CN113249127B - High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof - Google Patents

High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof Download PDF

Info

Publication number
CN113249127B
CN113249127B CN202110604412.6A CN202110604412A CN113249127B CN 113249127 B CN113249127 B CN 113249127B CN 202110604412 A CN202110604412 A CN 202110604412A CN 113249127 B CN113249127 B CN 113249127B
Authority
CN
China
Prior art keywords
rare earth
earth doped
time
resolved
fluorescent nanoparticle
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202110604412.6A
Other languages
Chinese (zh)
Other versions
CN113249127A (en
Inventor
杨海
李媛
胡军
杨祥良
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wuhan Nano Diagnosis For Health Biotechnology Co ltd
Huazhong University of Science and Technology
Original Assignee
Wuhan Nano Diagnosis For Health Biotechnology Co ltd
Huazhong University of Science and Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Wuhan Nano Diagnosis For Health Biotechnology Co ltd, Huazhong University of Science and Technology filed Critical Wuhan Nano Diagnosis For Health Biotechnology Co ltd
Priority to CN202110604412.6A priority Critical patent/CN113249127B/en
Publication of CN113249127A publication Critical patent/CN113249127A/en
Application granted granted Critical
Publication of CN113249127B publication Critical patent/CN113249127B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
    • C09K11/77—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals
    • C09K11/7783—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals containing two or more rare earth metals one of which being europium
    • C09K11/779—Halogenides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y20/00—Nanooptics, e.g. quantum optics or photonic crystals
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
    • C09K11/025—Use of particular materials as binders, particle coatings or suspension media therefor non-luminescent particle coatings or suspension media
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
    • G01N33/587—Nanoparticles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/18—Metal complexes
    • C09K2211/182—Metal complexes of the rare earth metals, i.e. Sc, Y or lanthanide
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08—RNA viruses
    • G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2469/00—Immunoassays for the detection of microorganisms
    • G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Nanotechnology (AREA)
  • Pathology (AREA)
  • Materials Engineering (AREA)
  • Organic Chemistry (AREA)
  • Virology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Optics & Photonics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Composite Materials (AREA)
  • Condensed Matter Physics & Semiconductors (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Inorganic Chemistry (AREA)
  • Biophysics (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

The invention discloses a high-sensitivity rare earth doped time-resolved fluorescence nanoparticle and preparation and application thereof, belonging to the technical field of chemistry and materials. The fluorescent nanoparticles are prepared from LaF 3 As hexagonal phase matrix material, Eu 3+ Doping as luminescent centers to hexagonal phase LaF 3 In the matrix, sensitizing agents of 2-thenoyltrifluoroacetone and 1, 10-phenanthroline and Eu 3+ Coordination occurs. The method is to react NH 4 Dropwise addition of aqueous solution of F to LaCl 3 And EuCl 3 Adding an alcoholic solution of 2-thenoyltrifluoroacetone and an alcoholic solution of 1, 10-phenanthroline. The invention adopts TTA as a sensitizing agent, Phen as a non-aqueous auxiliary ligand and utilizes TTA and Eu 3+ Resonance energy transfer between ions and Phen by substituting fluorescence quenching group-OH, with Eu 3+ The ions generate strong coordination, so that the fluorescence of the nanoparticles is effectively enhanced.

