CN108693172A - Ascorbic acid electrogenerated chemiluminescence assay method - Google Patents
Ascorbic acid electrogenerated chemiluminescence assay method Download PDFInfo
- Publication number
- CN108693172A CN108693172A CN201810893801.3A CN201810893801A CN108693172A CN 108693172 A CN108693172 A CN 108693172A CN 201810893801 A CN201810893801 A CN 201810893801A CN 108693172 A CN108693172 A CN 108693172A
- Authority
- CN
- China
- Prior art keywords
- electrode
- gold nano
- ascorbic acid
- nano cluster
- electrogenerated chemiluminescence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
Abstract
The present invention discloses a kind of ascorbic acid electrogenerated chemiluminescence assay method.The ascorbic acid electrogenerated chemiluminescence assay method of gold nano cluster probe of the present invention is to prepare high quantum production rate gold nano cluster electrogenerated chemiluminescence probe using reduction method, using nano material of manganese dioxide as electrogenerated chemiluminescence quencher, restore electrochemiluminescence signal using the redox reaction of ascorbic acid and manganese dioxide, realizes the measurement of Ascorbic Acid content.1.0 × 10-9~1.0×10-2 The logarithm of ascorbic acid concentrations and opposite electrogenerated chemiluminescence Strength Changes value are in a linear relationship in the concentration range of mol/L, and detection is limited to 2.56 × 10-10mol/L.Also, the present invention is easy to operate, high sensitivity, reproducibility and selectivity are good, thus has preferable application prospect.
Description
Technical field
The present invention relates to a kind of ascorbic acid electrogenerated chemiluminescence assay methods of gold nano cluster probe, belong to analysisization
And field of nanometer technology.
Background technology
Ascorbic acid(ascorbic acid)Also known as vitamin C, it is widely present in food and plant tissue, is one
Important antioxidant is planted, and one of the important vitamin for safeguarding body normal physiological function.Ascorbic acid wide participation body
A series of interior metabolism and redox reaction, and contribute to the cytoplasm such as rubber polymer member and glutinous polysaccharide, in adrenal cortex
The synthesis of hormone, vivo oxidation are also indispensable in restoring.The shortage of human body ascorbic acid can lead to scurvy, therefore fight bad
Hematic acid it is sensitive and selectively detect medicine and field of food all have significance.
The common method for measuring ascorbic acid has enzyme assay, spectrophotometry, red, orange, green, blue, yellow (ROGBY), chemoluminescence method and glimmering
Optical analysis etc..More demanding experiment condition and operating technology mostly, some method and steps are loaded down with trivial details, are unfavorable for quickly analyzing.
More in addition by the method that color is analyzed also due to sample to be tested is per se with color influences the result measured.Electricity
It is a kind of new analysis method for being combined chemiluminescence with electrochemistry and growing up to cause chemiluminometry, has the back of the body
The features such as scape signal is low, selectivity is good, high sensitivity, wide low with the detection limit range of linearity, be widely used in clinical diagnosis,
Drug, immune, food quality and other materials measurement.In recent years, the electrogenerated chemiluminescence based on quantum dot or nanocluster
Analytic approach, because with that background signal is low, high sensitivity, controllability are good, reagent can recycle and analyze test scope is wide etc.
Advantage is widely used in fields such as biology, medicine and environment.Gold nano cluster as novel nano illuminator,
With nontoxic, good water solubility, large specific surface area, preparation condition be mild, surface is easily modified, and with spies such as special photoelectric properties
Point, oneself is in the fields extensive use such as biological medicine, clinical analysis, sensing detection and catalysis.
The present invention is using gold nano cluster as electrogenerated chemiluminescence probe, using manganese dioxide nano-plates as quencher, based on anti-
Redox reaction between bad hematic acid and manganese dioxide establishes highly sensitive one kind, high selection, quick ascorbic acid and measures
New method.
Invention content
The purpose of the present invention is to provide a kind of using gold nano cluster as the ascorbic acid of electrogenerated chemiluminescence probe measurement
Method.
To achieve the goals above, the present invention uses following technical scheme:It is a kind of based on the anti-bad of gold nano cluster probe
Hematic acid electrogenerated chemiluminescence assay method, it is characterized in that:First in electrode face finish gold nano cluster, prepared by reduction method
High quantum production rate gold nano cluster electrogenerated chemiluminescence probe, and further in electrode face finish nano material of manganese dioxide,
As electrogenerated chemiluminescence quencher, so by modified electrode immerse the sample containing ascorbic acid in, ascorbic acid with
Redox reaction occurs for manganese dioxide, the electrochemiluminescence signal of modified electrode is acquired, according to electrochemiluminescence signal
Change realize ascorbic acid measurement.
