions, resulting in deterioration of the indicator phases or of sensors on prolonged use, could also be an important factor. Further developments should focus on improvement of the lifetime of this type of sensor and on improvement of selectivity by careful selection of suitable CL reaction systems. Miniaturization of CL sensors for special applications should also be studied to extend this new technique to life sciences as well as to specific areas.
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Appendix: Abbreviations
A/D |
Analog to digital |
AAS |
Atomic absorption spectrometry |
Ab |
Antibody |
ABAP |
2,2′-Azo-bis(2-aminodipropane)dihydrochloride |
ABCP |
2,2′-Azo-bis(2-cyanopropane) |
ABEI |
N-(4-Aminobutyl)-N-ethylisoluminol |
ABTS |
2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) |
AchE |
Acetylcholinesterase |
ACL |
Antioxidative capacity of lipid-soluble compounds |
ACP |
Antioxidative capacity of proteins |
ACR |
Rest of total antiradical capacity |
ACU |
Urate independent component of ACW |
ACW |
Antioxidative capacity of water-soluble compounds |
ADP |
Adenosine diphosphate |
AE |
Acridinium ester |
AFP |
α-Fetoprotein |
Ag |
Antigen |
ALP (or AP) |
Alkaline phosphatase |
AMP |
Adenosine monophosphate |
6-AMP |
6-Aminomethylphthalhydrazide |
AMPPD |
3-(2′-Spiroadamantane)-4-methoxy-4- |
|
(3″-phosphoryloxy)phenyl-1,2-dioxetane |
AMVN |
2,2′-Azo-bis (2,4-dimethylvaleronitrile) |
ANS |
8-Aniline-1-naphthalenesulfonic acid |
593
594 |
Appendix |
AP |
Aminophthalate anion |
APAN |
Atmospheric pressure active nitrogen |
APP-CLS |
Analyte pulse perturbation-chemiluminescence spectroscopy |
ARP |
Arthromyces rasomus peroxidase |
ASC |
Ascorbic acid |
ATP |
Adenosine triphosphate |
AVP |
Avalanche photodiode |
BCI |
5-Bromo-4-chloro-3-indolyl |
BHT |
2,6-Di-tert-butyl-4-methylphenol |
BL |
Bioluminescence |
Brij-35 |
Polyoxyethylene (23) dodecanol |
BSA |
Bovine serum albumin |
c.m.c. |
Critical micelle concentration |
CAP |
Calf alkaline phosphatase |
CAR |
Continuous-addition-of-reagent |
CAR-CLS |
Continuous-addition-of-reagent chemiluminescence |
|
spectroscopy |
CAS |
Catecholamines |
CAT |
Catechol |
CBI |
Cyanobenz[f] isoindole |
CCD |
Charge coupled device |
cDNA |
Complementary deoxyribonucleic acid |
CE |
Capillary electrophoresis |
CE-CL |
Capillary electrophoresis-chemiluminescence |
CEDAB |
Cetylethyldimethylammonium bromide |
CE-ECL |
Capillary electrophoresis-electrogenerated chemiluminescence |
CETAS |
Capillary electrophoresis micrototal analysis system |
CGE |
Capillary gel electrophoresis |
CGH |
Comparative genomic hybridization |
ChO |
Choline oxidase |
CIA |
Capillary ion analysis |
CID |
Charge injection device |
CIEEL |
Chemically initiated electron exchange luminescence |
CIEF |
Capillary isoelectric focusing |
CITP |
Capillary isotachophoresis |
CK |
Creatine kinase |
CL |
Chemiluminescence |
CMPI |
2-Chloro-1-methylpyridinium iodide |
CMV |
Cytomegalovirus |
CNN |
Computational neural network |
CPPO |
2,4,5-Trichloro-6-carbopentoxyphenyl oxalate |
CSTR |
Continuous stirring tank reactor |
Appendix |
595 |
CZE |
Capillary zone electrophoresis |
d(G)n |
Polydeoxyguanylic acid |
DBD-F |
4-(N,N-Dimethylaminosulphonyl)-7-fluoro-2,1,3- |
|
benzoxadiazole |
DBD-H |
4-(N,N-Dimethylaminosulphonyl)-7-hydrazino-2,1,3- |
|
benzoxadiazole |
DBPM |
N-[4-(6-Dimethylamino-2-benzofuranyl)phenyl]maleimide |
DCIA |
7-Dimethylamino-3-{4-[(iodoacetyl)amino]phenyl}-4- |
|
methylcoumarin |
DDDAB |
Didodecyldimethylammonium bromide |
DEA |
Diethanolamine |
DETBA |
1,3-Diethyl-2-thiobarbituric acid |
DFPO |
Bis(2,6-difluorophenyl)oxalate |
DIC |
Differential interference contrast |
DMF |
N,N′-Dimethylformamide |
DMQPH |
6,7-Dimethoxy-1-methyl-2(1H )-quinoxalinone-3- |
|
proprionylcarboxylic acid hydrazide |
DMS |
Dimethyl sulfide |
DMSO |
Dimethyl sulfoxide |
DNA |
Deoxyribonucleic acid |
DNPO |
Bis-(2,4-dinitrophenyl) oxalate |
DNS-Cl |
Dansyl chloride |
DNS-H |
Dansyl hydrazine |
DODAB |
Dioctadecyldimethylammonium bromide |
DPA |
9,10-Diphenylanthracene |
DPH |
4,5-Diaminophthalhydrazide |
DPPA |
4-[4,5-Di(2-pyridyl)-1 H-imidazol-2-yl]phenylboronic acid |
DSC |
N,N′-Disuccinimidyl carbodiimide |
DTDCI |
3,3′-Diethylthiadicarbocyanine iodide |
DVS |
Divinyl sulfone |
EC |
Electrochemical |
ECL |
Electrogenerated chemiluminescence |
ELISA |
Enzyme-linked immunosorbent assay |
EMMA |
Electrophoretically-mediated microanalysis |
EOF |
Electroosmotic flow |
EY |
Eosine Y |
FCLD |
Fluorine-induced chemiluminescence detector |
FHC |
Fluorohydrocarbon |
FIA |
Flow injection analysis |
FID |
Flame ionization detector |
Fl |
Fluorescein |
FL |
Fluorescence |