CN109682874A - Three-dimensional nitrogen-doped nanometer porous carbon and its electrochemical sensor and application - Google Patents
Three-dimensional nitrogen-doped nanometer porous carbon and its electrochemical sensor and application Download PDFInfo
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Abstract
The invention discloses a kind of three-dimensional nitrogen-doped nanometer porous carbon and its electrochemical aptamer sensor and applications, its three-dimensional nitrogen-doped nanometer porous carbon, it is prepared by the following method: dry plant biomass being ground 2 hours, is subsequently placed in tube furnace and in the case where being passed through air conditions with 1 DEG C of min‑1Heating rate to 300 DEG C heat 2 hours.Obtained corresponding black powder is immersed into 0.1mol L‑ 1In HCl solution overnight, it removes KCl and then other impurity obtains three-dimensional nitrogen-doped nanometer porous carbon finally, sediment is dried under vacuum 12 hours with ethyl alcohol and water washing.Three-dimensional nitrogen-doped nanometer porous carbon of the invention, manufacturing cost is low, environmental-friendly, with functional group abundant, aptamers chain has stronger binding ability, corresponding electrochemical sensor electro-chemical activity is excellent, and for detecting, the alpha-fetoprotein range of linearity is wide, detection limit is low, specificity is good, reproducible, stability is good.
Description
Technical field
The present invention relates to electrochemical sensor technology field, in particular to a kind of three-dimensional nitrogen-doped nanometer porous carbon and its electricity
Chemical aptamer sensor and application.
Background technique
Cancer seriously threatens the health of the whole world mankind as one of highest disease of the death rate.Therefore, one kind is found
The method that cancer can be early diagnosed is critically important for the mankind.It is reported that protein biomarker is as a kind of biology
Marker, low expression or high expression can be used as the indicant of morbid state in serum.Therefore, it develops and is able to detect cancer egg
The sensitive biological sensor of white biomarker is of great significance.Alpha-fetoprotein (AFP) is a kind of embryo's specificity glycoprotein,
It is widely used in the important protein biology mark of the Several Kinds of Malignancy such as mammary gland, stomach, Colon and rectum, lung, prostate, liver, bladder cancer
Will object.So the effective ways of detection trace AFP will play a significant role in the diagnosis of Cancerous disease.In general, most common
AFP detection method is enzyme linked immunosorbent assay (ELISA) (ELISA), radiommunoassay (RIA), fluorescence method, high performance liquid chromatography string
Join mass spectrography, mass spectrometric immunoassay method and electrochemical techniques.Most of in these methods expose some disadvantages, such as at high cost, behaviour
Make that process complicated, sensitivity and selectivity are limited, detection time is long and requires the qualification of operator high.
Electrochemical aptamer sensor is with its highly sensitive, low detection limit, highly selective, portability, easily operated and low
The significant advantages such as cost have attracted the concern of people.In general, polymer, metal oxide, quantum dot, metal organic framework or carbon
Material is used for electrode material.Wherein, carbon material (carbon cloth, carbon nanotube and graphene) is achieved as matrix in sensory field
It is most of satisfactory as a result, having big good mechanical performance, surface area, easy functionalization, good biocompatibility, fluorescence sudden
The advantages that ability of going out is strong.By covalent fixed test AFP, use GO as in conjunction with NH2The AFP specificity aptamers of functionalization
Platform provides 3pg mL-1Detection limit (LODs).Use receiving based on reduced form graphene oxide-thionine and nanogold particle
Nano composite material is as the sensor platform for constructing the unmarked electrochemical aptamer sensor for AFP detection, display
LODs is 0.050 μ gmL-1.It may be evident, however, that these carbon nanomaterials are combined by cumbersome method or with other components
And synthesize, this further causes the manufacturing cost height and multi-step process of aptamer sensor.Recently, a kind of to be derived from not
With the attention that the carbon nanomaterial (being named by biomass carbon) of biomass has attracted most of researchers, there are several reason phases
Than in synthesis carbon material.These biomass carbon nano materials have some internal characteristics, including abundance, cheap, ring
Border is friendly, and different elements or component (phosphorus, nitrogen, sulphur and some inorganic salts) coexist.Moreover, in further high-temperature process
Period, it can be easily converted to the carbon material with the dopant of N-shaped feature, to improve the electric conductivity of carbon material.Cause
This, biomass carbonization strategy allows the plant of a large amount of high abundances to be used as cheap and reproducible carbon precursor.Biggish surface area and
Excellent electric conductivity provides the application for these carbon materials in fields such as electro-catalysis, energy storage, supercapacitors to absolute guarantee.So
And platform of the biomass carbon as building target molecule detection electrochemical aptamer, but it is rarely reported.