The reduction method prepares high quantum production rate gold nano cluster electrogenerated chemiluminescence probe and uses following electrochemistry also
It is prepared by former method or chemical reduction method:
(1)Electrochemical reducing:Using three-electrode system, using gold nano cluster modified glassy carbon electrode as working electrode, platinum filament electricity
Extremely to electrode, Ag/AgCl is reference electrode, and above-mentioned electrode is inserted into 0.1 mol/L pH, 7.4 phosphate buffer solutions,
Apply negative potential voltage within the scope of -1.4 V of V ~ -2, carry out 5 h of min ~ 1 of constant potential reduction treatment, obtains electrochemical luminescence gold
Nanocluster probe;
(2)Chemical reduction method:Gold nano cluster modified glassy carbon electrode is immersed in 0.1 ~ 1 mol/L sodium borohydride solutions reaction 5
The min of min ~ 30 obtain the gold nano cluster probe modification electrode of chemical reduction method preparation.
The gold nano cluster is functional modification gold nano cluster, using N- acetylation-L-cysteines-gold nano group
Cluster or bovine serum albumin(BSA)-gold nano cluster.
The method in electrode face finish nano material of manganese dioxide is electrochemical deposition method:Using three electrode bodies
System handles gold nano cluster modified electrode as working electrode using reduction method, and platinum electrode is to electrode, and Ag/AgCl is reference electricity
Pole, using I-t methods, 1:The KMnO of 1 V/V4-H2SO4In solution, electrochemical deposition manganese dioxide, sedimentation potential is -0.1 V
~ -0.5 V, time are the s of 100 s ~ 600, cleaning, nitrogen drying.
The electrochemiluminescence signal method of the acquisition modified electrode is as follows:It is tested using three-electrode system, with
Modified electrode is working electrode, and platinum electrode is to electrode, and Ag/AgCl is reference electrode, and buffer solution is that phosphate-buffered is molten
Liquid or Tris-HCl buffer solutions, electrolyte used are KCl or KNO3;Above-mentioned electrode is inserted into anti-altogether containing over cure acid ion
In the buffer solution for answering agent, electrochemical luminescence test is carried out using step pulse method, initial potential is 0 V, burst length 10
S, termination current potential are -2 V, and the burst length is 1 s, and photomultiplier high pressure is set as 600 ~ 800 V, detects electrogenerated chemiluminescence
Radiation signal.
The pH value of the sample of the ascorbic acid is 8.0;Ascorbic acid and manganese dioxide occur redox reaction when
Between be 4 min.
The phosphate buffer solution or Tris-HCl buffer solutions of the electrochemiluminescence signal of above-mentioned acquisition modified electrode
PH value be 7.4, persulfuric acid ion concentration be 0.1 mol/L.
The logarithm of above-mentioned electrogenerated chemiluminescence Strength Changes value and ascorbic acid concentrations is 1.0 × 10-9~1.0×10-2
It is in good linear relationship in the range of mol/L, detection is limited to 2.56 × 10-10 mol/L。
Specifically, to achieve the goals above, the present invention is using technical solution in detail below:Gold nano cluster is modified
On glass-carbon electrode, high quantum production rate gold nano cluster electrogenerated chemiluminescence probe is prepared by reduction method, and then in electrode table
Face electro-deposition nano material of manganese dioxide, and further immerse modified electrode in the sample solution containing ascorbic acid, acquisition
The electrochemiluminescence signal of modified electrode restores electroluminescent chemistry using the redox reaction of ascorbic acid and manganese dioxide and sends out
Optical signal realizes the measurement of ascorbic acid according to the change of electrochemiluminescence signal.
Above-mentioned gold nano cluster probe is prepared by following methods:(1)Electrochemical reducing:It is gone back using three-electrode system
Original, using gold nano cluster modified glassy carbon electrode as working electrode, platinum electrode is to electrode, and Ag/AgCl is reference electrode, will be upper
It states electrode to be inserted into buffer solution, applies different negative potential voltages, carry out constant potential reduction treatment, obtain electrochemical luminescence Jenner
Rice cluster probe;(2)Chemical reduction method:Gold nano cluster solution is reacted to a timing with certain density sodium borohydride solution
Between, eccentric cleaning obtains gold nano cluster probe.Or gold nano cluster modified glassy carbon electrode is steeped in certain density hydroboration
Sodium solution reacts certain time, obtains the gold nano cluster probe modification electrode of chemical reduction method preparation.
The gold nano cluster material is functional modification gold nano cluster, such as:N- acetylations-L-cysteine-gold
Nanocluster, bovine serum albumin(BSA)-gold nano cluster etc..
The method in electrode surface electro-deposition nano material of manganese dioxide is:Using three-electrode system, with reduction
It is working electrode that method, which handles gold nano cluster modified electrode, and platinum electrode is to electrode, and Ag/AgCl is reference electrode, using I-T
Method, in KMnO4-H2SO4In solution, nano material of manganese dioxide modified electrode is made in potentiostatic electrodeposition.
