CN107421933A - Utilize the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin - Google Patents

Utilize the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin Download PDF

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CN107421933A
CN107421933A CN201710619645.7A CN201710619645A CN107421933A CN 107421933 A CN107421933 A CN 107421933A CN 201710619645 A CN201710619645 A CN 201710619645A CN 107421933 A CN107421933 A CN 107421933A
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hemoglobin
solution
quantum dot
carbon quantum
nitrogen phosphorus
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CN107421933B (en
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黄珊
肖琦
杨二利
刘义
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Wuhan Shute Biotechnology Co.,Ltd.
Nanning Normal University
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Guangxi Teachers College
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"

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Abstract

The invention discloses a kind of method using nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin, including：Step 1: preparing nitrogen phosphorus doping carbon quantum dot, it is specially：Hydrotalcite powder and Ludox are dissolved in deionized water, add phosphoric acid and the stirring of sulfamic acid sodium, cooling takes supernatant liquid filtering, take filtrate microwave treatment, drying, take powder, iminodiacetic acid and phosphoric acid solution to be dissolved in anaerobic high purity water, add methyliminodiacetic acid, stirring, supernatant infrared radiation is taken, is configured to solution, adds the hemoglobin standard solution of various concentrations thereto, exciting light is irradiated, establishes the relation of hemoglobin concentration and fluorescence intensity；Step 2: irradiation exciting light, detects fluorescence intensity corresponding to the solution to be measured containing hemoglobin, according to the hemoglobin concentration and fluorescence intensity detected in step 1, the concentration of corresponding hemoglobin in solution to be measured is read.The present invention, which has, detects the high beneficial effect of sensitive, strong antijamming capability, accuracy in detection.

Description

Utilize the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin
Technical field
The present invention relates to hemoglobin detection field.It is more particularly related to a kind of utilize nitrogen phosphorus doping carbon amounts The method of sub- point probe detection hemoglobin.
Background technology
Hemoglobin is the indispensable albumen of human body, and majority is stored in red blood cell, and it is one kind by 2 α and 2 The tetramer protein that beta polypeptides chain is combined into, wherein every peptide chain combines with a prosthetic heme group (ferroporphyrin prothetic group) again. Research shows, in organism many key activities related to oxygen, energy and metabolism have the participation of hemoglobin.And blood Lactoferrin is both a kind of redox protein, is a kind of allosteric protein again, the allosteric effect and oxidation-reduction quality of hemoglobin by To showing warm solicitude for for vast scientific researcher.And the concentration of hemoglobin in blood and many phases such as with blood, heart The disease of pass has close contact, so the measurement of hemoglobin concentration is quite important in blood of human body.
Science and technology is developed rapidly so that applications to nanostructures and research and development are swift and violent, and wherein fluorescent nano material is because having The general character and the outstanding fluorescence property of common nano material and be widely used in the field such as biology, chemistry, medical science.Often The fluorescent nano material seen have quantum dot (Quantum dots, QDs), metal-doped fluorescent nano material, metal nanometre cluster, Composite organic-inorganic material etc..Quantum dot is again because it has narrow transmitting peak shape, width and continuous absorption spectrum, easily controllable Launch wavelength the advantages that, it is widely used in fields such as biochemistry, biology, materia medica.But due to itself Containing heavy metal ion, very important injury can all be caused to environment and human body, limit its application in each field.2004 Year, a kind of fluorescent material is found, and thereafter, by a series of research, carbon point (Carbon dots) formally turns into this new The title of fluorescent nano material.Relative to the material of other same types, carbon point is so that raw material is cheap and easy to get, hypotoxicity, bio-compatible Property, the advantages that good water solubility, chemical property are stable and obtained extensively in fields such as biochemistry, biological medicine, analytical chemistry General application.In the recent period, the research day that the heavy metal ion and biomolecule using the property of carbon point fluorescence as foundation are measured Benefit increases.The detection in ion and molecule, cell imaging etc. are applied carbon point, in the field such as biology, chemistry, material There are good development prospect and wide development space.
The method of conventional detection hemoglobin uses colorimetric method more, but during the concentration of colorimetric determination hemoglobin, spirit Sensitivity is not high, and accuracy is not high enough, and antijamming capability has to be strengthened.
