CN111537578B - Electrochemical sensor material for detecting L-cysteine and preparation method thereof - Google Patents
Electrochemical sensor material for detecting L-cysteine and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a preparation method of an electrochemical sensor material for detecting L-cysteine, which comprises the following steps: (1) GF pretreatment; (2) preparing BNC; (3) preparing a PB growth solution; (4) preparing BNC/GF; (5) BNC @ PB/GF and provides the products obtained by the method. The method can obtain the electrode material with excellent electrochemical characteristics, overcomes the defects of low sensitivity and high detection limit of the conventional L-cysteine electrochemical sensor, realizes the advantages of low detection limit, high sensitivity, wide linear range and high stability, and is simple, rapid and easy to control.
Description
Technical Field
The invention belongs to the technical field of electrochemical sensors, and particularly relates to an electrochemical sensor material for detecting L-cysteine and a preparation method thereof.
Background
L-cysteine (L-cys), one of the most important thiol-containing amino acids, plays a crucial role in biological systems and can be used for the diagnosis of disease states.
L-cys has been used in the food and pharmaceutical industries as an antioxidant, antitoxin, radioprotectant, cancer indicator and free radical scavenger. Deficiency of L-cys is associated with slow growth, diabetes, alopecia, skin disorders, and susceptibility to fatigue.
At present, the detection method of L-cys mainly comprises chromatographic separation, atomic absorption spectroscopy, atomic emission spectroscopy, capillary electrophoresis, electrochemical method and the like. Among them, the electrochemical method is preferred because of its advantages such as easy operation and low cost.
Disclosure of Invention
The first purpose of the invention is to provide a preparation method of an electrochemical sensor material for detecting L-cysteine, by which an electrode material with uniform appearance and stable structure can be obtained, the electrode material has good conductivity and catalytic performance, the prepared sensor has the advantages of high sensitivity, low detection limit and the like, and the preparation method is simple and easy to control.
A second object of the present invention is to provide an electrochemical sensor material obtained according to the above method.
The purpose of the invention is realized by the following technical scheme:
a preparation method of an electrochemical sensor material for detecting L-cysteine comprises the following steps:
(1) GF pretreatment, cutting a graphite felt GF, alternately washing the graphite felt with ultrapure water and methanol to remove surface residues, vacuum-drying overnight, then activating with a sulfuric acid solution, washing with ultrapure water, and vacuum-drying overnight;
(2) Preparing BNC, namely dispersing boric acid, urea and polyethylene glycol into ultrapure water, ultrasonically dissolving, drying at 80 ℃ to obtain powder, placing the obtained powder into a porcelain boat, preserving heat at 900 ℃ in Ar atmosphere, and naturally cooling after heat preservation to obtain boron-nitrogen doped carbon nanotube BNC powder;
(3) Preparing a PB growth solution, wherein the PB growth solution comprises a solution A and a solution B, and the solution A contains K 3 [Fe(CN) 6 ]The solution B contains FeCl 3 KCl and HCl;
(4) Preparing BNC/GF, namely dissolving the boron-nitrogen doped carbon nanotube powder prepared in the step (2) in ultrapure water to obtain a BNC aqueous solution, adding the graphite felt pretreated in the step (1), carrying out ultrasonic treatment, then washing the graphite felt with the ultrapure water, and carrying out vacuum drying overnight to obtain BNC/GF;
(5) And (3) preparing BNC @ PB/GF, mixing the solution A and the solution B which are prepared in step (3), adding the BNC/GF obtained in step (4), growing at room temperature, cleaning the surface of the graphite felt with ultrapure water, and drying in vacuum overnight to obtain the nano-graphite/GF.
Preferably, in the step (1), the temperature of vacuum drying is 60 ℃, the sulfuric acid solution is formed by mixing 98% sulfuric acid and ultrapure water according to the volume ratio of 1:1, and the activation time is 1h.
Preferably, in the step (2), the weight ratio of boric acid to urea to polyethylene glycol is 0.15.
Preferably, in step (3), the solution A contains 2.0mmol/L K 3 [Fe(CN) 6 ]The solution B contains 2.0mmol/L FeCl 3 、0.2mol/L KCl、0.05mmol/L HCl。
Preferably, in the step (4), the concentration of the BNC aqueous solution is 0.1-1.0mg/mL, the ultrasonic time is 30min, and the vacuum drying temperature is 60 ℃.
Further preferably, the concentration of the aqueous BNC solution is 0.5mg/mL.
