CN109521072A - Detect heteroauxin and salicylic method and its electrochemical sensor used - Google Patents
Detect heteroauxin and salicylic method and its electrochemical sensor used Download PDFInfo
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Abstract
The invention discloses a kind of detection heteroauxin and salicylic method and its electrochemical sensors used, the electrochemical sensor includes three-electrode system and detection cell, three-electrode system includes working electrode, Ag/AgCl reference electrode and platinum filament are to electrode, the working electrode is the glass-carbon electrode of modification, there is the detection bottom liquid of buffer in the detection cell, Electrochemical Detection is carried out to IAA and SA using graphene hydrogel modified electrode, solves general chromatography, immunological detection method is complicated for operation, it is expensive, sample volume is high, the problems such as time-consuming, realize the instant detection of IAA and SA, with convenient and efficient, the high sensitivity of detection, IAA detection is limited down to 0.677 μ g/ml, SA detection is limited down to 0.569 μ g/ml, need sample size few, testing cost Extremely low, the advantages that interference is small, may be implemented to IAA and SA quickly, easy, accurate quantitative detection.
Description
Technical field
The invention belongs to food inspection and electrochemical sensor technology fields, and in particular to a kind of detection heteroauxin and water
The method of poplar acid and its electrochemical sensor used.
Background technique
By plant generate or artificial synthesized plant growth regulator indole-3-acetic acid (IAA) be change plant growth and
The small organic molecule of development, IAA stimulating growth process, similar many plant growth regulator such as abscisic acid (ABA), jasmonic
(JA) it is used widely in many countries such as Australia, Japan, the nations of China and India, which results in people to eat it
With more concerns of the toxicity and residues problem of plant.Legislative bodies it has been noted that protection health maximum residue limit, and
And reducing plant genetic resources debris concentration in commodity as far as possible is most important quality mark in market monitorings and international trade
One of standard.Studies have found that can express to a kind of rice leaf roller (harm by the dosage for adjusting gaultherolin MeSA
The pest of rice) the higher death rate.
Electrochemical sensor with measured matter by having an effect and generating the electric signal proportional with material concentration
Carry out work.Currently used for detecting the more modified electrodes of electrochemical sensor of IAA and SA, such as Suriyanarayanan S, Mandal
S, Ramanujam K,et al.Electrochemically synthesized molecularly imprinted
polythiophene nanostructures as recognition elements for an aspirin-
Chemosensor [J] .Sensors&Actuators B Chemical, 2017,253, using on gold plating quartz resonator
Alumina formwork is sacrificed to be included in by function monomer 3 thienylboronic acid (3-TBA) and 3- thiophene acetic acid (3-TAA) ingredient trace
Polymer (MIP) is electrode, and good effect is achieved in the detection of SA, using can be to microsphere or structure is similar therewith
As the molecule method of realizing specific recognition, these methods based on molecular engram require to carry out electrode prolonged pre-
Processing, and the type of function monomer is very little, is not able to satisfy the identification requirement of certain molecules；Such as Feng Z L, Yao Y Y, Xu J
K,et al.One-step co-electrodeposition of graphene oxide doped poly
(hydroxymethylated-3,4-ethylenedioxythiophene)film and its electrochemical
studies of indole-3-acetic acid[J].Chinese Chemical Letters,2014, 25(4):511-
516, using a step coelectrodeposition graphene oxide doped methylolation -3,4-rthylene dioxythiophene film as electrode, in IAA
Detection in achieve good result, but selected modified electrode material price is expensive, higher cost.
Summary of the invention
It is an object of that present invention to provide a kind of detection heteroauxin and salicylic method and its electrochemical sensings used
Device.
To achieve the above object, present invention provide the technical scheme that a kind of electrochemical sensor, including three electrode bodies
System, which includes working electrode, Ag/AgCl reference electrode and platinum filament to electrode, and the working electrode is modification
Glass-carbon electrode；The preparation method of the working electrode includes: the drop coating graphene hydrogel on glass-carbon electrode, is used after being dried with nitrogen
Distilled water elution is to get the glass-carbon electrode for arriving modification.
Preferred technical solution are as follows: it is 1.0 μm, 0.3 μm and 0.05 μ that the glass-carbon electrode successively uses partial size before modification
The Al of m2O3Powder polishing grinding glass-carbon electrode makes glassy carbon electrode surface to mirror surface, then clear with dehydrated alcohol and distilled water ultrasound
After washing, it is dried with nitrogen spare.
