CN107677650B - Dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles - Google Patents
Dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles Download PDFInfo
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- 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
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- G—PHYSICS
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- 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
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
Abstract
The invention discloses a dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles, wherein the particle size of the nanoparticles is concentrated at 5nm, the luminescence position is concentrated at 473nm, the dopamine detection method has better water solubility and stability, the synthesis method has simple steps, the dopamine detection method can be directly applied without marking, does not need any modification, is convenient and fast to operate, has simple steps, reduces the use of expensive instruments in the aspect of detection, has short time consumption, reduces the detection cost, and has the detection limit of 0.2 mu M.
Description
Technical Field
The invention belongs to the technical field of biological analysis and detection, and particularly relates to a synthetic method of up-conversion nanoparticles with good water solubility and small particle size and detection application of the up-conversion nanoparticles to biological micromolecular dopamine.
Background
Upconversion luminescence is a process of emitting high-energy light by excitation with low-energy light (typically 980nm near-infrared light) through successive multiphoton absorption and energy transfer processes, and is therefore also referred to as upconversion. The rare earth up-conversion luminescent material has excellent performance due to rich energy level structure of rare earth elements, such as large anti-stokes shift, narrow emission band, long fluorescence life, no background fluorescence, low toxicity, strong penetrating power and the like, which arouse extensive attention of scientific researchers. In recent years, rare earth doped up-conversion nanoparticles show important application prospects in the fields of biomolecular labeling, biological imaging and the like, and the up-conversion nanoparticles are used for detecting Zn2+And biological small molecules, so the method has good application prospect in the analysis and detection direction. Dopamine (dopamine) is a neurotransmitter in hypothalamic dopamine and pituitary gland, a neuronal substance of the midbrain. Dopamine is used for transferring information of excitement and pleasure, and can affect mood and endocrine function of human bodyThe presence of multiple dopaminergic amines can provide sustained excitement and thus can treat depression, but also cause addiction, resulting in dependence on addicting men. Insufficient dopamine in the endocrine of a human body can cause the loss of the control capability of muscles, and in severe cases, the involuntary vibration of hands and feet of the human body or the Parkinson's disease can be caused. Dopamine contributes to the improvement of memory and thus to the treatment of patients with alzheimer's disease. In addition, dopamine is of great importance for the treatment of shock syndromes of various shock types, such as myocardial infarction, endotoxemia, renal failure, etc., especially shock patients with impaired myocardial contractility, decreased urine volume and blood volume replenishment.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles.
The technical purpose of the invention is realized by the following technical scheme:
the dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles is carried out according to the following steps: uniformly mixing an upconversion material solution, high-purity water, a Tri-HCl buffer solution and a dopamine solution of uniformly dispersed ytterbium and thulium-doped sodium yttrium tetrafluoride upconversion nanoparticles, detecting fluorescence intensity by using a fluorescence photometer, fitting a linear equation of Y0.0039X +1.1563, wherein the linear range is 1-200 mu M, and the detection limit is 0.2 mu M.
During testing, an upconversion material solution, high-purity water, a Tri-HCl buffer solution and a dopamine solution, which are used for uniformly dispersing ytterbium and thulium doped sodium yttrium tetrafluoride upconversion nanoparticles, are uniformly mixed and placed at room temperature of 20-25 ℃ for at least 1 hour, preferably 1-2 hours.
In the test, the volume ratio of the upconverting material solution, high purity water, Tri-HCl buffer solution and dopamine solution was 2:1.5:0.4:0.1, preferably 2mL of upconverting solution, 1.5mL of high purity water, 400. mu.L of Tri-HCl buffer solution and 100. mu.L of dopamine solution.
The pH of the Tri-HCl buffer solution was 7-9.5 at the time of the test.
The upconverting material solution concentration was 50-120. mu.g/mL when tested.
In the invention, the particle size of ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles is concentrated at 4-5 nm and is concentrated at 473nm for luminescence, and the preparation method comprises the following steps:
step 2, adding 0.15-0.25 part by mass of polyethyleneimine, 4-5 parts by volume of ethylene glycol and 3-3.5 parts by volume of high-purity water into the substance prepared in the step 1, and performing ultrasonic treatment and stirring to uniformly disperse the substance;
step 3, weighing 0.2-0.6 part by mass of sodium hydroxide, 0.2-0.5 part by mass of ammonium fluoride, 4-5 parts by volume of high-purity water and 3-4 parts by volume of ethylene glycol, and carrying out ultrasonic treatment and stirring to uniformly disperse the high-purity water and the ethylene glycol;
and 4, adding the uniformly dispersed solution obtained in the step 2 into the uniformly dispersed solution obtained in the step 3, and reacting at 200-250 ℃ for at least 10 hours, preferably 12-24 hours after uniform dispersion.