Description

High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof
Technical Field
The invention belongs to the technical field of chemistry and materials, and particularly relates to a high-sensitivity rare earth doped time-resolved fluorescent nanoparticle, preparation and application thereof, and in particular relates to a TTA/Phen sensitized rare earth doped time-resolved fluorescent nanoparticle, a preparation method and application thereof.
Background
The time-resolved fluorescence immunoassay (TRFIA) relies on the unique fluorescence properties of long fluorescence life of rare earth elements, large Stokes shift, sharp emission spectrum signal peak and the like, and is widely applied to various fields of immunodiagnosis, food detection, environmental monitoring and the like by combining an immunoassay method. However, the conventional fluorescent probe using rare earth chelate as TRFIA has the disadvantages of weak light stability, low fluorescence quantum yield, etc., and thus results in low sensitivity of the detection result. The rare earth doped nanocrystals developed in recent years have excellent fluorescence performance and long fluorescence lifetime, but have no allowable absorption in an ultraviolet region, and are limited in practical application. Therefore, it is very important to explore and develop a rare earth doped time-resolved fluorescent nanoparticle which is highly sensitive and can absorb ultraviolet light.
In the invention patent with publication number CN 108134101A, LaCl is used 3 ·7H 2 O、EuCl 3 ·6H 2 O and NH 4 F is used as a raw material, and Eu is prepared through simple reaction 3+ Doped LaF 3 Fluorescent nanoparticles; 2-thenoyltrifluoroacetone (TTA) and 1, 10-phenanthroline (Phen) are used as sensitizing agents to realize sensitized luminescence of the rare earth doped fluorescent nanoparticles. However, the rare earth ion doped fluoride prepared by the method has certain limitations, the fluorescence quantum yield is low, and the fluoride does not have allowable absorption in an ultraviolet region, so that the fluoride has limitations in practical application.
Disclosure of Invention
Aiming at time division in the prior artThe invention provides a rare earth doped time-resolved fluorescent nanoparticle with high sensitivity and capability of absorbing ultraviolet light, and a preparation method and application thereof. The fluorescent nanoparticles are prepared from LaF 3 As matrix material for the hexagonal phase, Eu 3+ LaF doped to hexagonal phase as luminescent center 3 In the matrix, sensitizing agents of 2-thenoyltrifluoroacetone and 1, 10-phenanthroline and Eu 3+ Coordination occurs for sensitizing Eu 3+ And (4) emitting light.
According to the first aspect of the invention, the invention provides a sensitized rare earth doped time-resolved fluorescent nanoparticle which is prepared from LaF 3 As hexagonal phase matrix material, Eu 3+ Doping as luminescent centers to hexagonal phase LaF 3 In the matrix, sensitizing agents of 2-thenoyltrifluoroacetone and 1, 10-phenanthroline and Eu 3+ Coordination occurs for sensitizing Eu 3+ And (4) emitting light.
Preferably, the Eu 3+ The amount of the rare earth element-containing substance is 40 to 60 percent of the total amount of the rare earth element-containing substances.
Preferably, the particle size of the fluorescent nanoparticles is 30nm-50 nm.
According to another aspect of the invention, a preparation method of any one of the sensitized rare earth doped time-resolved fluorescent nanoparticles is provided, which comprises the following steps:
(1) reacting NH 4 Dropwise addition of aqueous solution of F to LaCl 3 And EuCl 3 To obtain Eu in the mixed aqueous solution of 3+ Doped LaF 3 Fluorescent nanoparticles;
(2) respectively adding an alcoholic solution of 2-thenoyltrifluoroacetone and an alcoholic solution of 1, 10-phenanthroline into the Eu obtained in the step (1) 3+ Doped LaF 3 In the fluorescent nano particles, 2-thenoyltrifluoroacetone, 1, 10-phenanthroline and Eu are reacted 3+ Coordination occurs to sensitize Eu 3+ And (3) emitting light, and centrifugally washing to obtain the sensitized rare earth doped time-resolved fluorescent nanoparticles.
Preferably, the LaCl 3 And EuCl 3 The ratio of the amounts of substances is 6: (4-9).
Preferably, the amount of the 2-thenoyltrifluoroacetone species is in a ratio to LaCl 3 With EuCl 3 The sum of the amounts of substances is (0.1-0.3): 1; the amount of the 1, 10-phenanthroline substances is LaCl 3 With EuCl 3 The sum of the amounts of substances is (0.1-0.3): 1.
preferably, the NH 4 The amount of F material is at least LaCl 3 With EuCl 3 30% of the sum of the amounts of the substances.