The content of the invention
It is an object of the invention to solve at least the above, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide a kind of side using nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin Method, can sensitively detect hemoglobin, accurately and reliably detect the concentration of hemoglobin in solution, antijamming capability By force.
In order to realize according to object of the present invention and further advantage, there is provided one kind utilizes nitrogen phosphorus doping carbon quantum dot The method of probe in detecting hemoglobin, comprises the following steps：
Step 1: preparing nitrogen phosphorus doping carbon quantum dot, nitrogen phosphorus doping carbon quantum dot solution is prepared, addition is different dense thereto The hemoglobin standard solution of degree, exciting light is irradiated, establish the relation of hemoglobin concentration and fluorescence intensity；
Step 2: irradiation exciting light, detects fluorescence intensity corresponding to the solution to be measured containing hemoglobin, according to step 1 The hemoglobin concentration and fluorescence intensity of middle detection, read the concentration of corresponding hemoglobin in solution to be measured.
Preferably, hemoglobin concentration is established in step 1 and the relation of fluorescence intensity is specially：Detect every part it is blood red Fluorescence spectrum corresponding to protein standard solution, and fluorescence intensity corresponding to every part of hemoglobin standard solution is recorded, using concentration as 0 Hemoglobin standard solution corresponding to fluorescence intensity fluorescence corresponding with the hemoglobin standard solution of the other concentration of any part The ratio of intensity is ordinate, and the concentration of every part of hemoglobin standard solution is abscissa, draws standard curve and accounting equation；
Step 2 is specially：Solution to be measured containing hemoglobin is added to a nitrogen phosphorus doping carbon quantum dot solution In, exciting light is irradiated, detects fluorescence spectrum corresponding to solution to be measured, and fluorescence intensity corresponding to solution to be measured is recorded, substitute into institute State equation and obtain the concentration of corresponding hemoglobin in solution to be measured.
Preferably, the concentration of the hemoglobin standard solution of more parts of various concentrations in step 1 is followed successively by：0、1× 10-9mol/L、6×10-9mol/L、2×10-8mol/L、4×10-8mol/L、6×10-8mol/L、2×10-7mol/L、4×10- 7mol/L、5×10-7mol/L、8×10-7mol/L、1.3×10-6mol/L、2.5×10-6mol/L、3.5×10-6mol/L、4× 10-6Mol/L and 6 × 10-6mol/L。
Preferably, the wavelength of exciting light is 380nm.
Preferably, use pH for 7.4 PBS buffer preparation nitrogen phosphorus doping carbon quantum dot solution and haemoglobin scale Quasi- solution, pH is used to adjust the pH value of solution to be measured to 7.4 for 7.4 PBS cushioning liquid.
Preferably, the concentration of nitrogen phosphorus doping carbon quantum dot solution is 0.5mg/mL.
Preferably, the preparation method of nitrogen phosphorus doping carbon quantum dot is specially：
S1,1.5g hydrotalcite powders and 0.5g Ludox be dissolved in 10mL deionized waters, adding 0.8mL mass fractions is 40% phosphoric acid and 0.2g sulfamic acids sodium stirs 3h under 60 DEG C of water bath conditions, is cooled to room temperature, takes supernatant in 0.45 μm Filtering with microporous membrane, take filtrate microwave treatment 30s, powder dried to obtain under the conditions of 30 DEG C, wherein, microwave power 900W, Microwave temperature is 180 DEG C；
S2, take the phosphoric acid that powder, 0.2g iminodiacetic acids and 50mL mass fractions in 0.1g steps S1 are 10% Solution is dissolved in 250mL anaerobic high purity waters, adds 1.5g methyliminodiacetic acids, and 1h is stirred with 200r/min speed, Supernatant infrared radiation 20min is taken, produces nitrogen phosphorus doping carbon quantum dot, wherein, infrared radiation power is 500W.
Preferably, preparing nitrogen phosphorus doping carbon quantum dot solution is specially：Nitrogen phosphorus doping carbon quantum dot and PBS bufferings is molten Liquid is 0.5mg by mass volume ratio:After 1mL mixing, 20min, supersonic frequency 40kHz are ultrasonically treated, ultrasonic temperature is 80 DEG C, Then it can only be respectively to be sealed in 10%, 40% and 80% container through the transmitance of blue light to be sequentially placed into, and is used Fluorescent lamp irradiates 40min respectively, pours into sealing frost 45min, the HIGH PRESSURE TREATMENT 30min under 150MPa in hermetic bag, then be placed in 30min is stood in camera bellows.
