CN104965014A - Preparation method of quantum dot/enzyme compounded carbon paste electrode for detecting SAM (S-Adenosylmethionine) - Google Patents
Preparation method of quantum dot/enzyme compounded carbon paste electrode for detecting SAM (S-Adenosylmethionine) Download PDFInfo
- Publication number
- CN104965014A CN104965014A CN201510389582.1A CN201510389582A CN104965014A CN 104965014 A CN104965014 A CN 104965014A CN 201510389582 A CN201510389582 A CN 201510389582A CN 104965014 A CN104965014 A CN 104965014A
- Authority
- CN
- China
- Prior art keywords
- quantum dot
- preparation
- carbon paste
- paste electrode
- 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
Abstract
The invention discloses a preparation method of a quantum dot/enzyme compounded carbon paste electrode for detecting SAM (S-Adenosylmethionine). The preparation method is characterized by comprising the following steps of doping water-soluble CdZnTe quantum dots and transmethylase in a carbon paste electrode; by taking 1-ethyl-3--methylimidazolium hexafluorophosphate as an adhesive, mixing a carbon nano-tube with graphite powder to prepare the carbon electrode, so as to obtain the quantum dot/enzyme compounded carbon paste electrode; compared with an ordinary carbon paste electrode, the quantum dot/enzyme compounded carbon paste electrode has the advantages of conducting property improved by 1 to 2 times, wide electrochemical window, simple preparation method, low cost, easy update for surface, low residual current and the like; the eelctrode can be used for quickly detecting the SAM in a sample; the method has the characteristics of high sensitivity, good selectivity, short response time and low interference, is superior to other detection methods, and is a determination method with simpleness, quickness, convenience and feasibility for the SAM. A fixed methyltransferase electrode sensor prepared by the preparation method disclosed by the invention is low in cost, simple in preparation process and good in specificity, and has the possibility of realizing automated field determination.
Description
Technical field
The present invention relates to a kind of preparation method of electrochemical sensor, particularly one is for detecting preparation method and the application of the quantum dot/enzyme composite carbon paste electrode of S-adenosylmethionine (SAM).
Background technology
S-adenosylmethionine (SAM), SAM contains active methyl, and in cell, nearly all methyl modified for methylating is all from SAM first sulphur energy-rich bond.Due to the popularity of methylation reaction, can say, SAM is a kind of coenzyme that the interior importance of participating in reaction of cell is only second to ATP, and the minor alteration of SAM concentration in cell, just can produce significant impact to the growth of cell, differentiation and function.SAM is mainly synthesized by methionine (Met) and ATP by SAM synzyme (MetK) in bacterial body.When
e.colisAM synzyme level decline, when to cause in cell methyl donor SAM to lack, cell would not proper splitting.If the AdoMet hydrolase gene from T3 bacteriophage is imported
e.colisomatic cells, when making SAM level decline in born of the same parents, colon bacillus also form nondividing long filament shape thalline.Further research shows, in thread thalline, causes
e.colifissional Z ring complex assembling can be normally initial, but can not complete, and when the SAM synzyme level of leucine adjustment recovers normal, when in cell, methyl donor SAM no longer lacks, cell division also recovers normal immediately.Clearly, in the growth division of bacterial cell and born of the same parents, SAM concentration is closely-related.
Universal bases donor SAM is by after methyl transferase catalytic demethyl, the universal product generated is adenosylhomocysteine (SAH), SAH is found to have general feedback inhibition to the methylation procedure of intracellular protein and nucleic acid, is the effective competition inhibitor of transmethylated reaction.In mammalian cell, SAH generates adenylate and homocysteine by SAH hydrolytic enzyme (SAHH) catalyzing hydrolysis, and in the cell of most of pathogenic microorganism, the metabolism of SAH then adopts diverse mode---generate adenine and S-ribosylhomocysteine by adenosylhomocysteine nucleosidase (SAHN) catalytic pyrolysis, SRH generates homocysteine and 4 further under the effect of S-Ribosylhomocysteinase (SRHH), 5 dihydroxies-2, 3-pentanedione (DPD), homocysteine is finally by several methionine synthetase (MetH, MetE) precursor of SAM is regenerated---methionine, or generate halfcystine through multistep enzymatic.
