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Method for highly-efficient detection of gas-producing bacteria in vinegar by combining bromcresol purple and Durham tube

A bromocresol violet and Duchenne tubule technology is applied in the directions of biochemical equipment and methods, and the determination/inspection of microorganisms. The effect of qualified products entering the market, reducing economic losses and simple operation steps

Inactive Publication Date: 2018-12-18
四川清香园调味品股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Vinegar is a condiment produced from rice through cooking, koji making, saccharification, alcoholic fermentation and other processes. Organic acids also contain amino acids, sugars, lipids, inorganic salts, vitamins, etc., so they provide good carbon sources, nitrogen sources, inorganic salts and other rich nutrients for the reproduction of microorganisms. In the process, there are still high-temperature-resistant and anaerobic gas-producing bacteria that survive, and the reproduction of gas-producing bacteria will cause food packaging bottles to swell, causing problems such as swell in the vinegar production process, so it is necessary to establish a rapid detection of food The detection method of gas-producing bacteria in vinegar minimizes the risk of vinegar quality in enterprises
[0003] The existing industry technology is that the MRS method is used as a culture medium to detect aerogenes. This is a general method, but it is not suitable for vinegar. With this method, the aerogenes in vinegar cannot be detected, and the production cannot be found in time. The unqualified products on the market lead to the phenomenon of bulging bottles and bags on the market
[0004] Chinese patent application document CN104313112A discloses a culture medium and a detection method for detecting gas-producing and film-forming bacteria in vinegar. The liquid culture medium is made from raw materials such as potatoes, glucose, yeast extract, beef extract, and ammonium sulfate. The test tube with Duchenne tubules is inoculated with the vinegar sample to be tested and then incubated for 72 hours for observation. It can detect the gas-producing and film-producing bacteria in the vinegar to a certain extent, but the incubation time is long, the efficiency is not high enough, and the production cannot be fully detected. Gas bacteria, when the content of gas-producing bacteria in vinegar is low, it cannot be detected, so the test results are not accurate enough
This method can accurately detect whether there are aerogenes in soy sauce in a relatively fast time, but when this method is used for the detection of aerogenes in vinegar, it is found that the detection limit is relatively high. When the content is low, it cannot be detected, which will affect the accurate judgment of the quality of vinegar. Based on this, it is necessary to establish a method that can quickly detect gas-producing bacteria in vinegar

Method used

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  • Method for highly-efficient detection of gas-producing bacteria in vinegar by combining bromcresol purple and Durham tube
  • Method for highly-efficient detection of gas-producing bacteria in vinegar by combining bromcresol purple and Durham tube
  • Method for highly-efficient detection of gas-producing bacteria in vinegar by combining bromcresol purple and Durham tube

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Raw materials and preparation of medium

[0039] Weigh 2g of beef extract, 4g of peptone, 0.58g of magnesium sulfate, 1.60g of ammonium sulfate, and 0.25g of dipotassium hydrogen phosphate, add these raw materials into a beaker, add distilled water and stir to dissolve, then set the volume to 1000mL, add 2mL of bromocresol purple to indicate agent, that is, the target medium. Another 13.5g of glucose was weighed and adjusted to 100mL with distilled water to prepare a glucose solution with a mass content of about 13.5% and placed in a conical flask for later use.

[0040] 2. Detection method

[0041] Distribute the prepared target medium into test tubes with Duchenne tubules, 8mL per tube, wipe the edge of the tube opening after removing the air bubbles in the Duchenne tubules, wrap and seal them, and sterilize at 0.1MPa at 121°C for 25min. At the same time, the glucose solution in the Erlenmeyer flask was also sterilized under the same conditions to obtain a steril...

Embodiment 2

[0049] 1. Raw materials and preparation of medium

[0050] Weigh 2g of beef extract, 4g of peptone, 0.58g of magnesium sulfate, 1.60g of ammonium sulfate, 0.25g of dipotassium hydrogen phosphate, and 13.5g of glucose into a beaker, stir and dissolve these raw materials with distilled water, and then set the volume to 1000mL, add 2mL of methyl bromide Phenol purple indicator, that is, the culture medium.

[0051] 2. Detection method

[0052] Distribute the prepared medium into test tubes with Duchenne tubules, 8 mL per tube, wipe the edge of the tube opening after removing the air bubbles in the Duchenne tubules, wrap and seal them, and sterilize at 0.1 MPa at 121°C for 25 minutes. Glucose and other nutrients are mixed together for sterilization.

[0053] Take 4 test tubes from the test tubes filled with the purpose culture medium, including one test tube for the blank sample and 3 test tubes for each test sample, and add 1 mL of the original sample to be tested prepared in E...

Embodiment 3

[0062] 1. Raw materials and preparation of medium

[0063] Weigh 2g of beef extract, 4g of peptone, 0.58g of magnesium sulfate, 1.60g of ammonium sulfate, 0.25g of dipotassium hydrogen phosphate and 13.5g of glucose into a beaker, stir and dissolve these raw materials with distilled water, and then set the volume to 1000mL, add 2mL of methyl bromide Phenol purple indicator, that is, the culture medium.

[0064] 2. Detection method

[0065] Distribute the prepared medium into test tubes with Duchenne tubules, 8 mL per tube, wipe the edge of the tube opening after removing the air bubbles in the Duchenne tubules, wrap and seal them, and sterilize at 0.1 MPa at 121°C for 25 minutes. Glucose and other nutrients are mixed together for sterilization.

[0066] Get 10ml of vinegar solution (different batch from Example 1) in a beaker, adjust its pH to 6.5-7.0 with saturated sodium carbonate solution to obtain the original sample to be tested.

[0067] Take 4 test tubes from the tes...

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Abstract

The invention discloses a method for highly-efficient detection of gas-producing bacteria in vinegar by combining bromcresol purple and a Durham tube. According to the invention, a culture medium suitable for the growth and reproduction of the gas-producing bacteria in the vinegar is prepared; a bromocresol purple indicator is added into the culture medium; in addition, a glucose solution with a content approximate to 13.5% is prepared; the culture medium and the glucose solution are separately sterilized and then combined for culturing of the gas-producing bacteria; after the pH value of a to-be-detected vinegar liquid is adjusted to 6.5 to 7.0, the to-be-detected vinegar liquid is inoculated onto the culture medium; observation is performed after culturing at 33+ / -1 DEG C for 48+ / -2 h,;if a culture solution is yellow, or air bubbles are generated in the Durham tube, or the culture solution is turbid, the gas-producing bacteria are proved to exist in a detected original sample; and if the culture solution is purple, and no air bubbles are generated in the Durham tube, the gas-producing bacteria are proved to not exist in the detected original sample. The method provided by the invention can accurately distinguish the content level of the gas-producing bacteria in the detected vinegar liquid, can accurately judge whether a vinegar liquid with low content of the gas-producing bacteria contains the gas-producing bacteria or not, and is accurate in detection results.

Description

technical field [0001] The invention relates to the field of vinegar quality detection, and more specifically, the invention relates to a method for efficiently detecting gas-producing bacteria in vinegar by combining bromocresol violet and Duchenne's tubules. Background technique [0002] Vinegar is a traditional condiment in various major Chinese cuisines. According to the literature records, the history of making vinegar is about 3,000 years. Vinegar not only has a sour taste, but also has a certain umami taste, sweet taste and aroma. It can increase appetite, help digestion and has good health care functions, preventing and curing many diseases. Vinegar is a condiment produced from rice through cooking, koji making, saccharification, alcoholic fermentation and other processes. Organic acids also contain amino acids, sugars, lipids, inorganic salts, vitamins, etc., so they provide good carbon sources, nitrogen sources, inorganic salts and other rich nutrients for the rep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04
CPCC12Q1/04C12Q1/045
Inventor 甘学锋马益孙洪瑶杨莉陈娜谭檑文旗
Owner 四川清香园调味品股份有限公司
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