CN101059480A - Glassy carbon electrode modified electrochemical detection method for diethylstilbestrol - Google Patents
Glassy carbon electrode modified electrochemical detection method for diethylstilbestrol Download PDFInfo
- Publication number
- CN101059480A CN101059480A CN 200710040472 CN200710040472A CN101059480A CN 101059480 A CN101059480 A CN 101059480A CN 200710040472 CN200710040472 CN 200710040472 CN 200710040472 A CN200710040472 A CN 200710040472A CN 101059480 A CN101059480 A CN 101059480A
- Authority
- CN
- China
- Prior art keywords
- electrode
- diethylstilbestrol
- glass
- polyethyleneimine
- carbon electrode
- 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.)
- Granted
Links
Images
Landscapes
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention relates to an electrochemical check method of glass-carbon electrode decoration of estromon, belonging to electrochemical analysis technical field, comprising that electrode pretreatment, nanometer gold sol preparation, polyethylene imine-nanometer gold decorated glass carbon electrode preparation, more particularly comprising that preparing polyethylene imine decorated glass-carbon electrode and preparaing polyethylene imine nanometer gold decorated electrode, and using the polyethylene imine nanometer gold decorated electrode to test the content of estromon. The inventive decoration of the working electrode is simple and repeatable, while the decorated electrode has strong selective collection on estromon, better test repeatability and high sensitivity, while the check limit can reach 0. 001mug/g.
Description
Technical field
What the present invention relates to is a kind of detection method of electrochemical analysis technical field, specifically the electrochemical detection method of a kind of glass-carbon electrode of diethylstilbestrol modification.
Background technology
(diethylstilbestrol, (E)-3,4-Bis (4-hydroxyphenyl)-3-hexene DES) is born in the forties in 20th century to diethylstilbestrol, is first oral activated synthetic estrogen.Diethylstilbestrol in artificial estrogen be act on stronger a kind of.Medically be mainly used in the treatment estrogen deficiency, but, once be widely used in livestock breeding industry as growth accelerator because it has the promotion growth of animal, increases effect proteins deposited and that improve food conversion ratio.Diethylstilbestrol is mainly used in promoting the sexual maturity of the not obvious animal that oestruses on veterinary clinic, also available treatment retention of afterbirth, discharge stillborn foetus, treatment hysteritis etc.Since it is found that animal because of the edible feed that contains DES in tissue residual bring harm to human health since, overwhelming majority of countries forbids using DES in food animal in the world.China Ministry of Agriculture regulation in 2002, DES and salt thereof, ester formulation are forbidden all purposes in all food animals.But the illegal DES situation of adding occurs frequently on livestock breeding industry, thus set up fast, the unusual necessity of the sensitive and method that can realize on-the-spot detection diethylstilbestrol.At present to be used for the method for DES residue detection a lot of for the domestic and foreign literature report, mainly contain immunological method: as radioimmunology (RIA) enzyme linked immunosorbent assay (ELISA) etc.; Chromatographia method: as vapor-phase chromatography (GC), GC-MS(gas chromatography-mass spectrography) (GC-MS), high performance liquid chromatography (HPLC), thin-layered chromatography (TLC), gas-liquid chromatography (GLC) etc.; Other method: as radiation spectrophotometric method, electrochemical process, bioanalysis etc.In these analytical approachs, the vapor-phase chromatography of gas chromatography-mass spectrometry machine method, radioimmunology and charged sub-acquisition detector is at present existingly can measure the horizontal content detecting method of DES ppb effectively, delicately.But these analytical approachs all need large-sized analytic instrument, and operating process is loaded down with trivial details, can not realize on-the-spot the detection.The modified electrode of the present invention's development can be directly used in long-pending the reaching with the portable electrochemical sensor of richness of DES in the on-the-spot sample and assemble laggard capable on-the-spot test.
Find by prior art documents, Shenghui Zhang, Kangbing Wu and Shengshui Hu.Voltammetric determination of diethylstilbestrol at carbonpaste electrode using cetrylpyridine bromide as medium.Talanta, 2002,58:747-754 (Zhang Shenghui, Wu Kangbin, Hu Shengshui was in 2002,747 pages-754 pages of " international pure " 58 phases, " being the electrochemical method that detects diethylstilbestrol on the carbon paste electrode of electron mediator with the bromohexadecane yl pyridines " of delivering).But the defective of this method is: owing to use carbon paste electrode, its repeatability is not good, and detectability is higher, is 0.00268 μ g/g, can not satisfy the requirement that detects the residual lowest detectable limit of hexene in the animal product.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, a kind of electrochemical detection method of glass-carbon electrode modification of diethylstilbestrol is provided.The modification of working electrode of the present invention is simple and can reuse, and this modified electrode is strong to the rich long-pending ability of the selectivity of diethylstilbestrol, and the reappearance of test is good to be reached highly sensitively, and detectability can reach 0.001 μ g/g.
The present invention is achieved by the following technical solutions, the present invention includes following steps:
The first step, the pre-service of electrode: glass-carbon electrode is used TiO on chamois leather
2Nano powder makes it become minute surface by figure of eight polishing, uses distilled water flushing, uses ultrasonic cleaning again 10 minutes～30 minutes; It is that 1: 1 nitric acid, acetone soln respectively soaked 3 minutes～15 minutes that glass-carbon electrode after cleaning is put into volume ratio successively, uses the distilled water flushing electrode then; Electrode is dried under infrared lamp.