Due to the high specificity of enzyme, the method has that selectivity is high, good stability, detection speed are fast, selectivity good, sensitivity high.Enzyme electrode research is started in the sixties in 20th century, since two thousand, the application of biosensor technology in environment measuring, food security, military affairs and medical science etc. is increasingly extensive, is to disclose the common fixed enzyme electrode preparation method and application that detect p-dihydroxy-benzene and catechol in the patent of 201410210210.3 at application number; Be disclose a kind of preparation of enzyme electrode in the patent of CN102435650 B and detect the method for peroxide value of vegetable oil fast at Authorization Notice No.; Be in the patent of CN102495115 B, disclose the electrochemical method utilizing biologic enzyme electrode method to detect malic acid in root exudates at Authorization Notice No..
Carbon paste electrode is the Carbon Materials utilizing electric conductivity, and the bonding agent as dag and hydrophobic nature is mixed and made into pastel, is then coated in the surface of electrode bar or is packed into the class electrode made in electrode tube.Due to advantages such as carbon paste electrode are nontoxic, electrochemical window is wide, preparation method is simple, cost is low, surface easily upgrades, aftercurrent is little, be widely used in electrochemical analysis, biology sensor preparation and environment measuring, food and medicine analyze in.But carbon paste electrode also exists some shortcomings, as poor electric conductivity, sensitivity is low, poor stability etc.In order to improve the performance of carbon paste electrode, dag is replaced with the carbon nano-tube conducted electricity very well in carbon paste electrode preparation, and in Carbon Materials, adding the excellent water-soluble quantum dot of electric conductivity, quantum dot, also known as semiconductor nano, is a kind of nanoparticle be made up of II-VI race or III-group Ⅴ element.Quantum dot, due to the special optical, electrical character had be different from block materials, makes it in the fields such as solar cell, biomarker, fluorescence probe, have huge using value.The quantum dot of synthesis in water is owing to having good water wettability and biocompatibility, and dissolubility quantum dot is widely used, and is mixed by water-soluble quantum dot in carbon paste electrode to improve the electric conductivity of electrode.The application utilizes the tackifier (as ionic liquid) of ion-type, and these attempt the performance changing electrode all to a certain extent.Carbon nano-tube has excellent electron transmission, shows excellent performance in bio-sensing and catalysis.Carbon nano-tube also has self lubricity and bio-compatibility, thus is widely used in analytical chemistry field.Ionic liquid is the salt be at room temperature in a liquid state be made up of ion completely, has the advantages such as the excellent and electrochemical window of stable in properties, electric conductivity is wide, is widely used in the fields such as material, synthesis, living things catalysis and separating and extracting.Ionic liquid accelerates electron transfer rate, improves electrode sensitivity and selectivity.
At present, the method of the mensuration SAM reported has HPLC, there is chromatographic column and easily pollute in the method, analyze expensive defect, spectrophotometric method, detection method (the paddy sturdy pines etc. of the first class transferase active that the research such as paddy sturdy pines S-adenosylmethionine relies on, the detection method of the first class transferase active that a kind of S-adenosylmethionine relies on, SCI, 2012, 33(3): 521 ~ 525), the method relies on transmethylase, SAM is decomposed into homocysteine by the catalytic action of adenosylhomocysteine nucleosidase and S-Ribosylhomocysteinase, again to homocysteine chromogenic reaction, operate more loaded down with trivial details, accuracy is also undesirable.Due to the matrix more complicated of sample, bring difficulty to detection.Therefore, set up a kind of sensitive, quick, easy, specificity is high, reproducible economy use detection method, to researchist, manufacturing enterprise, Quality Control personnel, import and export commodity inspection, government administration section etc. in the urgent need to, to in food, medicine, Environmental security, biological sample SAM content accurate quantitative analysis measure very necessary, for SAM produce and pharmacological research also tool be of great significance.
Biologic enzyme electrode sensor is that current exploitation has selectivity, stability, detection speed is fast, selectivity good, sensitivity high, be widely used in medicine clinical, food, environment and biology sample detection field, and transmethylase be modified at detection carbon paste electrode being used for SAM and have no report.
Summary of the invention
Transmethylase and water-soluble CdZnTe quantum dot is the object of the invention is to be embedded in carbon paste electrode, 1-ethyl-3-methylimidazole hexafluoro woods hydrochlorate is adopted to combine with galvanochemistry as a kind of carbon paste electrode of production of adhesive, provide the preparation method of quantum dot/enzyme composite carbon paste electrode, and application detects in SAM.
