CN108410449A - A kind of preparation and its application of the water soluble fluorescence nano silicon particles of detection of alkaline phosphatase - Google Patents
A kind of preparation and its application of the water soluble fluorescence nano silicon particles of detection of alkaline phosphatase Download PDFInfo
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Abstract
The invention discloses a kind of preparations and its application of the nano silicon particles of detection of alkaline phosphatase, specific preparation method includes being added to the water N [3 (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) under stiring, then it is separately added into hydroquinone, resorcinol and catechol, and stir at room temperature, at a normal, you can obtain yellow fluorescence nano silicon particles (yellow Si NPs), blue-fluorescence nano silicon particles (blue Si NPs) and orange fluorescence silicon nano particles (orange Si NPs).The water-soluble orange fluorescence silicon nano particles being prepared can be used in highly selective Sensitive Detection alkaline phosphatase (ALP).
Description
Technical field
The invention belongs to chemical fields, and in particular to a kind of water soluble fluorescence silicon nanometer of detection of alkaline phosphatase
The preparation and its application of grain.
Background technology
Alkaline phosphatase (ALP), which is one kind, being distributed widely in each organ of human body by the dephosphorylized enzyme of corresponding substrate
In, secondly it is kidney, the tissues such as bone, intestines and placenta wherein being most with liver.ALP detections are mainly used for obstructive Huang
The inspection of subcutaneous ulcer, primary carcinoma of liver, secondary carcinoma of liver, Cholestatic hepatitis etc..When suffering from these diseases, liver cell excessively manufactures
ALP enters blood through lymphatic channel and sinus hepaticus, simultaneously because biliary tract bile excretion obstacle, reflux enter blood and cause serum in liver
ALP is significantly raised.Therefore, the method for establishing ALP in accurate detection serum plays a crucial role human health.So far
Until, it has been developed and is examined based on CdS/CdTe/CdSe QDs, Au/Ag NCs, C dots, polymer probe and Small-molecule probe
The method for surveying ALP.There is the problems such as selectivity is relatively low, preparation is complicated, cost is higher or toxicity is higher in the above detection material,
In comparison nano silicon particles, which have, prepares that simple, toxicity is relatively low, stability is strong, good biocompatibility, biodegradable, former
The advantages such as abundant cheap, without complexity the modification of material.Therefore it is established using nano silicon particles as probe a kind of highly selective, highly sensitive
The new method of degree detection ALP is particularly necessary.So far, the preparation of nano silicon particles need to usually use high temperature and pressure or complexity
Instrument.Prepared nano silicon particles are mostly blue-fluorescence or green fluorescence at present, in relation to preparing long wavelength's fluorescent emission or tool
Having the nano silicon particles of fluorescence adjustability, there is not been reported.Therefore, it is quickly prepared at room temperature, at a normal in simple mild method
The nano silicon particles of fluorescence adjustability are realized very necessary from blue-fluorescence to yellow or red fluorescence regulation and control.
Invention content
In view of the deficiencies of the prior art and the above problem, it is prepared with multicolor fluorescence the purpose of the present invention is to provide a kind of
The new method of the water-soluble silicon nano particle of adjustability.On this basis, a kind of letter is established by fluorescence probe of nano silicon particles
The new method of single, highly selective, highly sensitive detection of alkaline phosphatase (ALP).
The present invention provides a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase, the water soluble fluorescences
Nano silicon particles are to synthesize to obtain by N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and benzenediol.
Further, the benzenediol is hydroquinone, resorcinol or catechol.
Further, when the benzenediol is hydroquinone, the preparation side of the water soluble fluorescence nano silicon particles
Method includes：
(1) N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added to the water；
(2) hydroquinone is added and stirs and can be obtained yellow fluorescence water soluble fluorescence nano silicon particles (yellow-Si
NPs)；
(3) bag filter for the water soluble fluorescence nano silicon particles 500Da being prepared is dialysed, you can obtain after purification
Water soluble fluorescence nano silicon particles.Wherein N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and hydroquinone rub
You are than being 1：10.
Further, when the benzenediol is resorcinol, the preparation side of the water soluble fluorescence nano silicon particles
Method includes：
(1) N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added to the water；
(2) resorcinol is added and stirs and can be obtained blue-fluorescence water soluble fluorescence nano silicon particles (blue-Si
NPs)；
(3) bag filter for the water soluble fluorescence nano silicon particles 500Da being prepared is dialysed, you can obtain after purification
Water soluble fluorescence nano silicon particles.Wherein N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and resorcinol rub
You are than being 1：140.
Further, when the benzenediol is catechol, the preparation side of the water soluble fluorescence nano silicon particles
Method includes：
(1) N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added to the water；
(2) catechol is added and stirs and can be obtained orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si
NPs)；
(3) bag filter for the water soluble fluorescence nano silicon particles 500Da being prepared is dialysed, you can obtain after purification
Water soluble fluorescence nano silicon particles.Wherein N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and catechol rub
You are than being 1：6.
