CN108490041B - A kind of urease biologic sensor - Google Patents

A kind of urease biologic sensor Download PDF

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CN108490041B
CN108490041B CN201810166839.0A CN201810166839A CN108490041B CN 108490041 B CN108490041 B CN 108490041B CN 201810166839 A CN201810166839 A CN 201810166839A CN 108490041 B CN108490041 B CN 108490041B
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platinum
carbon ball
chitosan
biologic sensor
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CN108490041A (en
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吴梦笔
曹锡忠
吴丽娜
董绍伟
周静珠
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JIANGSU ACADEMY OF SCIENCE AND TECHNOLOGY FOR INSPECTION AND QUARANTINE
JIANGSU ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU
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JIANGSU ACADEMY OF SCIENCE AND TECHNOLOGY FOR INSPECTION AND QUARANTINE
JIANGSU ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles

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Abstract

The invention discloses a urease biologic sensors, belong to analysis detection field.The sensor is to construct a kind of based on polyaniline film and chitosan/platinum carbon ball biosensor using screen printing electrode, realizes the quick detection of heavy metal ion.The urease biologic sensor that technical solution of the present invention is prepared has good reproducibility and time saving, laborsaving, cheap, reduction solvent, reduction are to pollution of environment etc..Sensor saves in the refrigerator that can be placed in 4 DEG C when not used, and the response of three weeks inner potential does not all decline, and the 95% of initial potential response is still able to maintain after saving 30 days, illustrates that chitosan can effectively keep the activity of urase, and can prevent enzyme from leaking.

Description

A kind of urease biologic sensor
The application is the applying date are as follows: on May 20th, 2016, application No. is 201610340251.3, titles are as follows: based on poly- Aniline modifies the divisional application of the urease biologic sensor of screen printing electrode and its patent of invention of application.
Technical field
The invention belongs to analysis detection fields, and in particular to a kind of urease biologic sensor and its application.
Background technique
With popularizing for green consumption idea, ecological textile increasingly becomes the mainstream in market, in recent years, in textile Index one of of the detection of extractable heavy metal as ecological textile is increasingly taken seriously.Heavy metal is main in textile It is dyestuff and auxiliary agent used in the process, such as various premetallized dyes, medium fuel, phthalein mountain valley with clumps of trees and bamboo structure dyestuff, solid Toner, catalyst, fire retardant, post-finishing agent etc. and for soft water hardening, desizing is concise, various in the bleaching processes such as stamp Metal chelating agent.It is absorption of human body inside human skin that extractable heavy metal can be entered by the sweat of human body in textile, Can cause health when heavy metal is accumulated to a certain extent in human organ huge in liver, bone, kidney, the heart and brain Damage.The detection method of extractable heavy metal is mainly GB/T 17593 in the textile of national regulation at present, and which specify spinnings The test method of the various heavies such as arsenic, cadmium, cobalt, chromium, copper, nickel, lead, antimony in fabric.Main analysis instrument to be used is atom Absorption spectrum and inductive coupling plasma emission spectrum.
Although existing analysis method precision is good, accuracy is high, and existing analysis method is not able to satisfy time saving, province Power, cheap, reduction solvent, reduction are to pollution of environment etc..
Summary of the invention
The present invention be directed to problems of the existing technology to provide a kind of urea based on Polyaniline-modified screen printing electrode Enzyme biologic sensor and its application.
