HPLC detection method of nisin

A technology of nisin and nisin, which is applied in the field of high-performance liquid chromatography detection of nisin, and can solve the problems of reproducibility, poor accuracy, long time course and difficult integration of sample determination results

Inactive Publication Date: 2010-07-14
SHANGHAI INST OF PHARMA IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The time course of analyzing samples by this method is long, nearly 1 hour, and the gradient elution baseline drifts, which makes integration difficult, and the reproducibility and accuracy of sample determination results are poor.
The inventor does not have the target peak in the HPLC spectrum by this method

Method used

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  • HPLC detection method of nisin
  • HPLC detection method of nisin
  • HPLC detection method of nisin

Examples

Experimental program
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Effect test

preparation example Construction

[0065] Solution preparation

[0066] Reference substance solution: Accurately weigh about 10 mg of Nisin reference substance, put it in a 10ml volumetric flask, dissolve it with 0.02N HCl and dilute to the mark, and use it as the reference substance solution.

[0067] Sample solution: Scrape Lactococcus lactis ssp.lactis from the inclined surface and insert it into the seed solution (the formula is 0.5% soybean protein isolate, 0.5% peptone, 0.25% yeast extract, 0.5% beef extract, 0.5% glucose , ascorbic acid 0.05%, Na 2 HPO 4 12H 2 O 1.0%, MgSO 4 ·7H 2 O 0.012%, pH6.8) at 37°C for 20h. Insert the seed liquid into the fermentation broth (the formula of the medium is: 2.0% sucrose, 3.0% yeast extract, 2.0% peptone, 0.02% magnesium sulfate, 1.0% potassium dihydrogen phosphate, 0.2% NaCl, Tween-800.5%, pH 6.8) Ferment at 30°C for 12-14 hours, stop the fermentation, take a certain amount of Nisin fermentation broth, adjust the pH to 2.0 with 0.02NHCL, centrifuge at 12000-140...

Embodiment 1

[0069] Initial mobile phase B: mobile phase A=20:80 isocratic elution for 3 minutes, then mobile phase B continuously increased from 20% to 40% elution for 27 minutes, and mobile phase B returned to 20% in the last 2 minutes ,balance. Under this method, the retention time of the reference substance is 16.38min, the resolution is 1.08, and the number of theoretical plates reaches 16738. See figure 2 with image 3 .

Embodiment 2

[0071] Initial mobile phase B: mobile phase A=20:80 isocratic elution for 3 minutes, then mobile phase B continuously increased from 20% to 40% for 12 minutes, and mobile phase B returned to 20% in the last 2 minutes ,balance. Under this method, the peak eluting time is about 10 minutes, the baseline drifts, and the number of theoretical plates is 12800.

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Abstract

The invention discloses a High Perfomance Liquid Chromatography (HPLC) detection method of nisin, the flowing phase contains acetonitrile and water, the initial concentration of the acetonitrile is 5-25 vol.% while the final concentration thereof is 35-100 vol.%.

Description

technical field [0001] The invention relates to the detection of target products in fermentation broth, in particular to a high performance liquid chromatography (High Performance Liquid Chromatography, HPLC) detection method for nisin. Background technique [0002] Nisin (Nisin, also known as lactin or nisin) is a small molecule peptide with strong antibacterial effect synthesized and secreted by Lactococcus lactis ssp. , One of the widely used bacteriocins. Nisin is currently one of the few natural food preservatives allowed to be produced commercially in the world. It is widely used in the preservation and preservation of dairy products, fermented beverages, canned foods, meat and meat products. It can effectively inhibit food spoilage. Gram-positive bacteria (G + ) and the reproduction of pathogenic bacteria. Compared with other preservatives, Nisin is produced by the naturally occurring Lactococcus bacteria and is particularly sensitive to proteases, and is quickly b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/34
Inventor 蒋家琛赵文杰魏晓东顾敏程晴华赵波
Owner SHANGHAI INST OF PHARMA IND CO LTD
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