Instrument and reagent
CHI660B electrochemical workstation (Shanghai Chen Hua instrument company), experiment adopts three-electrode system: platinum electrode is auxiliary electrode, and Ag/AgCl is contrast electrode (SCE), and quantum dot/enzyme composite carbon paste electrode (GCE) is working electrode; KQ-250E type ultrasonic cleaner (Kun Feng ultrasonic instrument company limited).
Dag, carbon nano-tube, tellurium powder, caddy, zinc chloride, sodium borohydride, halfcystine, 1-ethyl-3-methylimidazole hexafluorophosphate, NaOH, transmethylase (E.C.2.1.1.3), DNA, SAM; Sulfuric acid, nitric acid, hydrogen peroxide, phosphate buffered solution, it is pure that agents useful for same is analysis, and water is deionized water.
Object of the present invention is achieved through the following technical solutions.
A preparation method for quantum dot/enzyme composite carbon paste electrode, it is characterized in that, the method has following processing step:
(1) semiconductor precursor solution preparation: in the reactor, be in 25 ~ 50 times of deionized waters by the quality that adds of mol ratio 1:2 ~ 4 of tellurium powder and sodium borohydride, under argon shield, ultrasonic disperse 10 min, in 60 DEG C of constant temperature back flow reaction reaction 4 ~ 6h, be cooled to room temperature, be placed in ice-water bath, obtain semiconductor precursor solution;
(2) zinc cadmium precursor liquid preparation: in the reactor, caddy is added: 0.1 ~ 1.2% by following mass percent, zinc chloride: 0.1 ~ 1.0%, halfcystine: 0.2 ~ 1.5%, deionized water: 97 ~ 99%, each component sum is absolutely, be 10 with sodium hydrate regulator solution pH, pass into argon gas, in 80 ~ 90 DEG C of constant temperature back flow reaction 3 ~ 5h, be cooled to room temperature, obtained zinc cadmium precursor liquid;
(3) preparation of water-soluble CdZnTe quantum dot: semiconductor precursor solution step (1) prepared adds in zinc cadmium precursor liquid prepared by step (2) under argon shield, in 85 DEG C of constant temperature back flow reaction 4 ~ 6h, be cooled to room temperature, obtained water-soluble CdZnTe quantum dot;
(4) carbon nano-tube and dag pre-service: in the reactor, add dag by following mass percent: 20 ~ 30%, carbon nano-tube: 8 ~ 15%, the concentrated sulphuric acid: 40 ~ 50%, red fuming nitric acid (RFNA): 10 ~ 20%, each component sum is absolutely, in 80 ~ 90 DEG C of isothermal reaction 3 ~ 5h, be cooled to room temperature, filter, spend deionized water for neutral, dry in thermostatic drying chamber, be ground to powder, obtain pre-service carbon nano-tube and dag potpourri;
(5) preparation of quantum dot/enzyme composite carbon paste electrode: by water-soluble CdZnTe quantum dot: transmethylase: 1-ethyl-3-methylimidazole hexafluorophosphate: carbon nano-tube and dag potpourri mix for 1:0.2 ~ 0.4:4 ~ 10:80 ~ 90 in mass ratio, in agate mortar, grinding evenly, dry in 60 DEG C of warm drying boxes, obtain water-soluble CdZnTe quantum dot/transmethylase/1-ethyl-3-methylimidazole hexafluorophosphate/carbon nano-tube and dag potpourri carbon paste; Then its carbon paste being loaded is connected with in copper cash in the glass tube for Φ 4mm, compacting, and with abrasive paper for metallograph polishing, polishing, deionized water washs, and obtains quantum dot/enzyme composite carbon paste electrode.
It is as follows that quantum dot/enzyme composite carbon paste electrode sensor measures SAM step:
(1) standard solution preparation: prepare the SAM standard solution that a group comprises the variable concentrations of blank standard specimen, end liquid is the phosphate buffered solution of pH7.0 ~ 8.0;
(2) be contrast electrode by Ag/AgCl, platinum electrode is auxiliary electrode, quantum dot prepared by the present invention/enzyme composite carbon paste electrode is working electrode composition three-electrode system, connect CHI660B electrochemical workstation, end liquid is the phosphate buffered solution of pH7.0, in the potential range of-1.5 ~ 0.2V, with 50mV/s scan round 15min, take out washing.Then adopt chronoamperometry to scan this solution, operating voltage is-1.1V, gets the peak point current of SAM under variable concentrations and SAM concentration and to work curve;
(3) detection of SAM: replace the SAM standard solution in step (1) with testing sample, detects according to the method for step (2), according to the difference that response current reduces
△ Iand working curve, obtain the content of SAM in testing sample.