Therefore, it is necessary to improve to solve the above technical problems the prior art.
Summary of the invention
In view of this, can be used as electrochemistry the purpose of the present invention is to provide a kind of three-dimensional nitrogen-doped nanometer porous carbon
Sensor, the super quick detection for alpha-fetoprotein.It is realized in particular by following technical scheme:
Three-dimensional nitrogen-doped nanometer porous carbon of the invention, is prepared by the following method: by dry plant biomass grinding 2
Hour, it is subsequently placed in tube furnace and in the case where being passed through air conditions with 1 DEG C of min-1Heating rate to 300 DEG C heat 2 hours.It will
Obtained corresponding black powder immerses 0.1mol L-1In HCl solution overnight, KCl and other impurity are removed, then, with ethyl alcohol and
Water washing obtains three-dimensional nitrogen-doped nanometer porous carbon finally, sediment is dried under vacuum 12 hours.
In optimal technical scheme, the plant biomass is using grass, colored or peanut shell.
In optimal technical scheme, the grass uses herba setariae viridis.
In optimal technical scheme, the flower uses China rose.
The invention also discloses a kind of electrochemical sensors, by above-mentioned three-dimensional nitrogen-doped nanometer porous carbon modification to matrix
On electrode.
In optimal technical scheme, the method for modifying are as follows:
(1) base electrode pre-processes, and is polished with 0.05mm alumina slurry, successively with Piranha solution and ultrapure water ultrasound
Washing, then, in high purity N2It flows down under drying, which is v (H2SO4): v (H2O2The solution of)=7:3；
(2) the three-dimensional nitrogen-doped nanometer porous carbon is prepared into 0.5mg mL-1Unit for uniform suspension, will it is obtained uniformly
Suspension impregnation is on the surface of clean electrode and in N2Electrochemical sensor is dried to obtain in stream.
The invention also discloses a kind of applications of electrochemical sensor, and above-mentioned electrochemical sensor is used for alpha-fetoprotein
Super quick detection.
Beneficial effects of the present invention: three-dimensional nitrogen-doped nanometer porous carbon of the invention, manufacturing cost is low, environmental-friendly, tool
There is functional group abundant, aptamers chain has stronger binding ability, and corresponding electrochemical sensor electro-chemical activity is excellent, uses
In the detection alpha-fetoprotein range of linearity is wide, detection limit is low, specificity is good, reproducible, stability is good.
Other beneficial effects of the invention will be further detailed in conjunction with embodiment in detail below.