In the above technical scheme, 1 part by mass is 1g, and 1 part by volume is 1 ml.
In the technical scheme, after the reaction in the step 4, the solution in the reaction kettle after the reaction is uniformly stirred by a glass rod, then ethanol and the solution in the reaction kettle are mixed, wherein the volume ratio of the ethanol to the solution is 3:1, the volume ratio of the ethanol to the solution is 3mL, the volume ratio of the solution to the solution is 1mL, the mixture is transferred to a 5mL centrifuge tube after uniform mixing, centrifugation is carried out, and the supernatant is discarded. Washing for three times, washing with high-purity water, centrifuging for three times, and discarding supernatant to obtain ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material, and storing in water solution (i.e. uniformly dispersing in water).
In the invention, the prepared sodium yttrium tetrafluoride is doped with ytterbium and thulium NaYF4：Yb/TmHas better water solubility and stability, and ytterbium and thulium doped yttrium tetrafluorideSodium (NaYF)4：Yb/Tm) The synthesis method of the up-conversion nanoparticles is simple in steps, can be directly applied without a label when detecting dopamine, and does not need any modification. The operation is convenient and fast, and the steps are simple. The material is carried out under the excitation of near infrared light, the damage to a human body caused by ultraviolet excitation is avoided, and the light stability is good. Meanwhile, in the detection, complex processes such as target object and material treatment are not needed. The following table shows the analysis characteristic quantity of the detection method and the comparison with other methods, and the fitting linear equation of the technical scheme of the invention is that Y is 0.0039X +1.1563, the linear range is 1-200 mu M, and the detection limit is 0.2 mu M.
[1]Y.Kim,S.Bong,Y.Kang,Y.Yang,R.K.Mahajan,J.S.Kim,H.Kim，Electrochemical detection of dopamine in the presence of ascorbic acid usinggraphene modified electrodes,Biosens.Bioelectron.25(2010)2366–2369.
[2]X.Cao,X.Cai,N.Wang,Selective sensing of dopamine at MnOOH nanobeltmodified electrode,Sens.Actuators B 160(2011)771–776.
[3]M.Li,J.Zhu,L.Zhang,X.Chen,H.Zhang,F.Zhang,S.Xu,D.G.Evans,Facilesynthesis of NiAl-layered double hydroxide/graphene hybrid with enhancedelectrochemical properties for detection of dopamine,Nanoscale 3(2011)4240–4246.
[4]X.Liu,L.Cheng,J.Lei,H.Ju,Dopamine detection based on its quenchingeffect on the anodic electrochemiluminescence of CdSe quantum dots,Analyst133(2008)1161–1163.
[5]S.Liu,F.Shi,X.Zhao,L.Chen,X.Su,3-Aminophenyl boronic acid-functionalized CuInS2quantum dots as a near-infrared fluorescence probe forthe determination of dopamine,Biosens.Bioelectron.47(2013)79–384.
Drawings
FIG. 1 is a diagram of ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material (i.e., nanoparticle NaYF) used in the present invention4：Yb/Tm) Transmission Electron Microscopy (TEM) of (a), wherein the upper right image is a partial magnified view of the nanoparticles.
FIG. 2 is a diagram of ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material (i.e., nanoparticle NaYF) used in the present invention4：Yb/Tm) X-ray powder diffractogram (XRD).
FIG. 3 is a diagram of ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material (i.e., nanoparticle NaYF) used in the present invention4：Yb/Tm) Fourier transform infrared spectroscopy (FTIR).
FIG. 4 is a diagram of ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material (i.e., nanoparticle NaYF) used in the present invention4：Yb/Tm) The fluorescence spectrum of (a).
FIG. 5 is a sodium yttrium tetrafluoride up-conversion material (i.e., nanoparticle NaYF) doped with ytterbium and thulium4：Yb/Tm) Linear range plots for dopamine detection were performed.
Detailed Description
The technical scheme of the invention is further explained by combining specific examples.
Example 1: and (3) preparing and characterizing the ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material.