Advantage of the present invention and effect are:
(1) the present invention prepares quantum dot/enzyme composite carbon paste electrode, adulterate water-soluble CdZnTe quantum dot in carbon paste electrode, using 1-ethyl-3-methylimidazole hexafluorophosphate as tackifier, carbon paste electrode carbon nano-tube and dag are mixed with improves 1 ~ 2 times than common carbon paste electrode electric conductivity, and do not need the finishing of electrode, the advantages such as electrochemical window is wide, preparation method is simple, cost is low, surface easily upgrades, aftercurrent is little;
(2) this quantum dot/enzyme composite carbon paste electrode sensor shows very high selectivity and sensitivity to SAM, and the concentration of response current and SAM is in good linear relationship within the scope of 0.5 ~ 12 μm of ol/L, and coefficient R=0.998, detects and be limited to 8.92 × 10
-7mol/L;
(3) this quantum dot/enzyme composite carbon paste electrode does not use poisonous reagent, environment-friendly and green in the process of preparation;
(4) quantum dot the present invention prepared/enzyme composite carbon paste electrode sensor is used successfully in the detection of SAM in medicine, food, solves SAM detection difficult.
Embodiment
Embodiment 1
(1) semiconductor precursor solution preparation: in the reactor, add the tellurium powder of 1.0g respectively, the sodium borohydride of 0.9g, in the deionized water of 50mL, under argon shield, ultrasonic disperse 10 min, in 60 DEG C of constant temperature back flow reaction reaction 5h, be cooled to room temperature, be placed in ice-water bath, obtain semiconductor precursor solution;
(2) zinc cadmium precursor liquid preparation: in the reactor, add caddy respectively: 0.6g, zinc chloride: 0.5g, halfcystine: 0.9g, water: 98mL, be 10 with sodium hydrate regulator solution pH, pass into argon gas, in 85 DEG C of constant temperature back flow reaction 4h, be cooled to room temperature, obtained zinc cadmium precursor liquid;
(3) preparation of water-soluble CdZnTe quantum dot: semiconductor precursor solution step (1) prepared adds in zinc cadmium precursor liquid prepared by step (2) under argon shield, in 85 DEG C of constant temperature back flow reaction 5h, be cooled to room temperature, obtained water-soluble CdZnTe quantum dot;
(4) carbon nano-tube and dag pre-service: in the reactor, add 25g dag respectively, 10g carbon nano-tube, the 25mL concentrated sulphuric acid, 14mL red fuming nitric acid (RFNA), in 85 DEG C of isothermal reaction 4h, be cooled to room temperature, filter, spend deionized water for neutral, dry in thermostatic drying chamber, be ground to powder, obtain pre-service carbon nano-tube and dag potpourri;
(5) preparation of quantum dot/enzyme composite carbon paste electrode: in agate mortar, add the transmethylase of 0.2g respectively, the water-soluble CdZnTe quantum dot of 0.8g, the 1-ethyl-3-methylimidazole hexafluorophosphate of 2.0g, 18g carbon nano-tube and dag potpourri, in agate mortar, grinding evenly, dry in 60 DEG C of warm drying boxes, obtains CdZnTe quantum dot/transmethylase/1-ethyl-3-methylimidazole hexafluorophosphate/carbon nano-tube and dag potpourri carbon paste; Then its carbon paste being loaded is connected with in copper cash in the glass tube for Φ 4mm, compacting, and with abrasive paper for metallograph polishing, polishing, deionized water washs, and obtains quantum dot/enzyme composite carbon paste electrode.