Detailed description of the invention
The invention will be further described with reference to the accompanying drawings and examples:
Fig. 1 a is Fourier transform infrared (FT-IR) figure, and Fig. 1 b is X-ray diffraction (XRD) figure, and Fig. 1 c is Raman spectrum
Figure, Fig. 1 d are x-ray photoelectron spectroscopy (XPS) figure, (i) N-mC in figureg,(ii)N-mCp,and(iii)N-mCf；
Fig. 2 a1, a2, a3 are respectively N-mCgC 1s, N 1s and O 1s high-resolution XPS spectrum, Fig. 2 b1, b2, b3
Respectively N-mCpC 1s, N 1s and O 1s high-resolution XPS spectrum, Fig. 2 c1, c2, c3 are respectively N-mCfC 1s, N
The high-resolution XPS spectrum of 1s and O 1s；
Fig. 3 a, b N-mCgFE-SEM image, c, d N-mCpFE-SEM image, e, f N-mCfFE-SEM figure
Picture；
Fig. 4 a, b, c are respectively to be based on) N-mCg、N-mCpAnd N-mCf0.0001ngmL- under modified Au electrode and 0.1M
The CV figure of the electrochemistry aptamer sensor of modified electrode after 1AFP reaction；
Fig. 5 a, b, c are respectively to be based on N-mCg、N-mCpAnd N-mCfThe Au electrode and and 0.0001ngmL of modification-1Reaction
The EIS of the electrochemistry aptamer sensor of modified electrode afterwards schemes, and contains 5mM [Fe (CN) 6]3-/4-, 0.14M NaCl and 0.1M
AFP (equivalent circuit that the illustration in (a) is nyquist diagram) in the 0.1M PBS (pH7.4) of KCl, d are using based on N-
mCg、N-mCpAnd N-mCfThe Δ Rct value (n=3) of each step of the aptamer sensor of manufacture during AFP is detected；
Fig. 6 (a) is based on N-mCgAptamer sensor detection various concentration AFP (0.0001,0.001,0.01,
0.1,1,10,50 and 100ngmL-1) EIS Nyquist figure, (b) the respective alignment curve between Δ Rct and AFP concentration
(illustration: the Linear Fit Chart of Δ Rct, the function (n=3) of the logarithm as AFP concentration), (c) selectivity, (d) reproducibility, and
(e) it is based on N-mCgAptamer sensor to the stability of AFP；
Fig. 7 (a) is used based on N-mCgAptamer sensor detection various concentration AFP (0.0001,0.001,0.01,
0.1,1,10,50 and 100ngmL-1) DPV reaction, (b) the respective alignment curve between Δ Rct and AFP concentration (illustration:
The Linear Fit Chart of Δ Rct, the function of the logarithm (n=3) as AFP concentration).
Specific embodiment
Invention is further described combined with specific embodiments below.
One, N-mCg, N-mCfAnd N-mCpPreparation and its characterization
By dry grass, colored and peanut shell is ground 2 hours respectively with agate mortar, is subsequently placed in tube furnace and is being passed through
With 1 DEG C of min under air conditions-1Heating rate to 300 DEG C heat 2 hours.Obtained corresponding black powder is immersed into 0.1mol
L-1In HCl solution overnight, KCl and other impurity are removed.Then, for several times with ethyl alcohol and water washing.Finally, by sediment in vacuum
Lower drying obtains three-dimensional nitrogen-doped nanometer porous carbon for 12 hours.These materials are respectively designated as N-mCg, N-mCfAnd N-mCp, they
Respectively from grass, colored and peanut shell.Grass in the present embodiment uses herba setariae viridis, Hua Caiyong China rose, and grass and flower can also be with certainly
It using other grass and spends, without obvious sex differernce in obtained result and the present embodiment.
N-mC is studied by Fourier transform infraredg, N-mCfAnd N-mCpChemical structure, as shown in Figure 1a, due to them
There is similar characteristic peak in infrared spectroscopy, therefore observe that three samples have identical chemical structure.The result shows that
3100-3700cm-1Broad peak in range is attributed to the-OH stretching vibration of adsorbed water molecule.In 1000-1720cm-1Region Nei Guan
Additional strong band is observed, in 1032,1247,1605 and 1717cm-1Those of neighbouring peak and C-O, C-N, C=C and C=O group
Stretching vibration it is related.Furthermore, it is possible to will be in 2848 and 2926cm-1Distribute to the stretching mode of C-H in the peak at place.Therefore, it synthesizes
N-mC nano material series afterwards is made of functional group abundant, this can promote aptamers chain to fix and enhance the suction of target molecule
It is attached.
N-mC is studied by X-ray diffraction (XRD)g, N-mCfAnd N-mCpCrystal structure, as shown in Figure 1 b.All XRD
Figure clearly shows the diffraction maximum positioned at 15 °, is assigned to (101) of cellulosic regions complete ordering in three samples
Crystal face.As N-mCfAnd N-mCpThe comparison of nano material, it can be seen that N-mCgThe peak width of material is more sharp, shows its high-carbon knot
Brilliant degree.In addition, (002) face that the weak peak at 40 ° is designated as graphene nanometer sheet is micro- in turbostratic carbon in all three samples
In crystalline substance, and the peak at 25 ° is attributed to (200) face of carbon.
