CN101532961B - Method for detecting electrochemical luminescence - Google Patents
Method for detecting electrochemical luminescence Download PDFInfo
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- CN101532961B CN101532961B CN2009100668036A CN200910066803A CN101532961B CN 101532961 B CN101532961 B CN 101532961B CN 2009100668036 A CN2009100668036 A CN 2009100668036A CN 200910066803 A CN200910066803 A CN 200910066803A CN 101532961 B CN101532961 B CN 101532961B
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- pyridine ruthenium
- butyl diethanolamine
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Abstract
The present invention relates to a method for detecting electrochemical luminescence. A luminescence system of electrochemical luminescence detection is a pyridyl ruthenium/butyl diethanolamine luminescence system. Pyridyl ruthenium and butyl diethanolamine are mixed in phosphate buffer solution with pH between 7 and 9 to be simulated to be luminescent by an electrochemical method and analyzed. The concentration range of the pyridyl ruthenium is 1*10<-10> to 1mmol/L. The concentration range of the butyl diethanolamine is 0.05-50mmol/L. The electrochemical luminous efficiency of the pyridyl ruthenium/butyl diethanolamine luminescence system is high, and the concentration of co-reactant butyl diethanolamine can be very low. When the concentration of the pyridyl ruthenium is 1 mumol/L, and the concentration of the butyl diethanolamine is 25mmol/L, under the same condition, the electrochemical luminous intensity of the pyridyl ruthenium/butyl diethanolamine luminescence system at a gold electrode is approximately 1.3 times stronger than the electrochemical luminous intensity of the pyridyl ruthenium/butyl diethanolamine luminescence system. The invention can be widely applied to marker analysis.
Description
Technical field
The present invention relates to the method that a kind of electrochemiluminescence detects.
Background technology
The luminesceence analysis of pyridine ruthenium electrochemical is a labeled analysis technology of new generation.Pyridine ruthenium label generation electrochemiluminescence that prior art is based on the tripropyl amine (TPA) of high concentration and low concentration reacts and carries out bioanalysis.But there is following main weak point in pyridine ruthenium/tripropyl amine (TPA) electrochemiluminescsystem system.The reaction of first and third propylamine is slower, has limited the sensitivity that detects; The second, very the tripropyl amine (TPA) of high concentration could be realized highly sensitive detection; Three, tripropyl amine (TPA) solubleness in neutral solution is not so good, generally will adopt high concentrations of phosphoric acid salt very that concentration is dissolved in neutral phosphate buffered solution up to the tripropyl amine (TPA) of 100mmol/L; At last, the tripropyl amine (TPA) volatility is bigger, has bigger peculiar smell and certain toxicity.List of references (Angew.Chem.Int.Ed, 2007,46,421-424) compared the electrochemiluminescence intensity of tripropyl amine (TPA) and a series of amine, find the electrochemiluminescence intensity of the electrochemiluminescence intensity of di-n-butyl monoethanolamine and pyridine ruthenium much larger than tripropyl amine (TPA) and pyridine ruthenium, and advantage such as the di-n-butyl monoethanolamine also has environmental friendliness, solubleness is good, working concentration is low, the range of linearity of detection pyridine ruthenium concentration is wide.But the electrochemiluminescence intensity of di-n-butyl monoethanolamine and pyridine ruthenium is not the strongest, still needs to seek and the stronger co-reactant of pyridine ruthenium electrochemical luminous intensity.
Summary of the invention
The purpose of this invention is to provide the method that a kind of electrochemiluminescence detects.It is applied to the electrochemiluminescence labeled analysis.
