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Method for detecting cysteine content in real time by colorimetric method

A technology of cysteine ​​and colorimetry, which is applied in the field of real-time detection of cysteine ​​content by colorimetry, can solve problems such as inability to realize real-time detection, large steric hindrance of active groups, and slow reaction speed of thiols. High sensitivity, large absorption coefficient, and good reproducibility

Inactive Publication Date: 2010-12-15
DONGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the large steric hindrance of the active groups, the reaction speed of these dyes with thiols is slow, and instant detection cannot be realized; at the same time, the detection must be carried out under acidic or alkaline conditions, which cannot meet the physiological pH conditions.

Method used

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  • Method for detecting cysteine content in real time by colorimetric method
  • Method for detecting cysteine content in real time by colorimetric method
  • Method for detecting cysteine content in real time by colorimetric method

Examples

Experimental program
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Effect test

Embodiment 1

[0030] (1) Preparation of chromogenic agent N-methyl-N-hydroxyethylaniline squaraine dye

[0031] Weigh N-methyl-N-hydroxyethylaniline (302mg, 2mmol) and squaraine (114mg, 1mmol) into a 100mL three port equipped with a water separator, then add 15mL of n-butanol and toluene mixture (1: 1, v / v), reflux reaction at 120 DEG C for 8h, evaporate the organic solvent under reduced pressure, obtain the crude product, then separate through column chromatography [silica gel is the carrier, chloroform and methanol mixture (1: 8, v / v) is eluent] to obtain blue crystals with a yield of 89%;

[0032] IR(KBr), υ(cm -1 ): 3359(OH), 1585, 1508(Ar), 1373(Ar-N(R 1 R)), 1168(C-O). 1 H-NMR (DMSO-d 6 , 400Hz) δ (ppm): 8.11 (d, J = 9.3Hz, 2H, Ar-H), 7.02 (d, J = 9.3Hz, 2H, Ar-H), 3.65 (t, J = 4.8Hz, 4H , OCH 2 ), 3.18(t, J=4.7Hz, 4H, NCH 2 ), 2.09 (s, 6H, CH 3 ).

[0033] (2) Preparation of chromogen standard solution

[0034] Weigh 0.0353g of N-methyl-N-hydroxyethylaniline squaraine dye a...

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Abstract

The invention relates to a method for detecting cysteine content in real time by a colorimetric method. The method comprises the following steps of: preparing a color developing agent and preparing cysteine standard solution; adding 1.0mL of the color developing agent standard solution with a diluted concentration of 1.0*10<-6>mol / L, 1.0mL of 0.01mol / L NaCl solution, 1.0mL of 0.01mol / L sodium dodecyl sulfate, 1.0mL of mixed phosphate buffer solution with a pH value of 7.5 and 1mL of a sample to be detected into a 10mL volumetric flask in turn; fixing the volume and shaking uniformly; and taking a reagent blank as reference, measuring the absorbency A of the solution at a 646-nm wavelength by using a 1-cm cuvette, recording the absorbency difference delta A, and calculating the cysteine content of the sample to be detected. The method has the advantages of simple operation, high response speed, high selectivity, high sensitivity and the like, and can be quickly and accurately applied to the quantitative measurement of cysteine concentration in synthetic samples and biological samples.

Description

technical field [0001] The invention belongs to the field of cysteine ​​content detection, in particular to a colorimetric method for real-time detection of cysteine ​​content. Background technique [0002] Cysteine ​​is a thiol-containing natural amino acid that is an important building block of most peptides and proteins. Under normal physiological conditions, due to the presence of highly reactive sulfhydryl groups, cysteine ​​plays a very important role in many biological functions, such as achieving protein intramolecular cross-linking through disulfide bonds to maintain the secondary structure of protein molecules Structure and function; at the same time, cysteine ​​plays a role in maintaining intracellular redox activity, foreign body metabolism, intracellular signal transduction and gene regulation. However, epidemiological studies have shown that changes in intracellular cysteine ​​content will be accompanied by various diseases. For example, high cysteine ​​conten...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/31
Inventor 徐洪耀严正权光善仪柯福佑
Owner DONGHUA UNIV
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