Fig. 1 c is N-mCg, N-mCfAnd N-mCpRaman analysis figure, wherein in 1350 and 1580cm-1Place obtains two clearly
Peak, correspond respectively to the D band and G band of graphitic carbon.G band is attributed to 1582cm-1 plane stretch, indicates the graphite degree of order, and
In 1350cm-1The carbon content D at place unordered with expression.Intensity ratio (the I of D band and G bandD/IG) it is the unordered parameter of assessment material.
It can be seen that N-mCgThe I of nano materialD/IGAbout 0.82, it is higher than N-mCf(0.79) and N-mCp(0.70).The result shows that N-
mCgContain a large amount of disordered carbons in composite material, this is consistent with XRD result.
As shown in Figure 1 d, N-mCg, N-mCfAnd N-mCpX-ray photoelectron spectroscopy XPS measuring spectrum show, respectively position
In three strong peaks of 286.4,399.5 and 531.6eV, mainly with C 1s, N 1s and O 1s is related.In addition, summarizing C in table 1
The atomic percent of 1s, N 1s and O 1s:
Table 1
In order to further obtain N-mCg, N-mCfAnd N-mCpChemical constituent and environment, carry out XPS characterization, analyze C 1s,
The high-resolution XPS spectrum of N 1s and O 1s.
As shown in Fig. 2 a1, N-mCgThe high-resolution C 1s spectrum of nano material is combining energy (BE) C-C/C-H
For deconvolution at a main peak, five weak peaks at BE are 285.7,286.2,287.7,288.8 and 291.7eV at (284.6eV),
It is attributed to C-N, C-O, C=O, O-C=O and C-F.By to N-mCfAnd N-mCpHigh-resolution C 1s XPS spectrum analysis,
Similar result is obtained.Show their chemical structure very close to demonstrating this point by INFRARED SPECTRUM.As shown in Fig. 2 a2,
For the high-resolution N 1s spectrum of N-mCg nano material, it is fitted at the BE of 398.3,399.6,400.9 and 403.7eV
Four main peaks, this is attributed to C=N-C, N-N, N=C and NOx.As shown in Fig. 2 a3, it is due to containing that the peak value of BE, which is 532.7eV,
Oxygen intermediate, and the peak value of BE is the formation that 531eV indicates C-O-C key.This means that carbon and N-mCgCombination not only pass through π-
Pi accumulation interaction, but also pass through C-O-C key bridge.In addition, the BE of 534.2eV is attributed to O-N key, biomass carbon is come from
The intermediate of acidification.For N-mCp(Fig. 2 b3) and N-mCfThe XPS spectrum of the high-resolution O 1s of (Fig. 2 c3), is not observed C-
O-C key.Meanwhile the position of oxygen-containing intermediate and O-N key is transferred to 531.6 and 533.5eV respectively.It illustrates in three kinds of materials
The combining form of C and N is different.
The configuration of surface of carbon material derived from series of biologic matter is measured by FE-SEM, as shown in Figure 3.Such as Fig. 3 a and b institute
Show, N-mCgThe FE-SEM image of nano material shows direct carbonization treated in nanometer chip architecture and some nanometer sheets
Many nanometer chip architectures.However, N-mC is not observedf(figure c and d) and N-mCpThe clear knot of (figure e and f) nano material
Structure.For N-mCf, it can be seen that the structure is made of blocky and stack of sheets.This is mainly due to the accumulation of carbon material, cause
Different structures is formed in high-temperature burning process.Observe N-mCpNano material is similar to the structure of N-mCf, forms machine
It manages identical.With N-mCfAnd N-mCpNano material is compared, N-mCgUnique texture specific surface area can be enhanced, further promote suitable
The fixed absorption with target AFP molecule of ligand.It can also provide convenient internal environment for electronics transfer.