(1) Weighing Y with mass of 0.0750g, 0.0350g and 0.0005g respectively2O3、Yb2O3、Tm2O3Putting the three rare earth oxides into a small beaker A, respectively adding 1mL of high-purity water and 2mL of concentrated nitric acid (63 wt%) into the small beaker containing the rare earth oxides, putting the small beaker into an electric heating jacket for heating to nitrify for 2 hours, adding 4mL of high-purity water into the small beaker after the oxides are completely nitrified, continuously adding 4mL of high-purity water when the rare earth oxides are quickly evaporated to dryness, then evaporating to dryness until the pH value of the solution is close to neutral, and evaporating to dryness for later use.
(2) 0.45g of sodium hydroxide and 0.4g of ammonium fluoride were weighed in a B beaker, and 0.25g of polyethyleneimine (PEI, number average molecular weight 5000) was weighed in an A beaker, and 4.5mL of ethylene glycol and 3mL of high purity water were added to the A beaker. To beaker B was added 4.5mL of high purity water, 3mL of ethylene glycol. The A, B beakers were sonicated for 10 minutes and then stirred for 10 minutes, with the sonication alternating until the contents of the beakers were dissolved for 60 minutes. And transferring the transparent solution containing the rare earth particles and the Polyethyleneimine (PEI) in the beaker A to a beaker B under magnetic stirring, stopping stirring after the transfer is completed, standing, transferring to a reaction kettle, heating to 200 ℃ from room temperature of 20-25 ℃ at a speed of 3 ℃ per minute, preserving heat at 200 ℃ and reacting for 12 hours. And uniformly stirring the solution in the reaction kettle after the reaction by using a glass rod, then mixing ethanol and the solution in the reaction kettle, wherein the volume ratio of the ethanol to the solution is 3:1, the volume ratio of the ethanol to the solution is 3mL, the volume ratio of the solution to the solution is 1mL, transferring the mixture to a 5mL centrifuge tube after uniform mixing, centrifuging, and removing the supernatant. Washing for three times, washing with high-purity water, centrifuging for three times, and discarding supernatant to obtain ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material, and storing in water solution (i.e. uniformly dispersing in water).
The prepared ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material is characterized, as shown in the attached drawings 1-3, the ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material is successfully synthesized by the method, the particle size is concentrated in 4-5 nm, and polyethyleneimine is successfully coated on the up-conversion material, namely the up-conversion material is provided with PEI (UCNPS-PEI), so that the up-conversion material has amino. Fluorescence tests were performed on the above upconverting materials using agilent in usa, and the ytterbium and thulium doped sodium yttrium tetrafluoride upconverting material used in the present invention emitted light at 473.03nm, i.e. concentrated at 473nm, as shown in fig. 4.
Example 2: dopamine detection by using ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material prepared in the above way
Carrying out constant volume on the prepared aqueous solution of the uniformly dispersed ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material, measuring 1mL of the aqueous solution of the uniformly dispersed ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material, drying to obtain the mass of a solid phase substance, obtaining the concentration value of the uniformly dispersed upconversion material in the aqueous solution of the uniformly dispersed ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material through conversion, and taking the aqueous solution of the uniformly dispersed ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion material as the aqueous solution of the uniformly dispersed ytterbium andyb and thulium doped sodium yttrium tetrafluoride up-conversion material (NaYF)4：Yb/Tm) Taking 100 μ L NaYF as mother liquor (stock solution)4：Yb/TmDiluted to 50-120. mu.g/mL upconverter solution.
0.1mol/L, preparation of Tris-HCl buffer solution at 25 ℃: weighing 1.2114g Tris, dissolving in 100mL water; the solution was adjusted to pH 7-9.5 with (0.2moL/L) hydrochloric acid, and the volume was measured in a 100mL volumetric flask with high-purity water and stored at low temperature.
Preparing a dopamine solution: weighing 0.0120g of dopamine, dissolving the dopamine in 8mL of water to prepare 8mM Dopamine (DA) solution; 8mM DA solutions were diluted to 0.04, 0.4, 1.2, 2, 2.8, 4, 6mM solutions, respectively, and stored at low temperature.
In the test, the upconversion material solution, the high purity water, the Tri-HCl buffer solution and the dopamine solution are mixed uniformly, the mixture is placed at room temperature of 20-25 ℃ for 2 hours, and the fluorescence intensity is detected by a fluorescence photometer (agilent, usa). The volume of the up-conversion solution was 2mL, the volume of high purity water was 1.5mL, the volume of Tri-HCl buffer solution was 400. mu.L, and the volume of dopamine solution was 100. mu.L.
