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Mould and yeast microorganism limit detection method

A microbial limit, yeast technology, applied in the field of medicine, can solve the problems of accuracy interference, the total number of RS mold and yeast inspection results exceeding the standard, etc., and achieve the effect of improving accuracy and scientific quality control of product microbial limit.

Inactive Publication Date: 2018-06-12
XIAN DAQING PHARMA FACTORY JINHUA ENTERPRISE GROUP CORP
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Based on the nature of the SDA medium and the change in the counting principle of the 2015 edition of the Pharmacopoeia, the accuracy of the test results for the total number of RS molds and yeasts will be affected to a certain extent. We have done 10 groups of experiments. From the experimental results, the number of bacteria on the SDA plate accounted for More than 80% of the total number of colonies, some even as high as 100%, resulting in the total number of RS mold and yeast test results exceeding the standard

Method used

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  • Mould and yeast microorganism limit detection method
  • Mould and yeast microorganism limit detection method
  • Mould and yeast microorganism limit detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 1 Screen the concentration of gentamicin sulfate injection added in SDA

[0020] Such as figure 1 As shown, after preparing the test solution, prepare sample groups with different concentrations, wherein, each concentration group takes 1ml of the test solution, and adds 1mg / 100ml, 2mg / 100ml, 3mg / 100ml, 4mg / 100ml or 20ml of SDA medium with 5mg / 100ml gentamicin sulfate (injection) is used, and two plates are used for each concentration group. The sample group was cultured at 25° C. for 5 days, and the colony forming units (cfu / ml) of each concentration group were detected.

[0021] Mix 1ml of the test solution with 1ml of Escherichia coli CMCC (B) 44102 bacterial suspension (1:1) to obtain 2ml of an artificially polluted sample; then prepare artificially polluted sample groups with different concentration groups, each concentration group Contains 1 artificially contaminated sample, and according to different concentration requirements, add 20ml of SDA medium containing ...

Embodiment 2

[0027] Culture medium bacteriostasis test

[0028] This example mainly investigates whether SDA containing 4mg / 100mL gentamycin sulfate injection inhibits bacterial growth and can achieve the expected effect.

[0029] Such as figure 2 shown

[0030] Test group: Take 1ml of the bacterial suspensions of Staphylococcus aureus CMCC (B) 26003, Pseudomonas aeruginosa CMCC (B) 10104, and Bacillus subtilis CMCC (B) 63501 respectively, and add 4 mg / 20ml of the SDA medium of 100ml gentamicin sulfate injection was used as the test group, and two plates were selected for each group, cultured at 25°C for 5 days, and the colony forming units (cfu / ml) in each plate were detected.

[0031] Bacterial liquid group: Take 1ml of the bacterial suspensions of Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus subtilis respectively, add 20ml of SDA medium as the bacterial liquid group, select two plates for each group, and culture them at 25°C for 5 days , to detect the colony forming ...

Embodiment 3

[0036] Applicability test of counting method for the total number of mold and yeast

[0037] This example mainly investigates whether SDA containing 4mg / 100mL gentamycin sulfate (injection) inhibits bacterial growth and has inhibitory effect on mold and yeast.

[0038] Such as image 3 As shown, Candida albicans CMCC (F) 98001 (purchase 0 generation dry powder, no definite concentration) was cultured in SDB medium at 25°C for 3 days to obtain a suspension of Candida albicans CMCC (F) 98001; the concentration was made 4 cfu / ml bacterial suspension..

[0039] Aspergillus niger CMCC (F) 98003 (purchased 0 generation bacteria suspension, no definite concentration) was cultured in SDB medium at 25°C for 7 days to obtain Aspergillus niger CMCC (F) 98003 bacteria suspension; the concentration was made 4 cfu / ml bacterial suspension.

[0040] Preparation of the test solution: Weigh 10 g of proteanin yeast powder, add pH 7.0 sterile sodium chloride-peptone buffer to 100 ml, shake for 1...

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Abstract

The invention provides a mould and yeast microorganism limit detection method. The method comprises the steps of adding a tested liquid into a Sabouraud's dextrose agar culture medium, and conductingRS mould and yeast microorganism limit detection after culturing, wherein Sabouraud's dextrose agar culture medium contains gentamicin which is gentamicin sulphate, and the content of gentamicin sulphate in the Sabouraud's dextrose agar culture medium is 4 mg / 100 mL-5 mg / 100 mL. According to the mould and yeast microorganism limit detection method, the problem that due to excessive growth bacteriain the SDA culture medium, the overall quantity of RS moulds and yeasts exceeds a standard is solved by adding a gentamicin sulphate injection solution into the SDA culture medium, the accuracy of anRS mould and yeast detection result is improved, and thus microorganism limit quality control of a product is more scientific and reasonable.

Description

technical field [0001] The invention belongs to the technical field of medicine and relates to a method for detecting the limit of mold and yeast microorganisms. Background technique [0002] With the implementation of the 2015 edition of the "Chinese Pharmacopoeia", the microbial limit detection items have undergone tremendous changes. First, the medium has changed. The medium used for the detection of the total number of molds and yeasts has changed from Rose Bengal Sodium to "Sabouraud Dextrose Agar Medium (SDA) ", SDA is a broad-spectrum medium with high sensitivity, which is conducive to the growth of all surviving microorganisms in the product, thus increasing the risk of detection of microorganisms in products prone to contamination. The second is the change of the counting principle. The 2010 edition of the Pharmacopoeia stipulates that when there are molds and yeasts growing on the nutrient agar and rose bengal sodium agar medium, the results of the counting of the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/06
CPCC12Q1/06
Inventor 陈建华栗芬琴唐奕
Owner XIAN DAQING PHARMA FACTORY JINHUA ENTERPRISE GROUP CORP