Second step, the preparation of nano gold sol liquid;
The 3rd step, the preparation of polyethyleneimine-decorated by nano-gold glass-carbon electrode: can successively be divided into the preparation of polyethylene imine beautify glass-carbon electrode and two steps of preparation of polyethyleneimine-decorated by nano-gold electrode;
In the 4th step, use polyethyleneimine-decorated by nano-gold electrode test diethylstilbestrol content:
A. the phosphate buffer of 0.01M that adds 6 milliliters～10 milliliters of pH6.8 in different electro-chemical test cups respectively adds the diethylstilbestrol titer or the sample liquid to be measured of the 7% ethanol water preparation of 100 microlitres～300 microlitres respectively then as supporting solution in each small beaker.(when making typical curve, make the ultimate density of diethylstilbestrol in each beaker be respectively 0.06mg/L, 0.12mg/L, 0.3mg/L, 0.6mg/L, 1.2mg/, 2.4mg/L, 4.8mg/L, 6.0mg/L and 9.0mg/L.)
B. will modifying good polyethyleneimine-decorated by nano-gold glass-carbon electrode, Ag/AgCL contrast electrode (commercially available) and platinum electrode (commercially available), to be connected to the multi-functional voltamograph of MF-1A (commercially available, Jiangfen Electric Analytic Instrument Co., Ltd., Jiangsu) on, (regulate the test parameter of the multi-functional voltamograph of MF-1A: the scanning potential range is 0.0V～0.7V, 0.5 μ A/mV～2 μ A/mV are selected in sensitivity) the rich long-pending time is 60 seconds～120 seconds, oxidation peak current value with differentiated pulse voltammetry (be the DPV method, be routine techniques) record diethylstilbestrol.Every survey finishes a sample, places the sodium hydroxide solution of 0.2mol/L to clean 3 minutes～5 minutes to remove the residual of electrode surface diethylstilbestrol this modified electrode.
C. draw out the typical curve of diethylstilbestrol according to the concentration value of diethylstilbestrol with corresponding oxidation peak current, and calculate the content of diethylstilbestrol in the respective sample according to typical curve.
Described second step, the preparation method of nano gold sol liquid, specifically:
A. chlorauric acid solution is added in the large beaker that 200ml distilled water is housed;
B. beaker is placed on the electric furnace, big fire is boiled;
C. join while stirring in the large beaker that is seething with excitement with 1% the trisodium citrate of liquid-transfering gun the new preparation of 0.5ml～2ml;
D. use moderate heat instead and continue heating;
E. observing solution colour changes: to darkviolet, arrive aubergine (claret look) by colourless again;
In a single day f. note finding that solution colour changes to mauve trend, promptly pick up counting, continue to boil the back and close fire;
G. after treating that solution is cooled to room temperature, be stored in 4 ℃ of refrigerators, treat next step processing (mean diameter of gained colloidal gold particle can be observed and be obtained about 20nm) under electron microscope.
Described chlorauric acid solution, its amount of getting and concentration are: 1% chlorauric acid solution, 0.5ml～1ml.
Describedly continue to boil the back and close fire, its time that continues is: 5 minutes.
Described the 3rd step, the preparation of polyethylene imine beautify glass-carbon electrode, specifically: get commercially available mass percent concentration and be 0.5 milliliter～1 milliliter of 30%～50% polyethyleneimine stoste, correspondingly with 0.5 milliliter～1 milliliter isopropyl alcohol as cosolvent, and be slow 0.5% the polyethyleneimine: amine aqueous solution that is diluted to of phosphate of 0.01M with pH6.8 concentration.Getting this solution joins in the electro-chemical test pond for 5 milliliters.In the electro-chemical test pond, adhere to, form layer of even polyethyleneimine film on the glass-carbon electrode surface through deposition.
The described deposition of passing through in the electro-chemical test pond is adhered to, specifically: (profile diameter is 8.0mm with first glass carbon columnar electrode, the reaction surface diameter is 6.0mm) constitute three-electrode structure with Ag/AgCl contrast electrode and platinum electrode, be inserted in the electro-chemical test pond of containing 0.5% polyethyleneimine: amine aqueous solution, connect the multi-functional voltamograph of MF-1A type, carry out cyclic voltammetry scan, take out back oven dry (or drying naturally in the air) under infrared lamp, promptly form layer of even polyethyleneimine film on the glass-carbon electrode surface;
The described cyclic voltammetry scan that carries out, condition is: in the scope of-0.5V～0.5V, scanning 10 circles～30 circles.
Described the 3rd step, the preparation of polyethyleneimine-decorated by nano-gold electrode, specifically: the electrode (being the electrode that PEI-modifies) of above-mentioned polyethyleneimine-modification is immersed in the colloidal gold solution, after cleaning with tri-distilled water, air dry in air.Directly drip at last 5% the polyethyleneimine (being PEI) of 2 μ l～6 μ l at the electrode surface of having modified, stand-by behind air drying.
In the described immersion colloidal gold solution, the time is: 3 minutes～10 minutes.
The method principle of main invention is:
At first on pretreated glass-carbon electrode, modify the layer of polyethylene imine layer with electrochemical deposition method, then this modified electrode is immersed in the nm of gold ion that comes the adsorption band negative charge in the nm of gold colloidal solution and forms second decorative layer, on this modified electrode, form three layers of " sandwich " glass carbon modified electrode again by the absorption polyethyleneimine, because this method of modifying has increased the long-pending ability of the richness of electrode and increased the reaction area of electrode surface, thereby the sensitivity and the specificity of this method detection diethylstilbestrol have been increased.Core technology of the present invention is will (mainly to be included in TiO after the glass-carbon electrode pre-service earlier
2Polishing on the nano powder, ultrasonic cleaning and the cleaning in nitric acid, acetone and distilled water), with electrochemical deposition method at glass-carbon electrode surface deposition layer of polyethylene imines film, then will this film immerse in nano-Au solution and make the nanogold particle in the solution be deposited on polyethyleneimine film surface, at last the electro-deposition one strata Class B imines film again in the nano gold layer outside again.Wherein sodium rice gold solution is to make with sodium citrate reduction chlorauric acid solution.Should " sandwich " formula modified electrode and the assembling of portable electrochemical analyser after, just can be used for diethylstilbestrol residual in the specimen.
Application of the present invention:
Utilize diethylstilbestrol at polyethyleneimine---efficient absorption and characteristic redox reaction take place on the decorated by nano-gold electrode, content by diethylstilbestrol in the little testing current test sample of oxidation, this method of testing step is simple, analysis time is short, detect lower limit and can reach 0.001 μ g/g, detect applicable to the scene that estrogenic chemicals in the biological sample is residual.Can be applicable to quick, sensitive, the on-the-spot electrochemical sensor that detects diethylstilbestrol in animal product, the feed sample and detect, control the security of animal food from the source.
The invention has the beneficial effects as follows:
The estrogenic electrochemical method of existing relevant test is to adopt the unmodified carbon paste electrode directly to measure, and detectability reaches 0.4 μ g/g, thereby the sensitivity of this method is low and be not suitable for applying to estrogenic detection in the animal product.Method advantage set forth in the present invention is that the modification of working electrode is simple and can reuses that the rich long-pending ability of the selectivity to diethylstilbestrol of this modified electrode is strong, and the reappearance of test well reaches highly sensitive, and detectability can reach 0.001 μ g/g.This method has overcome to be needed large-scale instrument when chromatography detects DES and expends time in and the shortcoming of manpower, has also overcome the immune-gold labeled test strips false positive of existing detection diethylstilbestrol and the higher shortcoming of false negative simultaneously.
Description of drawings
Fig. 1 is the embodiment of the invention 1 " sandwich " formula polyethyleneimine---the tactic pattern of decorated by nano-gold electrode.
Embodiment
Below embodiments of the invention are elaborated: present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1
Present embodiment 1 is implemented under following implementation condition and technical requirement condition:
(1) preparation of polyethyleneimine-decorated by nano-gold electrode
(1) pre-service of electrode:
At first glass-carbon electrode is used TiO on chamois leather
2Nano powder is polished by the figure of eight, makes it become minute surface, behind distilled water flushing, uses ultrasonic cleaning again 10 minutes.It is that 1: 1 nitric acid, acetone soln respectively soaked 3 minutes that glass-carbon electrode after will cleaning is then put into volume ratio successively, uses the distilled water flushing electrode.At last electrode is dried under infrared lamp.
(2) preparation of nm of gold colloidal particle: the chlorauric acid solution with 0.5ml 1% is added in the large beaker that 200ml distilled water is housed earlier.Then beaker is placed on the electric furnace, big fire is boiled.Join while stirring in the large beaker that is seething with excitement with the trisodium citrate of liquid-transfering gun, use moderate heat instead and continue heating 0.5ml freshly prepared 1%.The discovery solution colour changes and is the trend of claret, promptly picks up counting to continue to boil to close fire after 5 minutes.After treating that solution is cooled to room temperature, be stored in 4 ℃ of refrigerators.The mean diameter of observing colloid gold particle is about 38nm under the electron microscope.
(3) preparation of polyethylene imine beautify glass-carbon electrode: get mass percent concentration and be 0.5 milliliter of 30% polyethyleneimine stoste, with 0.5 milliliter of isopropyl alcohol as cosolvent, being that the phosphate of 0.01M is slow with pH6.8 concentration is diluted to 0.5% polyethyleneimine: amine aqueous solution, gets this solution and joins in the electro-chemical test pond for 5 milliliters.(profile diameter is 8.0mm with glass carbon columnar electrode, the reaction surface diameter is 6.0mm) constitute three-electrode structure with Ag/AgCl contrast electrode and platinum electrode, be inserted in the electro-chemical test pond of containing 0.5% polyethyleneimine: amine aqueous solution, connect the multi-functional voltamograph of MF-1A type, in the scope of 0.5V, carry out cyclic voltammetry scan 10 circles at-0.5V, take out back oven dry (or drying naturally in the air) under infrared lamp, promptly form layer of even polyethyleneimine film on the glass-carbon electrode surface.
(4) preparation of polyethyleneimine-decorated by nano-gold electrode: the electrode that above-mentioned PEI-is modified immersed in the colloidal gold solution 3 minutes, after cleaning with tri-distilled water, and air dry in air.Directly drip at last 5% the PEI of 2 μ l at the electrode surface of having modified, stand-by behind air drying, finish the preparation of polyethyleneimine-decorated by nano-gold electrode, referring to " sandwich " shown in Figure 1 formula polyethyleneimine---the tactic pattern of decorated by nano-gold electrode, in this structure, the adsorbable electronegative nanogold particle of polyethyleneimine with cationic active agent character, the strange adsorbable again polyethyleneimine layer of nanogold particle.Owing to after nm of gold in the interlayer and the polyethyleneimine combination, improved adsorption area and reaction area greatly, make hundreds of times of amplifications of electric current like this, thereby the detectability that detects diethylstilbestrol is descended.
(2) foundation of the test diethylstilbestrol electrochemical analysis method of polyethyleneimine-decorated by nano-gold glass-carbon electrode.
(1) drafting of typical curve
1. the PBS of 0.01M that adds 6 milliliters of pH6.8 in different electro-chemical test cups (25ml) respectively is as supporting solution, the diethylstilbestrol titer that in each small beaker, adds the 7% ethanol water preparation of 100 microlitres then respectively, make the ultimate density of diethylstilbestrol in each beaker be respectively 0.06mg/L, 0.12mg/L, 0.3mg/L, 0.6mg/L, 1.2mg/, 2.4mg/L, 4.8mg/L, 6.0mg/L and 9.0mg/L.
2. will modify good polyethyleneimine-decorated by nano-gold glass-carbon electrode, Ag/AgCL contrast electrode and platinum electrode is connected on the multi-functional voltamograph of MF-1A (Jiangfen Electric Analytic Instrument Co., Ltd., Jiangsu).
3. regulate the instrument test parameter: the scanning potential range is 0.0V～0.7V.Instrumental sensitivity is selected 0.5 μ A/mV, and the rich long-pending time is 60 seconds, with the oxidation peak current value of differentiated pulse voltammetry (DPV method) record diethylstilbestrol.The test modulation parameter of this method is as follows: pulse height is 50mV, pulse width 20ms, and pulse period is 200ms, sweep velocity 50mV.s
-1Every survey finishes a sample, places the sodium hydroxide solution of 0.2mol/L to clean 3 minutes to remove the residual of electrode surface diethylstilbestrol this modified electrode.
4. draw out the typical curve that polyethyleneimine-decorated by nano-gold glass-carbon electrode method detects diethylstilbestrol according to the concentration value of diethylstilbestrol with corresponding oxidation peak current.
(2) test of actual sample
1. the pre-treatment of sample
Obtain 30 parts in the freezing sample of pork from Shanghai City food quality inspection department, under room temperature, thaw, accurately take by weighing the 5.0g tissue and place the 30mL plastic centrifuge tube, add the 10mL ether, vortex vibration 3min uses ultrasonic concussion 15 minutes again behind the mixing, the centrifugal 5min of 6000r/min, draw supernatant to the 30mL glass tube, add the 10mL ether in the residue again and repeat to extract once, merge ether nitrogen stream under 40 ℃ of water bath condition and dry up.With 2mL 70% dissolve with methanol solution residue, vortex mixing, add behind the 4mL sherwood oil vortex mixing again.Leave standstill and moments later absorb the upper strata sherwood oil, add sherwood oil again and repeat to extract 2 times.The methyl alcohol of lower floor nitrogen stream under 45 ℃ of water bath condition dries up.Remaining aqueous solution is used the 4mL extracted by ether again, abundant mixing, and behind the standing demix, the ether of drawing the upper strata repeats aforesaid operations once to 10mL glass centrifuge tube, merge the ether phase, and nitrogen stream dries up under 0 ℃ of water-bath.After 1 milliliter 1% methanol aqueous solution dissolving, refrigerate to be measured.
2. method of testing and result
Under the test condition identical with the testing standard sample, get sample liquid 100 microlitres to be checked, add the electro-chemical test pond, with the oxidation peak current value of differentiated pulse voltammetry record diethylstilbestrol.And calculate the content of diethylstilbestrol in the respective sample according to typical curve.Test result shows in 30 duplicate samples, have in 2 duplicate samples to detect diethylstilbestrol, and content is 0.025 μ g/g and 0.086 μ g/g respectively, and other sample is all negative.When high performance liquid chromatography conclusive evidence method detected, it was residual to have only portion to detect diethylstilbestrol, and content is 0.096 μ g/g.The sensitivity that this method is described will be higher than chromatography, and trace routine is short simple and detection time, and each sample detection time is 3 minutes, and it is residual to can be used in quick, the on-the-spot detection of biological sample diethylstilbestrol.
Embodiment 2
Present embodiment 2 is implemented under following implementation condition and technical requirement condition:
(1) preparation of polyethyleneimine-decorated by nano-gold electrode
(1) pre-service of electrode:
At first with glass-carbon electrode on chamois leather polishing and steam the water flushing process with embodiment 1.Used ultrasonic cleaning then 30 minutes, it is that 1: 1 nitric acid, acetone soln respectively soaked 15 minutes that the glass-carbon electrode after will cleaning is again put into volume ratio successively, uses the distilled water flushing electrode.At last electrode is dried in air naturally.
(2) preparation of nm of gold colloidal particle: the chlorauric acid solution with 1.0ml 1% is added in the large beaker that 200ml distilled water is housed earlier.Then beaker is placed on the electric furnace, big fire is boiled.Join while stirring in the large beaker that is seething with excitement with the trisodium citrate of liquid-transfering gun, use moderate heat instead and continue heating 2.0ml freshly prepared 1%.The discovery solution colour changes and is mauve trend, promptly picks up counting to continue to boil to close fire after 5 minutes.After treating that solution is cooled to room temperature, be stored in 4 ℃ of refrigerators.The mean diameter of observing colloid gold particle is about 22nm under the electron microscope.
(3) preparation of polyethylene imine beautify glass-carbon electrode: the preparation of 0.5% polyethyleneimine: amine aqueous solution is got this solution and is joined in the electro-chemical test pond for 5 milliliters with embodiment 1.Glass carbon columnar electrode, Ag/AgCl contrast electrode and platinum electrode are constituted three-electrode structure, be inserted in the electro-chemical test pond of containing 0.5% polyethyleneimine: amine aqueous solution, connect the multi-functional voltamograph of MF-1A type (Jiangfen Electric Analytic Instrument Co., Ltd., Jiangsu), in the scope of 0.5V, carry out cyclic voltammetry scan 30 circles at-0.5V, take out the back or air in dry naturally, can be at glass-carbon electrode surface formation layer of even polyethyleneimine film.
(4) preparation of polyethyleneimine-decorated by nano-gold electrode: the electrode that above-mentioned PEI-is modified immersed in the colloidal gold solution 10 minutes, after cleaning with tri-distilled water, and air dry in air.Directly drip at last 5% the PEI of 6 μ l at the electrode surface of having modified, stand-by behind air drying.
(2) foundation of the test diethylstilbestrol electrochemical analysis method of polyethyleneimine-decorated by nano-gold glass-carbon electrode.
(1) drafting of typical curve: the method for drafting of typical curve and step are with embodiment 1, and difference is that using the polyethyleneimine-decorated by nano-gold electrode of this modification is test electrode in this embodiment implements.Instrument parameter is regulated as follows: the scanning potential range is 0.0V～0.7V.Instrumental sensitivity is selected 2.0 μ A/mV, and the rich long-pending time is 120 seconds, with the oxidation peak current value of differentiated pulse voltammetry (DPV method) record diethylstilbestrol.The test modulation parameter of this method is as follows: pulse height is 100mV, pulse width 40ms, and pulse period is 400ms, sweep velocity 100mV.s
-1Every survey finishes a sample, places the sodium hydroxide solution of 0.2mol/L to clean 5 minutes to remove the residual of electrode surface diethylstilbestrol this modified electrode.
(2) test of actual sample
1. the pre-treatment of sample: sample collecting and pre-treatment process are with embodiment 1.
2. method of testing and result
Under the test condition identical with the testing standard sample, get sample liquid 100 microlitres to be checked, add the electro-chemical test pond, with the oxidation peak current value of differentiated pulse voltammetry record diethylstilbestrol.And calculate the content of diethylstilbestrol in the respective sample according to typical curve.Test result shows in 30 duplicate samples, have in 2 duplicate samples to detect diethylstilbestrol, and content is 0.016 μ g/g and 0.058 μ g/g respectively, and other sample is all negative.Explanation is compared with embodiment 1, and sensitivity has decline slightly, but still will be higher than method for chromatographic determination, and detection time is short, and operating process is simpler than chromatography.
Embodiment 3
Present embodiment 3 is implemented under following implementation condition and technical requirement condition:
(1) preparation of polyethyleneimine-decorated by nano-gold electrode
(1) pre-service of electrode:
At first with glass-carbon electrode on chamois leather polishing and steam the water flushing process with embodiment 1.Used ultrasonic cleaning then 20 minutes, it is that 1: 1 nitric acid, acetone soln respectively soaked 9 minutes that the glass-carbon electrode after will cleaning is again put into volume ratio successively, uses the distilled water flushing electrode.At last electrode is dried in air naturally.
(2) preparation of nm of gold colloidal particle: the chlorauric acid solution with 0.75ml 1% is added in the large beaker that 200ml distilled water is housed earlier.Then beaker is placed on the electric furnace, big fire is boiled.Join while stirring in the large beaker that is seething with excitement with the trisodium citrate of liquid-transfering gun, use moderate heat instead and continue heating 1.0ml freshly prepared 1%.The discovery solution colour changes and is wine-colored trend, promptly picks up counting to continue to boil to close fire after 5 minutes.After treating that solution is cooled to room temperature, be stored in 4 ℃ of refrigerators.The mean diameter of observing colloid gold particle is about 31nm under the electron microscope.
(3) preparation of polyethylene imine beautify glass-carbon electrode: the preparation of 0.5% polyethyleneimine: amine aqueous solution is got this solution and is joined in the electro-chemical test pond for 5 milliliters with embodiment 1.Glass carbon columnar electrode, Ag/AgCl contrast electrode and platinum electrode are constituted three-electrode structure, be inserted in the electro-chemical test pond of containing 0.5% polyethyleneimine: amine aqueous solution, connect the multi-functional voltamograph of MF-1A type (Jiangfen Electric Analytic Instrument Co., Ltd., Jiangsu), in the scope of 0.5V, carry out cyclic voltammetry scan 20 circles at-0.5V, take out the back or air in dry naturally, can be at glass-carbon electrode surface formation layer of even polyethyleneimine film.
(4) preparation of polyethyleneimine-decorated by nano-gold electrode: the electrode that above-mentioned PEI-is modified immersed in the colloidal gold solution 6 minutes, after cleaning with tri-distilled water, and air dry in air.Directly drip at last 5% the PEI of 4 μ l at the electrode surface of having modified, stand-by behind air drying.
(2) foundation of the test diethylstilbestrol electrochemical analysis method of polyethyleneimine-decorated by nano-gold glass-carbon electrode.
(1) drafting of typical curve: the method for drafting of typical curve and step are with embodiment 1, and difference is that using the polyethyleneimine-decorated by nano-gold electrode of this modification is test electrode in this embodiment implements.Instrument parameter is regulated as follows: the scanning potential range is 0.0V～0.7V.Instrumental sensitivity is selected 1.0 μ A/mV, and the rich long-pending time is 90 seconds, with the oxidation peak current value of differentiated pulse voltammetry (DPV method) record diethylstilbestrol.The test modulation parameter of this method is as follows: pulse height is 25mV, pulse width 10ms, and pulse period is 100ms, sweep velocity 25mV.s
-1Every survey finishes a sample, places the sodium hydroxide solution of 0.2mol/L to clean 4 minutes to remove the residual of electrode surface diethylstilbestrol this modified electrode.
(2) test of actual sample
1. the pre-treatment of sample: sample collecting and pre-treatment process are with embodiment 1.
2. method of testing and result
Under the test condition identical with the testing standard sample, get sample liquid 100 microlitres to be checked, add the electro-chemical test pond, with the oxidation peak current value of differentiated pulse voltammetry record diethylstilbestrol.And calculate the content of diethylstilbestrol in the respective sample according to typical curve.Test result shows, in 30 duplicate samples, still has in 2 duplicate samples and detects diethylstilbestrol, and content is 0.083 μ g/g and 0.117 μ g/g respectively, and other sample is still all negative.Explanation is compared with embodiment 1 and embodiment 2, and sensitivity is higher and detectability is lower.
Claims (9)
1. the electrochemical detection method of the glass-carbon electrode of diethylstilbestrol modification is characterized in that, comprises the steps:
The first step, the pre-service of electrode: glass-carbon electrode is used TiO on chamois leather
2Nano powder makes it become minute surface by figure of eight polishing, uses distilled water flushing, uses ultrasonic cleaning again 10 minutes～30 minutes; It is that 1: 1 nitric acid, acetone soln respectively soaked 3 minutes～15 minutes that glass-carbon electrode after cleaning is put into volume ratio successively, uses the distilled water flushing electrode then; Electrode is dried under infrared lamp;
Second step, the preparation of nano gold sol liquid;
The 3rd step, the preparation of polyethyleneimine-decorated by nano-gold glass-carbon electrode: successively be divided into the preparation of polyethylene imine beautify glass-carbon electrode and two steps of preparation of polyethyleneimine-decorated by nano-gold electrode;
The 4th goes on foot, and uses the content of polyethyleneimine-decorated by nano-gold electrode test diethylstilbestrol:
A. the phosphate buffer of 0.01M that adds 6 milliliters～10 milliliters of pH6.8 in different electro-chemical test cups respectively adds the diethylstilbestrol titer or the sample liquid to be measured of the 7% ethanol water preparation of 100 microlitres～300 microlitres respectively then as supporting solution in each small beaker;
B. will modify good polyethyleneimine-decorated by nano-gold glass-carbon electrode, Ag/AgCL contrast electrode and platinum electrode and be connected on the multi-functional voltamograph of MF-1A, rich long-pending 60 seconds～120 seconds, the oxidation peak current value of record diethylstilbestrol;
C. draw out the typical curve of diethylstilbestrol according to the concentration value of diethylstilbestrol with corresponding oxidation peak current, and calculate the content of diethylstilbestrol in the respective sample according to typical curve.
2. the electrochemical detection method of the glass-carbon electrode of diethylstilbestrol as claimed in claim 1 modification is characterized in that, described second step, and the preparation method of nano gold sol liquid, specifically:
A. chlorauric acid solution is added in the large beaker that 200ml distilled water is housed;
B. beaker is placed on the electric furnace, big fire is boiled;
C. join while stirring in the large beaker that is seething with excitement with 1% the trisodium citrate of liquid-transfering gun the new preparation of 0.5ml～2ml;
D. use moderate heat instead and continue heating;
E. observing solution colour changes: to darkviolet, arrive aubergine by colourless again;
F. find that solution colour changes to mauve trend, promptly pick up counting, continue to boil the back and close fire;
G. after treating that solution is cooled to room temperature, be stored in 4 ℃ of refrigerators, treat next step processing.
3. the electrochemical detection method of the glass-carbon electrode of diethylstilbestrol as claimed in claim 2 modification is characterized in that, described chlorauric acid solution, and its amount of getting and concentration are: 1% chlorauric acid solution, 0.5ml～1ml.
4. the electrochemical detection method modified of the glass-carbon electrode of diethylstilbestrol as claimed in claim 2 is characterized in that, describedly continues to boil the back and closes fire, and its time that continues is: 5 minutes.
5. the electrochemical detection method modified of the glass-carbon electrode of diethylstilbestrol as claimed in claim 1, it is characterized in that, described the 3rd step, the preparation of polyethylene imine beautify glass-carbon electrode, specifically: get mass percent concentration and be 0.5 milliliter～1 milliliter of 30%～50% polyethyleneimine stoste, correspondingly with 0.5 milliliter～1 milliliter isopropyl alcohol as cosolvent, and be slow 0.5% the polyethyleneimine: amine aqueous solution that is diluted to of phosphate of 0.01M with pH6.8 concentration, getting 5 milliliters of this solution joins in the electro-chemical test pond, in this electro-chemical test pond, adhere to, form layer of even polyethyleneimine film on the glass-carbon electrode surface through deposition.
6. the electrochemical detection method modified of the glass-carbon electrode of diethylstilbestrol as claimed in claim 5, it is characterized in that, the described deposition of passing through in the electro-chemical test pond is adhered to, specifically: with first glass carbon columnar electrode and Ag/AgCl contrast electrode and platinum electrode formation three-electrode structure, be inserted in the electro-chemical test pond of containing 0.5% polyethyleneimine: amine aqueous solution, connect the multi-functional voltamograph of MF-1A type, carry out cyclic voltammetry scan, take out the back and under infrared lamp, dry.
7. the electrochemical detection method modified of the glass-carbon electrode of diethylstilbestrol as claimed in claim 6 is characterized in that, the described cyclic voltammetry scan that carries out, and condition is: in the scope of-0.5V～0.5V, scanning 10 circles～30 circles.
8. the electrochemical detection method modified of the glass-carbon electrode of diethylstilbestrol as claimed in claim 1, it is characterized in that, described the 3rd step, the preparation of polyethyleneimine-decorated by nano-gold electrode, specifically: the electrode of above-mentioned polyethyleneimine-modification is immersed in the colloidal gold solution, after cleaning with tri-distilled water, air dry in air, directly drip at last 5% the polyethyleneimine of 2 μ l～6 μ l at modifying polyethyleneimine-decorated by nano-gold electrode surface, stand-by behind air drying.
9. the electrochemical detection method of the glass-carbon electrode of diethylstilbestrol as claimed in claim 8 modification is characterized in that in the described immersion colloidal gold solution, the time is: 3 minutes～10 minutes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200710040472XA CN100483123C (en) | 2007-05-10 | 2007-05-10 | Glassy carbon electrode modified electrochemical detection method for diethylstilbestrol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200710040472XA CN100483123C (en) | 2007-05-10 | 2007-05-10 | Glassy carbon electrode modified electrochemical detection method for diethylstilbestrol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101059480A true CN101059480A (en) | 2007-10-24 |
| CN100483123C CN100483123C (en) | 2009-04-29 |
Family
ID=38865699
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200710040472XA Expired - Fee Related CN100483123C (en) | 2007-05-10 | 2007-05-10 | Glassy carbon electrode modified electrochemical detection method for diethylstilbestrol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100483123C (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101966594A (en) * | 2010-10-18 | 2011-02-09 | 上海市七宝中学 | Electro-deposition gold nanoparticles, preparation method and application thereof |
| CN102262125A (en) * | 2011-07-28 | 2011-11-30 | 南京师范大学 | Electrochemical immune sensor for detecting diethylstilbestrol and preparation method and application of sensor |
| CN103698383A (en) * | 2013-12-18 | 2014-04-02 | 济南大学 | Preparation method and application of electrochemical immunosensor for detecting diethylstilbestrol |
| CN105699646A (en) * | 2016-02-25 | 2016-06-22 | 济南大学 | Manufacturing method and application of electrochemiluminescence nonyl phenol sensor based on titanium dioxide nanosheet composite |
| CN105717178A (en) * | 2016-02-25 | 2016-06-29 | 济南大学 | Preparation method and application of electrochemical diethylstilbestrol sensor based on titanium-dioxide-based two-dimensional composite nanomaterial |
| CN109046453A (en) * | 2018-07-03 | 2018-12-21 | 山东交通学院 | A kind of preparation method and application of CdSeS point complex composite material |
| CN109116015A (en) * | 2018-09-27 | 2019-01-01 | 深圳大学 | A kind of production method and application method of non-sealed ELISA ELISA Plate |
| CN109580758A (en) * | 2018-12-21 | 2019-04-05 | 湖南科技大学 | A kind of copper ion electrochemical sensor and its preparation method and application |
| CN111272842A (en) * | 2020-02-25 | 2020-06-12 | 衡阳师范学院 | Polyaniline modified electrode for quantitatively detecting estrone and preparation method and application thereof |
| CN111595915A (en) * | 2020-07-02 | 2020-08-28 | 盐城工学院 | Preparation method of diethylstilbestrol molecularly imprinted electrochemical sensor |
| CN115060766A (en) * | 2022-06-07 | 2022-09-16 | 湖北大学 | Titanium dioxide gas sensor and preparation method thereof |
-
2007
- 2007-05-10 CN CNB200710040472XA patent/CN100483123C/en not_active Expired - Fee Related
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101966594A (en) * | 2010-10-18 | 2011-02-09 | 上海市七宝中学 | Electro-deposition gold nanoparticles, preparation method and application thereof |
| CN102262125A (en) * | 2011-07-28 | 2011-11-30 | 南京师范大学 | Electrochemical immune sensor for detecting diethylstilbestrol and preparation method and application of sensor |
| CN103698383A (en) * | 2013-12-18 | 2014-04-02 | 济南大学 | Preparation method and application of electrochemical immunosensor for detecting diethylstilbestrol |
| CN105699646A (en) * | 2016-02-25 | 2016-06-22 | 济南大学 | Manufacturing method and application of electrochemiluminescence nonyl phenol sensor based on titanium dioxide nanosheet composite |
| CN105717178A (en) * | 2016-02-25 | 2016-06-29 | 济南大学 | Preparation method and application of electrochemical diethylstilbestrol sensor based on titanium-dioxide-based two-dimensional composite nanomaterial |
| CN109046453B (en) * | 2018-07-03 | 2020-11-20 | 山东交通学院 | Preparation method and application of CdSeS point complex composite material |
| CN109046453A (en) * | 2018-07-03 | 2018-12-21 | 山东交通学院 | A kind of preparation method and application of CdSeS point complex composite material |
| CN109116015A (en) * | 2018-09-27 | 2019-01-01 | 深圳大学 | A kind of production method and application method of non-sealed ELISA ELISA Plate |
| CN109116015B (en) * | 2018-09-27 | 2021-07-09 | 深圳大学 | Manufacturing method and using method of blocking-free ELISA (enzyme-linked immunosorbent assay) plate |
| CN109580758B (en) * | 2018-12-21 | 2020-10-13 | 湖南科技大学 | Copper ion electrochemical sensor and preparation method and application thereof |
| CN109580758A (en) * | 2018-12-21 | 2019-04-05 | 湖南科技大学 | A kind of copper ion electrochemical sensor and its preparation method and application |
| CN111272842A (en) * | 2020-02-25 | 2020-06-12 | 衡阳师范学院 | Polyaniline modified electrode for quantitatively detecting estrone and preparation method and application thereof |
| CN111272842B (en) * | 2020-02-25 | 2022-09-30 | 衡阳师范学院 | Polyaniline modified electrode capable of quantitatively detecting estrone and preparation method and application thereof |
| CN111595915A (en) * | 2020-07-02 | 2020-08-28 | 盐城工学院 | Preparation method of diethylstilbestrol molecularly imprinted electrochemical sensor |
| CN115060766A (en) * | 2022-06-07 | 2022-09-16 | 湖北大学 | Titanium dioxide gas sensor and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100483123C (en) | 2009-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100483123C (en) | Glassy carbon electrode modified electrochemical detection method for diethylstilbestrol | |
| CN102749373B (en) | Preparation method and application of environmental estrogen electrochemical immunosensor | |
| CN110308286B (en) | Enhanced thyroglobulin electrochemiluminescence immunosensor based on photothermal release signals | |
| CN103954669A (en) | Enzyme electrode, enzyme biosensor as well as preparation methods and application thereof | |
| CN106290514B (en) | A kind of TiO based on silicon phthalocyanine functionalization2It is situated between and sees the aflatoxin optical electro-chemistry detection method of crystal | |
| Deng et al. | Sensitive voltammetric determination of tryptophan using an acetylene black paste electrode modified with a Schiff's base derivative of chitosan | |
| CN101793862B (en) | L-cysteine/chitosan-modified electrochemical sensor and use thereof | |
| CN101915792A (en) | Preparation method of electrochemical immunosensor for detecting salbutamol quickly | |
| CN106324065A (en) | Preparation of chemiluminiscence light-induced electrochemical sensor and organic phosphorus pesticide detection | |
| CN114527113A (en) | Electrochemical luminescence aptamer sensor for specifically detecting enrofloxacin, preparation method and application | |
| CN102416313A (en) | Bisphenol A dummy template molecularly imprinted stir bar and preparation method thereof | |
| CN109085225A (en) | A kind of preparation method of the protein electrochemistry trace sensor of step sedimentation modification carbon electrode | |
| CN102998449B (en) | Preparation based on tumor marker sensor of sodamide group smectite and application | |
| CN110988067B (en) | Electrochemical luminescence method for detecting diethylstilbestrol | |
| CN103499620A (en) | Method for preparing current type aptamer sensor for detecting tetracycline residue | |
| CN112505024A (en) | Electrochemiluminescence aptamer sensor for detecting enrofloxacin, preparation method thereof and method for detecting enrofloxacin | |
| CN100339703C (en) | Biosensor electrode used for detecting aflatoxin and variegated aspergillin and its preparation method | |
| CN1675539A (en) | Electrochemical biosensor | |
| CN106124584B (en) | One kind is based on CdS@SnS2The preparation method and application of the unmarked type insulin photoelectricity immunosensors of@MWCNTs | |
| CN105301077A (en) | Preparation method of aptamer sensor for detecting chlorpyrifos | |
| CN1226614C (en) | Nano immunological biosensor | |
| CN114544720B (en) | Electrochemical sensor for ultra-sensitively detecting enrofloxacin and preparation method thereof | |
| CN110132946A (en) | A kind of aptamer sensor and its preparation method and application | |
| Wang et al. | A convenient electrochemiluminescent immunosensor for detecting methamphetamine antibody | |
| CN1710424A (en) | Collaurum marking method and quick-detection sevin test paper strip its preparation and use |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090429 Termination date: 20120510 |