Application of glutathione-based stable gold nano cluster particles to detection of sulfhydryl compound

A technology of sulfhydryl compounds and glutathione, which is applied in measuring devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of small probe Stokes shift, serious photobleaching, fluorescence self-quenching, etc., and achieve specificity Good, short analysis time, significant specific effect

Active Publication Date: 2015-07-01
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the disadvantage of this method is that some samples (especially blood samples) may have a large background absorption, which will cause detection interference
However, most of the detection reagents in this method are small organic molecules, the problem of photobleaching is serious, and the Stokes shift of the probe is small, which may cause serious fluorescence self-quenching and excitation background interference

Method used

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  • Application of glutathione-based stable gold nano cluster particles to detection of sulfhydryl compound
  • Application of glutathione-based stable gold nano cluster particles to detection of sulfhydryl compound
  • Application of glutathione-based stable gold nano cluster particles to detection of sulfhydryl compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of glutathione-stabilized gold nanoclusters

[0022] At 25 °C, take 5 mL of chloroauric acid aqueous solution with a concentration of 1.0-10.0 mmol / L into a 15 mL plastic centrifuge tube, add 5 mL of 0.8-10.0 mmol / L glutathione aqueous solution to the chloroauric acid solution and Mix well to ensure that the mol ratio of chloroauric acid: glutathione is between 1:0.5 and 1:1. After 10-15 min, when the color of the mixture becomes colorless, adjust the pH value to 2-3. And put it in a 60~80 oC water bath for 1~2 hours to get a dark yellow transparent solution. Cool the solution to 25 °C, filter it with a 0.22 micron filter membrane, add the same volume of absolute ethanol to the filtrate, mix well, centrifuge at 10000 rpm for 10-20 minutes, weigh the sediment after drying, add 5-100 mmol / L phosphate buffer solution, ultrasonic (40 W, 40 KHz) for 2 min to dissolve the sediment in the phosphate buffer solution to obtain a gold nanocluster solut...

Embodiment 2

[0023] Example 2 Sulfhydryl compounds trigger fluorescence enhancement of gold nanoclusters

[0024] Take 0.5 mL of the glutathione-stabilized gold nanocluster solution prepared in Example 1, dilute it to 1.0 mL with deionized water, and record the excitation light of different wavelengths from 350 to 490 nm, and the wavelength range is 500 to 680 nm. Fluorescence Spectroscopy. Such as figure 2 The emission spectra of different wavelengths of excitation light of 350, 370, 390, 410, 430, 450, 470, and 490 nm are shown. Under the excitation light of 450 nm, glutathione-stabilized gold nanoparticles The cluster solution has the strongest fluorescence intensity, and its emission peak is located at 570 nm.

[0025] Take two 0.5 mL glutathione-stabilized gold nanocluster solutions, add 0.5 mL deionized water and glutathione aqueous solution of known concentration respectively, mix well for 15 minutes, and record in the wavelength range 350-490 nm Under the excitation light, th...

Embodiment 3

[0026] Example 3 Drawing of standard relationship curve between fluorescence enhancement factor and glutathione concentration

[0027] Take 0.5 mL of the above-mentioned glutathione-stabilized gold nanocluster solution and add 0.5 mL of glutathione with concentrations of 0, 0.1, 0.25, 0.5, 0.75, 1.0, 2.0, 3.0, 4.0, and 5.0 mmol / L respectively. Peptide aqueous solution, numbered 1~10 in sequence, after mixing for 15 minutes, record the fluorescence intensity of samples 1~10 at the emission wavelength of 570 nm under the excitation light with a wavelength of 450 nm. Calculate the ratio of the fluorescence intensity of samples 2 to 10 to sample 1, that is, the fluorescence enhancement factor F. With the glutathione concentration [thiol] as the abscissa and the fluorescence enhancement factor F as the ordinate, draw a linear standard curve, and use linear fitting to obtain the calibration relation: F= 1.39+ 1.44×[thiol] (mmol / L), the linear correlation coefficient of the fitti...

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Abstract

The invention discloses application of glutathione-based stable gold nano cluster particles to detection of a sulfhydryl compound, wherein the glutathione-based stable gold nano cluster particles are prepared from a chloroauric acid solution and a glutathione solution, wherein a molar ratio of chloroauric acid to glutathione is between 1:0.5 and 1:1. By using an fluorescence enhancement characteristic of the glutathione-based stable gold nano cluster particles after the addition of the sulfhydryl compound, through detecting the change of fluorescence intensity before and after the sulfhydryl compound with an unknown concentration reacts with the glutathione-based stable gold nano cluster particles, a linear standard curve chart is drawn, and a calibration relation is obtained by adopting linear fitting, so that the sulfhydryl compound with the unknown concentration is detected.

Description

technical field [0001] The invention relates to the technical field of detection of mercapto compounds, in particular to the application of glutathione-based stabilized gold nano-cluster particles in the detection of mercapto compounds. Background technique [0002] Sulfhydryl compounds are important antioxidants for maintaining the physiological environment, responsible for scavenging free radicals in tissues and cells, and preventing free sulfhydryl groups in intracellular protein molecules from forming disulfide bonds. Sulfhydryl compounds such as glutathione can also inhibit the tyrosinase activity of the epidermis and reduce melanin production, so they are of great significance in cosmetic medicine. In view of its important practical value, accurate and rapid quantification of thiol compounds is of great significance. [0003] The detection methods of mercapto compounds mainly include colorimetry, mass spectrometry, fluorescence and so on. The classic Ellman reagent m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/531
Inventor 姜晖王雪梅苏小清
Owner SOUTHEAST UNIV
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