Preferably, the centrifugal rotating speed is at least 12000rpm, and the centrifugal time is at least 10 min.
Preferably, NH in step (1) 4 The dropping speed of the aqueous solution F is less than or equal to 1 mL/min.
According to another aspect of the invention, the application of any one of the sensitized rare earth doped time-resolved fluorescent nanoparticles in preparing the immune test paper for determining the new crown neutralizing antibody is provided, and the sensitized rare earth doped time-resolved fluorescent nanoparticles are used as a marker in time-resolved fluorescence immunoassay so as to detect the new crown neutralizing antibody.
Generally, compared with the prior art, the above technical solution conceived by the present invention mainly has the following technical advantages:
(1) the invention adopts TTA as a sensitizing agent, Phen as a non-aqueous auxiliary ligand, two ligands and Eu 3+ The ions generate strong coordination, so that the fluorescence of the nanoparticles is effectively enhanced. Wherein TTA is a sensitizer having a beta-diketone function, which upon absorption of UV light gives rise to a pi-pi' transition of electrons from the ground state S 0 Transition to excited state S 1 Or S 2 Then transits to the triplet state T again in a non-radiative transition manner 1 Finally, coupled to Eu by vibration of bond 3+ The energy level of the vibration state of the Eu is transferred to realize the Eu 3+ The sensitized light emitting of (2); phen is mainly used as a non-aqueous auxiliary ligand to replace the position of-OH, water or other molecules containing high-frequency vibration energy are easily coordinated with central ions only containing beta-diketone complexes to quench, and the photo-quenching factor can be eliminated by introducing the Phen ligand and filling the residual coordination sites, so that the sensitization effect is improved.The prepared TTA/Phen sensitized rare earth doped time-resolved fluorescent nanoparticles show excellent photophysical properties in a liquid phase system, and are suitable for the fields of analysis and detection, biomedicine, material science and the like.
(2) The method of the invention adjusts La 3+ And Eu 3+ The rare earth doped fluorescent nanoparticles with the fluorescence life of 0.63-4.14ms are obtained according to the proportion, the interference of background fluorescence is greatly reduced by utilizing the long fluorescence life and the time-resolved detection technology, and the high-sensitivity detection of the new crown neutralizing antibody is realized.
(3) According to the method, the ratio of TTA, Phen and the rare earth elements is adjusted, so that the fluorescence intensity of the rare earth doped fluorescent nanoparticles is improved by 1-5 times, the average particle size is 30-50 nm, the particle size distribution is uniform, and the fluorescence performance is excellent.
(4) In the prior art, a sensitizer is directly chelated on the surface of rare earth luminescence central ions to synthesize a rare earth chelate material, and the material has weak light stability and low fluorescence performance. The invention is based on rare earth doped fluorescent nanoparticle material for the first time, ligands TTA and Phen are connected on the surface of the rare earth doped fluorescent nanoparticle material and used as a sensitizer to sensitize a luminescence center Eu 3+ The material has excellent fluorescence performance and good stability, and is suitable for the field of time-resolved fluorescence immunoassay.
(5) The method directly prepares the TTA/Phen sensitized rare earth doped time-resolved fluorescent nanoparticles in the aqueous solution, and has the advantages of simple preparation process, easily obtained raw materials, environment-friendly used reagent, mild reaction conditions and strong controllability.
Drawings
FIG. 1 is a transmission electron microscope photograph of TTA/Phen sensitized rare earth doped fluorescent nanoparticles prepared by the invention.
FIG. 2 is X-ray diffraction pattern of TTA/Phen sensitized rare earth doped fluorescent nanoparticle prepared by the invention.
FIG. 3 is TTA/Phen sensitized rare earth doped fluorescent nanoparticle infrared spectrum prepared by the invention.
FIG. 4 is a TTA/Phen sensitized rare earth doped fluorescent nanoparticle fluorescence excitation spectrum prepared by the invention.
FIG. 5 is the fluorescence emission spectrum of TTA/Phen sensitized rare earth doped fluorescent nanoparticles prepared by the invention.
FIG. 6 is a fluorescent lifetime spectrum of TTA/Phen sensitized rare earth doped fluorescent nanoparticles prepared by the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
In order to realize the sensitization of the rare earth doped time-resolved fluorescence nanoparticles, the invention adopts rare earth europium ions (Eu) 3+ ) As a luminescence center, 2-thenoyltrifluoroacetone (TTA) as a sensitizer, and 1, 10-phenanthroline (Phen) as a non-aqueous ancillary ligand. TTA has high ultraviolet absorption coefficient, strong coordination ability with rare earth ions, and Eu as luminescence center 3+ The ions can be effectively transferred; phen can replace a fluorescence quenching group-OH to improve the sensitization effect. The prepared TTA/Phen sensitized rare earth doped fluorescent nanoparticle has stable fluorescence property, high fluorescence intensity and long fluorescence life. The method specifically comprises the following steps:
(1) preparation of LaCl 3 ·7H 2 O and EuCl 3 ·6H 2 A mixed aqueous solution of O;
(2) preparation of NH 4 Dropwise adding the aqueous solution F into the rare earth hydrochloride solution under stirring;
(3) preparing ethanol solution of 2-thenoyltrifluoroacetone (TTA) and ethanol solution of 1, 10-phenanthroline (Phen), and dropwise adding into NH under stirring 4 F and rare earth hydrochloride mixed solution;
(4) stirring at room temperature overnight, obtaining a white milky suspension of the reaction system, and centrifugally washing to obtain TTA/Phen sensitized rare earth doped fluorescent nanoparticles.
LaCl in the invention 3 ·7H 2 O and EuCl 3 ·6H 2 The molar ratio of O can be adjusted at will, the fluorescence quantum yield of the products is different when the molar ratio is different, and when Eu is used 3+ When the doping degree of the ions is 40%, the fluorescence quantum yield of the prepared fluorescent nanoparticles is highest.
Wherein, NH in the step (2) 4 Slowly dripping the aqueous solution F at constant speed, and adding NH by using a peristaltic pump 4 The solution F is pumped into the rare earth hydrochloride solution at a constant speed of 1 mL/min.
And (4) repeatedly washing for 4 times at the centrifugal rotation speed of 12000rpm for 10min in the step (4) to obtain the TTA/Phen sensitized rare earth doped fluorescent nanoparticles.
Example 1 TTA/Phen sensitized rare earth doped fluorescent nanoparticles
(1) 29.71mg (0.8mmol) of LaCl are weighed out 3 ·7H 2 O and 7.33mg (0.2mmol) of EuCl 3 ·6H 2 Dissolving O in 25mL of ultrapure water, and uniformly stirring;
(2) 11.11mg (3mmol) NH were weighed out 4 Dissolving the mixture in 6mL of ultrapure water, dropwise adding the mixture into a rare earth hydrochloride solution while stirring, controlling the rotating speed of a peristaltic pump, adjusting the dropwise adding speed to be 1mL/min, and gradually changing the color of a reaction solution from original colorless transparency to slightly light blue opalescence in the dropwise adding process;
(3) weighing 22.22mg (0.1mmol) of 2-thenoyltrifluoroacetone (TTA) and 19.82mg (0.1mmol) of 1, 10-phenanthroline (Phen), dissolving in 1mL of ethanol solution, and sequentially and slowly dripping into NH 4 F. In a mixed solution of rare earth hydrochloride;
(4) stirring and reacting overnight at room temperature, taking out the reaction solution, centrifuging at 12000rpm for 10min, repeatedly washing for 4 times, ultrasonically dispersing the precipitate with 5mL of ultrapure water to obtain TTA/Phen sensitized rare earth doped fluorescent nanoparticles, and storing in a refrigerator at 4 ℃ for later use.
Fig. 1 is a transmission electron microscope image of the rare earth-doped fluorescent nanoparticle prepared in example 1, wherein the nanoparticle has a hexagonal structure, an average particle diameter of 30nm to 50nm, and good dispersibility. FIG. 2 is an X-ray diffraction pattern of the rare earth-doped fluorescent nanoparticles prepared in example 1, wherein the diffraction peak position and relative intensity of the sample are equal to those of LaF 3 The standard card PDF #32-0483 matches well,no impurity peak appears, and the crystallinity is better, and is a hexagonal crystal phase.
Example 2 TTA/Phen sensitized rare earth doped fluorescent nanoparticles
(1) 22.28mg (0.6mmol) of LaCl are weighed out 3 ·7H 2 O and 14.66mg (0.4mmol) EuCl 3 ·6H 2 Dissolving O in 25mL of ultrapure water, and uniformly stirring;
(2) 11.11mg (3mmol) NH were weighed out 4 Dissolving the mixture in 6mL of ultrapure water, dropwise adding the mixture into a rare earth hydrochloride solution while stirring, controlling the rotating speed of a peristaltic pump, adjusting the dropwise adding speed to be 1mL/min, and gradually changing the color of a reaction solution from original colorless transparency to slightly light blue opalescence in the dropwise adding process;
(3) weighing 22.22mg (0.1mmol) of 2-thenoyltrifluoroacetone (TTA) and 19.82mg (0.1mmol) of 1, 10-phenanthroline (Phen), dissolving in 1mL of ethanol solution, and sequentially and slowly dripping into NH 4 F. In a mixed solution of rare earth hydrochloride;
(4) stirring and reacting overnight at room temperature, taking out the reaction solution, centrifuging at 12000rpm for 10min, repeatedly washing for 4 times, ultrasonically dispersing the precipitate with 5mL of ultrapure water to obtain TTA/Phen sensitized rare earth doped fluorescent nanoparticles, and storing in a refrigerator at 4 ℃ for later use.
FIG. 3 is an infrared absorption spectrum of the rare earth-doped fluorescent nanoparticle prepared in example 2, and several characteristic peaks appearing in the spectrum indicate that TTA and Phen are successfully adsorbed on the surface of the nanoparticle. Fig. 4 is an excitation spectrum of the rare earth-doped fluorescent nanoparticle prepared in example 2, which has a relatively wide excitation peak in an ultraviolet region, and an optimal excitation wavelength is 370 nm. FIG. 5 shows the emission spectrum of the rare earth-doped fluorescent nanoparticles prepared in example 2, in which TTA and Phen absorb ultraviolet light and transfer energy to the luminescence center Eu 3+ The fluorescence intensity of the rare earth doped fluorescent nanoparticles is effectively enhanced.
Example 3 TTA/Phen sensitized rare earth doped fluorescent nanoparticles
(1) 14.85mg (0.4mmol) of LaCl are weighed out 3 ·7H 2 O and 21.98mg (0.6mmol) of EuCl 3 ·6H 2 Dissolving O in 25mL of ultrapure water, and uniformly stirring;
(2) 11.11mg (3mmol) NH were weighed out 4 F, dissolved in 6mAdding the L ultrapure water into a rare earth hydrochloride solution dropwise under stirring, controlling the rotating speed of a peristaltic pump, adjusting the adding speed to be 1mL/min, and gradually changing the color of a reaction solution from original colorless transparency to slightly light blue opalescence in the adding process;
(3) weighing 22.22mg (0.1mmol) of 2-thenoyltrifluoroacetone (TTA) and 19.82mg (0.1mmol) of 1, 10-phenanthroline (Phen), dissolving in 1mL of ethanol solution, and sequentially and slowly dripping into NH 4 F. In a mixed solution of rare earth hydrochloride;
(4) stirring and reacting overnight at room temperature, taking out the reaction solution, centrifuging at 12000rpm for 10min, repeatedly washing for 4 times, ultrasonically dispersing the precipitate with 5mL of ultrapure water to obtain TTA/Phen sensitized rare earth doped fluorescent nanoparticles, and storing in a refrigerator at 4 ℃ for later use.
Fig. 6 is a fluorescence lifetime spectrum of the rare earth-doped fluorescent nanoparticle prepared in example 3, wherein the fluorescence lifetime can reach millisecond level, is significantly superior to other common biological fluorescent materials, and is suitable for a time-resolved fluorescence immunoassay technology.
Example 4 TTA/Phen sensitized rare earth doped fluorescent nanoparticles
(1) 7.43mg (0.2mmol) of LaCl are weighed out 3 ·7H 2 O and 29.31mg (0.8mmol) of EuCl 3 ·6H 2 Dissolving O in 25mL of ultrapure water, and uniformly stirring;
(2) 11.11mg (3mmol) NH were weighed out 4 Dissolving the mixture in 6mL of ultrapure water, dropwise adding the mixture into a rare earth hydrochloride solution while stirring, controlling the rotating speed of a peristaltic pump, adjusting the dropwise adding speed to be 1mL/min, and gradually changing the color of a reaction solution from original colorless transparency to slightly light blue opalescence in the dropwise adding process;
(3) weighing 22.22mg (0.1mmol) of 2-thenoyltrifluoroacetone (TTA) and 19.82mg (0.1mmol) of 1, 10-phenanthroline (Phen), dissolving in 1mL of ethanol solution, and sequentially and slowly dripping into NH 4 F. In a mixed solution of rare earth hydrochloride;
(4) stirring and reacting overnight at room temperature, taking out the reaction solution, centrifuging at 12000rpm for 10min, repeatedly washing for 4 times, ultrasonically dispersing the precipitate with 5mL of ultrapure water to obtain TTA/Phen sensitized rare earth doped fluorescent nanoparticles, and storing in a refrigerator at 4 ℃ for later use.
Luminescence center Eu 3+ The excessive doping of (1) causes an internal concentration quenching effect to some extent, lowering the luminescence intensity, but on the other hand, the excessive Eu 3+ More TTA and Phen can be adsorbed, so that the energy transfer efficiency is improved.
Example 5 TTA/Phen sensitized rare earth doped fluorescent nanoparticles
(1) 22.28mg (0.6mmol) of LaCl are weighed out 3 ·7H 2 O and 14.66mg (0.4mmol) EuCl 3 ·6H 2 Dissolving O in 25mL of ultrapure water, and uniformly stirring;