To achieve the above object, present invention provide the technical scheme that a kind of detect indole-3-acetic acid and salicylic acid simultaneously
Method, including the following steps:
Step 1: the drafting of indole-3-acetic acid and salicylic standard curve；
Step 2: measuring samples are pulverized in the presence of liquid nitrogen, then measuring samples powder is added to 4 DEG C of first
Alcoholic solution extracts overnight under the conditions of 4 DEG C, CHCl is then added into solution3, be then first vortexed 8~12s, then in 4 DEG C of conditions
Under, 3~5min is vibrated with 800~100rpm, at 4 DEG C, 4~6min is centrifuged with 12000rpm, the supernatant collected is in room
Under temperature, upper press proof product are dried to obtain with nitrogen, when detecting, disperses upper press proof product in detection bottom liquid and obtains system to be checked,
Then the three-electrode system of claims 1 or 2 is inserted into system to be checked, is detected using linear sweep voltammetry.
Preferred technical solution are as follows: the drafting of the indole-3-acetic acid and salicylic standard curve includes:
(1) preparation of standard solution:
Indole-3-acetic acid is dissolved in dehydrated alcohol, then uses PBS buffer solution by the ethanol solution of indole-3-acetic acid
It is diluted to the indole-3-acetic acid standard solution of various concentration in proportion with PBS buffer solution；Salicylic acid is dissolved in dehydrated alcohol
In, then salicylic ethanol solution is diluted to the water of various concentration in proportion with PBS buffer solution with PBS buffer solution
Poplar acid solution；By the salicylic acid standard solution of the indole-3-acetic acid standard solution of various concentration and various concentration respectively by down to
Height is added in detection bottom liquid, obtains gradient indole-3-acetic acid and salicylic acid standard mixed solution to be checked；Detection bottom liquid is
0.1M PBS buffer solution；
(2) it detects:
By in the three-electrode system insertion detection bottom liquid of claims 1 or 2, detection obtains current signal 1, by claim
1 or 2 three-electrode system successively gradient salicylic acid solution to be checked, using GHs/GCE as working electrode, Ag/AgCl reference electrode
The three-electrode system that electrode is constituted is inserted into gradient indole-3-acetic acid and salicylic acid standard mixed solution to be checked with platinum filament, inspection
Measure current signal 2；
(3) drafting of standard curve:
Using the difference of current signal 1 and current signal 2 as ordinate, the concentration of SA or IAA are abscissa, are drawn simultaneously
Obtain SA standard curve and IAA standard curve.
Detailed description of the invention
Fig. 1 is the preparation flow figure of the graphene hydrogel in the present invention.
Fig. 2 is the electrochemical sensor in the present invention while the detection schematic diagram to IAA and SA.
Fig. 3 is the concentration curve of the IAA and SA of various concentration and the canonical plotting of drafting.
Since above-mentioned technical proposal is used, the present invention has the advantage, that compared with prior art
Graphene hydrogel modified glassy carbon electrode (GHs/GCE) is a kind of using one step hydro thermal method redox graphene shape
At porous structure, to small molecule have very strong adsorption capacity, the electrochemical sensing that direct drop coating is prepared on glass-carbon electrode
Device has the unexistent many advantages of other modified electrodes.It is mainly shown as: while detecting IAA and SA and can effectively save
Duration is detected, used graphene hydrogel material is environmentally protective, flexible design, simple and convenient processing method, at low cost, stability
With it is reproducible, it is anti-that to interfere with each other ability good.
Specific embodiment
Embodiments of the present invention are illustrated by particular specific embodiment below, those skilled in the art can be by this explanation
Content disclosed by book is understood other advantages and efficacy of the present invention easily.
Ginseng as shown in figure 1 to 3, it should however be clear that this specification structure depicted in this specification institute accompanying drawings, ratio, size etc., only to
Cooperate the revealed content of specification, so that those skilled in the art understands and reads, being not intended to limit the invention can be real
The qualifications applied, therefore do not have technical essential meaning, the tune of the modification of any structure, the change of proportionate relationship or size
It is whole, in the case where not influencing the effect of present invention can be generated and the purpose that can reach, it should all still fall in disclosed skill
Art content obtains in the range of capable of covering.Meanwhile in this specification it is cited as "upper", "lower", "left", "right", " centre " and
The term of " one " etc. is merely convenient to being illustrated for narration, rather than to limit the scope of the invention, relativeness
It is altered or modified, under the content of no substantial changes in technology, when being also considered as the enforceable scope of the present invention.