The electrochemiluminescence signal is acquired by following methods:It is tested using three-electrode system, to modify glass
Carbon electrode is working electrode, and platinum electrode is to electrode, and Ag/AgCl is reference electrode, and the insertion of above-mentioned electrode is contained coreaction
In the buffer solution of agent.Using step pulse method, photomultiplier high pressure is set as the V of 600 V ~ 800, detects working electrode
The electrochemiluminescence signal that surface generates.
It is of the present invention a kind of based on the Methods For Quantitative Estimation of Ascorbic Acid that gold nano cluster is electrogenerated chemiluminescence probe, packet
Include following steps:1)By glass-carbon electrode Al2O3Powder polishes, polishes successively, until smooth mirror surface, then it is sequentially placed into HNO3Solution,
It is cleaned by ultrasonic in absolute ethyl alcohol and deionized water, N2Drying;2)Gold nano cluster probe solution is added dropwise in the glass carbon handled well
Electrode surface, drying at room temperature prepare high quantum production rate Jenner to get gold nano cluster probe modification glass-carbon electrode, by reduction method
Rice cluster electrogenerated chemiluminescence probe, and nano material of manganese dioxide/gold nano cluster modified electrode is further made;3)Using
Three-electrode system is tested, and using manganese dioxide/gold nano cluster probe modification glass-carbon electrode as working electrode, platinum electrode is
To electrode, Ag/AgCl is reference electrode, and above-mentioned electrode is inserted into the buffer solution containing ascorbic acid and coreagent, inspection
Survey its electrochemiluminescence signal value.Using step pulse method, photomultiplier high pressure is set as the V of 600 V ~ 800, detection
The electrochemiluminescence signal that working electrode surface generates;4)With electrochemiluminescence signal changing value Ascorbic Acid concentration
Logarithm maps to obtain standard curve, is 1.0 × 10 in ascorbic acid concentrations-9~1.0×10-2 Vitamin C in the range of mol/L
The logarithm of acid concentration is in good linear relationship with electrogenerated chemiluminescence intensity value, and detection is limited to 2.56 × 10-10 mol/L。
Above-mentioned coreagent is over cure acid ion, and concentration range is 0.01 ~ 1 mol/L, preferably 0.1 mol/L.
The buffer solution is phosphate buffer or Tris-HCl buffer solutions, the added electrolyte in buffer solution
For KCl or KNO3, a concentration of 0.01 ~ 1 mol/L.
The specific technical solution that the present invention uses is as follows:
(One)The preparation of N-acetyl-L-cysteine-gold nano cluster
N-acetyl-L-cysteine-gold nano cluster synthesis step is as follows:By the sodium hydroxide of a concentration of 0.1 ~ 0.8 mol/L
The N-acetyl-L-cysteine of a concentration of 0.02 ~ 0.18 mol/L is added to a concentration of 0.01 ~ 0.1 g/L chlorauric acid solutions
In solution, mixing is placed on 20 ~ 70 DEG C of water bath with thermostatic control isothermal reactions 0 ~ 3.5 hour.Wait for dialysis purification processing after reaction,
N-acetyl-L-cysteine-gold nano cluster aqueous solution is obtained, N-acetyl-L-cysteine-Jenner can be obtained after freeze-drying
Rice cluster powder.
(Two)The preparation of gold nano cluster modified electrode
By glass-carbon electrode with 1.0 μm, 0.3 μm, 0.05 μm of Al2O3Powder polishes, polishes successively, until smooth mirror surface, then according to
It is secondary to be put into HNO3Solution(1:1), be cleaned by ultrasonic 3 minutes in absolute ethyl alcohol and deionized water, N2Drying.Take 5 μ L gold nano clusters
Aqueous solution is added dropwise in the glassy carbon electrode surface handled well, and drying at room temperature is to get gold nano cluster modified glassy carbon electrode.
(Three)It is prepared by gold nano cluster probe
(1)Electrochemical reducing:It is restored using three-electrode system, using gold nano cluster modified glassy carbon electrode as work electricity
Pole, platinum electrode are to electrode, and Ag/AgCl is reference electrode, and above-mentioned electrode is inserted into buffer solution, apply negative potential voltage
(Within the scope of -1.4 V of V ~ -2), 5 h of min ~ 1 of constant potential reduction treatment are carried out, electrochemical luminescence gold nano cluster probe is obtained.
(2)Chemical reduction method:It is anti-that gold nano cluster modified glassy carbon electrode is immersed in 0.1 ~ 1 mol/L sodium borohydride solutions
Answer 5 ~ 30 min(It is preferred that 0.1 mol/L sodium borohydride solutions react 5 min), obtain the gold nano cluster of chemical reduction method preparation
Probe modification electrode.