The purpose of the present invention can be achieved through the following technical solutions:
A kind of urease biologic sensor based on Polyaniline-modified screen printing electrode, the sensor are by the following method It is prepared:
(1) screen printing electrode screen printing electrode electropolymerization polyaniline: is placed in the electricity containing hydrochloric acid and aniline first In polymeric solution, electropolymerization is carried out using cyclic voltammetry later, obtains PAIN modified electrode；
(2) platinum/carbon ball nanocomposite: carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to chloroplatinic acid and is uniformly mixing to obtain mixed liquor in carbon slurry, adjusts the mixed liquor to alkalinity using alkaline reagent, 100~150 DEG C of reductase 12~6h are warming up to, later by the washing of obtained black product, drying, obtain the nano combined material of platinum/carbon ball Material；
(3) urease biologic sensor: the platinum that chitosan and step (2) are prepared/carbon ball nanocomposite is added It into acetum and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution；By chitosan-platinum/carbon ball mixed solution with Urase solution is uniformly mixed, and obtains urase-chitosan/platinum/carbon ball mixed solution；In the PAIN modification that step (1) is prepared Electrode surface drop coating urase-chitosan/platinum/carbon ball mixed solution, is prepared urease biologic sensor.Drop coating amount is about 15 μ L/cm2。
A kind of urease biologic sensor preparation method based on Polyaniline-modified screen printing electrode, this method includes following Step:
(1) screen printing electrode screen printing electrode electropolymerization polyaniline: is placed in the electricity containing hydrochloric acid and aniline first In polymeric solution, electropolymerization is carried out using cyclic voltammetry later, obtains PAIN modified electrode；
(2) platinum/carbon ball nanocomposite: carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to chloroplatinic acid and is uniformly mixing to obtain mixed liquor in carbon slurry, adjusts the mixed liquor to alkalinity using alkaline reagent, 100~150 DEG C of reductase 12~6h are warming up to, later by the washing of obtained black product, drying, obtain the nano combined material of platinum/carbon ball Material；
(3) urease biologic sensor: the platinum that chitosan and step (2) are prepared/carbon ball nanocomposite is added It into acetum and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution；By chitosan-platinum/carbon ball mixed solution with Urase solution is uniformly mixed, and obtains urase-chitosan/platinum/carbon ball mixed solution；In the PAIN modification that step (1) is prepared Electrode surface drop coating urase-chitosan/platinum/carbon ball mixed solution, is prepared urease biologic sensor.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof: the electropolymerization process of step (1) It is to be carried out under conditions of nitrogen protection, cyclic voltammetry scanning range is -0.1V~0.7V, the rate of scanning is 40~ 60mV/s is always scanned 80~120 times.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof: the electropolymerization solution of step (1) The concentration of middle hydrochloric acid is 0.1~3.5mol/L, and the concentration of aniline is 0.1~3mol/L.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof: the carbon ball and chlorine platinum of step (2) The mass ratio of acid is 1~3:1.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof: the alkalinity of step (2) refers to mixed The pH value for closing liquid is 9~11.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof: chitosan-platinum of step (3)/ In carbon ball mixed solution the mass fraction of chitosan be 0.1~1%, platinum/carbon ball nanocomposite mass fraction be 0.5~ 1.5%.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof: the urase solution of step (3) is dense Spending is 5~15mg/ml, and the volume ratio of chitosan-platinum/carbon ball mixed solution and urase solution is 1~5:1~5.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof: the drop coating amount of step (3) is 10 ~30 μ L/cm2。
Application of the aforementioned PAIN modified electrode in measurement pH value of solution；It is preferred that the inhibition time measured be >= 20min, response time >=2min.
Application of the aforementioned urease biologic sensor in detection heavy metal ion；It is preferred that being detected in urea liquid Hg2+And Cd2+In application；Hg in textile is detected more preferably in urea liquid2+And Cd2+Application.The inhibition time of measurement For >=20min, response time >=2min；It is preferred that the concentration of urea liquid≤selection 40mmol/L.
Beneficial effects of the present invention:
The urease biologic sensor that technical solution of the present invention is prepared have good reproducibility and it is time saving, laborsaving, cheap, It reduces solvent, reduce to pollution of environment etc..Sensor saves in the refrigerator that can be placed in 4 DEG C when not used, in three weeks Potential response does not all decline, and the 95% of initial potential response is still able to maintain after saving 30 days, illustrates that chitosan can be effectively The activity of urase is kept, and can prevent enzyme from leaking.