Preferably, the compound method of hemoglobin standard solution is specially：Hemoglobin and PBS cushioning liquid are mixed Close, be placed in 4 DEG C of cold bath and stir 2min, then be sequentially placed into can only pass through red light transmitance be respectively 80%, 40%, And 10% container in seal, while be placed in 4 DEG C of cold bath, and 20min is irradiated using fluorescent lamp respectively, then be placed in dark 30min is stood in case；
Solution to be measured containing hemoglobin is also pre-processed, and is specially：Solution to be measured is adjusted with PBS cushioning liquid Save pH value be 7.4, be placed in 4 DEG C of cold bath and stir 2min, then be sequentially placed into can only pass through red light transmitance be respectively 10%th, seal, while be placed in 4 DEG C of cold bath in 40% and 80% container, and irradiated respectively using fluorescent lamp 20min, then it is placed in standing 30min in camera bellows.
The present invention comprises at least following beneficial effect：
Firstth, the advantages of detection method of the invention has high sensitivity, strong antijamming capability, detection accurately and reliably；
Secondth, can be learnt from fluorescence spectra, hemoglobin concentration is fluorescence intensity corresponding to 0 and solution pair to be detected The fluorescence intensity answered increases with the increase of the concentration of hemoglobin, and the concentration of fluorescence intensity ratio and hemoglobin has well Linear relationship；
3rd, hydrotalcite powder and Ludox are dissolved in deionized water, add phosphoric acid and amino that mass fraction is 40% Sodium sulfonate stirs 3h under 60 DEG C of water bath conditions, is cooled to room temperature, takes supernatant to take filtrate in 0.45 μm of filtering with microporous membrane Microwave treatment 30s, powder is dried to obtain under the conditions of 30 DEG C, carbon atom can be made to be formed about some microcellular structures, to accommodate nitrogen Or phosphorus, and the larger fixed substance of other particles is eliminated, less than 0.45 μm of the carbon mix with microcellular structure is left, It is dissolved in again with iminodiacetic acid and mass fraction for 10% phosphoric acid solution in anaerobic high purity water, adds methyl-imino two Acetic acid, 1h is stirred with 200r/min speed, supernatant infrared radiation is taken, nitrogen and phosphorus can be inlaid into microcellular structure, and Certain free degree is kept, when exciting light irradiation, can improve energy in microcellular structure internal rotation, slight fluctuation Conversion efficiency, strengthen emitted luminescence intensity；
4th, after mixing nitrogen phosphorus doping carbon quantum dot and PBS cushioning liquid, it is ultrasonically treated, liquid is constantly impacted micro- Inside pore structure, promote nitrogen, phosphorus activity, then be sequentially placed into can only pass through blue light transmitance be respectively 10%, 40% and Seal in 80% container, and irradiated respectively using fluorescent lamp, using blue visible light gradient illumination, nitrogen, phosphorus and carbon are carried out Gentle activation, laggard horizontal high voltage processing is freezed, HIGH PRESSURE TREATMENT can crush the ice crystal outside micropore again after frost, and micropore Internal ice crystal structure is constant, keeps the micro-activated state of nitrogen, phosphorus and carbon, makes to be placed in camera bellows again and stands, ice crystal inside micropore Melt, nitrogen, phosphorus and carbon activate completely；
5th, hemoglobin is mixed with PBS cushioning liquid, cold bath stirring, shrinks hemoglobin construct, then successively It can only be respectively to be sealed in 80%, 40% and 10% container through the transmitance of red light to be placed in, while is placed in 4 DEG C In cold bath, and irradiated respectively using fluorescent lamp, red visible can slowly induce hemoglobin α subunits and β subunits to Outer protrusion, specific structure is formed, then be placed in camera bellows and stand, the structural stretch of hemoglobin, but it is outside by α subunits and β subunits The specific structure that protrusion is formed remains in that protruding state, and this structure can just extend into the micro- of nitrogen phosphorus doping carbon quantum dot In pore structure, suppress the activity of nitrogen, phosphorus and carbon, significantly inhibit the fluorescent effect of nitrogen phosphorus doping carbon quantum dot.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this The research and practice of invention and be understood by the person skilled in the art.
