R2A agar medium (ready to use bagged) (200ml)

R2A agar medium (ready to use bagged) (200ml)(Pharmacopoeia) Usage:Used for the determination of total bacterial count in purified water (EP, USP, Chp standards)

Ingredients (g/L)

Inspection principle:

Yeast extract powder, peptone, and acid protein hydrolysate provide nitrogen sources, vitamins, and growth factors; Glucose is a fermentable sugar; Soluble starch can absorb harmful substances produced by bacteria during the recovery process; Sodium pyruvate can enhance bacterial recovery. Potassium dihydrogen phosphate is an acid-base buffering agent; Anhydrous sulfuric acid provides divalent cations; Agar is a coagulant.

Usage:

Step 1 [Re solution Heating]：Immerse the unopened HBBD culture medium (retaining the outer packaging bag) in a 100 ℃ boiling water bath.

Heating method:

Recommended to useInduction cooker+stainless steel pot/stainless steel bucketWait for the container (add water until the bag is fully submerged).Or use a laboratory water bath (note slow heating, it is recommended to preheat in advance)

·Container during heatingDo not fully sealTo avoid leakage of liquid inside the bag due to increased pressure.

·OptionalPerforatedContainer lid, orLeave some gapsExhaust - see video for details.

Step 2 [Dissolve]：After heating continuously for 20-30 minutes, gently shake the bag to confirm that the culture medium is completely dissolved (uniform liquid without clots).

Step 3 [Cooling]：Take out the bag and let it cool naturally to 45 ℃~50 ℃ (slightly hot to the touch but not too hot).

Step 4 ：In a sterile environment, tear open the outer packaging, unscrew the bottle cap, slowly pour the culture medium into a sterile petri dish, let it stand and solidify before use.

Storage conditions and expiration date

1.2-25℃ 保存。

2. This product is valid for six months under storage conditions.

Usage

Place the complete packaging of this product in a 100 ℃ boiling water bath and heat it for 20-30 minutes. After the culture medium is completely dissolved and cooled to 45 ℃~50 ℃, tear off the outer packaging and unscrew the lid in a sterile environment. Pour the culture medium into a sterile petri dish and let it solidify before use.

Precautions

If there is pollution, damage, discoloration, leakage, etc., it cannot be used.

During the water bath heating process, there is no need to open the outer packaging. It is normal for a small amount of gas inside the product to expand when heated.

3. After use, the culture medium should be disinfected.

4. The usage method of this product is not mandatory. If the user has specific reference materials, corresponding adjustments can be made.

Microbial detection methods and quality standards for purified water in different pharmacopoeias

Pharmacopoeia of the People's Republic of China

【 Inspection 】 Microbial limit: Take not less than 1ml of this product, process it by membrane filtration method, use R2A agar medium, culture at 30-35 ℃ for not less than 5 days, and check according to the general rule 1105. The total number of aerobic bacteria in every 1ml of the test sample should not exceed 100cfu.

Operation description:

Except for glucose and agar, take the above ingredients, mix them, dissolve at a low temperature, adjust the pH value to 7.2 ± 0.2 after heating at 25 ℃, add agar, heat and dissolve, then add glucose, shake well, pack and sterilize. The suitability test of R2A agar medium was conducted according to the suitability test method of pancreatic casein soy peptone agar medium in General Rule 1105 for "suitability test of counting medium". The test strains were Pseudomonas aeruginosa and Bacillus subtilis. Should comply with regulations.

USP Standard

In the United States Pharmacopeia, it is recommended to use 0.45 μ m membrane filtration method and simultaneously cultivate high nutrient medium (Plate Counting Agar (PCA)) at 30-35 ℃ for more than 72 hours and low nutrient medium (R2A agar) at 20-25 ℃ for more than 96 hours. The detection standard is: ≤ 100cfu/ml.

Operation description: (can be adjusted according to actual situation)

Take 10ml of thoroughly mixed water sample and add it to the installed membrane filter through aseptic operation for filtration. Wash the filter membrane with sterile sodium chloride peptone buffer at pH 7.0, with a volume of 100ml per wash per membrane, for a total of 1 wash. After filtration, remove the filter membrane and place it face up on an agar plate. Two filter membranes were prepared, one of which was attached to a agar plate (high nutrient medium) for counting and placed upside down in a biochemical incubator at 30 ° C~35 ° C for 2-3 days for counting. Another filter membrane was attached to an R2A agar plate (low nutrient medium) and incubated upside down in a biochemical incubator at 20 ℃~25 ℃ for 5-7 days, counting. Report the microbial count in 1ml of purified water based on the proportion of bacterial growth on the culture medium plate as the detection result.

EP standard

In the European Pharmacopoeia, membrane filtration method (0.45 μ m pore size filter membrane) is used, and R2A agar medium is used to culture at 30 ° C to 35 ° C for more than 5 days. Quality standard: ≤ 100cfu/mL.

Operation description: (can be adjusted according to the actual situation)

Take 10ml of thoroughly mixed water sample and add it to the installed membrane filter through aseptic operation for filtration. Wash the filter membrane with sterile sodium chloride peptone buffer at pH 7.0, with a volume of 100ml per wash per membrane, for a total of 1 wash. After filtration, remove the filter membrane and place it upside down on an R2A agar plate. Incubate it upside down in a biochemical incubator at 30 ° C to 35 ° C for 5 days and count. Convert the microbial count to 1ml of purified water in proportion and report.

R2A agar medium (ready to use bagged) (200ml)(Pharmacopoeia) Microbial Sensitivity Test:

Prepare the culture medium according to the label instructions, inoculate the following quality control strains, and incubate at 30-35 ℃ for 18 hours under aerobic conditions.