Detailed description of the invention
Fig. 1 powers on 100 resulting voltammograms of polymerisation loop in screen printing electrode for 1 aniline monomer of embodiment.
Before and after Fig. 2 is aniline polymerization, the circulation of screen printing electrode is bent over the desk figure.
Fig. 3 is the potential response of PAIN modified electrode and the relational graph of solution ph.
Fig. 4 is that urease biologic sensor is placed in the potential response of urea liquid and changes with time figure.
Fig. 5 is in Hg2+Or Cd2+In the presence of, inhibiting rate, which changes with time, sees Fig. 5
Fig. 6 is that urease biologic sensor is placed in the solution containing urea, urea concentration and potential change figure.
Fig. 7 is that urease biologic sensor is placed in the solution containing urea, the relational graph of inhibiting rate and heavy metal concentration.
Specific embodiment
Below with reference to embodiment, the present invention will be further described, and but the scope of the present invention is not limited thereto:
Carbon ball described in the embodiment of the present invention can be commercial product or be prepared with the following method: with first Aldehyde-resorcinol is precursor, is usedMethod, 0.1mL ammonium hydroxide (25w%) are added to 8mL ethyl alcohol and 20mL deionized water Mixed solution in, stir 1h after, be added 0.2g resorcinol, stir to being completely dissolved, it is molten that 0.284mL formaldehyde is then added dropwise again Liquid (37w%), mixed liquor stir for 24 hours at 30 DEG C, then move to mixed liquor in water heating kettle, react at 100 DEG C for 24 hours, mixture It is centrifuged, dry 48h at 100 DEG C.350 are heated to next, carrying out charing process with the heating rate of 1 DEG C/min DEG C, 2h is kept, is then heated to 600 DEG C again with the heating rate of 1 DEG C/min, calcining 4h is retained, then naturally cools to room Carbon ball is prepared in temperature.
Embodiment 1
Screen printing electrode electropolymerization polyaniline: screen printing electrode is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, wherein the molar concentration of HC1 is 1mol/L, and the molar concentration of aniline is 0.5mol/L；Cyclic voltammetry is used later Electropolymerization is carried out, cyclic voltammetry scanning range is -0.1V~0.7V, sweep speed 50mV/s, is scanned 100 times in total.It is real Electropolymerization solution need to lead to nitrogen l0min before testing beginning, and entire electropolymerization process is completed under a nitrogen atmosphere.After the completion of electropolymerization, use 1mol/L hydrochloric acid and secondary water are rinsed electrode, dry, and PAIN modified electrode is made；
Platinum/carbon ball nanocomposite: 80mg carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to 42mg chloroplatinic acid in carbon slurry and is uniformly mixing to obtain mixed liquor, adjusts the pH of mixed using NaOH solution To 10,130 DEG C of reduction 3h are warming up to, later by the washing of obtained black product, drying, obtain platinum/carbon ball nanocomposite；
Urease biologic sensor: platinum/carbon ball nanocomposite that chitosan and step (2) are prepared is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution, the mass fraction of chitosan is in the mixed liquor 0.5%, platinum/carbon ball nanocomposite mass fraction is 1%；Chitosan-platinum/carbon ball that volume ratio is 1:1 is mixed molten Liquid is that 10mg/ml urase solution is uniformly mixed with concentration, obtains urase-chitosan/platinum/carbon ball mixed solution；It is made in step (1) Standby obtained 15 μ L/cm of PAIN modified electrode surface drop coating2Urase-chitosan/platinum/carbon ball mixed solution, is prepared urase Biosensor.
