抗生素的微生物检测
2008-9-25 11:38:21 青岛海博
ANTIBIOTICS—MICROBIAL ASSAYS (抗生素的微生物检测)
MEDIA AND DILUENTS(培养基和稀释剂)
Media
The media required for the preparation of test organism inocula are made
from the ingredients listed herein. Minor modifications of the individual
ingredients, or reconstituted dehydrated media, may be substituted, provided
the resulting media possess equal or better growth-promoting properties and
give a similar standard curve response.
Dissolve the ingredients in water to make 1 L, and adjust the solutions
with either 1 N sodium hydroxide or 1 N hydrochloric acid as required, so that
after steam sterilization the pH is as specified.
MEDIUM 1
Peptone 6.0 g
Pancreatic Digest of Casein 4.0 g
Yeast Extract 3.0 g
Beef Extract 1.5 g
Dextrose 1.0 g
Agar 15.0 g
Water 1000 mL
pH after sterilization: 6.6 ± 0.1.
MEDIUM 2
Peptone 6.0 g
Yeast Extract 3.0 g
Beef Extract 1.5 g
Agar 15.0 g
Water 1000 mL
pH after sterilization: 6.6 ± 0.1.
MEDIUM 3
Peptone 5.0 g
Yeast Extract 1.5 g
Beef Extract 1.5 g
Sodium Chloride 3.5 g
Dextrose 1.0 g
Dibasic Potassium Phosphate 3.68 g
Monobasic Potassium Phosphate 1.32 g
Water 1000 mL
pH after sterilization: 7.0 ± 0.05.
MEDIUM 4
Same as Medium 2, except for the additional ingredient 1.0 g of Dextrose.
MEDIUM 5
Same as Medium 2, except that the final pH after sterilization is 7.9 ± 0.1.
MEDIUM 8
Same as Medium 2, except that the final pH after sterilization is 5.9 ± 0.1.
MEDIUM 9
Pancreatic Digest of Casein 17.0 g
Papaic Digest of Soybean 3.0 g
Sodium Chloride 5.0 g
Dibasic Potassium Phosphate 2.5 g
Dextrose 2.5 g
Agar 20.0 g
Water 1000 mL
pH after sterilization: 7.2 ± 0.1.
MEDIUM 10
Same as Medium 9, except to use 12.0 g of Agar instead of 20.0 g, and to add 10 mL of Polysorbate 80 after boiling the medium to dissolve the agar.
pH after sterilization: 7.2 ± 0.1.
MEDIUM 11
Same as Medium 1, except that the final pH after sterilization is 8.3 ± 0.1.
MEDIUM 13
Dextrose 20.0 g
Peptone 10.0 g
Water 1000 mL
pH after sterilization: 5.6 ± 0.1.
MEDIUM 19
Peptone 9.4 g
Yeast Extract 4.7 g
Beef Extract 2.4 g
Sodium Chloride 10.0 g
Dextrose 10.0 g
Agar 23.5 g
Water 1000 mL
pH after sterilization: 6.1 ± 0.1.
MEDIUM 32
Same as Medium 1, except for the additional ingredient 0.3 g of Manganese Sulfate.
MEDIUM 34
Glycerol 10.0 g
Peptone 10.0 g
Beef Extract 10.0 g
Sodium Chloride 3.0 g
Water 1000 mL
pH after sterilization: 7.0 ± 0.1.
MEDIUM 35
Same as Medium 34, except for the additional ingredient 17.0 g of Agar.
MEDIUM 36
Pancreatic Digest of Casein 15.0 g
Papaic Digest of Soybean 5.0 g
Sodium Chloride 5.0 g
Agar 15.0 g
Water 1000 mL
pH after sterilization: 7.3 ± 0.1.
MEDIUM 39
Same as Medium 3, except that the final pH after sterilization is 7.9 ± 0.1.
MEDIUM 40
Yeast Extract 20.0 g
Polypeptone 5.0 g
Dextrose 10.0 g
Monobasic Potassium Phosphate 2.0 g
Polysorbate 80 0.1 g
Agar 10.0 g
Water 1000 mL
pH after sterilization: 6.7 ± 0.2.
MEDIUM 41
Pancreatic Digest of Casein 9.0 g
Dextrose 20.0 g
Yeast Extract 5.0 g
Sodium Citrate 10.0 g
Monobasic Potassium Phosphate 1.0 g
Dibasic Potassium Phosphate 1.0 g
Water 1000 mL
pH after sterilization: 6.8 ± 0.1.
Phosphate Buffers and Other Solutions (磷酸盐缓冲液和其他的解决办法)
Prepare as follows, or by other suitable means, the potassium phosphate
buffers required for the antibiotic under assay. The buffers are sterilized
after preparation, and the pH specified in each case is the pH after
sterilization.
BUFFER NO. 1, 1 PERCENT, pH 6.0— Dissolve 2.0 g of dibasic potassium
phosphate and 8.0 g of monobasic potassium phosphate in 1000 mL of water.
Adjust with 18 N phosphoric acid or 10 N potassium hydroxide to 6.0 ± 0.05.
BUFFER NO. 3, 0.1 M, pH 8.0— Dissolve 16.73 g of dibasic potassium
phosphate and 0.523 g of monobasic potassium phosphate in 1000 mL of water.
Adjust the pH with 18 N phosphoric acid or 10 N potassium hydroxide to 8.0 ± 0.1.
BUFFER NO. 4, 0.1 M, pH 4.5— Dissolve 13.61 g of monobasic potassium
phosphate in 1000 mL of water. Adjust the pH with 18 N phosphoric acid or 10 N
potassium hydroxide to 4.5 ± 0.05.
BUFFER NO. 6, 10 PERCENT, pH 6.0— Dissolve 20.0 g of dibasic potassium
phosphate and 80.0 g of monobasic potassium phosphate in 1000 mL of water.
Adjust the pH with 18 N phosphoric acid or 10 N potassium hydroxide to 6.0 ± 0.05.
BUFFER NO. 10, 0.2 M, pH 10.5— Dissolve 35.0 g of dibasic potassium
phosphate in 1000 mL of water, and add 2 mL of 10 N potassium hydroxide.
Adjust the pH with 18 N phosphoric acid or 10 N potassium hydroxide to 10.5 ± 0.1.
BUFFER NO. 16, 0.1 M, pH 7.0— Dissolve 13.6 g of dibasic potassium phosphate and 4.0 g of monobasic potassium phosphate in 1000 mL of water.
Adjust with 18 N phosphoric acid or 10 N potassium hydroxide to a pH of 7.0 ±
0.2.
OTHER SOLUTIONS— Use the substances specified under Reagents, Indicators,
and Solutions. For water, use Purified Water. For saline, use Sodium Chloride
Injection. Dilute formaldehyde is Formaldehyde Solution diluted with water 1:3.