(Four)Nano material of manganese dioxide method of modifying
Nano material of manganese dioxide is modified using electrochemical deposition method, is as follows:Using three-electrode system, with reduction method
Processing gold nano cluster modified electrode is working electrode, and platinum electrode is to electrode, and Ag/AgCl is reference electrode, using I-t
Method, in KMnO4-H2SO4(1:1 V/V)In solution, electrochemical deposition manganese dioxide, sedimentation potential is -0.1 V of V ~ -0.5, is sunk
The product time is the s of 100 s ~ 600, and preferably 300 s are rinsed well with distilled water later, and nitrogen drying is spare.
(Five)Electrochemiluminescence signal acquisition method
By polishing electrode, then it is sequentially placed into HNO3It is cleaned by ultrasonic in solution, absolute ethyl alcohol and deionized water, N2Drying.Using three
Electrode system is tested, and using modified glassy carbon electrode as working electrode, platinum electrode is to electrode, and Ag/AgCl is reference electrode,
Above-mentioned electrode is inserted into the buffer solution containing coreagent.Using step pulse method, initial potential is 0 V, burst length
For 10 s, termination current potential is -2 V, and the burst length is 1 s.Photomultiplier high pressure is set as the V of 600 V ~ 800, detects work
Make the electrochemiluminescence signal of electrode surface generation.
The electrode is glass-carbon electrode, screen printing electrode or ITO electrode etc..Above-mentioned coreagent be over cure acid group from
Son, concentration range are 0.01 ~ 1 mol/L, preferably 0.1 mol/L.The buffer solution is phosphate buffer or Tris-HCl
Buffer solution, added electrolyte is KCl or KNO in buffer solution3, a concentration of 0.01 ~ 1 mol/L, preferably 0. 1 mol/
L。
(Six)The measurement of ascorbic acid
Above-mentioned manganese dioxide/gold nano cluster modified electrode is immersed and reacts 4 ~ 10 min in ascorbic acid solution, is used after taking-up
Distilled water is rinsed well, N2Drying.The electrochemiluminescence signal that collecting work electrode surface generates, is believed with electrogenerated chemiluminescence
Standard curve is drawn in the mapping of number Ascorbic Acid concentration.
Compared with prior art, beneficial effects of the present invention are:
The present invention is using high quantum production rate gold nano cluster probe as electrogenerated chemiluminescence material, with nano-manganese dioxide for electroluminescentization
Luminescence quenchers are learned, restore the detection that its electrochemiluminescence signal realizes ascorbic acid using ascorbic acid.The present invention is fought
The detection sensitivity of bad hematic acid is high, easy to operate, specificity is good, accuracy is high.Also, the present invention is at low cost, it is simple, steady to make
It is qualitative it is good, the range of linearity is wide(1.0×10-9~1.0×10-2 mol/L), detection limits low(2.56×10-10mol/L), have very
Good market value.
Description of the drawings
Fig. 1 is electrogenerated chemiluminescence-time plot of gold nano cluster probe modification glass-carbon electrode.
Fig. 2 is electrogenerated chemiluminescence-time plot of manganese dioxide/gold nano cluster probe modification glass-carbon electrode.
Fig. 3 is the electrogenerated chemiluminescence-of manganese dioxide/gold nano cluster probe modification glass-carbon electrode after ascorbic acid is added
Time plot.
Relational graphs of the Fig. 4 between electrogenerated chemiluminescence Strength Changes and buffer solution pH.
Fig. 5 is electrogenerated chemiluminescence Strength Changes and the relational graph between the reaction time.
Linear relationship charts of the Fig. 6 between electrogenerated chemiluminescence Strength Changes and ascorbic acid concentrations logarithm.
Specific implementation mode
The present invention is further elaborated in the following with reference to the drawings and specific embodiments, and the present invention is not limited thereto.
Embodiment 1
A concentration of 0.5 mol/L of 0.6 mL are added into the N-acetyl-L-cysteine solution of a concentration of 0.08 mol/L of 4 mL
Sodium hydroxide and a concentration of 20 mg/mL chlorauric acid solutions of 0.4 mL, mixing be placed in 37 DEG C of thermostatic water baths and be incubated 3 h.
Reaction solution after reaction carries out dialysis purification processing with the bag filter that retention molecule is 3500, obtains half Guangs of N- acetyl-L-
Propylhomoserin-gold nano cluster aqueous solution obtains N-acetyl-L-cysteine-gold nano cluster powder after freeze-drying.
Embodiment 2
By the glass-carbon electrode of diameter 3mm with 1.0 μm, 0.3 μm and 0.05 μm of Al2O3Powder polishes successively, polishing, until light
Sliding minute surface, then it is sequentially placed into HNO3Solution, absolute ethyl alcohol are cleaned by ultrasonic 3 minutes in deionized water, N2Drying.Take 5 μ L embodiments
1 N-acetyl-L-cysteine-gold nano cluster the solution prepared is added dropwise in the glassy carbon electrode surface handled well, drying at room temperature,
Obtain N-acetyl-L-cysteine-gold nano cluster modified glassy carbon electrode.By N-acetyl-L-cysteine-gold nano cluster modification
Glass-carbon electrode, which is immersed in 0.1 mol/L sodium borohydride solutions, reacts 5 min, obtains gold nano cluster probe modification electrode.It will
Above-mentioned gold nano cluster probe modification electrode is working electrode, and platinum filament is to electrode, and Ag/AgCl electrodes are reference electrode, are inserted into
In 0.1 mol/L pH, 7.4 phosphate buffer solutions containing 0.1 mol/L potassium peroxydisulfates and 0.1 mol/L KCl.Using
Step pulse method, initial potential are 0 V, and the burst length is 10 s, and termination current potential is -2 V, and the burst length is 1 s.Photomultiplier transit
Pipe high pressure is set as 750 V, and the electrochemiluminescence signal that detection working electrode surface generates obtains electrochemical luminescence signals
(See Fig. 1).
Embodiment 3
By the glass-carbon electrode of 3 mm of diameter with 1.0 μm, 0.3 μm, 0.05 μm of Al2O3Powder polishes successively, polishing, until light
Sliding minute surface, then it is sequentially placed into HNO3Solution, absolute ethyl alcohol are cleaned by ultrasonic 3 minutes in deionized water, N2Drying.Take 5 μ L embodiments
The gold nano cluster solution of the 1 N-acetyl-L-cysteine protection prepared is added dropwise in the glassy carbon electrode surface handled well, and room temperature is dry
It is dry, and further the electrode is immersed in 0.1 mol/L sodium borohydride solutions and is reacted 5 minutes, obtain gold nano cluster probe
Modified glassy carbon electrode.Using three-electrode system, using gold nano cluster probe modification glass-carbon electrode as working electrode, platinum electrode is
To electrode, Ag/AgCl is reference electrode, using chronoamperometry, in KMnO4-H2SO4(1:1 V/V)In solution, electro-deposition two
Manganese oxide, current potential are -0.2 V, and sedimentation time is 300 s, and obtained electrode is that manganese dioxide/gold nano cluster modifies glass carbon
Electrode is rinsed well with distilled water later, N2Gas gently dries up spare.Above-mentioned electrode is inserted into and contains 0.1 mol/L persulfuric acid
In 0.1 mol/L pH, 7.4 phosphate buffer solutions of potassium and 0.1 mol/L KCl.Using step pulse method, initial potential
For 0 V, the burst length is 10 s, and termination current potential is -2 V, and the burst length is 1 s.Photomultiplier high pressure is set as 750 V,
The electrochemiluminescence signal that working electrode surface generates is detected, electrochemical luminescence signals are obtained(See Fig. 2).
Embodiment 4
Manganese dioxide prepared by embodiment 3/gold nano cluster probe modification electrode is working electrode, and platinum filament is to electrode, Ag/
AgCl electrodes are reference electrode, are inserted into and contain 0.1 mol/L ascorbic acid, 0.1 mol/L potassium peroxydisulfates and 0.1 mol/L
0.1 mol/L, pH 8.0 of KCl is in phosphate buffer solution.Using step pulse method, initial potential is 0 V, burst length
For 10 s, termination current potential is -2 V, and the burst length is 1 s.Photomultiplier high pressure is set as 750 V, after reacting 4 min, inspection
Survey the electrochemiluminescence signal that working electrode surface generates, solution of the obtained electrochemical luminescence signals than not adding ascorbic acid
It significantly increases(See Fig. 3).
Embodiment 5
Manganese dioxide prepared by embodiment 3/gold nano cluster probe modification electrode is working electrode, and platinum filament is to electrode, Ag/
AgCl electrodes are reference electrode, are inserted into and contain 0.1 mol/L ascorbic acid, 0.1 mol/L potassium peroxydisulfates and 0.1 mol/L
In the 0.1 mol/L phosphate buffer solutions of KCl.Using step pulse method, initial potential is 0 V, and the burst length is 10 s, eventually
Only current potential is -2 V, and the burst length is 1 s.Photomultiplier high pressure is set as 750 V, after reacting 4 min, in phosphate-buffered
The pH value of solution detects working electrode surface generation respectively under conditions of being 5.0,6.0,7.0,7.4,8.0,8.4,9.0 and 10.0
Electrochemiluminescence signal, obtain optimal ph be 8.0(See Fig. 4).
Embodiment 6
Manganese dioxide prepared by embodiment 3/gold nano cluster probe modification electrode is working electrode, and platinum filament is to electrode, Ag/
AgCl electrodes are reference electrode, are inserted into and contain 0.1 mol/L ascorbic acid, 0.1 mol/L potassium peroxydisulfates and 0.1 mol/L
In the phosphate buffer solution of 0.1 mol/L, pH 7.4 of KCl.Using step pulse method, initial potential is 0 V, burst length
For 10 s, termination current potential is -2 V, and the burst length is 1 s.Photomultiplier high pressure is set as 750 V, respectively in reaction 0
Electroluminescentization that working electrode surface generates is detected after min, 1 min, 2 min, 3 min, 4 min, 6 min, 8 min and 10 min
Luminous signal is learned, it is 4 min to obtain optimum reacting time(See Fig. 5).
Embodiment 7
Manganese dioxide prepared by embodiment 3/gold nano cluster probe modification electrode is working electrode, and platinum filament is to electrode, Ag/
AgCl electrodes are reference electrode, are inserted into and contain various concentration ascorbic acid, 0.1 mol/L potassium peroxydisulfates and 0.1 mol/L KCl
0.1 mol/L, pH 7.4 phosphate buffer solution in.Using step pulse method, initial potential is 0 V, and the burst length is
10 s, termination current potential are -2 V, and the burst length is 1 s.Photomultiplier high pressure is set as 600 V, after reacting 4 min, slow
It rushes in solution and detects the electroluminescent chemistry hair of working electrode surface generation under the conditions of being added existing for various concentration ascorbic acid respectively
Optical signal.It is 1.0 × 10 in ascorbic acid concentrations-9~1.0×10-2 In the range of mol/L the logarithm of glutathione concentrations with
Electrogenerated chemiluminescence intensity value is in good linear relationship, and detection is limited to 2.56 × 10-10mol/L(See Fig. 6).
The foregoing is merely the exemplary embodiments of the present invention, are not intended to limit the invention, all essences in the present invention
Any modification made by within refreshing and principle, equivalent replacement and improvement etc., should all be included in the protection scope of the present invention.
Claims (8)
1. a kind of ascorbic acid electrogenerated chemiluminescence assay method based on gold nano cluster probe, it is characterized in that:First in electricity
Pole surface modifies gold nano cluster, prepares high quantum production rate gold nano cluster electrogenerated chemiluminescence probe by reduction method, goes forward side by side
One step is in electrode face finish nano material of manganese dioxide, as electrogenerated chemiluminescence quencher, and then by modified electrode
It immerses in the sample containing ascorbic acid, with manganese dioxide redox reaction occurs for ascorbic acid, acquires the electricity of modified electrode
Chemiluminescence signal is caused, the measurement of ascorbic acid is realized according to the change of electrochemiluminescence signal.
2. a kind of ascorbic acid electrogenerated chemiluminescence measurement side based on gold nano cluster probe according to claim 1
Method, it is characterized in that the reduction method, which prepares high quantum production rate gold nano cluster electrogenerated chemiluminescence probe, uses following electrochemistry
It is prepared by reduction method or chemical reduction method:
(1)Electrochemical reducing:Using three-electrode system, using gold nano cluster modified glassy carbon electrode as working electrode, platinum filament electricity
Extremely to electrode, Ag/AgCl is reference electrode, and above-mentioned electrode is inserted into 0.1 mol/L pH, 7.4 phosphate buffer solutions,
Apply negative potential voltage within the scope of -1.4 V of V ~ -2, carry out 5 h of min ~ 1 of constant potential reduction treatment, obtains electrochemical luminescence gold
Nanocluster probe;
(2)Chemical reduction method:Gold nano cluster modified glassy carbon electrode is immersed in 0.1 ~ 1 mol/L sodium borohydride solutions reaction 5
The min of min ~ 30 obtain the gold nano cluster probe modification electrode of chemical reduction method preparation.
3. a kind of ascorbic acid electrogenerated chemiluminescence measurement side based on gold nano cluster probe according to claim 1,2
Method, it is characterized in that the gold nano cluster is functional modification gold nano cluster, using N- acetylations-L-cysteine-Jenner
Rice cluster or bovine serum albumin(BSA)-gold nano cluster.
4. a kind of ascorbic acid electrogenerated chemiluminescence measurement side based on gold nano cluster probe according to claim 1
Method, it is characterized in that the method in electrode face finish nano material of manganese dioxide is electrochemical deposition method:Using three electrodes
System handles gold nano cluster modified electrode as working electrode using reduction method, and platinum electrode is to electrode, and Ag/AgCl is reference
Electrode, using I-t methods, 1:The KMnO of 1 V/V4-H2SO4In solution, electrochemical deposition manganese dioxide, sedimentation potential is -0.1
The V of V ~ -0.5, time are the s of 100 s ~ 600, cleaning, nitrogen drying.
5. a kind of ascorbic acid electrogenerated chemiluminescence measurement side based on gold nano cluster probe according to claim 1
Method, it is characterized in that the electrochemiluminescence signal method of the acquisition modified electrode is as follows:It is tested using three-electrode system,
Using modified electrode as working electrode, platinum electrode is to electrode, and Ag/AgCl is reference electrode, and buffer solution is phosphate-buffered
Solution or Tris-HCl buffer solutions, electrolyte used are KCl or KNO3;Above-mentioned electrode is inserted into and is total to containing over cure acid ion
In the buffer solution of reactant, electrochemical luminescence test is carried out using step pulse method, initial potential is 0 V, and the burst length is
10 s, termination current potential are -2 V, and the burst length is 1 s, and photomultiplier high pressure is set as 600 ~ 800 V, detects electroluminescent chemistry
Luminous radiation signal.
6. a kind of ascorbic acid electrogenerated chemiluminescence measurement side based on gold nano cluster probe according to claim 1
Method, it is characterized in that the pH value of the sample of the ascorbic acid is 8.0;With manganese dioxide redox reaction occurs for ascorbic acid
Time is 4 min.
7. a kind of ascorbic acid electrogenerated chemiluminescence measurement side based on gold nano cluster probe according to claim 5
Method, it is characterized in that the phosphate buffer solution or Tris-HCl buffer solutions of the electrochemiluminescence signal of acquisition modified electrode
PH value is 7.4, and persulfuric acid ion concentration is 0.1 mol/L.
8. a kind of ascorbic acid electrogenerated chemiluminescence measurement side based on gold nano cluster probe according to claim 1
Method, it is characterized in that the logarithm of electrogenerated chemiluminescence Strength Changes value and ascorbic acid concentrations is 1.0 × 10-9~1.0×10-2
It is in good linear relationship in the range of mol/L, detection is limited to 2.56 × 10-10 mol/L。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810893801.3A CN108693172A (en) | 2018-08-08 | 2018-08-08 | Ascorbic acid electrogenerated chemiluminescence assay method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810893801.3A CN108693172A (en) | 2018-08-08 | 2018-08-08 | Ascorbic acid electrogenerated chemiluminescence assay method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108693172A true CN108693172A (en) | 2018-10-23 |
Family
ID=63841566
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810893801.3A Pending CN108693172A (en) | 2018-08-08 | 2018-08-08 | Ascorbic acid electrogenerated chemiluminescence assay method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108693172A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110208233A (en) * | 2019-06-09 | 2019-09-06 | 福建医科大学 | The method of nano cupric oxide enhancing fluoremetry ascorbic acid |
| CN111157769A (en) * | 2020-01-06 | 2020-05-15 | 广州大学 | Electrochemiluminescence imaging system and imaging method thereof |
| CN112129747A (en) * | 2020-09-24 | 2020-12-25 | 广州大学 | Method for detecting L-cysteine by electrochemiluminescence |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102683044A (en) * | 2012-06-17 | 2012-09-19 | 兰州大学 | Combined electrode for super capacitor and preparation method thereof |
| CN102709058A (en) * | 2012-07-01 | 2012-10-03 | 吉林大学 | Method for preparing manganese dioxide-nickel hydroxide composite electrode materials of super capacitors |
| CN103337639A (en) * | 2013-06-24 | 2013-10-02 | 太原理工大学 | Preparation method of carbon nano tube array/carbon fiber fabric integrated three-dimensional porous air electrode |
| CN104597019A (en) * | 2015-01-26 | 2015-05-06 | 郑州大学 | In-situ composite system based on carbon quantum dot/manganese dioxide nanometer sheet layer and using method for detecting content of glutathione |
| CN106706607A (en) * | 2017-02-07 | 2017-05-24 | 福建医科大学 | High-quantum-yield electrochemiluminescence gold nano-cluster probe and preparation method of high-quantum-yield electrochemiluminescence gold nano-cluster probe |
| CN106872546A (en) * | 2017-02-07 | 2017-06-20 | 福建医科大学 | Electrochemical reducing prepares high quantum production rate electrochemical luminescence gold nano cluster probe |
-
2018
- 2018-08-08 CN CN201810893801.3A patent/CN108693172A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102683044A (en) * | 2012-06-17 | 2012-09-19 | 兰州大学 | Combined electrode for super capacitor and preparation method thereof |
| CN102709058A (en) * | 2012-07-01 | 2012-10-03 | 吉林大学 | Method for preparing manganese dioxide-nickel hydroxide composite electrode materials of super capacitors |
| CN103337639A (en) * | 2013-06-24 | 2013-10-02 | 太原理工大学 | Preparation method of carbon nano tube array/carbon fiber fabric integrated three-dimensional porous air electrode |