Embodiment 2
Screen printing electrode electropolymerization polyaniline: screen printing electrode is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, wherein the molar concentration of HC1 is 2mol/L, and the molar concentration of aniline is 1mol/L；Later using cyclic voltammetry into Row electropolymerization, cyclic voltammetry scanning range are -0.1V~0.7V, sweep speed 40mV/s, are scanned 80 times in total.Experiment is opened Electropolymerization solution need to lead to nitrogen l0min before beginning, and entire electropolymerization process is completed under a nitrogen atmosphere.After the completion of electropolymerization, 1mol/L is used Hydrochloric acid and secondary water are rinsed electrode, dry, and PAIN modified electrode is made；
Platinum/carbon ball nanocomposite: 50mg carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to 42mg chloroplatinic acid in carbon slurry and is uniformly mixing to obtain mixed liquor, adjusts the pH of mixed using NaOH solution To 9,100 DEG C of reduction 6h are warming up to, later by the washing of obtained black product, drying, obtain platinum/carbon ball nanocomposite；
Urease biologic sensor: platinum/carbon ball nanocomposite that chitosan and step (2) are prepared is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution, the mass fraction of chitosan is in the mixed liquor 1%, platinum/carbon ball nanocomposite mass fraction is 1.5%；Chitosan-platinum/carbon ball that volume ratio is 1:3 is mixed molten Liquid is that 5mg/ml urase solution is uniformly mixed with concentration, obtains urase-chitosan/platinum/carbon ball mixed solution；It is made in step (1) Standby obtained 10 μ L/cm of PAIN modified electrode surface drop coating2Urase-chitosan/platinum/carbon ball mixed solution, is prepared urase Biosensor.
Embodiment 3
Screen printing electrode electropolymerization polyaniline: screen printing electrode is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, wherein the molar concentration of HC1 is 3mol/L, and the molar concentration of aniline is 1.5mol/L；Cyclic voltammetry is used later Electropolymerization is carried out, cyclic voltammetry scanning range is -0.1V~0.7V, sweep speed 60mV/s, is scanned 120 times in total.It is real Electropolymerization solution need to lead to nitrogen l0min before testing beginning, and entire electropolymerization process is completed under a nitrogen atmosphere.After the completion of electropolymerization, use 1mol/L hydrochloric acid and secondary water are rinsed electrode, dry, and PAIN modified electrode is made；
Platinum/carbon ball nanocomposite: 120mg carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to 42mg chloroplatinic acid in carbon slurry and is uniformly mixing to obtain mixed liquor, adjusts the pH of mixed using NaOH solution To 11,150 DEG C of reductase 12 h are warming up to, later by the washing of obtained black product, drying, obtain platinum/carbon ball nanocomposite；
Urease biologic sensor: platinum/carbon ball nanocomposite that chitosan and step (2) are prepared is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution, the mass fraction of chitosan is in the mixed liquor 0.4%, platinum/carbon ball nanocomposite mass fraction is 1.5%；Chitosan-platinum/carbon ball that volume ratio is 3:1 is mixed Solution is that 15mg/ml urase solution is uniformly mixed with concentration, obtains urase-chitosan/platinum/carbon ball mixed solution；In step (1) The 30 μ L/cm of PAIN modified electrode surface drop coating being prepared2Urase-chitosan/platinum/carbon ball mixed solution, is prepared urea Enzyme biologic sensor.
Performance detection
The preparation of 1 PAIN modified electrode
Fig. 1 powers on 100 resulting voltammograms of polymerisation loop in screen printing electrode for 1 aniline monomer of embodiment, follows Ring voltammogram shows that its electrode process has invertibity.Spike potential is aoxidized respectively in 0.22V, restores spike potential in 0.08V.The peak The process of radical cation is oxidized to for the aniline of protonation.With the increase of cycle-index, peak current increases, this is because Aniline is once in electrode surface polymerization film formation, it may occur that self-catalyzed reaction, while the film thickness of polymer also gradually increases.Preparation Obtained polyaniline film modified electrode is blue-green.Before and after aniline polymerization, the circulation of screen printing electrode figure of bending over the desk is shown in Fig. 2, bent Line a is before electropolymerization starts, and the cyclic voltammogram of screen printing electrode, curve b is after the completion of electropolymerization, and polyaniline film covers The screen printing electrode of lid recycles figure of bending over the desk.Figure it is seen that the Polyaniline-modified being prepared is electric after electropolymerization There are a pair of apparent redox peaks in pole, and peak current significantly increases, and illustrates that polyaniline film cathode is successfully prepared.