(2) 11.11mg (3mmol) NH were weighed out 4 Dissolving the mixture in 6mL of ultrapure water, dropwise adding the mixture into a rare earth hydrochloride solution while stirring, controlling the rotating speed of a peristaltic pump, adjusting the dropwise adding speed to be 1mL/min, and gradually changing the color of a reaction solution from original colorless transparency to slightly light blue opalescence in the dropwise adding process;
(3) weighing 44.44mg (0.2mmol) of 2-thenoyltrifluoroacetone (TTA) and 39.64mg (0.2mmol) of 1, 10-phenanthroline (Phen), dissolving in 1mL of ethanol solution, and sequentially and slowly dripping into NH 4 F. In a mixed solution of rare earth hydrochloride;
(4) stirring and reacting overnight at room temperature, taking out the reaction solution, centrifuging at 12000rpm for 10min, repeatedly washing for 4 times, ultrasonically dispersing the precipitate with 5mL of ultrapure water to obtain TTA/Phen sensitized rare earth doped fluorescent nanoparticles, and storing in a refrigerator at 4 ℃ for later use.
The dosage of TTA and Phen is increased properly, so that the fluorescence intensity of the rare earth doped fluorescent nanoparticles is improved by about 3 times.
Example 6 TTA/Phen sensitized rare earth doped fluorescent nanoparticles
(1) 22.28mg (0.6mmol) of LaCl are weighed out 3 ·7H 2 O and 14.66mg (0.4mmol) EuCl 3 ·6H 2 Dissolving O in 25mL of ultrapure water, and uniformly stirring;
(2) 11.11mg (3mmol) NH were weighed out 4 Dissolving the mixture in 6mL of ultrapure water, dropwise adding the mixture into the rare earth hydrochloride solution while stirring, controlling the rotating speed of a peristaltic pump, adjusting the dropwise adding speed to be 1mL/min, and gradually changing the color of the reaction solution from original colorless transparency in the dropwise adding processA slight bluish opalescence;
(3) weighing 66.66mg (0.3mmol) of 2-thenoyltrifluoroacetone (TTA) and 59.46mg (0.3mmol) of 1, 10-phenanthroline (Phen), dissolving in 1mL of ethanol solution, and sequentially and slowly dripping into NH 4 F. In a mixed solution of rare earth hydrochloride;
(4) stirring and reacting overnight at room temperature, taking out the reaction solution, centrifuging at 12000rpm for 10min, repeatedly washing for 4 times, ultrasonically dispersing the precipitate with 5mL of ultrapure water to obtain TTA/Phen sensitized rare earth doped fluorescent nanoparticles, and storing in a refrigerator at 4 ℃ for later use.
And the excessive addition of TTA and Phen improves the fluorescence intensity of the rare earth doped fluorescent nanoparticles by about 1 time.
Example 7 immunological application of rare earth-doped fluorescent nanoparticles
(1) Respectively dispersing the TTA/Phen sensitized rare earth doped fluorescent nanoparticles prepared in the embodiments 1-6 in ultrapure water with a certain volume, adding 5mL of polyacrylic acid (PAA) aqueous solution, reacting for 5h at room temperature, taking out the reaction solution, centrifuging at 12000rpm for 10min, repeatedly washing for 4 times, ultrasonically dispersing the precipitate with 5mL of ultrapure water to obtain PAA modified rare earth doped fluorescent nanoparticles, and storing in a refrigerator at 4 ℃ for later use.
(2) And (2) taking a proper amount of the PAA modified rare earth doped fluorescent nanoparticles prepared in the step (1), activating by adopting NHS solution and EDC solution with the concentration of 10mg/mL, adding a certain amount of RBD antigen, placing on a blood mixing instrument for reacting at room temperature for 2h, and sealing by adopting 10% BSA solution for 1 h. And (4) after sealing, centrifuging at 14000rpm for 15min to remove supernatant, redissolving the precipitate by using redissolution, fixing the volume to 250 mu L, ultrasonically mixing uniformly, and storing in a refrigerator at 4 ℃ for later use.
(3) And (3) spraying the fluorescent nanoparticle immune complex prepared in the step (2) on a combination pad, and assembling the fluorescent nanoparticle immune complex, a PVC bottom plate, a sample pad, an NC membrane and a water absorption pad into an immunochromatography test strip for measuring a new crown neutralizing antibody in a serum sample.
The rare earth doped fluorescent nanoparticles are used as markers, the characteristics of long fluorescence life of the fluorescent nanoparticles are utilized, the time-resolved fluorescence immunoassay technology is adopted, the new crown neutralizing antibody in a serum sample is detected, and the detection result has high sensitivity and strong specificity.
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.