A kind of electrochemical luminous detection method, be applicable to all electrochemical light-emitting detectors, it is characterized in that, described electrochemical light-emitting detector comprises optical instrument and electrochemical apparatus two large divisions, optical instrument is the chemiluminescence intensity analyzer, and electrochemical apparatus is to use CH series electrochemical analyser; The luminescence system that electrochemiluminescence detects is to be luminescent substance with the pyridine ruthenium, the pyridine ruthenium also is a detected material simultaneously, the normal-butyl diethanolamine is the pyridine ruthenium/normal-butyl diethanolamine luminescence system of co-reactant, pyridine ruthenium and normal-butyl diethanolamine are mixed in buffer solution, analyze with the electrochemical method stimulated luminescence; The working concentration scope of described pyridine ruthenium is 1 * 10
-10～1mmol/L, its structural formula is:
The working concentration scope of described normal-butyl diethanolamine is 0.05～50mmol/L; Described buffer solution is the phosphate buffered solution of pH 7～9, and the structural formula of described normal-butyl diethanolamine is as follows:
Beneficial effect: the method that a kind of electrochemiluminescence provided by the invention detects is applied to the electrochemiluminescence labeled analysis.The electrochemiluminescence efficient height of normal-butyl diethanolamine and pyridine ruthenium, employed co-reactant normal-butyl diethanolamine concentration can be very low.When pyridine ruthenium concentration is that the concentration of 1 μ mol/L and normal-butyl diethanolamine is under the situation of 25mmol/L, under the same terms, pyridine ruthenium on gold electrode/normal-butyl diethanolamine luminescence system is than the electrochemiluminescence intensity strong about 1.3 times (as Fig. 3) of the pyridine ruthenium of reporting in the document/di-n-butyl monoethanolamine luminescence system.
Description of drawings
Embodiment
Embodiment 1: use electrochemical workstation and BPCL Weak-luminescence measuring instrument to detect.Fixedly normal-butyl diethanolamine concentration is 0.05mmol/L, detected material pyridine ruthenium concentration is 1 μ mol/L, buffer solution is the 0.1mol/L phosphate of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.5V, apply luminous detection voltage 800V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect, the luminous signal mean intensity that obtains is 22.33.
Embodiment 2: use electrochemical workstation and BPCL Weak-luminescence measuring instrument to detect.Fixedly normal-butyl diethanolamine concentration is 15mmol/L, and buffer solution is the 0.1mol/L phosphate of pH 7.5, adds 4.0 * 10
-10The detected material pyridine ruthenium of mol/L is a working electrode with the gold electrode, applies the step potential of 1.35V, applies luminous detection voltage 700V simultaneously, detects pyridine ruthenium luminous signal, finishes electrochemiluminescence and detects, and the mean intensity that obtains pyridine ruthenium luminous signal is 68.
Embodiment 3: use electrochemical workstation and BPCL Weak-luminescence measuring instrument to detect.Fixedly the concentration of normal-butyl diethanolamine is 1mmol/L, detected material pyridine ruthenium concentration is 1mmol/L, be dissolved in the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.5V, apply luminous detection voltage 500V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect, the luminous signal mean intensity that obtains is 6751.
Embodiment 4: use electrochemical workstation and BPCL Weak-luminescence measuring instrument to detect.Fixedly normal-butyl diethanolamine concentration is 25mmol/L, detected material pyridine ruthenium concentration is 1 μ mol/L, buffer solution is in the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, and current potential is swept to+1.5V from 0, and speed is 0.1V/s, apply luminous detection voltage 700V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect, the luminous signal mean intensity that obtains is 3123.33.
Embodiment 5: use electrochemical workstation and BPCL Weak-luminescence measuring instrument to detect.Fixedly normal-butyl diethanolamine concentration is 50mmol/L, detected material pyridine ruthenium concentration is 1 μ mol/L, buffer solution is in the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.35V, apply luminous detection voltage 700V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect, the luminous signal mean intensity that obtains is 5204.
Embodiment 6: use MPI-A type capillary electrophoresis electrochemical light-emitting detector to detect.Fixedly normal-butyl diethanolamine concentration is 10mM, detected material pyridine ruthenium concentration is 1 μ M, buffer solution is in the 0.1M phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.5V, apply suitable luminous detection voltage 800V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect, the luminous signal mean intensity that obtains is 8820.67.
