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Detecting method of epsilon-polylysine

A polylysine and detection method technology, which is applied in the field of qualitative and quantitative detection of ε-polylysine, can solve the problems of poor repeatability, low sensitivity of ε-polylysine, and high cost of high-performance liquid chromatography. problems, to achieve the effect of simple operation, high sensitivity and short detection time

Inactive Publication Date: 2011-09-07
NANJING TECH UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Itzhaki colorimetric method is based on the reaction of ε-polylysine and methyl orange (an acid dye), and the content of ε-polylysine is quickly quantified by colorimetry. Although this method has a short detection time, However, the repeatability is poor, and because most of the food has certain turbidity, this method is not suitable for determining the content of ε-polylysine in food
Although high-performance liquid chromatography has good repeatability and rapid detection, due to the similar properties of ε-polylysine and protein, it is difficult to completely separate it from protein substances. The inventors found that high-performance liquid chromatography can detect ε in food -Polylysine is not very sensitive
In addition, the high-performance liquid chromatography has a high cost and is not suitable for most enterprise users.

Method used

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  • Detecting method of epsilon-polylysine
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Examples

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Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Detection of ε-polylysine content in fermentation broth

[0028] (1) Preparation of methylene blue agar plate: prepare methylene blue agar plate according to the following weight percentages: 0.002% methylene blue and 1.0% agar, and the rest is distilled water. After sterilization, take 15ml of the plate solution and add it to a sterilized Petri dish (100×15mm). After cooling and solidifying, it becomes a methylene blue agar plate.

[0029] (2) Preparation of the standard curve: Dilute the fermentation medium by 4 times with distilled water, adjust the pH to 2.0, heat-treat at 60°C for 15 minutes, and centrifuge at 10,000 rpm for 10 minutes. , 4000, 6000, 8000mg / L ε-polylysine solution, as the standard solution when detecting ε-polylysine content in the fermentation broth. Use a sterile puncher to punch holes on the methylene blue agar plate, add 50 μl of the above-mentioned standard solutions with different concentration gradients into the holes, cover the ...

Embodiment 2

[0036] Embodiment 2: the detection of ε-polylysine content in the beverage

[0037] (1) Preparation of methylene blue agar plate: prepare methylene blue agar plate according to the following weight percentage: 0.002% methylene blue and 0.75% agar, and the rest is distilled water. After sterilization, take 10ml of the plate solution and add it to a sterilized Petri dish (100×15mm). After cooling and solidifying, it becomes a methylene blue agar plate.

[0038](2) Preparation of the standard curve: Dilute the beverage without ε-polylysine by 2 times with distilled water, adjust the pH to 2.0, heat-treat at 60°C for 15 minutes, centrifuge at 10,000 rpm for 10 minutes, and use the supernatant as a solvent, and prepare the concentration of 2 , 3, 5, 10, 15, 20, 25mg / L ε-polylysine solution, as a standard solution for detecting ε-polylysine content in beverages. Place a sterile Oxford cup on the methylene blue agar plate, add 250 μl of the above-mentioned standard solutions with di...

Embodiment 3

[0045] Example 3: Detection of ε-polylysine content in solid food samples

[0046] (1) Preparation of methylene blue agar plate: prepare methylene blue agar plate according to the following weight percentage: 0.002% methylene blue and 0.75% agar, and the rest is distilled water. After sterilization, take 10ml of the plate solution and add it to a sterilized Petri dish (100×15mm). After cooling and solidifying, it becomes a methylene blue agar plate.

[0047] (2) Preparation of the standard curve: dilute the solid food without ε-polylysine 20 times with distilled water, after homogenization, adjust the pH to 2.0, heat-treat at 60°C for 15 minutes, centrifuge at 10,000rpm for 10 minutes, and use the supernatant as a solvent. Prepare ε-polylysine solutions with concentrations of 2, 3, 5, 10, 15, 20, and 25 mg / L, respectively, as standard solutions for detecting the content of ε-polylysine in solid food. Place a sterile Oxford cup on the methylene blue agar plate, add 250 μl of t...

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Abstract

The invention discloses a detecting method of epsilon-polylysine. The method comprises the following steps of: calculating a standard curve of a numerical value according to the diameter of a diffusing ring of a standard solution and the concentration of the epsilon-polylysine, then measuring the diameter of the diffusing ring generated by the sample solution, and calculating the concentration of the epsilon-polylysine in the unknown sample from the standard curve. Compared with the current colorimetric method and liquid phase method, the method of the invention is simpler in operation, lower in cost and stronger in specificity; furthermore, the purpose of fast detecting the content of the epsilon-polylysine in the fermentation liquor and detecting the content of the epsilon-polylysine in the food in high sensitivity can be realized by respectively adjusting the detecting conditions.

Description

technical field [0001] The invention relates to a qualitative and quantitative detection method of ε-polylysine. Background technique [0002] ε-polylysine (structural formula I) is a lysine polymer synthesized and secreted by actinomycetes in the metabolic process. into, so it is called ε-polylysine. ε-polylysine is one of the natural food preservatives allowed to be produced and used in the world at present, and is widely used in the preservation of pastries, beverages, aquatic products, meat products, dairy products and other foods. As a preservative, ε-polylysine has the characteristics of good water solubility, no peculiar smell, high safety, broad antibacterial spectrum, good thermal stability and wide applicable pH range. [0003] [0004] ε-polylysine is usually produced by microbial fermentation. At present, the yield of ε-polylysine in the fermentation process and the content of ε-polylysine in the product are mainly detected by Itzhaki colorimetry and high p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N11/00
Inventor 徐虹张全景冯小海李莎欧阳平凯
Owner NANJING TECH UNIV