A method of including the detection of alkaline phosphatase of water soluble fluorescence nano silicon particles, which is characterized in that including with
Lower step：
(1) vitamin C phosphoric ester (ascorbic of 0.7mol/L is added in the Tris-HCl buffer solutions that pH is 7.4
Acid 2-phosphate, AAP) and various concentration alkaline phosphatase (ALP), be placed in 37 DEG C of water-baths and hatch 25 minutes；
(2) prepared orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si is sequentially added in above-mentioned solution
NPs) the KMnO of solution and a concentration of 10mM4Solution；
(3) excitation wavelength is set as 481nm, is to measure fluorescence intensity at 558nm in launch wavelength, it is bent to make standard work
Line, and according to the content of fluorescence intensity and standard curve calculating ALP.
Further, the method for above-mentioned detection of alkaline phosphatase can be applied to the detection of serum alkaline phosphatase.
The beneficial effects of the present invention are：
(1) it is simple, at low cost that the present invention was developed prepares water soluble fluorescence nano silicon particles method.
(2) the water soluble fluorescence nano silicon particles toxicity that is prepared of the present invention is low, has that water-soluble, stability is good, tool
There are stronger salt tolerance, anti-light Bleachability, pH stability and good biocompatibility.
(3) present invention establishes a kind of new method of the detection of alkaline phosphatase in serum, and this method is selectively good, sensitive
Degree is high.
Description of the drawings
Fig. 1：The TEM for the nano particle being prepared schemes and size distribution plot：
Wherein A is yellow fluorescence water soluble fluorescence nano silicon particles (Y-Si NPs), B is orange fluorescent water-soluble fluorescence silicon
Nano particle (O-Si NPs), C are blue-fluorescence water soluble fluorescence nano silicon particles (B-Si NPs)；
Fig. 2：Fourier transform infrared spectroscopy (FT-IR) figure for the nano particle being prepared：
Wherein a lines indicate that orange fluorescent water-soluble fluorescence silicon nano particles (O-Si NPs), b lines indicate that yellow fluorescence is water-soluble
Property fluorescence silicon nano particles (Y-Si NPs), c lines indicate blue-fluorescence water soluble fluorescence nano silicon particles (B-Si NPs)；
Fig. 3：The excitation spectrum for the nano particle being prepared, emission spectrum, ultraviolet-visible absorption spectroscopy figure：
Wherein A is the excitation spectrum (expression of a lines) of orange fluorescent water-soluble fluorescence silicon nano particles (O-Si NPs), transmitting
Spectrum (expression of b lines), ultraviolet-visible absorption spectroscopy (expression of c lines)；B is yellow fluorescence water soluble fluorescence nano silicon particles (Y-Si
NPs excitation spectrum (expression of a lines), emission spectrum (expression of b lines), ultraviolet-visible absorption spectroscopy (expression of c lines))；C is that blue is glimmering
Excitation spectrum (expression of a lines), the emission spectrum (expression of b lines) of light water soluble fluorescence nano silicon particles (B-Si NPs)；
Fig. 4：The fluorescence spectra and standard curve of the water soluble fluorescence nano silicon particles detection of alkaline phosphatase of preparation：
Wherein A is the alkaline phosphorus that various concentration is added in orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si NPs)
The fluorescence spectra of sour enzyme；B is standard working curve；
Specific implementation mode
Below in conjunction with Figure of description, technical scheme in the embodiment of the invention is clearly and completely described, shows
So, described embodiment is only the part of the present invention, rather than the whole invented.Based on the embodiments of the present invention,
Every other embodiment obtained by those of ordinary skill in the art without making creative efforts, belongs to this hair
The range of bright protection.
The preparation of 1 water soluble fluorescence nano silicon particles of embodiment simultaneously carries out structural characterization to it
(1) preparation of yellow fluorescence water soluble fluorescence nano silicon particles
1mL N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added under stiring in 5mL water.Then add
Enter 5.0mg hydroquinones and stir 25 minutes at room temperature, at a normal, you can obtains yellow fluorescence water soluble fluorescence nano silicon particles
(yellow-Si NPs)1.13g.The prepared nano silicon particles bag filter of 500Da is dialysed 10 hours, you can obtain pure
Nano silicon particles after change.
(2) preparation of blue-fluorescence water soluble fluorescence nano silicon particles
1mL N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added under stiring in 5mL water.Then add
Enter 70.0mg resorcinols and stir 25 minutes at room temperature, at a normal, you can obtains blue-fluorescence nano silicon particles (blue-Si
NPs)0.97g.The prepared nano silicon particles bag filter of 500Da is dialysed 10 hours, you can the silicon obtained after purification is received
Rice grain.
(3) preparation of orange fluorescent water-soluble fluorescence silicon nano particles
1mL N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added under stiring in 5mL water.Then add
After entering 3.0mg catechols and stirring 2 hours at room temperature, at a normal, you can obtain orange fluorescence silicon nano particles (orange-
Si NPs)1.17g.The prepared nano silicon particles bag filter of 500Da is dialysed 10 hours, you can obtain silicon after purification
Nano particle.
The TEM for the nano particle being prepared as shown in Figure 1 schemes and size distribution plot, and wherein A is that yellow fluorescence is water-soluble
Fluorescence silicon nano particles (yellow-Si NPs), B be orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si NPs),
C is blue-fluorescence water soluble fluorescence nano silicon particles (blue-Si NPs)；
Fourier transform infrared spectroscopy (FT-IR) figure for the nano particle being prepared as shown in Figure 2, wherein a lines indicate
Orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si NPs), b lines indicate yellow fluorescence water soluble fluorescence silicon nanometer
Grain (yellow-Si NPs), c lines indicate blue-fluorescence water soluble fluorescence nano silicon particles (blue-Si NPs)；
The excitation spectrum for the nano particle being prepared as shown in Figure 3, emission spectrum, ultraviolet-visible absorption spectroscopy figure,
Middle A is excitation spectrum (expression of a lines), the emission spectrum of orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si NPs)
(expression of b lines), ultraviolet-visible absorption spectroscopy (expression of c lines)；B is yellow fluorescence water soluble fluorescence nano silicon particles (yellow-
Si NPs) excitation spectrum (expression of a lines), emission spectrum (expression of b lines), ultraviolet-visible absorption spectroscopy (expression of c lines)；C is indigo plant
Excitation spectrum (expression of a lines), the emission spectrum (expression of b lines) of color fluorescent water-soluble fluorescence silicon nano particles (blue-Si NPs).
The fluorescence of the alkaline phosphatase for the water soluble fluorescence nano silicon particles detection various concentration being prepared as shown in Figure 4
Spectrogram (A expressions) and standard working curve figure (B expressions).
A kind of method of the detection of alkaline phosphatase comprising water soluble fluorescence nano silicon particles of embodiment 2
A kind of method of the detection of alkaline phosphatase comprising water soluble fluorescence nano silicon particles includes：
(1) 30 μ L 0.7mol/L are added in 2.83mL, the Tris-HCl buffer solutions of pH 7.4, a concentration of 10mM
Vitamin C phosphoric ester (ascorbic acid 2-phosphate, AAP) and concentration be respectively 0U/L, 0.01U/L, 0.1U/
L, the alkaline phosphatase (ALP) of 1U/L, 5U/L, 50U/L and 500U/L is placed in 37 DEG C of water-baths and hatches 25 minutes.
(2) the orange fluorescent water-soluble fluorescence silicon nanometer prepared by 20 μ L embodiments 1 is sequentially added in above-mentioned solution
The KMnO of grain (orange-Si NPs) solution and 90 a concentration of 10mM of μ L4Solution.
(3) it is 481nm in excitation wavelength, launch wavelength is that fluorescence intensity is measured at 558nm
(4) it makes standard working curve and calculates the content of ALP：With orange-Si NPs+KMnO4The fluorescence of solution is strong
Degree is F1；orange-Si NPs+KMnO4The fluorescence intensity of+AAP+ALP solution is F2；With F2-F1For ordinate, logcALPFor cross
Coordinate；The equal 5nm of the slit width of excitation wavelength and launch wavelength, and according to fluorescence intensity and standard curve F2-F1=
116.17logcALP- 100.52 calculate the content of ALP.
A kind of method of the detection of alkaline phosphatase comprising water soluble fluorescence nano silicon particles of embodiment 3 is to serum neutral and alkali
The detection of phosphatase
(1) by the human serum sample of purchase with Tris-HCl buffer solutions (pH 7.4；A concentration of 10mM) 100 times of dilution,
The vitamin C phosphoric ester (ascorbic acid 2-phosphate, AAP) and concentration point of 30 a concentration of 0.7mol/L of μ L is added
Not Wei 50U/L, 100U/L, 150U/L ALP, final total volume 2.89mL, be placed in 37 DEG C of water-baths and hatch 25 minutes.
(2) the orange-Si NPs solution prepared by 20 μ L embodiments 1,90 μ L concentration are sequentially added in above-mentioned solution
For the KMnO of 10mM4Solution.
(3) excitation wavelength is set as 481nm, is that fluorescence intensity is measured at 558nm in launch wavelength.
(4) it makes standard working curve and calculates the content of ALP：With orange-Si NPs+KMnO4The fluorescence of solution is strong
Degree is F1；orange-Si NPs+KMnO4The fluorescence intensity of+AAP+ALP solution is F2；With F2-F1For ordinate, logcALPFor cross
Coordinate；The equal 5nm of the slit width of excitation wavelength and launch wavelength, and according to measuring fluorescence intensity F1、F2With standard curve F2-F1
=116.17logcALP- 100.52 calculate the content of ALP.
(5) experimental result：The water soluble fluorescence nano silicon particles being prepared such as 1 embodiment 1 of table are used for serum neutral and alkali
Shown in the measurement result of phosphatase：
The measurement result of ALP in 1. human serum sample of table
The above-mentioned the results show present invention can with the content of Sensitive Detection serum alkaline phosphatase, meet it is clinical in
The requirement for detecting serum alkaline phosphatase has certain directive significance to clinical monitoring analysis；And the present invention detects alkaline phosphorus
Sour enzyme result is more accurate, and it is consistent with PNPP colorimetric determination results that experiment obtains result, it was demonstrated that this method is reliable feasible.
Claims (10)
1. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase, which is characterized in that the water soluble fluorescence silicon
Nano particle is to synthesize to obtain by N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and benzenediol.
2. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase as described in claim 1, which is characterized in that institute
The benzenediol stated is hydroquinone, resorcinol or catechol.
3. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase as described in claim 1, which is characterized in that institute
The benzenediol stated is hydroquinone, and the preparation method of the water soluble fluorescence nano silicon particles includes：
(1) N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added to the water；
(2) hydroquinone is added and stirs and can be obtained yellow fluorescence water soluble fluorescence nano silicon particles (yellow-Si NPs)；
(3) bag filter for the water soluble fluorescence nano silicon particles 500Da being prepared is dialysed, you can obtain water after purification
Dissolubility fluorescence silicon nano particles.
4. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase as claimed in claim 3, which is characterized in that institute
The molar ratio of N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and hydroquinone stated is 1：10.
5. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase as described in claim 1, which is characterized in that institute
The benzenediol stated is resorcinol, and the preparation method of the water soluble fluorescence nano silicon particles includes：
(1) N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added to the water；
(2) resorcinol is added and stirs and can be obtained blue-fluorescence water soluble fluorescence nano silicon particles (blue-Si NPs)；
(3) bag filter for the water soluble fluorescence nano silicon particles 500Da being prepared is dialysed, you can obtain water after purification
Dissolubility fluorescence silicon nano particles.
6. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase as claimed in claim 5, which is characterized in that institute
The molar ratio of N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and resorcinol stated is 1：140.
7. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase as described in claim 1, which is characterized in that institute
The benzenediol stated is catechol, and the preparation method of the water soluble fluorescence nano silicon particles includes：
(1) N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine (DAMO) is added to the water；
(2) catechol is added and stirs and can be obtained orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si NPs)；
(3) bag filter for the water soluble fluorescence nano silicon particles 500Da being prepared is dialysed, you can obtain water after purification
Dissolubility fluorescence silicon nano particles.
8. a kind of water soluble fluorescence nano silicon particles of detection of alkaline phosphatase as claimed in claim 7, which is characterized in that institute
The molar ratio of N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines (DAMO) and catechol stated is 1：6.
9. a kind of method of the detection of alkaline phosphatase comprising water soluble fluorescence nano silicon particles, which is characterized in that including following
Step：
(1) vitamin C phosphoric ester (the ascorbic acid of 0.7mol/L are added in the Tris-HCl buffer solutions that pH is 7.4
2-phosphate, AAP) and various concentration alkaline phosphatase (ALP), be placed in 37 DEG C of water-baths and hatch 25 minutes；
(2) prepared orange fluorescent water-soluble fluorescence silicon nano particles (orange-Si is sequentially added in above-mentioned solution
NPs) the KMnO of solution and a concentration of 10mM4Solution；
(3) excitation wavelength is set as 481nm, is to measure fluorescence intensity at 558nm in launch wavelength, is made standard working curve,
And the content of ALP is calculated according to fluorescence intensity and standard curve.
10. a kind of method of the detection of alkaline phosphatase comprising water soluble fluorescence nano silicon particles as claimed in claim 9,
It is characterized in that, the method can be applied to the detection of serum alkaline phosphatase.
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| CN108949163A (en) * | 2018-09-18 | 2018-12-07 | 中国科学院兰州化学物理研究所 | A kind of blue, yellow, the photoemissive Fluorescent silicon nanoparticle of orange the method for one kettle way preparation |
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| CN116891737A (en) * | 2023-02-28 | 2023-10-17 | 中国科学院深圳先进技术研究院 | Synthesis method of fluorescent silicon dot, fluorescent silicon dot and application |
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