Two, it is based on N-mCg, N-mCfAnd N-mCpElectrochemical sensor building and its performance test
All electro-chemical tests carry out in CHI660E electrochemical workstation (Chinese Shanghai morning China).It is using diameter
Working electrode of the naked gold electrode (AE) of 3mm as electrochemical measurement.And Pt and Ag/AgCl respectively as to electrode and ginseng
It than electrode, is polished with 0.05mm alumina slurry, successively uses Piranha solution (v (H2SO4): v (H2O2)=7:3) and ultrapure water
Supersound washing.Then, in high purity N2Drying is flowed down, AEs is in 0.5M H2SO4In, from -0.2V to 1.6V (with Ag/AgCl phase
Than) carry out electrochemical redox circulation.In order to compare the biosensor efficiency of different materials, by N-mCgNano material point
It is dispersed in fresh Milli-Q water (Milli-pore company, U.S. Mill-Q ultrapure water production system water produced) to prepare
Concentration is the unit for uniform suspension of 0.5mg/mL.Then, the suspension prepared (5 μ L) is immersed in the surface of clean Au electrode
Above and in N2Dry in stream, obtained result is expressed as N-mCg/Au.Then by N-mCg/ Au and the AFP that concentration is 100nM are special
Property adaptation liquid solution be incubated with 2 hours.These aptamer sensors Apt/N-mCg/ Au is indicated.It is prepared with same method
N-mCfAnd N-mCpThe sensor of type.
Three kinds of biomass carbon nano materials, i.e. N-mCg、N-mCfAnd N-mCpIt is used as the bracket for combining adaptation ligand chain simultaneously,
Then AFP detection is assessed using these aptamer sensors.Using EIS, CV and DPV technology, research contains 0.14M
5mM [the Fe (CN) of NaCl and 0.1M KCl6]3-/4-In entire AFP detect program, induce naked AE modified biological matter carbon nanometer material
Material, aptamers immobilization and AFP detection.CV curve such as Fig. 4 institute of AFP detection is carried out using the carbon-based aptamer sensor of biomass
Show.As expected, naked AE shows specific redox peaks, shows its excellent chemical property.With N-
mCg、N-mCfAnd N-mCpAfter nano-material modified AE, the reduction of anode and cathode peak point current, Δ E are observedpRespectively
0.267,0.281 and 0.283V.The result shows that the electro-chemical activity of these three samples is lower, it is possible to reduce electrode surface and electricity
Solve the electron transmission between matter.After fixed adaptation body chain, the peak point current of electrode is further slightly reduced, and after AFP detection
It is gradually reduced.All these results are attributable to the negative electrical charge and [Fe (CN) of the phosphate group contained in aptamers chain6]3-/4-
The combination of repulsive force and AFP between redox probe and aptamers.
In order to which quantitative analysis is based on N-mCg、N-mCfAnd N-mCpThe detection efficiency of the aptamer sensor of nano material, also
EIS measurement has been carried out to study the detection of AFP.As shown in figure 5, it, which is shown in use, contains 5mM [Fe (CN)6]3-/4-、0.14M
N-mC in the 0.1M PBS solution of NaCl and 0.1M KClg, N-mCfAnd N-mCpCharge turns during nanometer-material-modified Au electrode
Resistance (Rct) is moved to dramatically increase.All EIS curves are analyzed by software.The software gives the equivalent electricity of Randles of simulation
Road, by solution resistance (Rs), charge transfer resistance (Rct), (Fig. 5 a is inserted for constant phase element and Warburg impedance (W) composition
Figure).It is based on N-mCgAptamer sensor for (Fig. 5 a), due to its good electro-chemical activity and low electronics transfer electricity
Resistance, naked AE show 132.2 ohm of small Rct value.With N-mCgIt is modified, N-mCgThe Rct value of/AE increases to 301.7 Europe
Nurse.Compared with polymer, metal organic framework, carbon nanomaterial etc. are as the nano material of carrier bracket, N-mCgIt is relatively small
Rct show good chemical property.It is mainly due to graphene sample nanostructure, biggish specific surface area.However,
N-mCf/ AE and N-mCpThe Rct value of/AE is respectively 401 and 632.9 ohm, is much larger than N-mCg/AE.Show and N-mCgIt compares,
Its chemical property is poor.After the aptamers immobilization on three kinds of N-mC/AEs, biggish Rct value has been obtained, this is because
Repulsion caused by the aptamers chain of introducing hinders electronics transfer.When detecting AFP, Rct value is continuously increased.In AFP and fit
Tetra- serobila of G- is formed between ligand chain will further prevent electronics to shift between electrode surface and electrolyte solution.For three kinds
Aptamer sensor observes similar chemical property trend.
Sensitivity is an important parameter of evaluation sensor quality, can also be indicated with detection limit (LODs).Here,
N-mC is based on using the detection of EIS and DPV technologygAptamer sensor to the analysis performance of AFP.By measurement Δ Rct to AFP
The dependence of concentration is assessed based on N-mCgAptamer sensor quantitative analysis.As shown in Figure 6 a, it illustrates EIS pairs
The reaction of various concentration AFP.It can be clearly seen that Rct value increases with the raising of AFP concentration.This may be because of AFP
Molecule is fixed more with specific aptamers, hinders the electronics transfer of electrode surface.When AFP is at concentrations up to 10ng mL-1,
EIS response shows the saturation absorption of AFP close to balance.It is rung by drawing Δ Rct (Δ Rct=Rct, AFP-Rct, aptamers)
It should be with the alignment of relationships curve (Fig. 6 b) of the logarithm of AFP concentration.Regression equation is Δ Rct (k Ω)=0.145logCAFP+
0.627, the correlation of R2=0.997 in 0.0001 to the 100ng mL-1 wide range of linearity.LODs is calculated as 61.8fgmL-
1 (s/n=3).
Specificity is to develop an extremely important parameter of aptamer sensor.It is passed to assess proposed aptamers
For sensor to the selectivity of AFP, determining PSA, CEA, IgG, BSA, VEGF, EGFR, MUC1 isoconcentration with same method is AFP
The chaff interferent that 100 times of concentration.The variation of the Δ Rct value as caused by each ingredient, mixed solution and AFP solution, as fig. 6 c.As a result
Show compared with AFP, high concentration interference shows significant low signal value, and (AFP concentration is 0.1pg mL-1, and other substances are dense
Degree is 10pg mL-1).Compared with AFP, the mixed solution of interference will not cause the substantial variations of Δ Rct value.Therefore, the base of preparation
In N-mCgAptamer sensor excellent selectivity is shown to AFP.
By detecting 0.1pg mL-1AFP determines the reproducibility of developed aptamer sensor.In fig. 6d it is shown that
Under the same conditions, we are prepared for 5 electrodes newly modified simultaneously, to assess developed aptamer sensor for detecting
AFP(0.1ng mL-1) reproducibility.As can be seen that the Δ Rct value between five electrodes there is no variation, show electrode
All show similar electrochemical response.Meanwhile also by measuring EIS daily come another important parameter of repercussion study, i.e. institute
The stability of the aptamer sensor of proposition.The AE electrode of modification is stored in 15 days (dried forms) in refrigerator at 4 DEG C.Figure
6e shows 15 Δ Rct values of measurement and not changing significantly for first time, sufficiently shows based on N-mCgAptamers pass
Sensor has excellent stability.Thus, it could be seen that prepared aptamer sensor shows excellent selection in AFP detection
Property, reproducibility and stability.
N-mC is based on by DPV researchgDetection performance of the aptamer sensor of preparation to AFP.Firstly, to Apt/N-
mCg/ AE aptamer sensor carries out DPV measurement, and records peak point current, is considered as I0.As shown in Figure 7a, show peak value
Electric current increases with the increase of AFP concentration.Calibration curve is as shown in Figure 7b, it can be seen from the figure that the variation of peak point current
It is directly proportional to AFP concentration, in 0.0001-100ng mL-1Between good linear relationship is presented.Regression equation is represented by Δ I
=3.93logCAFP+ 18.75, the coefficient of determination (R2) it is 0.985, show that LODs is 60.8fgmL-1(s/n=3).Obtain from
EIS and DPV infer based on N-mCgThe aptamer sensor for being suitable for AFP LOD, further demonstrate developed aptamer
The high sensitivity of sensor.
Compare (table 2) with the AFP detection method that electrochemistry and other technologies are reported, EIS and DPV detect based on N-mCg
AFP detection method LODs value it is much lower.There is the above results satisfactory sensing capabilities and EIS and DPV to realize
Low LODs, it may be possible to due to having the advantage that it is fixed most that (1) functional group (- OH ,-COOH) and large surface area facilitate
The specific aptamers of number, can be further in conjunction with a large amount of target molecules；(2) big specific surface area is to adsorb more aptamers
Guarantee is provided with target molecule.
Table 2
It is prepared based on N-mC in order to characterizegAptamer sensor clinical application, We conducted recovery experiments.