Embodiment 2
(1) semiconductor precursor solution preparation: in the reactor, add the tellurium powder of 0.5g respectively, the sodium borohydride of 0.6g, in the deionized water of 30mL, under argon shield, ultrasonic disperse 10 min, in 60 DEG C of constant temperature back flow reaction reaction 4h, be cooled to room temperature, be placed in ice-water bath, obtain semiconductor precursor solution;
(2) zinc cadmium precursor liquid preparation: in the reactor, add caddy respectively: 1.0g, zinc chloride: 0.8g, halfcystine: 1.2g, water: 97mL, be 10 with sodium hydrate regulator solution pH, pass into argon gas, in 80 DEG C of constant temperature back flow reaction 5h, be cooled to room temperature, obtained zinc cadmium precursor liquid;
(3) preparation of water-soluble CdZnTe quantum dot: semiconductor precursor solution step (1) prepared adds in zinc cadmium precursor liquid prepared by step (2) under argon shield, in 85 DEG C of constant temperature back flow reaction 4h, be cooled to room temperature, obtained water-soluble CdZnTe quantum dot;
(4) carbon nano-tube and dag pre-service: in the reactor, add 20g dag respectively, 15g carbon nano-tube, the 27mL concentrated sulphuric acid, 11mL red fuming nitric acid (RFNA), in 80 DEG C of isothermal reaction 5h, be cooled to room temperature, filter, spend deionized water for neutral, dry in thermostatic drying chamber, be ground to powder, obtain pre-service carbon nano-tube and dag potpourri;
(5) preparation of quantum dot/enzyme composite carbon paste electrode: in agate mortar, add the transmethylase of 0.1g respectively, the water-soluble CdZnTe quantum dot of 0.5g, the 1-ethyl-3-methylimidazole hexafluorophosphate of 1.2g, 9.5g carbon nano-tube and dag potpourri, in agate mortar, grinding evenly, dry in 60 DEG C of warm drying boxes, obtains CdZnTe quantum dot/transmethylase/1-ethyl-3-methylimidazole hexafluorophosphate/carbon nano-tube and dag potpourri carbon paste; Then its carbon paste being loaded is connected with in copper cash in the glass tube for Φ 4mm, compacting, and with abrasive paper for metallograph polishing, polishing, deionized water washs, and obtains quantum dot/enzyme composite carbon paste electrode.
Embodiment 3
(1) semiconductor precursor solution preparation: in the reactor, add the tellurium powder of 0.8g respectively, the sodium borohydride of 0.5g, in the deionized water of 40mL, under argon shield, ultrasonic disperse 10 min, in 60 DEG C of constant temperature back flow reaction reaction 6h, be cooled to room temperature, be placed in ice-water bath, obtain semiconductor precursor solution;
(2) zinc cadmium precursor liquid preparation: in the reactor, add caddy respectively: 0.2g, zinc chloride: 0.2g, halfcystine: 0.6g, water: 99mL, be 10 with sodium hydrate regulator solution pH, pass into argon gas, in 85 DEG C of constant temperature back flow reaction 4h, be cooled to room temperature, obtained zinc cadmium precursor liquid;
(3) preparation of water-soluble CdZnTe quantum dot: semiconductor precursor solution step (1) prepared adds in zinc cadmium precursor liquid prepared by step (2) under argon shield, in 85 DEG C of constant temperature back flow reaction 6h, be cooled to room temperature, obtained water-soluble CdZnTe quantum dot;
(4) carbon nano-tube and dag pre-service: in the reactor, add 30g dag respectively, 8g carbon nano-tube, the 24mL concentrated sulphuric acid, 13mL red fuming nitric acid (RFNA), in 90 DEG C of isothermal reaction 4.5h, be cooled to room temperature, filter, spend deionized water for neutral, dry in thermostatic drying chamber, be ground to powder, obtain pre-service carbon nano-tube and dag potpourri;
(5) preparation of quantum dot/enzyme composite carbon paste electrode: in agate mortar, add the transmethylase of 0.1g respectively, the water-soluble CdZnTe quantum dot of 0.2g, the 1-ethyl-3-methylimidazole hexafluorophosphate of 0.6g, 8.5g carbon nano-tube and dag potpourri, in agate mortar, grinding evenly, dry in 60 DEG C of warm drying boxes, obtains CdZnTe quantum dot/transmethylase/1-ethyl-3-methylimidazole hexafluorophosphate/carbon nano-tube and dag potpourri carbon paste; Then its carbon paste being loaded is connected with in copper cash in the glass tube for Φ 4mm, compacting, and with abrasive paper for metallograph polishing, polishing, deionized water washs, and obtains quantum dot/enzyme composite carbon paste electrode.
Embodiment 4
(1) semiconductor precursor solution preparation: in the reactor, add the tellurium powder of 0.6g respectively, the sodium borohydride of 0.4g, in the deionized water of 50mL, under argon shield, ultrasonic disperse 10 min, in 60 DEG C of constant temperature back flow reaction reaction 5.5h, be cooled to room temperature, be placed in ice-water bath, obtain semiconductor precursor solution;
(2) zinc cadmium precursor liquid preparation: in the reactor, add caddy respectively: 1.0g, zinc chloride: 1.0g, halfcystine: 1.0g, water: 97mL, be 10 with sodium hydrate regulator solution pH, pass into argon gas, in 85 DEG C of constant temperature back flow reaction 4.5h, be cooled to room temperature, obtained zinc cadmium precursor liquid;
(3) preparation of water-soluble CdZnTe quantum dot: semiconductor precursor solution step (1) prepared adds in zinc cadmium precursor liquid prepared by step (2) under argon shield, in 85 DEG C of constant temperature back flow reaction 5.5h, be cooled to room temperature, obtained water-soluble CdZnTe quantum dot;
(4) carbon nano-tube and dag pre-service: in the reactor, add 28g dag respectively, 12g carbon nano-tube, the 26mL concentrated sulphuric acid, 9mL red fuming nitric acid (RFNA), in 85 DEG C of isothermal reaction 4.5h, be cooled to room temperature, filter, spend deionized water for neutral, dry in thermostatic drying chamber, be ground to powder, obtain pre-service carbon nano-tube and dag potpourri;
(5) preparation of quantum dot/enzyme composite carbon paste electrode: in agate mortar, add the transmethylase of 0.1g respectively, the water-soluble CdZnTe quantum dot of 0.3g, the 1-ethyl-3-methylimidazole hexafluorophosphate of 0.8g, 9g carbon nano-tube and dag potpourri, in agate mortar, grinding evenly, dry in 60 DEG C of warm drying boxes, obtains CdZnTe quantum dot/transmethylase/1-ethyl-3-methylimidazole hexafluorophosphate/carbon nano-tube and dag potpourri carbon paste; Then its carbon paste being loaded is connected with in copper cash in the glass tube for Φ 4mm, compacting, and with abrasive paper for metallograph polishing, polishing, deionized water washs, and obtains quantum dot/enzyme composite carbon paste electrode.
Embodiment 5
By the quantum dot prepared by above-described embodiment 1 ~ 4/enzyme composite carbon paste electrode sensor, for the detection of SAM in medicine, step is as follows:
(1) standard solution preparation: prepare the SAM standard solution that a group comprises the variable concentrations of blank standard specimen, end liquid is the phosphate buffered solution of pH 7.5;
(2) working curve is drawn: be contrast electrode by Ag/AgCl, platinum electrode is auxiliary electrode, electrode prepared by the present invention is working electrode composition three-electrode system, connect CHI660B electrochemical workstation, chronoamperometry is adopted to scan this solution, operating voltage is-1.1V, the peak point current of SAM under variable concentrations and SAM concentration is gone to work curve, the regression equation of working curve is I=0.017+0.409c (μm ol/L), coefficient R=0.998, the range of linearity detected is 0.5 ~ 12 μm of ol/L, detection limit 18.92 × 10
-7mol/L;
(3) detection of SAM: get 20, Transmetil tablet, after grinding, 1 hour is gone with deionized water leaching, filter, filtrate constant volume, in 250 mL volumetric flasks, is diluted to during mensuration within the scope of working curve, the SAM standard solution in step (1) is replaced with testing sample, detect according to the method for step (2), according to response current value and working curve, obtain the content of SAM in testing sample; The recovery is between 96.21 ~ 104.65%.
Immobilized transmethylase electrode sensor prepared by the present invention is used successfully in the detection of SAM in medicine, food, biological sample, the recovery is between 96.21 ~ 104.65%%, therefore the molecular engram sensor that prepared by the present invention can be widely used in the association areas such as chemical industry, biological medicine, food, environmental protection tests, solves the difficulty that SAM detects.