(1) The concentration of the upconversion material solution is 100 mug/mL, and the pH value of the Tri-HCl buffer solution is 7.4;
(2) the concentration of the upconversion material solution is 120 mug/mL, and the pH value of the Tri-HCl buffer solution is 8.0;
(3) the concentration of the up-conversion material solution is 60 mug/mL, and the pH value of the Tri-HCl buffer solution is 9.0;
(4) the concentration of the upconversion material solution is 50 mug/mL, and the pH value of the Tri-HCl buffer solution is 7.0;
in the four cases, the DA solution concentration was changed to obtain the relationship between the fluorescence intensity and the concentration, and in the linear range of 1 to 200 μ M, the fitted linear equation was 0.0039X +1.1563 in each case, and the detection limit was 0.2 μ M on average.
The patent of the invention is funded by a project 21375095 on the national science foundation, special funding fund FANEDD-201023 of national excellent doctor academic paper authors, an application basic research plan major project 12JCZDJC21700 in Tianjin City and an innovative talent culture project first-level project ZX110185 in Tianjin City of '131'.
The invention has been described in an illustrative manner, and it is to be understood that any simple variations, modifications or other equivalent changes which can be made by one skilled in the art without departing from the spirit of the invention fall within the scope of the invention.
Claims (4)
1. The dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles is characterized by comprising the following steps: uniformly mixing an upconversion material solution, high-purity water, a Tri-HCl buffer solution and a dopamine solution of uniformly dispersed ytterbium and thulium-doped sodium yttrium tetrafluoride upconversion nanoparticles, detecting fluorescence intensity by using a fluorescence photometer, fitting a linear equation of Y & lt 0.0039X & gt +1.1563, wherein the linear range is 1-200 mu M, the detection limit is 0.2 mu M, when in test, uniformly mixing the upconversion material solution, the high-purity water, the Tri-HCl buffer solution and the dopamine solution of uniformly dispersed ytterbium and thulium-doped sodium yttrium tetrafluoride upconversion nanoparticles, placing the mixture at room temperature of 20-25 ℃ for at least 1h, wherein the volume ratio of the upconversion material solution, the high-purity water, the Tri-HCl buffer solution and the dopamine solution is 2:1.5:0.4:0.1, the pH of the Tri-HCl buffer solution is 7-9.5, and the concentration of the upconversion material solution is 50-120 mu g/mL, the particle size of ytterbium and thulium doped sodium yttrium tetrafluoride up-conversion nanoparticles is concentrated at 4-5 nm, and the particles concentrate at 473nm to emit light, and the particles are prepared according to the following steps:
step 1, 0.07-0.08 part by mass of Y2O30.03 to 0.05 part by mass of Yb2O30.0004 to 0.0008 mass portion of Tm2O3Putting the mixture into a nitric acid solution for nitration reaction, repeatedly evaporating the mixture to dryness by using high-purity water after the reaction is finished until the pH value is neutral, and evaporating the mixture to dryness for later use;
step 2, adding 0.15-0.25 part by mass of polyethyleneimine, 4-5 parts by volume of ethylene glycol and 3-3.5 parts by volume of high-purity water into the substance prepared in the step 1, and performing ultrasonic treatment and stirring to uniformly disperse the substance;
step 3, weighing 0.2-0.6 part by mass of sodium hydroxide, 0.2-0.5 part by mass of ammonium fluoride, 4-5 parts by volume of high-purity water and 3-4 parts by volume of ethylene glycol, and carrying out ultrasonic treatment and stirring to uniformly disperse the high-purity water and the ethylene glycol;
and 4, adding the uniformly dispersed solution obtained in the step 2 into the uniformly dispersed solution obtained in the step 3, and reacting at 200-250 ℃ for at least 10 hours after uniform dispersion.
2. The dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride upconversion nanoparticles according to claim 1, characterized in that, in the test, an upconversion material solution, high purity water, Tri-HCl buffer solution and dopamine solution, in which ytterbium and thulium doped sodium yttrium tetrafluoride upconversion nanoparticles are uniformly dispersed, are mixed and placed at room temperature of 20-25 ℃ for 1-2 hours.
3. The dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride upconversion nanoparticles according to claim 1, wherein in the test, the upconversion solution is 2mL, the high purity water is 1.5mL, the Tri-HCl buffer solution is 400 μ L, and the dopamine solution is 100 μ L.
4. The dopamine detection method based on ytterbium and thulium doped sodium yttrium tetrafluoride upconversion nanoparticles according to claim 1, wherein 1 part by mass is 1g and 1 part by volume is 1ml when the preparation of ytterbium and thulium doped sodium yttrium tetrafluoride upconversion nanoparticles is performed.
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