Brief description of the drawings
Fig. 1 is the fluorescence spectra of the hemoglobin standard solution of the present invention；
Fig. 2 is the fluorescence intensity ratio of the present invention and the canonical plotting of hemoglobin concentration.
Embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to specification Word can be implemented according to this.
<Embodiment 1>
A kind of method using nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin, comprise the following steps：
Step 1: the nitrogen phosphorus doping carbon quantum dot for using pH to be 0.5mg/mL for 7.4 PBS buffer preparation concentration is molten Liquid, uses pH to configure the hemoglobin standard solution of more parts of various concentrations for 7.4 PBS cushioning liquid, and hemoglobin standard is molten The concentration of liquid is followed successively by：0、1×10-9mol/L、6×10-9mol/L、2×10-8mol/L、4×10-8mol/L、6×10-8mol/ L、2×10-7mol/L、4×10-7mol/L、5×10-7mol/L、8×10-7mol/L、1.3×10-6mol/L、2.5×10- 6mol/L、3.5×10-6mol/L、4×10-6mol/L、6×10-6mol/L、7×10-6mol/L、8×10-6mol/L、1×10- 5Mol/L, the nitrogen phosphorus doping carbon quantum dot solution that 18 parts of volumes are 3ml is measured respectively, in a nitrogen phosphorus doping carbon quantum dot solution A hemoglobin standard solution is added respectively, and illumination wavelength is 380nm exciting light, obtains transmitting light as shown in Figure 1 Fluorescence spectra, and record the fluorescence intensity of 480nm corresponding to every part of hemoglobin standard solution, using concentration as 0 blood red egg 480nm fluorescence intensity 480nm corresponding with the hemoglobin standard solution of the other concentration of any part corresponding to white standard liquid The ratio of fluorescence intensity be ordinate, the concentration of every part of hemoglobin standard solution is abscissa, as shown in Figure 2, blood red The concentration of protein standard solution is 0~6 × 10-6During mol/L, the concentration of fluorescence intensity ratio and hemoglobin has good line Sexual intercourse, draw standard curve and accounting equation；
Step 2: pH is used to arrive the pH value regulation of the solution to be measured containing hemoglobin for 7.4 PBS cushioning liquid 7.4, solution to be measured is added in the nitrogen phosphorus doping carbon quantum dot solution that a volume is 3ml, illumination wavelength is swashing for 380nm It is luminous, fluorescence spectrum corresponding to solution to be measured is detected, and the fluorescence intensity of 480nm corresponding to solution to be measured is recorded, described in substitution Equation obtains the concentration of corresponding hemoglobin in solution to be measured.
<Embodiment 2>
Determination step with embodiment 1, wherein, unlike the preparation method of nitrogen phosphorus doping carbon quantum dot, nitrogen in step 1 The compound method of phosphorus doping carbon quantum dot solution, the compound method of hemoglobin standard solution and treated containing hemoglobin It is different to survey the pretreatment of solution, it is described in detail below：
The preparation method of nitrogen phosphorus doping carbon quantum dot is specially：
S1,1.5g hydrotalcite powders and 0.5g Ludox be dissolved in 10mL deionized waters, adding 0.8mL mass fractions is 40% phosphoric acid and 0.2g sulfamic acids sodium stirs 3h under 60 DEG C of water bath conditions, is cooled to room temperature, takes supernatant in 0.45 μm Filtering with microporous membrane, take filtrate microwave treatment 30s, powder dried to obtain under the conditions of 30 DEG C, wherein, microwave power 900W, Microwave temperature is 180 DEG C；
S2, take the phosphoric acid that powder, 0.2g iminodiacetic acids and 50mL mass fractions in 0.1g steps S1 are 10% Solution is dissolved in 250mL anaerobic high purity waters, adds 1.5g methyliminodiacetic acids, and 1h is stirred with 200r/min speed, Supernatant infrared radiation 20min is taken, produces nitrogen phosphorus doping carbon quantum dot, wherein, infrared radiation power is 500W.
Preparing nitrogen phosphorus doping carbon quantum dot solution is specially：Nitrogen phosphorus doping carbon quantum dot and PBS cushioning liquid are pressed into quality Volume ratio is 0.5mg:After 1mL mixing, 20min, supersonic frequency 40kHz are ultrasonically treated, ultrasonic temperature is 80 DEG C, then successively It can only be respectively to be sealed in 10%, 40% and 80% container through the transmitance of blue light to be placed in, and using fluorescent lamp point 40min is not irradiated, pours into sealing frost 45min, the HIGH PRESSURE TREATMENT 30min under 150MPa in hermetic bag, then be placed in quiet in camera bellows Put 30min.
The compound method of hemoglobin standard solution is specially：Hemoglobin is mixed with PBS cushioning liquid, is placed in 4 DEG C Cold bath in stir 2min, then be sequentially placed into can only pass through red light transmitance be respectively 80%, 40% and 10% Seal, while be placed in 4 DEG C of cold bath in container, and 20min is irradiated using fluorescent lamp respectively, then be placed in camera bellows and stand 30min；
Solution to be measured containing hemoglobin is also pre-processed, and is specially：Solution to be measured is adjusted with PBS cushioning liquid Save pH value be 7.4, be placed in 4 DEG C of cold bath and stir 2min, then be sequentially placed into can only pass through red light transmitance be respectively 10%th, seal, while be placed in 4 DEG C of cold bath in 40% and 80% container, and irradiated respectively using fluorescent lamp 20min, then it is placed in standing 30min in camera bellows.
<Comparative example 1>
Using the concentration of hemoglobin in colorimetric determination solution to be measured, wherein, prepare 16 parts and treated containing hemoglobin Solution is surveyed, the concentration of hemoglobin is respectively 2 × 10 in the solution to be measured containing hemoglobin-9mol/L、3×10-8mol/L、6 ×10-7Mol/L and 5 × 10-6Mol/L, it is divided into totally 4 groups of A, B, C and D, every group 4 parts, virtue is added into B groups solution to be measured Base ruthenium, the concentration for making aryl ruthenium in solution to be measured are finally 1 × 10-7Mol/L, 4 parts are obtained containing hemoglobin and aryl ruthenium Solution to be measured, Cr VI is added into C groups solution to be measured, the concentration for making concentration Cr VI in solution to be measured is finally 1.2 × 10- 5Mol/L, 4 parts of solution to be measured containing hemoglobin and Cr VI are obtained, aryl ruthenium and sexavalence are added into D groups solution to be measured Chromium, it is respectively 1 × 10 to make the concentration of aryl ruthenium and Cr VI in solution to be measured-7Mol/L and 1.2 × 10-5Mol/L, contained The solution to be measured of hemoglobin, aryl ruthenium and Cr VI.
<Comparative example 2>
Determination step with embodiment 1, wherein, solution to be measured containing hemoglobin is different, the compound method of solution to be measured With comparative example 1.
<Comparative example 3>
Determination step with embodiment 2, wherein, solution to be measured containing hemoglobin is different, the compound method of solution to be measured With comparative example 1.
<Testing result>
It is as shown in table 1 hemoglobin testing result：
The hemoglobin detection limit of embodiment 1 and embodiment 2 is significantly lower than comparative example 1 as can be seen from Table 1, and explanation is adopted Method with nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is lower than the detection limit of conventional colorimetric method, and detection is sensitiveer, real The Detection accuracy of the A group hemoglobins of example 1 and embodiment 2 is applied apparently higher than comparative example 1, illustrates to use nitrogen phosphorus doping carbon quantum The degree of accuracy of the method for point probe detection hemoglobin is higher than conventional colorimetric method；
The hemoglobin Detection accuracy of the B groups of comparative example 3, C groups and D groups is all remarkably higher than comparative example 1 and comparative example 2, the interference of the detection of aryl ruthenium and Cr VI to hemoglobin can be avoided by illustrating the method for comparative example 3, improve accuracy rate；
The method that the hemoglobin Detection accuracy of the B groups of comparative example 2, C groups and D groups is below comparative example 1, explanation When have contain aryl ruthenium or Cr VI in solution to be measured when, the degree of accuracy of the method for comparative example 2 can be reduced, on the contrary without routine side Method is accurate.
Number of devices and treatment scale described herein are the explanations for simplifying the present invention.To the present invention application, Modifications and variations will be readily apparent to persons skilled in the art.
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited In specific details and shown here as the legend with description.

Claims (9)

  1. A kind of 1. method using nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin, it is characterised in that comprise the following steps：
    Step 1: preparing nitrogen phosphorus doping carbon quantum dot, nitrogen phosphorus doping carbon quantum dot solution is prepared, adds various concentrations thereto Hemoglobin standard solution, exciting light is irradiated, establish the relation of hemoglobin concentration and fluorescence intensity；
    Step 2: irradiation exciting light, detect the solution to be measured containing hemoglobin corresponding to fluorescence intensity, examined according in step 1 The hemoglobin concentration and fluorescence intensity of survey, read the concentration of corresponding hemoglobin in solution to be measured.
  2. 2. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 1, it is characterised in that The relation that hemoglobin concentration and fluorescence intensity are established in step 1 is specially：Detect corresponding to every part of hemoglobin standard solution Fluorescence spectrum, and record fluorescence intensity corresponding to every part of hemoglobin standard solution, using concentration as 0 hemoglobin standard solution The ratio of corresponding fluorescence intensity fluorescence intensity corresponding with the hemoglobin standard solution of the other concentration of any part is ordinate, The concentration of every part of hemoglobin standard solution is abscissa, draws standard curve and accounting equation；
    Step 2 is specially：Solution to be measured containing hemoglobin is added in a nitrogen phosphorus doping carbon quantum dot solution, shone Exciting light is penetrated, detects fluorescence spectrum corresponding to solution to be measured, and fluorescence intensity corresponding to recording solution to be measured, substitute into the equation Obtain the concentration of corresponding hemoglobin in solution to be measured.
  3. 3. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 1, it is characterised in that The concentration of the hemoglobin standard solution of more parts of various concentrations in step 1 is followed successively by：0、1×10-9mol/L、6×10- 9mol/L、2×10-8mol/L、4×10-8mol/L、6×10-8mol/L、2×10-7mol/L、4×10-7mol/L、5×10- 7mol/L、8×10-7mol/L、1.3×10-6mol/L、2.5×10-6mol/L、3.5×10-6mol/L、4×10-6Mol/L, with And 6 × 10-6mol/L。
  4. 4. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 2, it is characterised in that The wavelength of exciting light is 380nm.
  5. 5. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 1, it is characterised in that Use pH for 7.4 PBS buffer preparation nitrogen phosphorus doping carbon quantum dot solution and hemoglobin standard solution, use pH for 7.4 PBS cushioning liquid adjusts the pH value of solution to be measured to 7.4.
  6. 6. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 1, it is characterised in that The concentration of nitrogen phosphorus doping carbon quantum dot solution is 0.5mg/mL.
  7. 7. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 1, it is characterised in that The preparation method of nitrogen phosphorus doping carbon quantum dot is specially：
    S1,1.5g hydrotalcite powders and 0.5g Ludox be dissolved in 10mL deionized waters, it is 40% to add 0.8mL mass fractions Phosphoric acid and 0.2g sulfamic acids sodium stir 3h under 60 DEG C of water bath conditions, are cooled to room temperature, take supernatant in 0.45 μm of micropore Membrane filtration, filtrate microwave treatment 30s is taken, powder is dried to obtain under the conditions of 30 DEG C, wherein, microwave power 900W, microwave temperature Spend for 180 DEG C；
    S2, take the phosphoric acid solution that powder, 0.2g iminodiacetic acids and 50mL mass fractions in 0.1g steps S1 are 10% It is dissolved in 250mL anaerobic high purity waters, adds 1.5g methyliminodiacetic acids, 1h is stirred with 200r/min speed, taken Clear liquid infrared radiation 20min, nitrogen phosphorus doping carbon quantum dot is produced, wherein, infrared radiation power is 500W.
  8. 8. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 5, it is characterised in that Preparing nitrogen phosphorus doping carbon quantum dot solution is specially：It is by mass volume ratio by nitrogen phosphorus doping carbon quantum dot and PBS cushioning liquid 0.5mg:After 1mL mixing, 20min, supersonic frequency 40kHz are ultrasonically treated, ultrasonic temperature is 80 DEG C, and being then sequentially placed into can only Transmitance through blue light is respectively to be sealed in 10%, 40% and 80% container, and is irradiated respectively using fluorescent lamp 40min, sealing frost 45min, the HIGH PRESSURE TREATMENT 30min under 150MPa in hermetic bag is poured into, then be placed in camera bellows and stand 30min .
  9. 9. the method for nitrogen phosphorus doping carbon quantum dot probe in detecting hemoglobin is utilized as claimed in claim 5, it is characterised in that The compound method of hemoglobin standard solution is specially：Hemoglobin is mixed with PBS cushioning liquid, is placed in 4 DEG C of cold bath Middle stirring 2min, then be sequentially placed into can only pass through red light transmitance be respectively 80%, 40% and 10% container in it is close Envelope, while be placed in 4 DEG C of cold bath, and 20min is irradiated using fluorescent lamp respectively, then it is placed in standing 30min in camera bellows；
    Solution to be measured containing hemoglobin is also pre-processed, and is specially：Solution to be measured is adjusted into pH with PBS cushioning liquid Be worth for 7.4, be placed in 4 DEG C of cold bath and stir 2min, then be sequentially placed into can only pass through red light transmitance be respectively 10%, Seal, while be placed in 4 DEG C of cold bath in 40% and 80% container, and 20min is irradiated using fluorescent lamp respectively, then It is placed in standing 30min in camera bellows.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108956552A (en) * 2018-05-02 2018-12-07 中国科学院化学研究所 A kind of preparation method of carbon quantum dot and method with carbon quantum dot detection dichromate ion
CN110132917A (en) * 2019-05-14 2019-08-16 山西大学 A kind of cobalt nitrogen codope carbon dots and its preparation method and application
CN111591975A (en) * 2020-06-04 2020-08-28 山东丰益泰和科技有限公司 Method for synthesizing carbon quantum dots based on diethylenetriamine penta (methylene phosphonic acid)
CN112694887A (en) * 2020-12-07 2021-04-23 黑龙江省农业科学院植物保护研究所 Light-emitting sensor, construction method thereof and application of light-emitting sensor in detection of salicylic acid content in plants

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106315557A (en) * 2016-08-18 2017-01-11 常州大学 Preparation method for nitrogen and phosphorus-doped carbon quantum dots
CN106770100A (en) * 2016-12-05 2017-05-31 上海大学 A kind of method that hemoglobin is detected based on graphene quantum dot

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106315557A (en) * 2016-08-18 2017-01-11 常州大学 Preparation method for nitrogen and phosphorus-doped carbon quantum dots
CN106770100A (en) * 2016-12-05 2017-05-31 上海大学 A kind of method that hemoglobin is detected based on graphene quantum dot

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEN-I WANG ET AL.: "Electrochemical synthesis of photoluminescent carbon nanodots from glycine for highly sensitive detection of hemoglobin", 《GREEN CHEMISTRY》 *
JIN ZHOU ET AL.: "Carbon dots doped with heteroatoms for fluorescent bioimaging: a review", 《MICROCHIM ACTA》 *
SHAN HUANG ET AL.: "A carbon dots based fluorescent probe for selective and sensitive detection of hemoglobin", 《SENSORS AND ACTUATORS B: CHEMICAL》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108956552A (en) * 2018-05-02 2018-12-07 中国科学院化学研究所 A kind of preparation method of carbon quantum dot and method with carbon quantum dot detection dichromate ion
CN108956552B (en) * 2018-05-02 2020-10-27 中国科学院化学研究所 Preparation method of carbon quantum dots and method for detecting dichromate by using carbon quantum dots
CN110132917A (en) * 2019-05-14 2019-08-16 山西大学 A kind of cobalt nitrogen codope carbon dots and its preparation method and application
CN111591975A (en) * 2020-06-04 2020-08-28 山东丰益泰和科技有限公司 Method for synthesizing carbon quantum dots based on diethylenetriamine penta (methylene phosphonic acid)
CN112694887A (en) * 2020-12-07 2021-04-23 黑龙江省农业科学院植物保护研究所 Light-emitting sensor, construction method thereof and application of light-emitting sensor in detection of salicylic acid content in plants
CN112694887B (en) * 2020-12-07 2024-04-26 黑龙江省农业科学院植物保护研究所 Luminous sensor, construction method thereof and application thereof in detecting salicylic acid content of plants

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