The blood of four kinds of different types of cancer patients's (osteocarcinoma, gastric cancer, breast cancer and lung cancer) is collected from the first affiliated hospital, Zhengzhou University
Final proof sheet.With 0.01M PBS solution (pH7.4) by 100 times of serum samples diluted.Actual sample is determined using EIS measurement,
In the Δ Rct values of four kinds of different actual samples be respectively 0.467,0.821,0.535 and 0.893k Ω.It can be according to Δ Rct (k
The AFP concentration contained in four kinds of blood serum samples of Ω)=0.145logCAFP+0.627 regression equation calculation, as the result is shown in table
In 3.The result shows that aptamer sensor performance of the building for AFP detection is good, and pass through clinical measurement and identical serum
The quantitative matching of sample.As a result the potential practical applicability for the aptamer sensor developed at present is disclosed.
Table 3
In conclusion applicant has synthesized a series of three-dimensional nitrogen extracted from different plants (including grass, flower, peanut shell)
Dopen Nano hole carbon nanomaterial, and be configured to be used to detect the novel no label electrochemical aptamer sensor of AFP
Bracket.In different biomass carbon nano materials, from grass in N-mCgShow richer functional group, biggish ratio
Surface area and excellent electro-chemical activity.In conjunction with peculiar property (high-affinity and the spy of biomass carbon and AFP specificity aptamers
It is anisotropic), the aptamer sensor based on N-mCg of exploitation has excellent performance, has highly selective, reproducibility, long-term steady
Qualitative and Clinical practicability.It was found that the LODs from EIS and DPV is respectively in 0.1pg mL-1To 100ng mL-1Wide scope in
For 61.8 and 60.8fg mL-1.Meanwhile we are also to building based on N-mCgAptamer sensor for analyzing cancer patient
True serum sample, result and clinical measures coincide preferable.Therefore, current work highlights biomass carbon nanometer
Potential application of the material in cancer markers early stage measurement.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to compared with
Good embodiment describes the invention in detail, those skilled in the art should understand that, it can be to skill of the invention
Art scheme is modified or replaced equivalently, and without departing from the objective and range of technical solution of the present invention, should all be covered at this
In the scope of the claims of invention.
Claims (7)
1. a kind of three-dimensional nitrogen-doped nanometer porous carbon, it is characterised in that be prepared by the following method: by dry plant biomass
Grinding 2 hours, is subsequently placed in tube furnace and in the case where being passed through air conditions with 1 DEG C of min-1Heating rate it is 2 small to 300 DEG C of heating
When.Obtained corresponding black powder is immersed into 0.1mol L-1In HCl solution overnight, it removes KCl and then other impurity uses second
Pure and mild water washing obtains three-dimensional nitrogen-doped nanometer porous carbon finally, sediment is dried under vacuum 12 hours.
2. three-dimensional nitrogen-doped nanometer porous carbon shown according to claim 1, it is characterised in that: the plant biomass uses
Grass, colored or peanut shell.
3. the three-dimensional nitrogen-doped nanometer porous carbon according to shown in claim 2, it is characterised in that: the grass uses herba setariae viridis.
4. the three-dimensional nitrogen-doped nanometer porous carbon according to shown in claim 2, it is characterised in that: the flower uses China rose.
5. a kind of electrochemical sensor, sign are: three-dimensional N doping described in claim 1-4 any claim is received
In rice porous carbon modification to base electrode.
6. electrochemical sensor according to claim 5, it is characterised in that the method for modifying are as follows:
(1) base electrode pre-processes, and is polished with 0.05mm alumina slurry, is successively washed with Piranha solution and ultrapure water ultrasound
It washs, then, in high purity N2It flows down under drying, which is v (H2SO4): v (H2O2The solution of)=7:3；
(2) the three-dimensional nitrogen-doped nanometer porous carbon is prepared into 0.5mg mL-1Unit for uniform suspension, by even suspension obtained
Liquid is immersed on the surface of clean electrode and in N2Electrochemical sensor is dried to obtain in stream.
7. a kind of application of electrochemical sensor, it is characterised in that: by electrification described in claim 5 or 6 any claims
Learn the super quick detection that sensor is used for alpha-fetoprotein.
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