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Detection method for in-situ measuring content of phenyllactic acid in fermentation samples

A medium phenyllactic acid and detection method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low sensitivity, small capillary diameter, inconvenient on-site detection, etc., and achieve the effect of high detection sensitivity

Inactive Publication Date: 2011-09-07
HENAN BUSINESS SCI RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Capillary electrophoresis uses a small capillary diameter, which tends to make the optical path too short, and the sensitivity is low, and it will change due to the composition of the sample during electroosmosis, which will affect the reproducibility of the results
Due to the limitation of equipment, high performance liquid phase method is not convenient for on-site detection
There is no literature report on the use of gas chromatography to detect the content of phenyllactic acid in fermented products

Method used

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  • Detection method for in-situ measuring content of phenyllactic acid in fermentation samples
  • Detection method for in-situ measuring content of phenyllactic acid in fermentation samples
  • Detection method for in-situ measuring content of phenyllactic acid in fermentation samples

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1 phenyllactic acid standard curve

[0023] A standard solution of phenyllactic acid with a concentration of 2 mg / mL was prepared with phenyllactic acid produced by Alfa Aesar. Take 2mL of the standard solution and heat it to remove water, add 0.3mL of 3.0mol / L concentrated hydrochloric acid-n-butanol mixed solution, shake it and heat it to 70°C for 15 min, N 2 Blow dry the remaining solution. Then add 0.3 mL of trifluoroacetic anhydride-ethyl acetate mixed solution with a volume ratio of 0.3:1, shake it and heat it to 80°C for 15 min in an airtight manner. 2 Blow dry the remaining solution. Add 1 mL of ethyl acetate, shake and centrifuge for 5-7 minutes, take the supernatant and dilute it with ethyl acetate to a concentration of 100, 400, 700, 1000, 1400 μg / mL (based on phenyllactic acid), and take 1 μl (that is, phenyllactic acid Contents were 100, 400, 700, 1000, 1400ng) sample injection, the gas chromatographic results of the first standard (that is, c...

Embodiment 2

[0028] Example 2 Take the on-site detection of the solid-state fermentation sample a of Mugoniella as an example.

[0029] Sample pretreatment: take 2g of pulverized Mugnittum solid-state fermentation sample, add 2mL of deionized water, shake it upside down several times to fully mix with water, heat and evaporate to dryness, add 3mL of deionized water for shaking (for Dissolve phenyllactic acid) and then centrifuge, take the supernatant and heat to remove water, add 0.3mL of 3.0mol / L concentrated hydrochloric acid-n-butanol mixed solution, vibrate, seal and heat to 70°C for 15 min, N 2 Blow dry the remaining solution. Then add 0.3 mL of trifluoroacetic anhydride-ethyl acetate mixed solution with a volume ratio of 0.3:1, shake it and heat it to 80°C for 15 min in an airtight manner. 2 Blow dry the remaining solution. Add 1 mL of ethyl acetate, shake and centrifuge for 6 min, take the clear and transparent supernatant for gas chromatography analysis, and the injection volume ...

Embodiment 3

[0033] Example 3 Take the on-site detection of Lactobacillus plantarum liquid fermentation sample b as an example.

[0034] Sample pretreatment: Take 2 mL of liquid fermentation sample, heat and evaporate to dryness, add deionized water to shake and dissolve, then centrifuge, take the supernatant and heat to remove water, add 0.3 mL of 3.0 mol / L concentrated hydrochloric acid-n-butanol mixed solution, shake and heat to 70°C for 15 minutes, N 2 Blow dry the remaining solution. Then add 0.3 mL of trifluoroacetic anhydride-ethyl acetate at a volume ratio of 0.3:1, shake and heat in a sealed container to 80°C for 15 min, N 2 Blow dry the remaining solution. Add 1mL of ethyl acetate, shake and centrifuge for 6min, take the clear and transparent supernatant for gas chromatography analysis, the injection volume is 1μl.

[0035] Chromatographic conditions: Chromatographic column DB-1, quartz capillary column with an inner diameter of 15m×0.53mm and a film thickness of 3.0μm, coated w...

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Abstract

The invention relates to a detection method for in-situ measuring content of phenyllactic acid in fermentation samples, wherein a sample is measured by gas chromatography after the sample is pre-processed; the pre-processing of the sample comprises the steps of: evenly mixing a liquid fermentation sample or a smashed solid fermentation sample with deionized water; heating up the sample for evaporating the sample to dryness; adding the deionized water into the sample and oscillating, and centrifuging the solution; heating up the supernatant for removing water; adding 0.2-0.5mL of 3.0-3.4 mol / L concentrated hydrochloric acid-normal butanol mixed liquor; maintaining solution at 65-72 DEG C for 12-15 minutes; drying the solution by nitrogen; adding 0.2-0.5mL of a mixed solution of trifluoroacetic anhydride and ethyl acetate mixed solution at a ratio of 0.15-0.3:1; maintaining the temperature of the solution at 78-85 DEG C for 12-15 minutes; drying the solution by nitrogen; dissolving by ethyl acetate; and centrifuging and taking the supernatant for detection. The detection sensitivity of the method is high; the lowest detection limit of phenyllactic acid is 4.2-7g / mL.

Description

technical field [0001] The invention belongs to the technical field of detection of biological preservatives, and in particular relates to a detection method suitable for measuring the content of phenyllactic acid in fermented samples on the industrial site. Background technique [0002] Phenyllactic acid (PLA), also known as 3-phenyllactic acid or β-phenyllactic acid, that is, 2-hydroxy-3-phenylpropionic acid, is a new type of small molecule produced by microorganisms such as lactic acid bacteria discovered in recent years. Antibacterial substances have a strong antibacterial effect on a variety of food-borne pathogens, spoilage bacteria that cause food deterioration, including toxin-producing filamentous fungi. Phenyllactic acid is a new type of natural and broad-spectrum biological food preservative. [0003] The food preservatives used by most food companies are still chemical preservatives represented by sodium benzoate and potassium sorbate. However, due to the conti...

Claims

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Application Information

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IPC IPC(8): G01N30/12G01N30/28
Inventor 王永李向力王法云田春华吕微
Owner HENAN BUSINESS SCI RES INST
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