Preferably, in the step (5), the volume ratio of the solution A to the solution B is 1, the volume ratio of the solution A to the solution B to the BNC aqueous solution in the step (4) is 1:1, the room-temperature growth time is 5-60min, and the vacuum drying temperature is 60 ℃.
Further preferably, the room temperature growth time is 30min.
An electrochemical sensor material for the detection of L-cysteine obtained by any of the above methods.
Prussian Blue (PB) is a typical hexacyanoferrate coordination compound, and has attracted wide attention in the fields of medicines and materials due to the characteristics of no pollution, low cost, easy preparation, excellent electrochemical performance and the like. Fe in PB 2+ And Fe 3+ Reversible electrochemical redox promotes electron transport and enhances the electrochemical performance of the electrode material.
Carbon nanotubes are typical carbon materials and have high specific surface area, excellent electrical conductivity, excellent stability, and characteristics of simple preparation, low price, and corrosion resistance. Furthermore, heteroatom doping is one of the effective methods to modulate the properties of carbon nanostructures. Of the various heteroatoms, nitrogen and boron are excellent dopants for carbon-based materials because the atomic sizes of boron, carbon and nitrogen are the closest.
The invention combines the advantages of heteroatom-doped carbon nanotubes and Prussian blue for the first time, and the graphite felt GF is used as an electrode substrate material to prepare the composite electrode, so that the defects of high detection limit, poor reproducibility, low sensitivity and narrow linear range of the conventional electrochemical detection of L-cys are overcome, and the efficient detection of the L-cys is realized.
The invention has the following beneficial effects:
according to the preparation method of the electrochemical sensor material for detecting the L-cysteine, the obtained BNC @ PB/GF composite electrode can exert the advantages of the three components, and the conductivity, the stability and the electrochemical response of the composite material are enhanced by the synergistic effect of the components; the method can prepare the nanometer material with uniform appearance; the combination of BNC and GF and the in-situ growth process of PB reduces the consumption of raw materials and greatly improves the preparation efficiency; the composite electrode pair L-cys can be carried out under neutral medium and low potential, and has the advantages of good detection sensitivity, circulation stability, anti-interference performance, high sample recovery rate and the like.
Drawings
FIG. 1 is an SEM image of BNC (left panel) and BNC @ PB (right panel) prepared in example 1.
FIG. 2 is a graph showing the potential optimization and quantification curve of the BNC @ PB/GF electrode prepared in example 1 for L-cys detection, wherein A is a CV curve of the electrode BNC @ PB/GF with or without L-cys; b is an i-t curve of the response of the electrode BNC @ PB/GF to L-cys under different potentials; c is a current response curve of the electrode BNC @ PB/GF continuously adding L-cys with different concentrations; d is a current concentration linear curve.
FIG. 3 is the anti-interference ability and long-term stability test of the BNC @ PB/GF electrode prepared in example 1 on L-cys detection, wherein A is the anti-interference detection curve of the electrode BNC @ PB/GF on L-cys detection; b is an electrode BNC @ PB/GF detection L-cys stability test.
FIG. 4A is the CV curve of GF, PB/GF, BNC @ PB/GF of the different electrodes prepared in example 1; b is a partial enlarged view of A; c and D are CV curves of different BNC concentrations and different growth times in the preparation process of the electrode BNC @ PB/GF.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for the purpose of illustration and description, and is in no way intended to limit the invention.
The raw materials in the following examples are all commercially available products. Wherein the graphite felt GF is purchased from Hedgeon carbon fiber of Gansu, inc., and has a thickness of 3 mm; boric acid was purchased from Tianjin optical science and technology development Limited; urea was purchased from tianjin shin science and technology development ltd; polyethylene glycol was purchased from Shanghai Aladdin science and technology, inc.
Example 1
A preparation method of a BNC @ PB/GF composite electrode material comprises the following steps:
(1) GF pretreatment
Cutting GF into 2 cm-1 mm sheets, washing away surface scum of the cut GF with ultrapure water, washing with methanol and ultrapure water alternately until no scum is formed, and vacuum drying at 60 deg.c overnight; selecting 30-50 cleaned GF, and adding the GF into 200ml sulfuric acid: water =1:1 (v/v), then washed to neutrality with ultrapure water, and dried overnight under vacuum at 60 ℃.
(2) Preparation of BNC
0.15g of boric acid, 5g of urea and 0.5g of polyethylene glycol are dispersed in 50mL of ultrapure water, dissolved by ultrasonic treatment for 10min, placed in an oven at 80 ℃ for 10h to obtain white powder, the white powder is placed in a porcelain boat, kept for 4h under Ar atmosphere and 900 ℃ (the heating rate is 5 ℃/min), and naturally cooled under Ar atmosphere to obtain black BNC powder.
(3) Preparation of PB growth liquid
The A solution contains 2mmol/L K 3 [Fe(CN) 6 ]The solution B contains 2.0mmol/L FeCl 3 Mixing 0.2mol/LKCl and 0.05mmol/L HCl, and mixing the solution A and the solution B in equal volume to obtain the PB growth solution.
(4) Preparation of BNC/GF
And (3) putting 1.6mL of BNC aqueous solution with the concentration of 0.5mg/mL into a 7mL centrifuge tube, putting the GF pretreated in the step (1), carrying out ultrasonic bath for 30min, washing the surface of the GF with ultrapure water, and carrying out vacuum drying at 60 ℃ overnight.
(5) Preparation of BNC @ PB/GF
Mixing the solution A and the solution B in the step (3) in equal volume to obtain PB growth solution, and taking 1.6mL of freshly prepared PB growth solution (containing 1.0mmol/L K) 3 [Fe(CN) 6 ]，1.0mmol/LFeCl 3 0.1mol/L KCl and 0.025mmol/L HCl) into a 7ml centrifuge tube, putting the BNC/GF prepared in the step (4), growing for 30min at room temperature, washing the surface of the GF with ultrapure water, and drying in vacuum at 60 ℃ overnight to obtain the product.
Example 2
The preparation method of the BNC/GF and PB/GF composite electrode material comprises the following steps:
(1) GF pretreatment was carried out in the same manner as in step (1) of example 1.
(2) BNC/GF was prepared according to the same procedure as in step (4) of example 1.
(3) PB/GF was prepared according to the same procedure as in step (5) of example 1, except that BNC/GF was not incorporated and GF pretreated in step (1) was incorporated.
Example 3
A preparation method of BNC @ PB/GF composite electrode material is characterized in that 1.6mL of 0.1mg/mL BNC solution is taken in the step (4), and the rest is the same as that in the example 1.
Example 4
A preparation method of BNC @ PB/GF composite electrode material is characterized in that 1.6mL of 1.0mg/mL BNC solution is taken in the step (4), and the rest is the same as that in the example 1.
Example 5
A preparation method of a BNC @ PB/GF composite electrode material is characterized in that the PB nano particle growth time in the step (5) is 5min, and the rest is the same as that in the example 1.
Example 6
A preparation method of a BNC @ PB/GF composite electrode material is characterized in that the PB nano particle growth time in the step (5) is 15min, and the rest is the same as that in the example 1.
Example 7
A preparation method of a BNC @ PB/GF composite electrode material is characterized in that in the step (5), the growth time of PB nanoparticles is 60min, and the rest is the same as that in the example 1.
The effect test of the composite electrode material prepared in the above example has the following results:
electrochemical performance tests are carried out on the electrodes GF, BNC/GF, PB/GF and BNC @ PB/GF by Cyclic Voltammetry (CV), which are shown in fig. 4A and 4B, wherein GF and BNC/GF have no peak due to CV, so that GF and BNC/GF are overlapped under the current icon, and fig. 4B can be distinguished after being locally enlarged. The result shows that the BNC @ PB/GF electrode has stronger current response signals, better electrochemical reversibility and cycle stability.
In order to optimize the preparation conditions of the composite electrode, electrochemical performance tests were performed using Cyclic Voltammetry (CV), see fig. 4C, 4D. The result shows that when the BNC solution concentration is 0.5mg/ml and the PB growth time is 30min, the prepared electrode has better electrochemical performance.
The detection potential of the BNC @ PB/GF composite electrode for L-cys detection was optimized by Chronoamperometry (CA), see FIG. 2B. Experiments prove that BNC @ PB/GF shows stronger current response to L-cys at the potential of 0.24V.
The linear range, the sensitivity and the lowest detection limit of the L-cys detection of the BNC @ PB/GF composite electrode are researched by a timing current method (CA) and a standard addition method. The optimized result is linear range of 0.0055-4177 mu M, sensitivity of 0.21A/M and minimum detection limit of 1.1nM.
In conclusion, the electrochemical sensor material for detecting L-cysteine obtained by the method has the linear response range of 0.0055-4177 mu M to L-cys, the sensitivity of 0.21A/M and the minimum detection limit of 1.1nM (S/N = 3).
The electrochemical sensor material prepared by the invention realizes the high-efficiency detection of L-cys and provides a feasible idea for the detection of L-cys.
Detection test
The electrochemical sensor of the composite electrode material of the embodiment 1 of the invention is used for detecting the L-cys in serum (fetal bovine serum, hangzhou Biotechnology GmbH in Zhejiang and Tian, zhejiang) and tablets (cysteine capsule brand: puritan's Pride), and the method is a standard addition method and is specifically operated as follows: (1) diluting serum, preparing L-cys serum solution, and detecting the content of L-cys in the L-cys serum solution by using an electrode BNC @ PB/GF; (2) quantitatively weighing the L-cys capsules, preparing an aqueous solution with a certain concentration, detecting the L-cys content in the L-cys tablet solution by using an electrode BNC @ PB/GF, and comparing the detected content with the label content to calculate the recovery rate. The results are shown in the following table.
| Sample (I) | Label content (μ M) | Assay (μ M) | Recovery (%) |
| #1 (serum) | 25 | 24.6 | 98.5 |
| #2 (serum) | 25 | 24.3 | 97.2 |
| #3 (serum) | 25 | 24.6 | 98.5 |
| #4 (tablet) | 25 | 25.9 | 103.6 |
| #5 (tablet) | 25 | 24.4 | 97.6 |
| #6 (tablet) | 25 | 24.9 | 99.6 |
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A preparation method of an electrochemical sensor material for detecting L-cysteine is characterized by comprising the following steps:
(1) Pretreating a graphite felt GF, cutting the graphite felt, alternately washing the graphite felt with ultrapure water and methanol to remove surface residues, carrying out vacuum drying overnight, then activating with a sulfuric acid solution, washing with ultrapure water, and carrying out vacuum drying overnight;
(2) Preparing boron-nitrogen doped carbon nanotube BNC, dispersing boric acid, urea and polyethylene glycol into ultrapure water, ultrasonically dissolving, drying at 80 ℃ to obtain powder, placing the obtained powder into a porcelain boat, preserving heat at 900 ℃ in Ar atmosphere, and naturally cooling after heat preservation to obtain boron-nitrogen doped carbon nanotube powder;
(3) Preparing Prussian blue PB growth solution, wherein the Prussian blue PB growth solution comprises solution A and solution B, and the solution A contains K 3 [Fe(CN) 6 ]The solution B contains FeCl 3 KCl and HCl;
(4) Preparing BNC/GF, namely dissolving the boron-nitrogen doped carbon nanotube powder prepared in the step (2) in ultrapure water to obtain a BNC aqueous solution, adding the graphite felt pretreated in the step (1), performing ultrasonic treatment, then washing the graphite felt with the ultrapure water, and performing vacuum drying overnight to obtain BNC/GF;
(5) Preparing BNC @ PB/GF, mixing the solution A and the solution B which are prepared in step (3), adding the BNC/GF obtained in step (4), growing at room temperature, cleaning the surface of the graphite felt with ultrapure water, and drying in vacuum overnight to obtain the product;
in the step (1), the vacuum drying temperature is 60 ℃, the sulfuric acid solution is formed by mixing 98 percent sulfuric acid and ultrapure water according to the volume ratio of 1:1, and the activation time is 1h;
in the step (2), the weight ratio of boric acid to urea to polyethylene glycol is 0.15;
in the step (3), the solution A contains 2.0mmol/L K 3 [Fe(CN) 6 ]The solution B contains 2.0mmol/L FeCl 3 0.2mol/L KCl and 0.05mmol/L HCl.
2. The method for preparing an electrochemical sensor material for L-cysteine detection according to claim 1, wherein in the step (4), the BNC aqueous solution has a concentration of 0.1-1.0mg/mL, an ultrasonic time of 30min, and a vacuum drying temperature of 60 ℃.
3. The method for preparing an electrochemical sensor material for L-cysteine detection according to claim 2, wherein the concentration of the BNC aqueous solution is 0.5mg/mL.
4. The method for preparing the electrochemical sensor material for L-cysteine detection according to claim 1, wherein in the step (5), the volume ratio of the solution A and the solution B is 1, the volume ratio of the solution A and the solution B to the BNC aqueous solution in the step (4) is 1:1, the room temperature growth time is 5-60min, and the vacuum drying temperature is 60 ℃.
5. The method for preparing an electrochemical sensor material for L-cysteine detection according to claim 4, wherein in the step (5), the growth time at room temperature is 30min.
6. An electrochemical sensor material for L-cysteine detection obtained by the production method according to any one of claims 1 to 5.
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