| CN104597019A (en) * | 2015-01-26 | 2015-05-06 | 郑州大学 | In-situ composite system based on carbon quantum dot/manganese dioxide nanometer sheet layer and using method for detecting content of glutathione |
| CN106706607A (en) * | 2017-02-07 | 2017-05-24 | 福建医科大学 | High-quantum-yield electrochemiluminescence gold nano-cluster probe and preparation method of high-quantum-yield electrochemiluminescence gold nano-cluster probe |
| CN106872546A (en) * | 2017-02-07 | 2017-06-20 | 福建医科大学 | Electrochemical reducing prepares high quantum production rate electrochemical luminescence gold nano cluster probe |
Non-Patent Citations (4)
| Title |
|---|
| GAIZKA GARAI-IBABE ET AL: "《Peroxidase-Mimicking DNAzyme Modulated Growth of CdS Nanocrystalline Structures in Situ through Redox Reaction: Application to Development of Genosensors and Aptasensors》", 《ANALYTICAL CHEMISTRY》 * |
| 周伯劲: "《试剂化学》", 30 June 1980, 广东省韶关市科学技术协会 * |
| 姜香宇: "《基于内滤效应和功能核酸的荧光传感体系设计》", 《中国优秀硕士学位论文全文数据库》 * |
| 曲绵域，高云秋主编: "《现代运动医学诊疗手册》", 31 May 1997, 北京：北京医科大学、中国协和医科大学联合出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110208233A (en) * | 2019-06-09 | 2019-09-06 | 福建医科大学 | The method of nano cupric oxide enhancing fluoremetry ascorbic acid |
| CN111157769A (en) * | 2020-01-06 | 2020-05-15 | 广州大学 | Electrochemiluminescence imaging system and imaging method thereof |
| CN112129747A (en) * | 2020-09-24 | 2020-12-25 | 广州大学 | Method for detecting L-cysteine by electrochemiluminescence |
| CN112129747B (en) * | 2020-09-24 | 2022-10-04 | 广州大学 | Method for detecting L-cysteine by electrochemiluminescence |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106706607B (en) | High quantum production rate electrogenerated chemiluminescence gold nano cluster probe and preparation method thereof | |
| CN106872546B (en) | Electrochemical reducing prepares high quantum production rate electrochemical luminescence gold nano cluster probe | |
| CN108663357A (en) | A kind of atriphos electrogenerated chemiluminescence assay method | |
| CN105675689B (en) | A kind of preparation method and application of the hydrogen peroxide without enzyme sensor based on vulcanization molybdenum composite material structure | |
| CN108802139A (en) | A kind of electrogenerated chemiluminescence method of detection glutathione | |
| CN108693172A (en) | Ascorbic acid electrogenerated chemiluminescence assay method | |
| CN108827948A (en) | Acid phosphatase electrogenerated chemiluminescence measuring method based on gold nano cluster probe | |
| CN107044978B (en) | Glutathione electrogenerated chemiluminescence measuring method based on gold nano cluster probe | |
| CN110057801A (en) | A kind of ratio fluorescent probe and its hydrogen peroxide and glucose detection application based on aggregation-induced emission property | |
| CN110243891A (en) | A kind of label-free homogeneous electrochemical biosensor method detecting cancer cell | |
| WO2022120923A1 (en) | Single-cell electrochemical sensor based on functionalized nanoprobe, and application thereof | |
| CN109142748A (en) | Human prostate specific antigen detection method and its kit | |
| CN108414599A (en) | Preparation method of the copper nano particles without enzyme electrochemical glucose sensor | |
| CN110823970A (en) | Electrochemical detection method for rapidly determining content of L-cystine in acidic solution | |
| CN109164090A (en) | The electrochemiluminescdetection detection method and its kit of tumor necrosis factor α | |
| CN105738451A (en) | Direct electron transfer type glucose biosensor and preparation method and application | |
| CN108872209A (en) | Alkaline phosphatase assay method based on nanogold cluster electrogenerated chemiluminescence probe | |
| CN113155917A (en) | Construction method of photo-assisted bipolar self-powered sensor for detecting ochratoxin A or aflatoxin B1 | |
| CN106959323B (en) | A method of detection lysozyme | |
| CN105806832A (en) | Preparation method and application of hydrogen peroxide sensor based on double functions of electrochemiluminescence and photoelectrochemistry | |
| CN107037021A (en) | A kind of fluorescence copper nano-particle of poly- adenine dna template and its preparation method and application | |
| CN105784687A (en) | Preparation method and application of autoluminescence-exictation-based hydrogen peroxide electrochemical sensor | |
| CN110095518B (en) | Preparation method and application of hydrogen sulfide photoelectrochemical sensor | |
| CN113267544A (en) | Method for monitoring hydrogen peroxide concentration in living cell based on three-dimensional sensing and electrode | |
| CN113237929A (en) | Construction method of flexible sensing electrode modified by nano particles |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181023 |
|
| RJ01 | Rejection of invention patent application after publication |