Method for determining antibacterial mechanism of zeolite molecular sieve antibacterial agent by high performance liquid chromatography

A technology of zeolite molecular sieve and antibacterial agent, which is applied in the biological field, can solve the problems of slow analysis of antibacterial mechanism and high material consumption, and achieve the effects of high sensitivity, simple operation, and less sample consumption

Active Publication Date: 2017-09-19
GUILIN UNIVERSITY OF TECHNOLOGY
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Problems solved by technology

The traditional antibacterial mechanism analysis is slow and...

Method used

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  • Method for determining antibacterial mechanism of zeolite molecular sieve antibacterial agent by high performance liquid chromatography
  • Method for determining antibacterial mechanism of zeolite molecular sieve antibacterial agent by high performance liquid chromatography
  • Method for determining antibacterial mechanism of zeolite molecular sieve antibacterial agent by high performance liquid chromatography

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Experimental program
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Embodiment 1

[0021] Embodiment 1: zeolite molecular sieve antimicrobial agent is to escherichia coli bacteriostasis mechanism

[0022] (1) Amplification of Escherichia coli strains: Prepare 200 mL of liquid medium, adjust the pH to 7.0-7.2, and sterilize with high-pressure steam. Then inoculate the bacteria according to the amount of inoculating one ring per 100mL and shake well. Transfer to a shaking incubator at 37°C for 24 hours.

[0023] (2) Prepare zeolite molecular sieve antibacterial agent and potassium diformate solution with a concentration of 0mg / mL, 6mg / mL, 12mg / mL, 24mg / mL, 48mg / mL, 96mg / mL containing potassium diformate, each 3 Group.

[0024] (3) Expanded culture and antibacterial experiment: According to (2), after adding 3 blank control groups, a total of 21 100mL ordinary liquid culture media need to be prepared and sterilized. Then inoculate (1) bacterial suspension at 2 mL per bottle, add 100 mg of cefoxitin, transfer to a shaking incubator at 37°C for a total of 12 h...

Embodiment 2

[0029] Embodiment 2: zeolite molecular sieve antibacterial agent is to Staphylococcus aureus bacteriostasis mechanism

[0030] (1) Prepare potassium diformate solutions with concentrations of 0 mg / mL, 6 mg / mL, 12 mg / mL, 24 mg / mL, 48 mg / mL and 96 mg / mL, 3 groups for each.

[0031] (2) According to the number of groups in (1), add 3 blank control groups, prepare 100 mL of ordinary liquid medium each, and sterilize. Then add 2 mL of Staphylococcus aureus suspension in the logarithmic growth phase, and then add (1) 5 mL of antibacterial agent, mark it and transfer to 37°C for shaking culture for 18-24 hours. Take the Staphylococcus aureus suspension cultured for 24 hours and dilute it 100 times with liquid medium, continue shaking culture at 37°C for 12-18 hours, grow until the OD600 is about 3.6, take it out and centrifuge at 5000r / min for 15 minutes. Add 0.05mol / L pH 7.0 potassium phosphate buffer solution to the precipitate, and then sonicate (120W 15min). Centrifuge the bact...

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Abstract

The invention discloses a method for determining an antibacterial mechanism of a zeolite molecular sieve antibacterial agent by high performance liquid chromatography. The zeolite molecular sieve antibacterial agent is taken as a research object, escherichia coli and staphylococcus aureus are taken as indicator bacteria, and content changes of staphylococcus aureus mycoprotein and content of beta-lactamase in escherichia coli after action of the antibacterial agent are determined by means of high performance liquid chromatography to realize expounding of the antibacterial mechanism of the zeolite molecular sieve antibacterial agent. The method is simple in operation, high in sensitivity, low in sample consumption and quick in detection. Along with increase of the concentration of the antibacterial agent, HPLC (high performance liquid chromatography) peak area is reduced, and protein content is decreased, so that the bacterial structure is changed and even completely damaged, bacteria fail to grow normally since cell metabolism and growth are affected, and bacterium inhibition and killing are realized.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for measuring the antibacterial mechanism of a zeolite molecular sieve antibacterial agent by using high-performance liquid phase. Background technique [0002] Nowadays, the development of antibacterial technology is very fast. For the endless antibacterial substances, we need to study their antibacterial mechanism one by one. The traditional antibacterial mechanism analysis is slow and consumes a lot of materials, which needs to be optimized urgently. Therefore, in the study of the antibacterial mechanism of antibacterial agents, we conduct research on the proteins required for bacterial metabolism, establish a method combined with high-performance liquid phase analysis of protein content, and study the impact of antibacterial agents on the structure and quantity of proteins contained in bacteria from the side. Contents of the invention [0003] The invention utilizes a h...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 王秀丽沈阳阳陈南春李子院李海云
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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