The pH of 2 PAIN modified electrodes is responded
PAIN modified electrode has response to the current potential of solution, the pH value variation for the solution that can be surveyed.The potential response of electrode It is obtained with the relationship of solution ph by detecting the open circuit potential of itself and Ag/AgC1 reference electrode.Different pH value solution, can be with Different potential values is obtained, as a result as shown in Figure 3.The response range of this Polyaniline-modified screen printing electrode be pH 1~ PH12 shows preferable linear relationship, linearly dependent coefficient 0.999, slope are as follows: 60.8mV/pH (T:25 DEG C) is shown in Fig. 3. From the figure 3, it may be seen that polyaniline modified electrode has good potential response.
3 response times and the selection for inhibiting the time
The hydrolysis of urease biologic sensor catalyzing urea needs certain response time, should ensure that when testing identical Response time.Urease biologic sensor is placed in urea liquid, its potential response is tested and changes with time, concrete outcome See Fig. 4, from fig. 4, it can be seen that potential response increase at any time and increase, tend towards stability after the 2 minutes.Therefore in detection electricity When position, selecting the response time i.e. time of enzymatic reacting of electrode is 2min.
In the presence of having heavy metal ion in the solution, the activity of urase will receive inhibition.With the growth of time, Inhibiting rate will increase, and inhibiting rate reaches balance within a certain period of time.Sensor is placed in containing Hg2+Or Cd2+With the solution of urea In, inhibiting rate, which changes with time, sees Fig. 5, it can be seen that from 0-20min, with the growth of time, inhibiting rate increases, After 20min, inhibiting rate reaches a platform, therefore when carrying out heavy metal analysis, the inhibition time selected is 20min.
The selection of 4 urea concentrations
The urease biologic sensor that embodiment 1 is prepared is placed in the solution containing urea, urase can catalyzing urea Hydrolysis, generate CO2And NH3, cause the potential change of solution.The concentration of urea liquid will affect the degree of potential change, urea Concentration is shown in Fig. 6 from the relationship of 10-100mmol/L and potential change.From fig. 6, it can be seen that urea concentration is from 10 to 40mmol/L When, potential difference is directly proportional to urea concentration, urea concentration from 50mmol/L to 100mmol/L potential difference with urea concentration Variation, speedup slows down, this is because the enzyme that electrode surface is fixed, is only capable of the urea of catalysis Finite Concentration, when urea concentration mistake Gao Shi, the catalytic capability of enzyme can tend to be saturated.According to Fig.6, selection≤40mmol/L urea liquid, the most properly.
5 urease biologic sensors are used for the detection of heavy metal ion
The urease biologic sensor that embodiment 1 is prepared is placed in the urea liquid of 40mmol/L, measurement enzymatic is anti- Potential change Δ E caused by answering.The Hg of various concentration is separately added into urea liquid2+(10、50、100、500、1000、2000 μ g/L) and Cd2+(50,100,500,1000,2000,5000 μ g/L) solution, after twenty minutes, the current potential after test inhibition calculates The potential change Δ E' of enzymatic reaction after inhibition.Heavy metal to the inhibiting rate of urease activity be equal to [(Δ E- Δ E')/Δ E] × 100%.
The relationship of inhibiting rate and heavy metal concentration is as shown in fig. 7, it can be seen from figure 7 that Hg2+And Cd2+Inhibiting rate Increase with the increase of concentration, the inhibiting rate of enzymatic activity and the negative logarithm of concentration are in good linear relationship, linear correlation system Number is in 0.99 or more, Hg2+The range of linearity is 10-2000 μ g/L, Cd2+The range of linearity be 500-5000 μ g/L, calculate detection It is limited to Hg2+3.89 μ g/L, Cd2+5.41 μ g/L (are calculated) by inhibiting rate 10%.