The present invention provides the electrification of a kind of quick, easy, inexpensive detection indole-3-acetic acid (IAA) and salicylic acid (SA)
Sensor is learned, is quickly detected while realization to the detection respectively of IAA, SA or IAA and SA.It is prepared by a step hydrothermal reduction method
Graphene hydrogel modified electrode, pH 2.5,0.1M phosphate buffer solution be buffering bottom liquid, electrode enrichment time be 150
s.This method is 6~200 μM to the linear detection range of IAA and SA, and it is quick to IAA and SA, easy, accurately fixed to may be implemented
Amount detection.
Embodiment 1: a kind of to detect indole-3-acetic acid and salicylic method and its electrochemical sensor used simultaneously
(1) electrode pre-processes: respectively with 1.0,0.3 and 0.05 μm of Al2O3Powder polishing grinding is to mirror surface, then with anhydrous
N after ethyl alcohol and distilled water ultrasonic cleaning2It dries up spare.
(2) preparation of buffer: the NaH of 0.1M is prepared respectively2PO4The H of solution and 0.1M3PO4Solution, with 0.1M's
H3PO4The NaH of solution adjusting 0.1M2PO4PH value of solution is to 2.5.
(3) preparation of graphene hydrogel (GHs):
10mL GO dispersion (3mg/mL) is ultrasonically treated 20 minutes, and 66mg sodium ascorbate is added under ultrasonic treatment, adds
Enter NaOH or H2SO4Solution adjusts the pH to 3.47 of GO dispersion, and mixture is sealed in autoclave, is heated at 90 DEG C
3h is to form GH.
(4) preparation of modified glassy carbon electrode:
The GHs that on glass-carbon electrode prepared by 5 μ L step (1) of drop coating, is dried with nitrogen, is sufficiently eluted with secondary water to get arriving
The glass-carbon electrode GHs/GCE of GHs modification.
(5) standard curve is as shown in Figure 3.
(6) processing of sample:
The Sheep's-parsley vegetable sample bought from local market (Chinese Hefei) is cut into small pieces, and is ground into the presence of liquid nitrogen
Fine powder, the methanol (4 DEG C, 80%) of 2mL pre-cooling is added in the powder after 1g grinding, and after stirring, powder is extracted overnight at 4 DEG C.So
1mL CHCl is added in the solution afterwards3, by its vortex 10s, 4min is vibrated with 900rpm at 4 DEG C, at 4 DEG C, 12000 rpm
It is centrifuged 5min, collects supernatant, and nitrogen is dry at room temperature, sample is dissolved in 1mL 0.1M PBS.
(7) preparation of bottom liquid is detected:
With the H of 0.1M3PO4The NaH of solution adjusting 0.1M2PO4PH value of solution is obtained to 2.5.
(8) detection of sample:
By 0.1mL, the sample is added in the liquid of preprepared detection bottom, carries out Electrochemical Detection using LSV.Initially
Current potential is 0.3V, and termination current potential is 1.3V, and it is 0.80V that IAA, which goes out peak position, and it is 1.05V that SA, which goes out peak position,.By measurement IAA and
SA peak point current is respectively 2.82 μ A and 1.54 μ A, by having obtained IAA the and SA content point in standard curve calculating sample
It Wei not be 8.52 μM and 7.99 μM.
Embodiment 2: a kind of to detect indole-3-acetic acid and salicylic method and its electrochemical sensor used simultaneously
(1) electrode pre-processes: respectively with 1.0,0.3 and 0.05 μm of Al2O3Powder polishing grinding is to mirror surface, then with anhydrous
N after ethyl alcohol and distilled water ultrasonic cleaning2It dries up spare.
(2) preparation of buffer: the NaH of 0.1M is prepared respectively2PO4The H of solution and 0.1M3PO4Solution, with 0.1M's
H3PO4The NaH of solution adjusting 0.1M2PO4PH value of solution is to 2.5.
(3) preparation of graphene hydrogel (GHs):
10mL GO dispersion (3mg/mL) is ultrasonically treated 20 minutes, and 66mg sodium ascorbate is added under ultrasonic treatment, adds
Enter NaOH or H2SO4Solution adjusts the pH to 3.47 of GO dispersion, and mixture is sealed in autoclave, is heated at 90 DEG C
3h is to form GH.
(4) preparation of modified glassy carbon electrode:
The GHs that on glass-carbon electrode prepared by 5 μ L step (1) of drop coating, is dried with nitrogen, is sufficiently eluted with secondary water to get arriving
The glass-carbon electrode GHs/GCE of GHs modification.
(5) standard curve is as shown in Figure 3.
(6) processing of sample:
The tomato vegetable sample bought from local market (Chinese Hefei) is cut into small pieces, and is ground into the presence of liquid nitrogen
Fine powder, the methanol (4 DEG C, 80%) of 2mL pre-cooling is added in the powder after 1g grinding, and after stirring, powder is extracted overnight at 4 DEG C.So
1mL CHCl is added in the solution afterwards3, by its vortex 10s, 4min is vibrated with 900rpm at 4 DEG C, at 4 DEG C, 12000 rpm
It is centrifuged 5min, collects supernatant, and nitrogen is dry at room temperature, sample is dissolved in 1mL 0.1M PBS.
(7) preparation of bottom liquid is detected:
With the H of 0.1M3PO4The NaH of solution adjusting 0.1M2PO4PH value of solution is obtained to 2.5.
(8) detection of sample:
By 0.1mL, the sample is added in the liquid of preprepared detection bottom, carries out Electrochemical Detection using LSV.Initially
Current potential is 0.3V, and termination current potential is 1.3V, and it is 0.80V that IAA, which goes out peak position, and it is 1.05V that SA, which goes out peak position,.By measurement IAA and
SA peak point current is respectively 1.56 μ A and 0.95 μ A, by having obtained IAA the and SA content point in standard curve calculating sample
It Wei not be 4.00 μM and 4.42 μM.
Embodiment 3: a kind of to detect indole-3-acetic acid and salicylic method and its electrochemical sensor used simultaneously
(1) electrode pre-processes: respectively with 1.0,0.3 and 0.05 μm of Al2O3Powder polishing grinding is to mirror surface, then with anhydrous
N after ethyl alcohol and distilled water ultrasonic cleaning2It dries up spare.
(2) preparation of buffer: the NaH of 0.1M is prepared respectively2PO4The H of solution and 0.1M3PO4Solution, with 0.1M's
H3PO4The NaH of solution adjusting 0.1M2PO4PH value of solution is to 2.5.
(3) preparation of graphene hydrogel (GHs):
10mL GO dispersion (3mg/mL) is ultrasonically treated 20 minutes, and 66mg sodium ascorbate is added under ultrasonic treatment, adds
Enter NaOH or H2SO4Solution adjusts the pH to 3.47 of GO dispersion, and mixture is sealed in autoclave, is heated at 90 DEG C
3h is to form GH.
(4) preparation of modified glassy carbon electrode:
The GHs that on glass-carbon electrode prepared by 5 μ L step (1) of drop coating, is dried with nitrogen, is sufficiently eluted with secondary water to get arriving
The glass-carbon electrode GHs/GCE of GHs modification.
(5) standard curve is as shown in Figure 3.
(6) processing of sample:
The celery vegetable sample bought from local market (Chinese Hefei) is cut into small pieces, and is ground into the presence of liquid nitrogen
Fine powder, the methanol (4 DEG C, 80%) of 2mL pre-cooling is added in the powder after 1g grinding, and after stirring, powder is extracted overnight at 4 DEG C.So
1mL CHCl is added in the solution afterwards3, by its vortex 10s, 4min is vibrated with 900rpm at 4 DEG C, at 4 DEG C, 12000 rpm
It is centrifuged 5min, collects supernatant, and nitrogen is dry at room temperature, sample is dissolved in 1mL 0.1M PBS.
(7) preparation of bottom liquid is detected:
With the H of 0.1M3PO4The NaH of solution adjusting 0.1M2PO4PH value of solution is obtained to 2.5.
(8) detection of sample:
By 0.1mL, the sample is added in the liquid of preprepared detection bottom, carries out Electrochemical Detection using LSV.Initially
Current potential is 0.3V, and termination current potential is 1.3V, and it is 0.80V that IAA, which goes out peak position, and it is 1.05V that SA, which goes out peak position,.Measure the peak IAA and SA
Current value is respectively 1.85 μ A and 0.88 μ A, is respectively by having obtained IAA the and SA content in standard curve calculating sample
5.03 μM and 3.99 μM.
The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe
The personage for knowing this technology all without departing from the spirit and scope of the present invention, carries out modifications and changes to above-described embodiment.Cause
This, institute is complete without departing from the spirit and technical ideas disclosed in the present invention by those of ordinary skill in the art such as
At all equivalent modifications or change, should be covered by the claims of the present invention.
Claims (4)
1. a kind of electrochemical sensor, it is characterised in that: including three-electrode system, which includes working electrode, Ag/
To electrode, the working electrode is the glass-carbon electrode of modification for AgCl reference electrode and platinum filament；The preparation method of the working electrode
Include: the drop coating graphene hydrogel on glass-carbon electrode, is eluted after being dried with nitrogen with distilled water to get electric to the glass carbon modified
Pole.
2. electrochemical sensor according to claim 1, it is characterised in that: the glass-carbon electrode is successively used before modification
The Al that partial size is 1.0 μm, 0.3 μm and 0.05 μm2O3Powder polishing grinding glass-carbon electrode makes glassy carbon electrode surface to mirror surface, then uses
After dehydrated alcohol and distilled water ultrasonic cleaning, it is dried with nitrogen spare.
3. a kind of detection heteroauxin and salicylic method, it is characterised in that: include the following steps:
Step 1: the drafting of indole-3-acetic acid and salicylic standard curve；
Step 2: measuring samples are pulverized in the presence of liquid nitrogen, the methanol that measuring samples powder is then added 4 DEG C is molten
Liquid extracts overnight under the conditions of 4 DEG C, CHCl is then added into solution3, be then first vortexed 8~12s, then under the conditions of 4 DEG C,
3~5min is vibrated with 800~100rpm, at 4 DEG C, 4~6min is centrifuged with 12000rpm, the supernatant collected is in room temperature
Under, upper press proof product are dried to obtain with nitrogen, when detecting, disperses upper press proof product in detection bottom liquid and obtains system to be checked, so
The three-electrode system of claims 1 or 2 is inserted into system to be checked afterwards, is detected using linear sweep voltammetry.
4. detection heteroauxin according to claim 3 and salicylic method, it is characterised in that: the indoles -3- second
The drafting of sour and salicylic standard curve includes:
(1) preparation of standard solution:
Indole-3-acetic acid is dissolved in dehydrated alcohol, then with PBS buffer solution by the ethanol solution PBS of indole-3-acetic acid
Buffer solution is diluted to the indole-3-acetic acid standard solution of various concentration in proportion；Salicylic acid is dissolved in dehydrated alcohol, then
Salicylic ethanol solution is diluted to the salicylic acid solution of various concentration in proportion with PBS buffer solution with PBS buffer solution；
The salicylic acid standard solution of the indole-3-acetic acid standard solution of various concentration and various concentration is added to inspection from low to high respectively
It surveys in the liquid of bottom, obtains gradient indole-3-acetic acid and salicylic acid standard mixed solution to be checked；The detection bottom liquid is slow for 0.1M PBS
Rush solution；
(2) it detects:
By in the three-electrode system insertion detection bottom liquid of claims 1 or 2, detection obtains current signal 1, by claims 1 or 2
Three-electrode system successively gradient salicylic acid solution to be checked, using GHs/GCE as working electrode, Ag/AgCl reference electrode and platinum
Silk is inserted into gradient indole-3-acetic acid and salicylic acid standard mixed solution to be checked the three-electrode system that electrode is constituted, and detects
To current signal 2；
(3) drafting of standard curve:
Using the difference of current signal 1 and current signal 2 as ordinate, the concentration of SA or IAA are abscissa, while drawing and obtaining
SA standard curve and IAA standard curve.
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| CN111307890A (en) * | 2019-09-24 | 2020-06-19 | 天津理工大学 | Manganese oxide/graphene modified electrode and method for detecting salicylic acid in plant tissue based on manganese oxide/graphene modified electrode |
| CN113030208A (en) * | 2021-03-01 | 2021-06-25 | 广西壮族自治区农业科学院 | Indoleacetic acid porous molecularly imprinted sensor prepared by electrochemistry and application thereof |
| CN113567528A (en) * | 2021-06-03 | 2021-10-29 | 同济大学 | Composite material modified nicotine sensor, preparation method thereof and method for detecting nicotine content in tobacco leaves by using composite material modified nicotine sensor |
| CN114878659A (en) * | 2022-05-06 | 2022-08-09 | 南通大学 | High-sensitivity multi-flux detection method for disease-resistant related hormones IAA and SA of arabidopsis thaliana |
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