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Protamine detection method

A protamine and detection method technology, applied in the field of chemical detection, can solve problems such as detection limit and sensitivity not meeting requirements, complicated operation, etc., and achieve the effect of low detection limit, simple operation, fast and accurate detection

Active Publication Date: 2017-05-10
ANHUI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there are many detection methods for protamine, but the detection limit and sensitivity cannot meet the requirements, and the operation is complicated

Method used

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Embodiment 1

[0049] A detection method for protamine, comprising the following steps:

[0050] (1) GO–NH 2 preparation of

[0051] In this paper, graphene oxide was synthesized by low, medium and high temperature ultrasonic-assisted Hummers method:

[0052] A. Measure 23m L of concentrated sulfuric acid and pour it into a beaker, put the beaker in an ice bath and cool it down to below 3.5-4°C, weigh 1g of graphite powder and 0.5g of sodium nitrate into the beaker, mix with concentrated sulfuric acid, turn on the ultrasound, after 1h Slowly add 3g of potassium permanganate, after the addition is complete, turn off the ultrasound; control the temperature at 9°C while stirring, and react for 2 hours;

[0053] B. Then move the mixed solution prepared in step A to a water bath, turn on the ultrasound, control the temperature of the water bath at 38-39°C for 0.5h; then increase the temperature to 98°C and add 60mL of deionized water to stop the reaction;

[0054] C. Then add 25mL of hydrogen ...

Embodiment 2

[0078] Explore the effect of pH on fluorescence recovery efficiency;

[0079] Change system pH, other experimental process is with embodiment 1 step (4), and ANS-PAA final concentration is 1mg.L in the system -1 , GO-NH 2 The final concentration is 65mg L -1 , the final concentration of protamine was 2 μg mL- 1 ;

[0080] The result is as Figure 4 , 10 HEPES buffer solutions with a pH value of 2-7.5 to choose when the fluorescence recovery efficiency of the system is F / F 0 (where F 0 and F are the fluorescence intensity of ANS-PAA / GO-NH2 system and ANS-PAA / GO-NH 2 Add protamine 2μg mL to the system -1 The optimal pH value when the fluorescence intensity) is maximum, the results are as follows Figure 4 shown. It can be seen from the figure that as the pH value increases, the fluorescence recovery efficiency of the system first increases and then decreases, and the fluorescence recovery efficiency of the system is the largest at pH=5.2. Therefore, the HEPES buffer so...

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Abstract

The invention provides a protamine detection method. When amination oxidized graphene is added into an ANS-PAA system, fluorescence of ANS-PAA is quenched. After protamine is added into the system, as the protamine has strong positive charge, ANS-PAA fluorescent polymer with negative charge can be captured from an amination GO surface, the fluorescent strength of the ANS-PAA can be improved, the improved strength is in direct proportion to the concentration of the added protamine, and a method for rapidly detecting the protamine is built. Compared with the prior art, the detection method is simple to operate, low in detection limit and capable of rapidly and accurately detecting the protamine.

Description

technical field [0001] The invention belongs to the field of chemical detection, and in particular relates to a protamine detection method. Background technique [0002] In the last 20 years, with the advancement of science and technology and people's active efforts in finding natural food preservatives, protamine has attracted widespread attention because of its good antibacterial activity and natural activity. Protamine is a polycationic peptide extracted from fresh mature fish sperm. It has the characteristics of high efficiency, safety and functionality. It is a natural food preservative worth developing. [0003] As we all know, protamine has many medical applications, such as: it is used to treat bleeding caused by excessive injection of heparin, and it is currently the only drug against heparin in heart surgery with cardiopulmonary bypass. The complex of protamine and DNA-nucleosomes is considered medically to improve liver function, effectively relieve fatigue, and ...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 许美姣刘金水
Owner ANHUI NORMAL UNIV
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