The reproducibility and stability of 6 urease biologic sensors
The urease biologic sensor for selecting embodiment 1 to be prepared is to 500 μ g/L Hg2+With 500 μ g/L Cd2+Detection Relative standard deviation is respectively 7.2%, 8.9%.Show that the sensor has good reproducibility.Sensor can set when not used It is saved in 4 DEG C of refrigerator, the response of three weeks inner potential does not all decline, and initial potential response is still able to maintain after saving 30 days 95%, illustrate that chitosan can effectively keep the activity of urase, and can prevent enzyme from leaking.
Detection of 7 sensors to textile actual sample
The preparation of textile samples solution: textile samples first shred into 5mm × 5mm fragment, weigh 2g, and 80mL acid is added Property sweat, be put into thermostatic control oscillator vibration and vibrate after sixty minutes, it is cooling stand-by.Acidic sweat is according to GB/T17593.2-2007 Requirement prepare.
The urease biologic sensor for selecting embodiment 1 to be prepared carries out mark-on reclaims in textile samples extracting solution, Calculate its rate of recovery.Particular content is shown in Table 1, as it can be seen from table 1 the rate of recovery range of sample is between 80%-120%, it can With the detection for actual sample.
The recovery of standard addition of 1 actual sample of table

Claims (5)

1. a kind of urease biologic sensor, it is characterised in that: the sensor is to be prepared via a method which to obtain:
(1) screen printing electrode screen printing electrode electropolymerization polyaniline: is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, electropolymerization is carried out using cyclic voltammetry later, obtains PANI modified electrode；
(2) platinum/carbon ball nanocomposite: carbon ball being added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon slurry, Chloroplatinic acid is slowly added in carbon slurry and is uniformly mixing to obtain mixed liquor, which is adjusted to alkalinity, heating using alkaline reagent Platinum/carbon ball nanocomposite is obtained to 100 ~ 150 DEG C of reductase 12 ~ 6h later by the washing of obtained black product, drying；
(3) platinum/carbon ball nanocomposite that chitosan and step (2) are prepared urease biologic sensor: is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution；By chitosan-platinum/carbon ball mixed solution and urase Solution is uniformly mixed, and obtains urase-chitosan/platinum/carbon ball mixed solution；In the PANI modified electrode that step (1) is prepared Surface liquid-transfering gun drop coating urase-chitosan/platinum/carbon ball mixed solution, is prepared urease biologic sensor；
Wherein: the electropolymerization process of step (1) is carried out under conditions of nitrogen protection, cyclic voltammetry scanning range be- 0.1V ~ 0.7V, the rate of scanning are 40 ~ 60 mV/s, are always scanned 80 ~ 120 times；The carbon ball of step (2) and the mass ratio of chloroplatinic acid For 1 ~ 3:1.
2. urease biologic sensor according to claim 1, it is characterised in that: hydrochloric acid in the electropolymerization solution of step (1) Concentration be 0.1 ~ 3.5 mol/L, the concentration of aniline is 0.1 ~ 3mol/L.
3. urease biologic sensor according to claim 1, it is characterised in that: the alkalinity of step (2) refers to mixed liquor PH value is 9 ~ 11.
4. urease biologic sensor according to claim 1, it is characterised in that: chitosan-platinum/carbon ball of step (3) is mixed The mass fraction for closing chitosan in solution is 0.1 ~ 1%, and platinum/carbon ball nanocomposite mass fraction is 0.5 ~ 1.5%.
5. urease biologic sensor according to claim 1, it is characterised in that: the urase solution concentration of step (3) be 5 ~ The volume ratio of 15mg/ml, chitosan-platinum/carbon ball mixed solution and urase solution is 1 ~ 5:1 ~ 5, and drop coating amount is 10 ~ 30 μ L/ cm2 。
CN201810166839.0A 2016-05-20 2016-05-20 A kind of urease biologic sensor Expired - Fee Related CN108490041B (en)

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