Claims (7)

1. A preparation method of sensitized rare earth doped time-resolved fluorescence nanoparticles is characterized by comprising the following steps:
(1) reacting NH 4 Dropwise addition of aqueous solution of F to LaCl 3 And EuCl 3 To obtain Eu in the mixed aqueous solution of (1) 3+ Doped LaF 3 Fluorescent nanoparticles;
(2) respectively adding an alcoholic solution of 2-thenoyltrifluoroacetone and an alcoholic solution of 1, 10-phenanthroline into the Eu obtained in the step (1) 3+ Doped LaF 3 In the fluorescent nano particles, 2-thenoyltrifluoroacetone, 1, 10-phenanthroline and Eu are reacted 3+ Coordination occurs to sensitize Eu 3+ Luminescence, centrifugal washing to obtain sensitized rare earth doped time-resolved fluorescent nanoparticles;
the LaCl 3 And EuCl 3 The ratio of the amounts of substances is 6: (4-9);
the amount of the 2-thenoyltrifluoroacetone substance is in a ratio of LaCl 3 With EuCl 3 The sum of the amounts of substances is (0.1-0.3): 1; the amount of the 1, 10-phenanthroline substances is LaCl 3 With EuCl 3 The sum of the amounts of substances is (0.1-0.3): 1;
NH in step (1) 4 The dropping rate of the F aqueous solution is less than or equal to 1 mL/min.
2. The method for preparing the sensitized rare earth doped time-resolved fluorescent nanoparticle according to claim 1, wherein the NH is 4 The amount of F material is at least LaCl 3 With EuCl 3 30% of the sum of the amounts of the substances.
3. The method for preparing the sensitized rare earth doped time-resolved fluorescent nanoparticle according to claim 1, wherein the centrifugation rotation speed is at least 12000rpm, and the centrifugation time is at least 10 min.
4. A sensitized rare earth doped time-resolved fluorescent nanoparticle prepared by the method of any one of claims 1 to 3, wherein the fluorescent nanoparticle is prepared by LaF 3 As hexagonal phase matrix material, Eu 3+ Doping as luminescent centers to hexagonal phase LaF 3 In the matrix, sensitizing agents of 2-thenoyltrifluoroacetone and 1, 10-phenanthroline and Eu 3+ Coordination occurs for sensitizing Eu 3+ And (4) emitting light.
5. The sensitized rare earth doped time-resolved fluorescent nanoparticle according to claim 4, wherein said Eu is 3+ The amount of the rare earth element-containing substance is 40 to 60 percent of the total amount of the rare earth element-containing substances.
6. The sensitized rare earth doped time-resolved fluorescent nanoparticle according to claim 4, wherein the fluorescent nanoparticle has a particle size of 30nm to 50 nm.
7. The use of the sensitized rare earth doped time-resolved fluorescent nanoparticle according to any one of claims 4 to 6 for preparing an immunoassay paper for detecting a new corona neutralizing antibody, wherein the sensitized rare earth doped time-resolved fluorescent nanoparticle is used as a marker for time-resolved fluorescence immunoassay to detect a new corona neutralizing antibody.
CN202110604412.6A 2021-05-31 2021-05-31 High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof Active CN113249127B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110604412.6A CN113249127B (en) 2021-05-31 2021-05-31 High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110604412.6A CN113249127B (en) 2021-05-31 2021-05-31 High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof

Publications (2)

Publication Number Publication Date
CN113249127A CN113249127A (en) 2021-08-13
CN113249127B true CN113249127B (en) 2022-08-16

Family

ID=77185813

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110604412.6A Active CN113249127B (en) 2021-05-31 2021-05-31 High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof

Country Status (1)

Country Link
CN (1) CN113249127B (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101362944A (en) * 2008-09-25 2009-02-11 湖南大学 Method for preparing long-term durability luminous nano granule of core-shell rare-earth complexes
CN112175620A (en) * 2020-09-30 2021-01-05 厦门稀土材料研究所 COVID-19IgG/IgM detection kit, detection card, rare earth nano probe and preparation method
CN112342014A (en) * 2020-10-28 2021-02-09 安徽为臻生物工程技术有限公司 Preparation method of monodisperse polymer fluorescent microspheres

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101362944A (en) * 2008-09-25 2009-02-11 湖南大学 Method for preparing long-term durability luminous nano granule of core-shell rare-earth complexes
CN112175620A (en) * 2020-09-30 2021-01-05 厦门稀土材料研究所 COVID-19IgG/IgM detection kit, detection card, rare earth nano probe and preparation method
CN112342014A (en) * 2020-10-28 2021-02-09 安徽为臻生物工程技术有限公司 Preparation method of monodisperse polymer fluorescent microspheres

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Broadband UV Excited LaF3:Eu3+ Based Inorganic–Organic Mixed Hybrid Nanoparticles for Strong Luminescence;Xiaojing Wang et al.;《Journal of Nanoscience and Nanotechnology》;20181231;第18卷;第1876-1881页 *
Sensitized luminescence from water-soluble LaF3:Eu nanocrystals via partially-capped 1,10-phenanthroline: time-gated emission and multiple lifetimes;Mir Irfanullah et al.;《Dalton Transactions》;20160629;第45卷;第12483-12495页 *

Also Published As

Publication number Publication date
CN113249127A (en) 2021-08-13

Similar Documents

Publication Publication Date Title
Huang et al. Rare earth ion–and transition metal ion–doped inorganic luminescent nanocrystals: from fundamentals to biodetection
Li et al. Classification, synthesis, and application of luminescent silica nanoparticles: a review
Yang et al. Nanometer fluorescent hybrid silica particle as ultrasensitive and photostable biological labels
CN102134253B (en) Photoluminescent nano particle as well as preparation method and application thereof
CN101486903B (en) Preparation of rare earth luminous nanoparticle based on pyridine dicarboxylic acid
CN112898965B (en) Preparation and use method of ratiometric fluorescent probe for visual differential detection
CN107356570B (en) Solid-state up-conversion fluorescent probe and preparation method and application thereof
CN110776916A (en) Quantum dot dual-emission-ratio fluorescent probe and preparation method and application thereof
CN110609133B (en) Fluorescence ratio type spectral analysis method for detecting carcinoembryonic antigen and application thereof
WO2021109057A1 (en) Long-afterglow luminescent organic microspheres, and method for preparation thereof and application thereof
Wang et al. Preparation of ethanediamine-doped carbon quantum dots and their applications in white LEDs and fluorescent TLC plate
CN102504808A (en) Preparation method of rare-earth fluorescent silica nano particle
CN101957319B (en) Chemical preparation method of CaMoO4: Tb3+fluorescent probe for detecting trace amount of TNT (Tri-Nitro-Toluene)
Nabika et al. Surface-enhanced luminescence from Eu 3+ complex nearby Ag colloids
Wu et al. ZIF-8 encapsulated upconversion nanoprobes to evaluate pH variations in food spoilage
JP2012032263A (en) Reagent for measuring immune containing fluorescent fine particle
Liu et al. H2O2‐Induced persistent luminescence signal enhancement applied to biosensing
CN113249127B (en) High-sensitivity rare earth doped time-resolved fluorescent nanoparticle and preparation and application thereof
CN1265197C (en) Beta-diketone-trivalent europium complex nano fluorescent probe, its preparation and use thereof
CN102172497B (en) Preparation method of fluorescent coding microspheres based on up-conversion luminous nanocrystalline
CN105754600B (en) A kind of core-shell structure copolymer-shell structural nano fluorescence probe of europium ion activation and its preparation method and application
CN110244044B (en) Rare earth dyed magnetic bead and preparation and application thereof
CN109321245A (en) A kind of rare earth codope functional nanocomposite and preparation method thereof
CN114181695B (en) Preparation method of composite quantum dot for detecting concentration of hydrogen peroxide and glucose
CN111044493B (en) Near infrared excitation and emission-based lateral flow test strip for luminescent detection of serum marker and preparation and use methods thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant