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Cysteine detection method

A cysteine ​​and solution technology, used in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of mutual interference and indistinguishable detection, and achieve the effects of low detection cost, good selectivity and fast detection speed.

Inactive Publication Date: 2013-02-13
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since homocysteine ​​(Hcy) and cysteine ​​are very similar in structure (Hcy has only one more -CH in structure than Cys 2 ), most compounds cannot distinguish between
Therefore, there is mutual interference for their respective detections, so it is of great significance to design a fluorescent chemical sensor that can distinguish between Hcy and Cys

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1: Using this method to differentiate cysteine ​​and homocysteine

[0023] Prepare 2*10 with DMF -3 M's Ir 2 (ppy) 4 Cl 2 Solution, and prepare HEPES (0.2M) buffer solution with pH 7.0 with triple distilled water; respectively take 1750 μl DMF and 200 μl HEPES buffer solution and add them to the two fluorescence pools, and use a micro-sampler to absorb the above-mentioned Ir 2 (ppy) 4 Cl 2 Add 50 μl of the solution to a fluorescent dish and detect it on a fluorescent instrument. There is emission at 505 nm; take 4×10 -3 100 μl of cysteine ​​and homocysteine ​​solutions were gradually added to the above two fluorescent pools with a micro-injector, placed at room temperature for 20 minutes and then detected on a fluorescence instrument. The emission peak of the system added with cysteine ​​was determined by 505nm red shifted to 576nm ( Figure 1A ). Irradiated with 365nm ultraviolet light from a portable ultraviolet lamp, the color of the emitted light chan...

Embodiment 2

[0025] Prepare 2×10 in DMF -3 M's Ir 2 (ppy) 4 Cl 2 solution, and prepare HEPES (0.2M) buffer solution with pH 7.0 with triple distilled water; take 1750 μl DMF and 200 μl HEPES buffer solution and add them to the fluorescent cell, and use a micro-sampler to absorb the above-mentioned Ir 2 (ppy) 4 Cl 2 Add 50 μl of the solution to the fluorescence dish and detect it on the fluorescence instrument. There is emission at 505 nm; take the cysteine ​​solution and gradually add it to the above-mentioned fluorescence pool with a micro-injector, and detect it on the fluorescence instrument while adding the sample. The emission peak of the gradually adding cystine system is red-shifted from 505nm to 576nm ( figure 2 ).

Embodiment 3

[0027] Prepare 2*10 with DMF -3 M's Ir 2 (ppy) 4 Cl 2 solution, and prepare HEPES (0.2M) buffer solution with pH 7.0 with triple distilled water; take 900 μl DMF and 500 μl HEPES buffer solution and add them to the fluorescent cell, and use a micro-sampler to absorb the above-mentioned Ir 2 (ppy) 4 Cl 2 Add 50 μl of the solution to the fluorescence dish and detect it on the fluorescence instrument. There is emission at 505 nm; take the cysteine ​​solution and gradually add it to the above-mentioned fluorescence pool with a micro-injector, and detect it on the fluorescence instrument while adding the sample. The emission peak of the gradually added system of cystine shifted from 505nm to 570nm ( image 3 ).

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Abstract

The invention provides a method capable of detecting cysteine in a half-aqueous solution. Importantly, the method can distinguish cysteine from homocysteine. Based on a complex dichlorotetra[2-(2-pyridyl)phenyl]diiridium(III)[Ir2(ppy)4Cl2], in the half-aqueous solution of DMF (dimethylformamide) or DMSO (dimethylsulfoxide), the cysteine is directly detected by a fluorescent method. The determination process is simple and convenient and free from the interference of other amino acids including homocysteine and anions in the solution; and the method can be used for detecting whether cysteine exists or not.

Description

technical field [0001] The invention relates to a method for detecting thiol amino acids, in particular to a method for detecting cysteine ​​in a semi-aqueous solution, and the method can distinguish homocysteine ​​(Hcy) from cysteine ​​(Cys). Background technique [0002] Sulfhydryl amino acids include homocysteine ​​(Hcy) and cysteine ​​(Cys), which are small biomolecules necessary for the human body and play an important role in physiological processes. The increase of Hcy concentration in plasma can lead to many diseases, such as cardiovascular disease, Alzheimer's disease and osteoporosis. Deficiency of Cys can lead to stunted growth, liver damage, muscle and fat loss, skin damage, and physical weakness in children. Common methods for testing sulfhydryl groups include electrochemistry, ultraviolet-visible absorption spectroscopy, high-performance liquid chromatography (HPLC), capillary electrophoresis, and enzyme-linked immunosorbent assay. [0003] Due to the advanta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 陈绘丽李晓凯
Owner SHANXI UNIV
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