Method for quantitatively detecting molds and saccharomycetes in eye masks

A qualitative detection and yeast technology, applied in the field of microbial detection, can solve the problems of raw and auxiliary material production environment unable to meet sterility index, enterprise storage, transportation and production efficiency problems, long period from inspection to release, and achieve high specificity. , easy operation, low detection limit effect

Inactive Publication Date: 2017-07-25
杨晓莉
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is required to control the production process in accordance with the GMP standard of pharmaceuticals. The production process and its environment are extremely strict on the control of microorganisms, so that the product can finally meet the requirements of the sterility index. However, health care products are non-sterile products, and the raw materials, auxiliary materials, production environment and their products cannot. To meet the requirements of sterility index, from the analysis of actual product production and control, eye patch health care products are not suitable for sterility testing
The medium used in the sterility test method is tryptone soy liquid medium, which cannot rule out the interference caused by bacteria, which may easily cause false positives
In addition, the sterility inspection method has an inspection cycle of 14 days, which is a long cycle, and the product has a long cycle from production, inspection to release, which brings troubles to the storage, transportation and production efficiency of the enterprise.

Method used

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  • Method for quantitatively detecting molds and saccharomycetes in eye masks
  • Method for quantitatively detecting molds and saccharomycetes in eye masks
  • Method for quantitatively detecting molds and saccharomycetes in eye masks

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Specificity Verification

[0036] Take the freshly prepared Candida albicans solution and Aspergillus niger test bacteria solution not greater than 100cfu, and at the same time add Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus subtilis with a bacterial content greater than 100cfu and mix well. The growth-promoting ability and inhibitory ability of the mold liquid medium, Bengal red agar medium, and triphenyltetrazolium chloride-Sapauer agar medium were tested respectively. At the same time, add the sample and operate with the above method to investigate the influence of the sample. The results are shown in Table 1.

[0037] Table 1 The growth-promoting ability and inhibitory ability of the culture medium

[0038]

[0039] It can be seen from Table 1 that the target bacteria grew well in the selective medium, and the interfering bacteria did not grow, and the addition of samples for interference did not affect the results. It shows that the...

Embodiment 2

[0040] Embodiment 2: detection limit verification

[0041] Sample preparation: Test group: Weigh 10g sample, add 90mL 0.9% sterile sodium chloride solution, mix well, and make 1:10 (g / g) test solution. At the same time, the Candida albicans bacteria solution and the Aspergillus niger test bacteria solution with a bacterial content of not more than 5cfu (g / mL) were added respectively, and mixed evenly. Bacterial liquid control group: add Candida albicans bacterial liquid and Aspergillus niger test bacterial liquid with a bacterial content not greater than 5cfu (g / mL) to 90mL 0.9% sterile sodium chloride solution, and mix well. Sample group: Weigh 10g sample, add 90mL 0.9% sterile sodium chloride solution to make 1:10 (g / g) test solution.

[0042] Separation of bacteria enrichment: take 10mL of the above-mentioned homogeneous solution, add 90mL mold liquid culture medium, incubate at 28°C±1°C for 3 days, take 2mL of the homogeneous solution, mix thoroughly with a vortex mixer, ...

Embodiment 3

[0047] Example 3: Reproducibility Verification

[0048] Take samples of the same batch number, and change the experimental conditions (experimental location, experimenter, instrument, medium batch) for the test. At the same time, five batches of samples were selected for testing.

[0049] Condition A: Room 910, HFsafe-1200LC biological safety cabinet, Shanghai Lishen Scientific Instrument Co., Ltd.; mold liquid culture medium, Qingdao High-tech Industrial Park Haibo Biotechnology Co., Ltd., batch number: 20160730; Bengal red agar medium, Beijing Land Bridge Technology Co., Ltd., batch number: 160628; triphenyltetrazolium chloride-Sapauer culture medium, Beijing Land Bridge Technology Co., Ltd., batch number: 161009;

[0050] Condition B: Room 910, HFsafe 760S biological safety cabinet, Shanghai Lishen Scientific Instrument Co., Ltd.; mold liquid culture medium, Guangdong Huankai Biotechnology Co., Ltd., batch number: 3104619; Bengal red agar medium, Beijing Aoboxing Biotechno...

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Abstract

The invention belongs to the field of microbial detection and particularly discloses a method for quantitatively detecting molds and saccharomycetes in eye masks; the method comprises the steps of (1) weighing eye mask, and adding diluent of suitable volume to obtain a test liquid; (2) adding a mold liquid medium of suitable volume into the test liquid, and performing enrichment culture at 28+ / - DEG C for 48-72 h to obtain homogenous liquid; (3) collecting a suitable amount of the homogenous liquid, applying respectively to rose Bengal agar plate and triphenyl tetrazolium chloride-Sabouraud agar plate, standing, and culturing at 28+ / -1 DEG C for 3-5 days; (4) judging results, to be specific, if no colonies grow on the plates or if there are colonies grown on the plates while no suspected colonies are present, determining zero match; if it is confirmed that saccharomycetes or molds grown on any one of the plates, determining that a target bacterium is detected on the sample. The method has the advantages of high specificity, low detection limit, good reproducibility, good operational convenience and the like, and meets the requirement of manufacturers for product quality control.

Description

technical field [0001] The invention belongs to the field of microorganism detection, and in particular relates to a qualitative detection method for mold and yeast in eye patches. Background technique [0002] Health-care eye patch refers to the absorption of active ingredients such as extracts of Chinese herbal medicines with auxiliary materials (such as cotton, silk, brocade, non-woven fabrics, etc.), and sticking them on or around the eyes, so that the active ingredients can be absorbed locally to improve skin health. , A product that relieves discomfort symptoms such as eye fatigue. Mold and yeast widely exist in nature, and the main growth environment of yeast is a humid or liquid environment. The liquid environment of eye patch preparations provides favorable conditions for the reproduction and growth of mold and yeast. Mold and yeast belong to the fungus phylum, and many fungi can produce toxins. The use of polluted and deteriorated health care eye patches will caus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12R1/685C12R1/725
CPCC12Q1/04G01N2333/38G01N2333/40
Inventor 杨晓莉李翠周志云杨静李林樾
Owner 杨晓莉
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