Multi-wavelength excited fluorescent nano-plastic as well as synthesis method and application thereof

Abstract

The invention relates to the technical field of nano-plastics, and discloses a multi-wavelength excited fluorescent nano-plastic and a synthetic method and application thereof.The synthetic method comprises the steps that an emulsion polymerization method is adopted, styrene serves as a monomer, deionized water serves as a continuous phase, an emulsifier and an initiator are added, a polymerization reaction is conducted, and polystyrene nano-particles are synthesized; the preparation method further comprises the following steps: mixing styrene with other components, and then adding Nile red; and carrying out europium ion doping on the synthesized polystyrene nanoparticles. According to the invention, through dual effects of Nile red fluorescence labeling and europium doping, the imaging signal intensity of the polystyrene nanoparticles in a living body is obviously improved; the europium ions doped by the acetone swelling method enable the migration path and the accumulation process of the nanoparticles in a living body to be more accurately and quantitatively analyzed.

Description

Technical Field

[0001] The present invention relates to the technical field of nano plastics, and specifically relates to a fluorescent nano plastic excited by multiple wavelengths, a synthesis method thereof, and an application thereof. Background Art

[0002] Nano plastics are tiny in size, chemically stable, and have a large specific surface area. They can enter organisms through various routes (such as ingestion, inhalation, or skin contact). After entering the body, nano plastics can penetrate biological barriers and accumulate in tissues or organelles, thereby inducing oxidative stress, inflammatory responses, or genotoxicity, posing a potential threat to the health of organisms. This not only makes the migration and diffusion behavior of nano plastics in the environment difficult to predict, but also interferes with the metabolism and functions of organisms. Therefore, it is very necessary to develop precise and sensitive technical means for tracking the migration path, distribution pattern, and cumulative effect of nano plastics in organisms.

[0003] However, for existing nano plastic labeling and tracking methods, there are generally problems such as weak imaging signals and difficulty in quantitative analysis, which greatly limit the in-depth study of the behavior of nano plastics in the organism environment. Summary of the Invention

[0004] (I) Technical Problems to be Solved Aiming at the deficiencies of the prior art, the present invention provides a fluorescent nano plastic excited by multiple wavelengths, a synthesis method thereof, and an application thereof, which have the advantages of being able to effectively enhance the imaging signal intensity and improve the quantitative analysis accuracy.

[0005] (II) Technical Solutions To achieve the above purpose of effectively enhancing the imaging signal intensity and improving the quantitative analysis accuracy, the present invention provides the following technical solutions: A synthesis method of a fluorescent nano plastic excited by multiple wavelengths, which uses the emulsion polymerization method, uses styrene as a monomer, deionized water as a continuous phase, adds an emulsifier and an initiator, and performs a polymerization reaction to synthesize polystyrene nanoparticles, further including: Adding Nile Red after mixing styrene with other components; Doping the synthesized polystyrene nanoparticles with europium ions.

[0006] As a preferred technical solution of the present invention, the addition amount of Nile Red is 0.1-0.3% of the mass of styrene.

[0007] As a preferred technical solution of the present invention, the styrene concentration is 0.2-4.0 wt.%.

[0008] As a preferred technical solution of the present invention, the dosage of the initiator is 0.04 wt.%.

[0009] As a preferred technical solution of the present invention, the initiator is KPS or AIBA.

[0010] As a preferred technical solution of the present invention, the synthesized polystyrene nanoparticles are doped with europium ions by the acetone swelling method.

[0011] As a preferred technical solution of the present invention, the europium ion doping specifically includes the following steps: A1. Suspend the synthesized Nile red-labeled polystyrene nanoparticles in an acetone solution to swell the particle structure; A2. Add a rare earth metal europium solution to the swollen nanoparticle system, and make the europium ions uniformly distributed inside the particles through electrostatic interaction or pore adsorption; A3. Gradually evaporate the acetone solvent to restore the integrity of the nanoparticle structure and seal the europium ions.

[0012] (III) Beneficial effects Compared with the prior art, the present invention provides a fluorescent nanoplastic with multi-wavelength excitation, its synthesis method and application, and has the following beneficial effects: 1. Through the dual effects of Nile red fluorescence labeling and europium doping, the signal intensity of polystyrene nanoparticles in vivo imaging is significantly improved.

[0013] 2. The europium ions doped by the acetone swelling method enable the migration path and accumulation process of the nanoparticles in vivo to be more accurately quantitatively analyzed.

[0014] 3. It can be used for imaging and quantitative analysis of nanoparticles in various organisms, and is particularly suitable for the behavior research of environmental pollutants. Description of the drawings

[0015] Figure 1 It is the TEM characterization of 50 nm PS (left) and 300 nm PS (right); Figure 2 It is the Zeta potential of PS with different sizes and surface modifications; Figure 3 It is the PS-Eu particle image and elemental composition; Figure 4 It is the fluorescence emission spectrum of Eu-PS / Nile Red; Figure 5 It is the tracing of fluorescent polystyrene nanoparticles applied to nanoplastics in zebrafish. Detailed implementation manners

[0016] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0017] Embodiment 1: Please refer to Figures 1 - 5 , a synthesis method of fluorescent nanoplastics excited by multiple wavelengths, which adopts the emulsion polymerization method. Using styrene as the monomer and deionized water as the continuous phase, an emulsifier and an initiator are added to carry out the polymerization reaction to synthesize polystyrene nanoparticles. During the synthesis process, Nile red is added after the styrene is mixed with other components, and the addition amount is 0.1-0.3% of the mass of styrene to ensure that the nanoparticles have the function of fluorescent labeling. The concentration of Nile red within this range is sufficient to be evenly distributed in the particles, avoiding the too weak signal caused by insufficient fluorescent molecules, and also being able to avoid the non-radiative energy transfer (fluorescence quenching) between fluorescent molecules and the reduction of fluorescent signals caused by too high a concentration of Nile red.

[0018] The particle size and charge are regulated by adjusting the amount of styrene and the type of polymerization initiator. The concentration of styrene is 0.2 - 4.0 wt.%, and the types of initiators include KPS (potassium sulfate) and AIBA (2,2'-diaminohexanoic acid dihydrochloride), and the dosage of the initiator is 0.04 wt.%.

[0019] By changing the addition amount of styrene, the particle size of the nanoplastics can be adjusted within the range of 50nm - 300nm; Through TEM, the morphology of the prepared 50nm and 300nm polystyrene nanoparticles was analyzed. As Figure 1 shown, the 50nm PS particles are uniformly spherical and compactly distributed; the 300nm PS particles have a larger particle size and still maintain a high monodispersity, and no obvious aggregation phenomenon is observed, which indicates that the emulsion polymerization method can effectively control the particle size.

[0020] As Figure 2 shown, the Zeta potentials of the 50nm and 300nm positively charged particles are about +25mV and +35mV respectively, which indicates that the positively charged particles have good stability in the aqueous phase; the Zeta potentials of the 50nm and 300nm negatively charged particles are both about -35mV, which shows that the negatively charged particles have a strong electrical repulsion in the aqueous phase, thus maintaining good dispersibility.

[0021] In this embodiment, the emulsifier can be selected from sodium dodecyl sulfate (SDS) or polyvinyl alcohol (PVA).

[0022] Afterwards, the synthesized polystyrene nanoparticles were doped with europium ions by the acetone swelling method. The acetone swelling method can not only ensure the uniform distribution of europium ions, but also effectively maintain the morphology and dispersibility of the particles. At the same time, the fluorescence intensity of the particles doped with europium ions at the excitation wavelength of 450 nm was significantly enhanced, which was complementary to the fluorescence of Nile red at 610 nm, thus realizing multi-wavelength excitation and improving the imaging sensitivity and quantitative analysis accuracy.

[0023] The europium ion doping specifically includes the following steps: A1. Suspend the synthesized Nile red-labeled polystyrene nanoparticles in an acetone solution to swell the particle structure; A2. Add a rare earth metal europium solution to the swollen nanoparticle system, and through electrostatic interaction or pore adsorption, make the europium ions evenly distributed inside the particles; A3. Gradually evaporate the acetone solvent to restore the integrity of the nanoparticle structure and seal the europium ions.

[0024] In this example, the concentration of the acetone solution is 80%-100%, the swelling temperature is 25°C-50°C, and the swelling time is 30 minutes to 2 hours.

[0025] In the present invention, the visible light fluorescence signal of Nile red is used for imaging localization, and the rare earth element signal of europium is used for quantitative analysis. Through the dual effects of Nile red fluorescence labeling and europium doping, the signal intensity of polystyrene nanoparticles in vivo imaging is significantly improved.

[0026] The europium ions doped by the acetone swelling method enable the migration path and accumulation process of the nanoparticles in vivo to be more accurately quantitatively analyzed, and can be used for imaging and quantitative analysis of various nanoparticles in vivo, especially suitable for the behavior research of environmental pollutants.

[0027] Example Two: In this example, a fluorescent nanoplastics with multi-wavelength excitation is provided, which is prepared by using the synthesis method of a fluorescent nanoplastics with multi-wavelength excitation provided in Example One.

[0028] Example Three: This example discloses the application of the fluorescent nanoplastics prepared in Example One in in vivo imaging and migration quantification of organisms, such as zebrafish; Zebrafish embryos are transparent, develop rapidly, and their genomes are also highly homologous to humans. Through the imaging and quantitative analysis of the fluorescent nanoplastics with multi-wavelength excitation in zebrafish ( Figure 5 ), not only can the migration path and quantity of the nanoplastics be accurately traced, but also their accumulation patterns and potential toxic effects in specific organs can be revealed.

[0029] Although embodiments of the present invention have been shown and described, those of ordinary skill in the art will appreciate that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for synthesizing multi-wavelength excited fluorescent nanoplastics, which adopts emulsion polymerization, uses styrene as a monomer, deionized water as a continuous phase, adds an emulsifier and an initiator, performs polymerization reaction, and synthesizes polystyrene nanoparticles, characterized in that: Also includes: Nile Red is added after the styrene is mixed with the other ingredients; The synthesized polystyrene nanoparticles were doped with europium ions.

2. The method for synthesizing a multi-wavelength excited fluorescent nanoplastic according to claim 1, characterized in that: The amount of Nile Red added is 0.1-0.3% of the mass of styrene.

3. The method for synthesizing a multi-wavelength excited fluorescent nanoplastic according to claim 1, characterized in that: The styrene concentration is 0.2-4.0 wt.%.

4. The method for synthesizing a multi-wavelength excited fluorescent nanoplastic according to claim 1, characterized in that: The initiator dosage is 0.04 wt.%.

5. The method for synthesizing a multi-wavelength excited fluorescent nanoplastic according to claim 4, characterized in that: The initiator is KPS or AIBA.

6. The method for synthesizing a multi-wavelength excited fluorescent nanoplastic according to claim 1, characterized in that: The synthesized polystyrene nanoparticles were doped with europium ions using the acetone swelling method.

7. The method for synthesizing a multi-wavelength excited fluorescent nanoplastic according to claim 6, characterized in that: Europium ion doping specifically includes the following steps: A1. Suspending the synthesized Nile red labeled polystyrene nanoparticles in an acetone solution to expand the particle structure; A2. Adding a rare earth metal europium solution into the expanded nanoparticle system, and making the europium ions evenly distributed inside the particles through electrostatic action or pore adsorption; A3. Gradually evaporate the acetone solvent to restore the integrity of the nanoparticle structure and seal the europium ions.

8. A multi-wavelength excited fluorescent nanoplastic, characterized in that: The nanoplastic is prepared by the synthesis method of a multi-wavelength excited fluorescent nanoplastic as described in any one of claims 1 to 7.

9. Use of fluorescent nanoplastic prepared by the method as described in any one of claims 1 to 7 in in vivo imaging and migration quantification.

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