CN101824191A - Macromolecular fluorescent microspheres and preparation method thereof - Google Patents

Abstract

The invention discloses macromolecular fluorescent microspheres and a preparation method thereof. The macromolecular fluorescent microspheres use monodisperse sulfonated polystyrene microspheres as core microspheres, and the surfaces of the macromolecular fluorescent microspheres are coated with a shell layer of a p-phenylene vinylene (PPV) fluorescent conjugated macromolecular polymer. The preparation method of the macromolecular fluorescent microspheres comprises the following steps of: synthesizing a positively charged PPV fluorescent conjugated macromolecular polymer precursor in solution by using a Wessling sulfosalt polymer precursor method; adding the monodisperse sulfonated polystyrene microspheres into the precursor, and then processing the mixture for 0.5 to 2 hours under a vacuum condition that the temperature is between 90 and 150 DEG C; and washing and drying the mixture to obtain PPV conjugated macromolecular fluorescent microspheres. An inhomogeneous reaction for synthesizing the conjugated polymer in the method is simple and controllable, and the stability of the fluorescence of the microspheres is effectively improved.

Description

A kind of high molecular fluorescent microballoons and preparation method thereof

Technical field

The present invention relates to a kind of high molecular fluorescent microballoons and preparation method thereof, specifically is polystyrene fluorescent microsphere of area load polystyrene support (PPV) class conjugated polymer and preparation method thereof, belongs to the fluorescent sensing material preparation field.

Background technology

Fluorescent microsphere be meant diameter at nano level to the micron order scope, load has fluorescent substance, stimulated by outside energy to inspire the solia particle of fluorescence.Fluorescent microsphere more and more receives publicity in the application of biomedical sector in recent years, and this is because itself form stable, specific surface area is big, adsorptivity is strong, agglutination is big, the surface reaction ability by force and stablize and luminous efficiency efficiently.In addition, the various proterties that can give according to the different purposes of microballoon such as biospecific associativity, hollow porousness, magnetic etc., thereby be widely used, especially at immunoassay, high-flux medicaments sifting, pharmaceutical carrier, immobilized enzyme, bacterium and viral diagnosis, single nucleotide polymorphisms genotype, all there is very important use aspects such as cytokine evaluation and cell analysis.Therefore, the development several different methods prepares biological mark microballoon, fluorescence-encoded micro-beads or fluorescence sense microballoon with different fluorescence property features and different size not only good theory significance, also development and national economy is had actual application value.

The preparation method of existing fluorescent microsphere mainly contains: 1, be main raw with the fluorescence quantum, surface coverage certain protection layer or further introduce modification group " contains silica fluorescent microballoon of cadmium telluride fluorescence quantum and preparation method thereof " (CN1782020A) as Chinese invention patent; 2, fluorescence dye is introduced in the polymer or the microballoon of silicon-dioxide, (CN1475805A) as Chinese invention patent " magnetic fluorescent microspheres and preparation method thereof with adopt this magnetic fluorescent microspheres to carry out the method for biomolecule detection "; But because the toxicity of quantum dot and fluorescence dye excites and the unicity of emission wavelength makes them be subjected to certain restriction in the application aspect certain.

In recent years, fluorescence conjugated high molecular research has caused extensive interest, for the small molecules fluorescent substance, one of its advantage can be passed through the main chain or the side group of synthetic different structure, and the adjustment aggregated structure reaches the adjusting to the conjugated polymer fluorescence property; Two of its advantage is that conjugated polymer is otherwise known as " molecular wire ", and external interference can effectively be transmitted on macromolecular main chain the influence of its electron energy state, thereby the sensitivity that causes its fluorescence property to stimulate to external world is much higher than the small molecules chromophoric group.Bibliographical information has been arranged utilized electrostatic adsorption conjugated polymer preparation of electrolyte fluorescent microsphere in microsphere surface absorption, and inquired into its application (Langmuir2007 aspect transmitter, 23,112-115), though this method is fairly simple, but also there are some problems, before absorption, must be dissolved in certain solvent as conjugated polymer well, yet, the conjugated polymer solvability is relatively poor to be well-known, also may not necessarily reach good solubility property even introduce the hydrotropy group at side group; In addition, whether the resolvability of microsphere surface conjugated polymer can bring the conjugated polymer can stable existence, the i.e. problem of the fluorescence property stability of microballoon at microsphere surface again.Also have document to disclose with the microsphere surface functionalization, react with the conjugation high polymer monomer, carrying out cross-linked method at microsphere surface forms the method that the surface has the microballoon of conjugated polymer (Langmuir 2007,23,4541-4548), this method can make conjugated polymer link to each other by covalent linkage and microsphere surface are stable, but belongs to non-homogeneous polymerization at the cross-coupling reaction generation conjugated polymer of microsphere surface, has certain degree of difficulty.Chinese invention patent " copolymerization type cross-linking polymer fluorescent microspheres and preparation method thereof " (CN101177607A) in, employing fluorescent monomer and other multifunctional monomers are disclosed under the initiator effect, free-radical polymerized method prepares polymer fluorescent microspheres, the fluorescent microsphere stability that this method prepares is better, but to the also relative difficulty with controllability of the controllability of its fluorescence property.

Summary of the invention

The object of the invention provides a kind of synthesis technique simple controllable, high molecular fluorescent microballoons that fluorescence property is stable and preparation method thereof.

The technical solution used in the present invention is: a kind of high molecular fluorescent microballoons, and it is made up of core microballoon and surface coated shell, and wherein, the core microballoon is monodispersed and the sulfonated polystyrene microsphere; Surface coated shell is the fluorescence conjugated high molecular polymer of polystyrene support class.

The fluorescence conjugated high molecular polymer of described polystyrene support class, the molecular structural formula of its presoma is:

Wherein, R ₁With R ₂Identical or different, R ₁And R ₂Be H, OCH ₂COOH, OCH ₂COOCH ₃, OC ₃H ₆SO ₃A kind of among Na, F, Br or the CxHyO, in CxHyO, y=2x+1, x are 1～12 integer; N is a number of repeat unit.

A kind of preparation method of high molecular fluorescent microballoons with the Wessling salt polymer precursor method of giving birth, syntheticly in solution obtains the positively charged fluorescence conjugated high molecular polymer presoma of polystyrene support class, carries out the processing of following steps again:

(1) polystyrene is supportted the fluorescence conjugated high molecular polymer presoma of class and in solvent, dissolves after, add monodispersed, sulfonated polystyrene microsphere again;

(2) regulate pH value 〉=7 of above-mentioned solution, carry out dispersion treatment after, through washing, drying, obtained polystyrene core microsphere surface electrostatic adhesion polystyrene support the microballoon of the fluorescence conjugated high molecular polymer presoma of class;

(3) microballoon that step (2) is obtained is to handle 0.5～2 hour under 90～150 ℃ the vacuum condition in temperature, through washing, drying, obtains high molecular fluorescent microballoons again.

Principle of the present invention is: adopt the Wessling salt polymer precursor method of giving birth, the synthetic flexible macromolecule main chain that has positive charge, this flexible macromolecule is easy to dissolving in solution, salt precursor body electrostatic adhesion has on the polystyrene microsphere of negative charge on the surface with giving birth, by the control experiment condition, generate conjugated inflexible high polymer main chain at microsphere surface, this segmental solvability is relatively poor, makes that obtaining to stablize the conjugated polymer fluorescent microsphere becomes possibility.And, the reaction that generates conjugated polymer among the present invention is simple and controlled relatively in the high molecular inhomogeneous reaction of the direct synthesis of conjugate of microsphere surface, aftertreatment and also relatively easily carry out to the sign of high polymer main chain and to the adjusting of the fluorescence property of conjugated polymer.

Because the utilization of technique scheme, the present invention compared with prior art has following advantage:

(1) solvability of flexible polymer presoma is better, and each repeating unit all has electric charge, than the easier microsphere surface that is adsorbed on by electrostatic interaction of the conjugated polymer of small molecules or ionic with opposite charges, effectively improve the stability of microballoon fluorescence property, and had easily further machining characteristics.

(2) polyreaction formation macromolecular chain homogeneous phase in solution carries out, form conjugated polymer and eliminate the available more manageable heating means of reaction, therefore, the reaction of the generation conjugated polymer among the present invention is in the high molecular inhomogeneous reaction simple controllable of the direct synthesis of conjugate of microsphere surface, aftertreatment and the sign of high polymer main chain also relatively easily carried out.

Description of drawings

Fig. 1 is the nuclear-magnetism figure by polystyrene support (PPV) presoma of the embodiment of the invention 1 technical scheme preparation;

Fig. 2 is the fluorescence emission spectrogram by area load (PPV) fluorescent microsphere of the embodiment of the invention 1 technical scheme preparation;

Fig. 3 is the fluorescence emission spectrogram by area load (PPV) fluorescent microsphere of the embodiment of the invention 2 technical schemes preparation.

Embodiment

Below in conjunction with drawings and Examples the present invention is further described:

Embodiment one:

1; adopt the Wessling salt polymer precursor method of giving birth: in two mouthfuls of flasks of 50ml; use nitrogen protection; adding 1.0g phenyl ring has the polymerization single polymerization monomer of dodecyloxy side chain; be dissolved in the tetrahydrofuran solution of 10ml; magnetic agitation is dissolved it fully; put into ice-water bath; make reaction system under nitrogen protection, keep 0～5 ℃; the NaOH/MeOH solution 2.4ml that slowly adds 1.25M; react after 1 hour; add the HCl termination reaction, it is 14000 the dialysis tubing presoma at water dialysis preparation in 24 hours polymer P PV that reaction solution is put into molecular weight, and its synthetic route is as follows:

Wherein, n is a number of repeat unit.

Referring to accompanying drawing 1, it is the nuclear-magnetism figure (HNMR) of above-mentioned polystyrene support (PPV) presoma, is solvent with the deuterochloroform, at 7.0～6.7ppm, and 4.5ppm, 3.9ppm, 2.9ppm, 1.9ppm and 0.9ppm etc. locates to be the characteristic peak of hydrogen in the presoma.

2, the presoma 0.5g that makes in the step 1 is joined in the Erlenmeyer flask, with tetrahydrofuran (THF) and water by the mixed solution 12ml dissolving of making at 2: 1, adding the monodispersed particle diameter of 0.5g is the polystyrene microsphere of 25 microns sulfonating surface, drip the NaOH aqueous solution of 1.25M again in Erlenmeyer flask, regulating the pH value is 8; Ultrasonic concussion is 1 hour in ultrasonic concussion instrument, keeps the water temperature of ultrasonic concussion instrument to be lower than 30 ℃, and then uses the whizzer centrifugal treating, behind the removal supernatant liquid, washes microballoon repeatedly with tetramethylene sulfide and is washed off until polymer precursor.

3, the polystyrene microsphere that the surface electrostatic for preparing in the step 2 has been adsorbed the PPV presoma is put into vacuum drying oven, vacuumize, post-heating to 120 ℃, reacted 1 hour, naturally cooling at room temperature after the reaction, wash repeatedly until the PPV polymkeric substance that will not load to microballoon with tetramethylene sulfide and to wash off, obtain the fluorescent microsphere of clean polystyrene support (PPV) load.

Referring to accompanying drawing 2, it is the fluorescence emission spectrogram by area load (PPV) fluorescent microsphere of present embodiment technical scheme preparation.Can see that by Fig. 2 under the 410nm exciting light, the maximum absorption wavelength of this fluorescent microsphere is 560nm.

Embodiment two:

1, in two mouthfuls of flasks of 50ml, use nitrogen protection, adding 0.5g does not have the side group polymerization single polymerization monomer, is dissolved in the water of 10ml, and magnetic agitation is dissolved it fully, puts into ice-water bath, makes reaction system keep 0～5 ℃ under nitrogen protection, adds the NaOH/H of 1.25M ₂O solution 1ml reacted after 1 hour, added the HCl termination reaction, and it is 14000 the dialysis tubing presoma at water dialysis preparation in 24 hours polymer P PV that reaction solution is put into molecular weight, and its synthetic route is as follows:

2, the presoma 0.5g that makes in the step 1 is joined in the Erlenmeyer flask, use the 10ml water dissolution, adding the monodispersed particle diameter of 0.5g is the polystyrene microsphere of 25 microns sulfonating surface, NaOH adjusting pH value with 1.25M is 9, ultrasonic concussion is 1 hour in ultrasonic concussion instrument, centrifugal after keeping the water temperature of ultrasonic concussion instrument to be lower than 30 ℃ with whizzer, remove supernatant liquid, and water washes microballoon repeatedly.

3, the polystyrene microsphere that the surface electrostatic for preparing in the step 2 has been adsorbed the PPV presoma is put into vacuum drying oven, vacuumize, post-heating to 120 ℃, reacted 1 hour, naturally cooling, wash repeatedly until the PPV polymkeric substance that will not load to microballoon with tetramethylene sulfide and to wash off, obtain the fluorescent microsphere of clean polystyrene support (PPV) load.

Referring to accompanying drawing 2, it is the fluorescence emission spectrogram by area load (PPV) fluorescent microsphere of present embodiment technical scheme preparation.Can see that by Fig. 3 under the 410nm exciting light, the maximum absorption wavelength of this fluorescent microsphere is 510nm.

Claims (3)

1. high molecular fluorescent microballoons, it is characterized in that: it is made up of core microballoon and surface coated shell, and wherein, the core microballoon is monodispersed and the sulfonated polystyrene microsphere; Surface coated shell is the fluorescence conjugated high molecular polymer of polystyrene support class.

2. a kind of high molecular fluorescent microballoons according to claim 1 is characterized in that: the fluorescence conjugated high molecular polymer of described polystyrene support class, and the molecular structural formula of its presoma is:

Wherein, R ₁With R ₂Identical or different, R ₁And R ₂Be H, OCH ₂COOH, OCH ₂COOCH ₃, OC ₃H ₆SO ₃A kind of among Na, F, Br or the CxHyO, in CxHyO, y=2x+1, x are that 1～12 Integer n is a number of repeat unit.

3. the preparation method of a high molecular fluorescent microballoons with the Wessling salt polymer precursor method of giving birth, syntheticly in solution obtains the positively charged fluorescence conjugated high molecular polymer presoma of polystyrene support class, it is characterized in that carrying out the processing of following steps again:

(1) polystyrene is supportted the fluorescence conjugated high molecular polymer presoma of class and in solvent, dissolves after, add monodispersed, sulfonated polystyrene microsphere again;

(2) regulate pH value 〉=7 of above-mentioned solution, carry out dispersion treatment after, through washing, drying, obtained polystyrene core microsphere surface electrostatic adhesion polystyrene support the microballoon of the fluorescence conjugated high molecular polymer presoma of class;

(3) microballoon that step (2) is obtained is to handle 0.5～2 hour under 90～150 ℃ the vacuum condition in temperature, through washing, drying, obtains high molecular fluorescent microballoons again.

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