Claims (7)
1. an electrochemical luminous detection method is characterized in that, electrochemical light-emitting detector is to use electrochemical workstation and BPCL Weak-luminescence measuring instrument, or uses MPI-A type capillary electrophoresis electrochemical light-emitting detector; The luminescence system that electrochemiluminescence detects is to be luminescent substance with the pyridine ruthenium, and the normal-butyl diethanolamine is the pyridine ruthenium/normal-butyl diethanolamine luminescence system of co-reactant, and the pyridine ruthenium also is a detected material simultaneously; Detected material pyridine ruthenium and normal-butyl diethanolamine are mixed in buffer solution, analyze with the electrochemical method stimulated luminescence; The working concentration scope of described pyridine ruthenium is 1 * 10
-10～1mmol/L, its structural formula is:
The working concentration scope of described normal-butyl diethanolamine is 0.05～50mmol/L; Described buffer solution is the phosphate buffered solution of pH 7～9, and the structural formula of described normal-butyl diethanolamine is as follows:
2. electrochemical luminous detection method according to claim 1 is characterized in that, electrochemical light-emitting detector is to use electrochemical workstation and BPCL Weak-luminescence measuring instrument; Normal-butyl diethanolamine concentration is 0.05mmol/L, detected material pyridine ruthenium concentration is 1 μ mol/L, buffer solution is the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.5V, apply luminous detection voltage 800V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect.
3. electrochemical luminous detection method according to claim 1 is characterized in that, electrochemical light-emitting detector is to use electrochemical workstation and BPCL Weak-luminescence measuring instrument; Fixedly normal-butyl diethanolamine concentration is 15mmol/L, and buffer solution is the 0.1mol/L phosphate buffered solution of pH 7.5, adds 4.0 * 10
-10The detected material pyridine ruthenium of mol/L is a working electrode with the gold electrode, applies the step potential of 1.35V, applies luminous detection voltage 700V simultaneously, detects pyridine ruthenium luminous signal, finishes electrochemiluminescence and detects.
4. electrochemical luminous detection method according to claim 1 is characterized in that, electrochemical light-emitting detector is to use electrochemical workstation and BPCL Weak-luminescence measuring instrument; Fixedly the concentration of normal-butyl diethanolamine is 1mmol/L, detected material pyridine ruthenium concentration is 1mmol/L, be dissolved in the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.5V, apply luminous detection voltage 500V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect.
5. electrochemical luminous detection method according to claim 1 is characterized in that, electrochemical light-emitting detector is to use electrochemical workstation and BPCL Weak-luminescence measuring instrument; Fixedly normal-butyl diethanolamine concentration is 25mmol/L, detected material pyridine ruthenium concentration is 1 μ mol/L, be dissolved in the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, current potential is swept to+1.5V from 0, and speed is 0.1V/s, applies luminous detection voltage 700V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect.
6. electrochemical luminous detection method according to claim 1 is characterized in that, electrochemical light-emitting detector is to use electrochemical workstation and BPCL Weak-luminescence measuring instrument; Fixedly normal-butyl diethanolamine concentration is 50mmol/L, detected material pyridine ruthenium concentration is 1 μ mol/L, be dissolved in the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.35V, apply luminous detection voltage 700V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect.
7. electrochemical luminous detection method according to claim 1, it is characterized in that, electrochemical light-emitting detector is to use MPI-A type capillary electrophoresis electrochemical light-emitting detector, fixedly normal-butyl diethanolamine concentration is 10mmol/L, pyridine ruthenium concentration is 1 μ mol/L, be dissolved in the 0.1mol/L phosphate buffered solution of pH 7.5, with the gold electrode is working electrode, apply the step potential of 1.5V, apply luminous detection voltage 800V simultaneously, detect pyridine ruthenium luminous signal, finish electrochemiluminescence and detect.
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