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Ratiometric fluorescent probe and preparation thereof, and application of ratiometric fluorescent probe in detection of glutathione

A fluorescent probe and ratiometric technology, which is applied in the field of rapid visual ratiometric fluorescent probe preparation, can solve the problems of weak fluorescence intensity and fluorescent probe toxicity, and achieve the effects of good stability, simple synthesis method and simple preparation

Inactive Publication Date: 2018-01-19
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above fluorescent probes still have problems such as toxicity and weak fluorescence intensity.

Method used

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  • Ratiometric fluorescent probe and preparation thereof, and application of ratiometric fluorescent probe in detection of glutathione
  • Ratiometric fluorescent probe and preparation thereof, and application of ratiometric fluorescent probe in detection of glutathione
  • Ratiometric fluorescent probe and preparation thereof, and application of ratiometric fluorescent probe in detection of glutathione

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Preparation of green quantum dots

[0037] Add 50 mL of deionized water into the three-neck flask, start stirring, and add 0.80 mL of CdCl with a concentration of 0.20 M 2 Aqueous solution, 72.5 mg sodium hexametaphosphate (SHMP, 0.12 mmol) and 34.6 μL 3-mercaptopropionic acid (MPA, 0.32 μmol). Then adjust the pH to 9.0 with 1.0M NaOH aqueous solution, and then add 0.80 mL of NaOH with a concentration of 0.020M 2 SeO 3 aqueous solution, after reflux at 100°C for 10 minutes, add 3.70mL hydrazine hydrate (N 2 h 4 ·H 2 O, 0.072mol), after continuing to reflux for 9h, stop heating, obtain the CdSe quantum dot solution (i.e. green quantum dot solution) of green fluorescence, emission peak is at 522 nanometers, and concentration is 9.21 * 10 -6 mol / L.

[0038] Take 10mL of CdSe quantum dot solution, add 12mL of absolute ethanol, let it stand, centrifuge, and wash the precipitate with absolute ethanol for 1-2 times to obtain 2mg of solid-phase cadmium selenide quantum...

Embodiment 2

[0047] 1. Silica nanoparticles embedded with green quantum dots

[0048] Disperse 2 mg of green quantum dots obtained by the method in Example 1 in 40 ml of ethanol, and stir at 25 ° C for 30 min to form a homogeneous mixed solution; add 0.7 ml of ammonia water and 0.5 ml of tetraethyl orthosilicate to the mixed solution, and stir at 20 ° C for 12 h; Add 100 μL of 3-aminopropyltriethoxysilane, stir at 25°C for 12 hours; centrifuge, and wash the precipitate with absolute ethanol and deionized water for 2 to 3 times in turn to obtain surface amino-modified silica embedded with green quantum dots Nanospheres (i.e. CdSe@SiO 2 -NH 2 ) 0.120g, particle size 40 ~ 60nm (morphology TEM picture is the same as figure 1 , the fluorescence spectrum is the same as image 3 Middle curve b). CdSe@SiO 2 -NH 2 Disperse in deionized water and set aside.

[0049] 2. Preparation of ratiometric fluorescent probes

[0050] The N-acetyl-L-cysteine ​​modified CdTe quantum dot solution prepared...

Embodiment 3

[0052] 1. Silica nanoparticles embedded with green quantum dots

[0053] Disperse 2 mg of green quantum dots obtained by the method in Example 1 in 40 ml of ethanol, and stir at 25 ° C for 10 min to form a uniform mixed solution; add 1 ml of ammonia water and 1 ml of tetraethyl orthosilicate to the mixed solution, and stir at 25 ° C for 12 h; then add 3 - Aminopropyltriethoxysilane 200 μL, stirred at 25°C for 12 hours; centrifuged, and the precipitate was washed with absolute ethanol and deionized water for 2 to 3 times in sequence to obtain surface amino-modified silica nanoparticle embedded with green quantum dots. Balls (i.e. CdSe@SiO 2 -NH 2 ) 0.150g, particle size 40-60nm. CdSe@SiO 2 -NH 2 Disperse in deionized water and set aside.

[0054] 2. Preparation of ratiometric fluorescent probes

[0055]The N-acetyl-L-cysteine ​​modified CdTe quantum dot solution prepared by the method in Example 1 was centrifuged, and 2 mg of the precipitate (i.e. N-acetyl-L-cysteine ​​mo...

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Abstract

The invention discloses a ratiometric fluorescent probe and a preparation thereof, and an application of the ratiometric fluorescent probe in detection of glutathione. The ratiometric fluorescent probe is prepared through the following steps: with green quantum dots as cores, carrying out embedding in silica nanoparticles, then subjecting the surface of the silica nanoparticles to amination modification, and covalently coupling red quantum dots onto the amination modified surface of the silica nanoparticles so as to obtain the ratiometric fluorescent probe, wherein the green quantum dots are cadmium telluride quantum dots modified by N-acetyl-L-cysteine. According to the invention, a St ber method is utilized to prepare quantum dot embedded silica nanoparticles, so the synthetic method issimple and has less time consumption. The probe provided by the invention has low detection limit and can realize specific detection during detection of GSH; meanwhile, the probe has advantages like simple preparation and good stability, reduces the detection limit of glutathione to 45 nM, and provides convenience for rapid visual detection.

Description

(1) Technical field [0001] The invention relates to a preparation and detection method of a fast visualized ratio fluorescent probe for glutathione detection. (2) Background technology [0002] Glutathione (GSH) is a natural active peptide with important physiological functions, composed of glutamic acid, cysteine ​​and glycine, and exists in almost all biological cells. The cysteine ​​on glutathione contains a sulfhydryl group as an active group, which can effectively remove free radicals and other toxins that damage DNA and RDA (iodoacetic acid, mustard gas, lead, mercury, arsenic and other heavy metals). Studies have found that the abnormal behavior of glutathione concentration in the human body is related to human diseases such as cancer, liver disease, AIDS and heart disease. Although the human body can produce glutathione, according to research, the content of glutathione will decrease with age, and the decrease of glutathione will also be affected by the surrounding ...

Claims

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Application Information

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IPC IPC(8): C09K11/02C09K11/88B82Y30/00B82Y20/00G01N21/64
Inventor 陈爱民彭晓汪晶陈琦潘再法
Owner ZHEJIANG UNIV OF TECH
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