Claims (3)
1. a preparation method for quantum dot/enzyme composite carbon paste electrode, it is characterized in that, the method has following processing step:
(1) semiconductor precursor solution preparation: in the reactor, be in 25 ~ 50 times of deionized waters by the quality that adds of mol ratio 1:2 ~ 4 of tellurium powder and sodium borohydride, under argon shield, ultrasonic disperse 10 min, in 60 DEG C of constant temperature back flow reaction reaction 4 ~ 6h, be cooled to room temperature, be placed in ice-water bath, obtain semiconductor precursor solution;
(2) zinc cadmium precursor liquid preparation: in the reactor, caddy is added: 0.1 ~ 1.2% by following mass percent, zinc chloride: 0.1 ~ 1.0%, halfcystine: 0.2 ~ 1.5%, deionized water: 97 ~ 99%, each component sum is absolutely, be 10 with sodium hydrate regulator solution pH, pass into argon gas, in 80 ~ 90 DEG C of constant temperature back flow reaction 3 ~ 5h, be cooled to room temperature, obtained zinc cadmium precursor liquid;
(3) preparation of water-soluble CdZnTe quantum dot: semiconductor precursor solution step (1) prepared adds in zinc cadmium precursor liquid prepared by step (2) under argon shield, in 85 DEG C of constant temperature back flow reaction 4 ~ 6h, be cooled to room temperature, obtained water-soluble CdZnTe quantum dot;
(4) carbon nano-tube and dag pre-service: in the reactor, add dag by following mass percent: 20 ~ 30%, carbon nano-tube: 8 ~ 15%, the concentrated sulphuric acid: 40 ~ 50%, red fuming nitric acid (RFNA): 10 ~ 20%, each component sum is absolutely, in 80 ~ 90 DEG C of isothermal reaction 3 ~ 5h, be cooled to room temperature, filter, spend deionized water for neutral, dry in thermostatic drying chamber, be ground to powder, obtain pre-service carbon nano-tube and dag potpourri;
(5) preparation of quantum dot/enzyme composite carbon paste electrode: by water-soluble CdZnTe quantum dot: transmethylase: 1-ethyl-3-methylimidazole hexafluorophosphate: carbon nano-tube and dag potpourri mix for 1:0.2 ~ 0.4:4 ~ 10:80 ~ 90 in mass ratio, in agate mortar, grinding evenly, dry in 60 DEG C of warm drying boxes, obtain water-soluble CdZnTe quantum dot/transmethylase/1-ethyl-3-methylimidazole hexafluorophosphate/carbon nano-tube and dag potpourri carbon paste; Then its carbon paste being loaded is connected with in copper cash in the glass tube for Φ 4mm, compacting, and with abrasive paper for metallograph polishing, polishing, deionized water washs, and obtains quantum dot/enzyme composite carbon paste electrode.
2. the preparation method of a kind of quantum dot according to claim 1/enzyme composite carbon paste electrode, is characterised in that, the transmethylase described in step (5) is E.C.2.1.1.3 type transmethylase.
3. the quantum dot prepared by the preparation method/enzyme composite carbon paste electrode sensor of a kind of quantum dot according to claim 1/enzyme composite carbon paste electrode, is characterised in that, prepared quantum dot/enzyme composite carbon paste electrode sensor is used for the mensuration of SAM in sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510389582.1A CN104965014B (en) | 2015-07-06 | 2015-07-06 | The preparation method of the compound carbon paste electrode of quantum dot/enzyme for detecting SAM |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510389582.1A CN104965014B (en) | 2015-07-06 | 2015-07-06 | The preparation method of the compound carbon paste electrode of quantum dot/enzyme for detecting SAM |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104965014A true CN104965014A (en) | 2015-10-07 |
| CN104965014B CN104965014B (en) | 2017-06-23 |
Family
ID=54219061
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510389582.1A Expired - Fee Related CN104965014B (en) | 2015-07-06 | 2015-07-06 | The preparation method of the compound carbon paste electrode of quantum dot/enzyme for detecting SAM |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104965014B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548107B (en) * | 2015-12-16 | 2018-05-25 | 黄淮学院 | A kind of preparation method of luminous precursor and its application in iron ion detection and method |
| CN108663421A (en) * | 2018-05-14 | 2018-10-16 | 济南大学 | A kind of preparation of nano TiC/graphene composite paste electrode sensor |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1496411A (en) * | 2001-01-12 | 2004-05-12 | ��Ī������ | Method and apparatus for simultaneous quantification of different radionuclides on the surface of biological microarray |
| US20060024722A1 (en) * | 2004-07-30 | 2006-02-02 | Mark Fischer-Colbrie | Samples for detection of oncofetal fibronectin and uses thereof |
| WO2007032788A2 (en) * | 2005-05-24 | 2007-03-22 | Sandia National Laboratories | Method and apparatus for gel electrophoretic immunoassay |
| CN102879442A (en) * | 2012-10-12 | 2013-01-16 | 上海交通大学 | Graphene quantum dot modified electrochemical sensor, and preparation method and application thereof |
| CN103454325A (en) * | 2013-09-04 | 2013-12-18 | 上海移宇科技有限公司 | Photocatalysed glucose microelectrode sensor and preparation method thereof |
-
2015
- 2015-07-06 CN CN201510389582.1A patent/CN104965014B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1496411A (en) * | 2001-01-12 | 2004-05-12 | ��Ī������ | Method and apparatus for simultaneous quantification of different radionuclides on the surface of biological microarray |
| US20060024722A1 (en) * | 2004-07-30 | 2006-02-02 | Mark Fischer-Colbrie | Samples for detection of oncofetal fibronectin and uses thereof |
| WO2007032788A2 (en) * | 2005-05-24 | 2007-03-22 | Sandia National Laboratories | Method and apparatus for gel electrophoretic immunoassay |
| CN102879442A (en) * | 2012-10-12 | 2013-01-16 | 上海交通大学 | Graphene quantum dot modified electrochemical sensor, and preparation method and application thereof |
| CN103454325A (en) * | 2013-09-04 | 2013-12-18 | 上海移宇科技有限公司 | Photocatalysed glucose microelectrode sensor and preparation method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548107B (en) * | 2015-12-16 | 2018-05-25 | 黄淮学院 | A kind of preparation method of luminous precursor and its application in iron ion detection and method |
| CN108663421A (en) * | 2018-05-14 | 2018-10-16 | 济南大学 | A kind of preparation of nano TiC/graphene composite paste electrode sensor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104965014B (en) | 2017-06-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105044171A (en) | Production method and application of nano-platinum doped/enzyme modified carbon paste electrode | |
| Danhel et al. | Amalgam electrodes in organic electrochemistry | |
| CN107064264B (en) | A kind of construction method of the optical electro-chemistry sensor for no enzyme detection dopamine | |
| Tashkhourian et al. | Simultaneous determination of tyrosine and tryptophan by mesoporous silica nanoparticles modified carbon paste electrode using H-point standard addition method | |
| Amiri et al. | Electrocatalytic determination of sumatriptan on the surface of carbon-paste electrode modified with a composite of cobalt/Schiff-base complex and carbon nanotube | |
| CN105784796B (en) | A kind of sensitive determination method of the aptamer sensor based on gold/molybdenum disulfide/graphene nanocomposite material to lysozyme | |
| CN105510420B (en) | A kind of method based on Beads enrichment and DNA marker gold nanoparticle probe detection ATP contents | |
| CN103954675B (en) | A kind of preparation method of S-adenosylmethionine molecular engram sensor and application | |
| CN109001276B (en) | Construction and application of electrochemical sensor based on COFs (chemical-organic frameworks) material | |
| Mao et al. | Copper-enhanced gold nanoparticle tags for electrochemical stripping detection of human IgG | |
| CN105954336A (en) | Enzyme-free superoxide anion electrochemical sensor, and production method and application thereof | |
| Hwa et al. | Development of biocompatible cellulose microfiber stabilized carbon nanofiber hydrogel for the efficient electrochemical determination of nicotinamide adenine dinucleotide in physiological fluids | |
| CN102192942B (en) | Method for rapidly determining electron transfer capacity of DOM (dissolved organic matter) | |
| CN107044978B (en) | Glutathione electrogenerated chemiluminescence measuring method based on gold nano cluster probe | |
| Arul et al. | Graphene quantum dots-based nanocomposite for electrocatalytic application of L-cysteine in whole blood and live cells | |
| CN109187687B (en) | Preparation of conjugated organic microporous material modified electrode and application of modified electrode as peroxynitroso anion electrochemical sensor | |
| CN104965014A (en) | Preparation method of quantum dot/enzyme compounded carbon paste electrode for detecting SAM (S-Adenosylmethionine) | |
| CN109580731A (en) | The preparation method and the application in detection Polychlorinated biphenyls of DNA micro-capsule and gold electrode-DNA dendritic macromole sensor | |
| Qin et al. | Plant tissue-based chemiluminescence flow biosensor for urea | |
| Benvidi et al. | Developing a nanostructure electrochemical sensor for simultaneous determination of cysteine and tryptophan | |
| CN102830147B (en) | Modified electrode and applications of modified electrode on detecting micro/trace nitro aromatic compound | |
| CN104132919A (en) | Method for detecting nano-silver particle by using fluorescence method based on G-tetramer | |
| CN103344684B (en) | Method for manufacturing magnetic field induction self-assembled creatinine molecular imprinting electrochemical sensor | |
| CN110441373A (en) | A kind of preparation method of oxidation of ethanol enzyme modification composite paste electrode sensor | |
| CN106596676A (en) | Electrochemical method for microRNAs detection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170623 Termination date: 20200706 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |