Preparation method for N-methylisatoic anhydride rapidly-labeled polysaccharide

A technology of methyl isatonic anhydride and polysaccharide, which is applied in the field of fluorescent labeling, can solve problems such as adverse reactions and inconvenient polysaccharides, and achieve the effects of low price, short reaction time and mild reaction conditions.

Inactive Publication Date: 2015-04-22
NANCHANG UNIV
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, because radioactive isotopes will cause adverse reactions in experimental organisms, it is not convenient to apply to the activity research of polysaccharides.

Method used

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  • Preparation method for N-methylisatoic anhydride rapidly-labeled polysaccharide
  • Preparation method for N-methylisatoic anhydride rapidly-labeled polysaccharide
  • Preparation method for N-methylisatoic anhydride rapidly-labeled polysaccharide

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Embodiment 1

[0021] (1) Sample treatment: Dissolve 5-10 mL of 0.1 mol / L, pH 8.0 borate buffer solution and 50% w / v dimethyl sulfoxide solution to prepare a 40-80 mg / mL gum arabic solution.

[0022] (2) Preparation of fluorescent labeling stock solution: N-methylisatoic anhydride was dissolved in dimethyl sulfoxide to prepare a 20 mg / mL solution.

[0023] (3) Add the prepared fluorescent labeling stock solution to the gum arabic solution according to the mass ratio of the fluorescent label N-methyl isatoic anhydride to gum arabic at a ratio of 1:50 and incubate at 35°C for 15 minutes.

[0024] (4) Separation of fluorescently labeled polysaccharides: N-methylisatoic anhydride-labeled gum arabic was separated by organic solvent precipitation, absolute ethanol was added to the labeled polysaccharide solution, and polysaccharides were separated by alcohol precipitation. Fluorescence-labeled polysaccharide solution: volume ratio of absolute ethanol 1:4, centrifuge the alcohol-precipitated polysa...

Embodiment 2

[0027] (1) Sample treatment: Dissolve 5-10 mL of 0.1 mol / L, pH 8.0 borate buffer solution and 50% w / v dimethyl sulfoxide solution to prepare a 40-80 mg / mL maltodextrin solution.

[0028] (2) Preparation of fluorescent labeling stock solution: N-methylisatoic anhydride was dissolved in dimethyl sulfoxide to prepare a 20 mg / mL solution.

[0029] (3) Add the prepared fluorescent marker stock solution to the maltodextrin solution at a mass ratio of 1:55 of the fluorescent marker N-methylisatoic anhydride to maltodextrin and incubate at 35°C for 18 minutes.

[0030] (4) Separation of fluorescently labeled polysaccharides: N-methylisatoic anhydride-labeled gum arabic was separated by organic solvent precipitation, absolute ethanol was added to the labeled polysaccharide solution, and polysaccharides were separated by alcohol precipitation. Fluorescence-labeled polysaccharide solution: volume ratio of absolute ethanol 1:6, centrifuge the alcohol-precipitated polysaccharide at 6000×g ...

Embodiment 3

[0033] (1) Sample treatment: Dissolve 5-10 mL of 0.1 mol / L, pH 8.0 borate buffer solution and 50% w / v dimethyl sulfoxide solution to prepare a 40-80 mg / mL maltodextrin solution.

[0034] (2) Preparation of fluorescent labeling stock solution: N-methylisatoic anhydride was dissolved in dimethyl sulfoxide to prepare a 20 mg / mL solution.

[0035] (3) Add the prepared fluorescent labeling stock solution to the maltodextrin solution according to the mass ratio of fluorescent label N-methylisatoic anhydride to maltodextrin of 1:60 and incubate at 35°C for 20 minutes.

[0036] (4) Separation of fluorescently labeled polysaccharides: N-methylisatoic anhydride-labeled gum arabic was separated by organic solvent precipitation, absolute ethanol was added to the labeled polysaccharide solution, and polysaccharides were separated by alcohol precipitation. Fluorescence-labeled polysaccharide solution: volume ratio of absolute ethanol 1:8, alcohol-precipitated polysaccharides were centrifuge...

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Abstract

A preparation method for N-methylisatoic anhydride rapidly-labeled polysaccharide belongs to the technical field of fluorescent labeling. The preparation method employs a fluorescence tracer N-methylisatoic anhydride for labeling polysaccharide, and the method comprises: performing incubating on a N-methylisatoic anhydride stock solution and a polysaccharide solution under the condition of 35 DEG C for 15-20 min, and performing separation purification on the fluorescence-labeled polysaccharide through an organic solvent precipitation process. The reaction is mild in reaction conditions and short in reaction time, and the fluorescence-labeled polysaccharide separation purification process is simple. Polysaccharide labeled by fluorescence can be detected by using a routine fluorescence detector, and research on polysaccharide functions and mechanism is facilitated.

Description

technical field [0001] The invention belongs to the technical field of fluorescent labeling, and in particular relates to a preparation method for rapidly labeling polysaccharides with N-methylisatoic anhydride. technical background [0002] In life sciences, polysaccharides are indispensable biomacromolecules, but their study is hampered by the lack of luminescent groups that make them difficult to detect. Fluorescent markers can chemically bind their fluorescent luminescent groups to sugar chain molecules, so that the polysaccharides have fluorescent groups, which is convenient for the detection of polysaccharides by conventional detection methods, and is conducive to further research on their mechanism of action. [0003] According to literature reports, radioactive isotopes are often used to label polysaccharides, so as to trace the biological activity of polysaccharides, and then study their mechanism of action. However, because radioactive isotopes can cause adverse r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N21/64
CPCG01N33/52G01N21/6486
Inventor 涂宗财李如一石燕王辉杨文华李雪常海霞
Owner NANCHANG UNIV
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