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 |
Peptone | 6.0 g |
Yeast Extract | 3.0 g |
Beef Extract | 1.5 g |
Agar | 15.0 g |
Water | 1000 mL |
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 |
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 |
Dextrose | 20.0 g |
Peptone | 10.0 g |
Water | 1000 mL |
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 |
Glycerol | 10.0 g |
Peptone | 10.0 g |
Beef Extract | 10.0 g |
Sodium Chloride | 3.0 g |
Water | 1000 mL |
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 |
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 |
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 |
Stock Solution | Test Dilution | ||||
Antibiotic and Type of Assay [Cylinder-plate (CP) or Turbidimetric (T)] |
Initial Solvent (and initial concentration where specified); Further Diluent, if different | Final Stock Concentration per mL |
Use Within | Final Diluent | Median Dose (µg of activity or Units per mL) |
Amikacin (T) | Water | 1 mg | 14 days | Water | 10 µg |
Amphotericin B (CP) | Dimethyl sulfoxide | 1 mg | Same day | B. 10 | 1.0 µg |
Bacitracin Zinc (CP) | 0.01 N hydrochloric acid | 100 U | Same day | B. 1 | 1.0 U |
Bleomycin (CP) | B. 16 | 2 U | 14 days | B. 16 | 0.04 U |
Candicidin (T) | Dimethyl sulfoxide | 1 mg | Same day | Water | 0.06 µg |
Capreomycin (T) | Water | 1 mg | 7 days | Water | 100 µg |
Carbenicillin (CP) | B. 1 | 1 mg | 14 days | B. 1 | 20 µg |
Cephalothin (CP) | B. 1 | 1 mg | 5 days | B. 1 | 1.0 µg |
Cephapirin (CP) | B. 1 | 1 mg | 3 days | B. 1 | 1.0 µg |
Chloramphenicol (T) | Alcohol (10 mg/mL); [Water] | 1 mg | 30 days | Water | 2.5 µg |
Chlortetracycline (T) | 0.01 N hydrochloric acid | 1 mg | 4 days | Water | 0.06 µg |
Cloxacillin (CP) | B. 1 | 1 mg | 7 days | B. 1 | 5.0 µg |
Colistimethate Sodium (CP) | Water (10 mg/mL); [ B. 6] |
1 mg | Same day | B. 6 | 1.0 µg |
Colistin (CP) | Water (10 mg/mL); [ B. 6] |
1 mg | 14 days | B. 6 | 1.0 µg |
Cycloserine (T) | Water | 1 mg | 30 days | Water | 50 µg |
Demeclocycline (T) | 0.1 N hydrochloric acid | 1 mg | 4 days | Water | 0.1 µg |
Dihydrostreptomycin (CP) | B. 3 | 1 mg | 30 days | B. 3 | 1.0 µg |
Dihydrostreptomycin (T) | Water | 1 mg | 30 days | Water | 30 µg |
Doxycycline (T) | 0.1 N hydrochloric acid | 1 mg | 5 days | Water | 0.1 µg |
Erythromycin (CP) | Methanol (10 mg/mL); [ B. 3] | 1 mg | 14 days | B. 3 | 1.0 µg |
Gentamicin (CP) | B. 3 | 1 mg | 30 days | B. 3 | 0.1 µg |
Gramicidin (T) | Alcohol 95% | 1 mg | 30 days | Alcohol 95% | 0.04 µg |
Kanamycin (T) | Water | 1 mg | 30 days | Water | 10 µg |
Methacycline (T) | Water | 1 mg | 7 days | Water | 0.06 µg |
Nafcillin (CP) | B. 1 | 1 mg | 2 days | B. 1 | 2.0 µg |
Natamycin (CP) | Dimethyl sulfoxide | 1 mg | Same day | B. 10 | 5.00 µg |
Neomycin (CP) | B. 3 | 1 mg | 14 days | B. 3 | 1.0 µg |
Neomycin (T) | B. 3 | 100 µg | 14 days | B. 3 | 1.0 µg |
Netilmicin (CP) | B. 3 | 1 mg | 7 days | B. 3 | 0.1 µg |
Novobiocin (CP) | Alcohol (10 mg/mL); [ B. 3] | 1 mg | 5 days | B. 6 | 0.5 µg |
Nystatin (CP) | Dimethylformamide | 1,000 U | Same day | B. 6 | 20 U |
Oxytetracycline (T) | 0.1 N hydrochloric acid | 1 mg | 4 days | Water | 0.24 µg |
Paromomycin (CP) | B. 3 | 1 mg | 21 days | B. 3 | 1.0 µg |
Penicillin G (CP) | B. 1 | 1,000 U | 4 days | B. 1 | 1.0 U |
Polymyxin B (CP) | Water; [ B. 6] | 10,000 U | 14 days | B. 6 | 10 U |
Rolitetracycline (T) | Water | 1 mg | 1 day | Water | 0.24 µg |
Sisomicin (CP) | B. 3 | 1 mg | 14 days | B. 3 | 0.1 µg |
Streptomycin (T) | Water | 1 mg | 30 days | Water | 30 µg |
Tetracycline (T) | 0.1 N hydrochloric acid | 1 mg | 1 day | Water | 0.24 µg |
Thiostrepton (T) | Dimethyl sulfoxide | 1 U | Same day | Dimethyl sulfoxide |
0.80 U |
Ticarcillin (CP) | B. 1 | 1 mg | 1 day | B. 1 | 5.0 µg |
Tobramycin (T) | Water | 1 mg | 14 days | Water | 2.5 µg |
Troleandomycin (T) | Isopropyl alcohol-water (4:1) | 1 mg | Same day | Water | 25 µg |
Tylosin (T) | Methanol (10 mg/mL); [ B. 16] | 1 mg | 30 days | B.3: methanol (1:1) | 4 µg |
Vancomycin (CP) | Water | 1 mg | 7 days | B. 4 | 10 µg |
NOTESB denotes buffer, and the number following refers to the potassium phosphate buffers defined in this chapter.
For amphotericin B, colistimethate sodium, and nystatin, prepare the USP Reference Standard solutions and the sample test solution simultaneously.
For amphotericin B, further dilute the stock solution with dimethyl sulfoxide to give concentrations of 12.8, 16, 20, 25, and 31.2 µg per mL prior to making the test dilutions. The Test Dilution of the sample should contain the same amount of dimethyl sulfoxide as the test dilutions of the USP Reference Standard.
For bacitracin zinc, each of the Standard test dilutions should contain the same amount of hydrochloric acid as the Test Dilution of the sample.
For neomycin turbidimetric assay, dilute the 100-µg-per-mL stock solution quantitatively with Buffer No. 3 to obtain a solution having a concentration equivalent to 25.0 µg of neomycin per mL. To separate 50-mL volumetric flasks add 1.39, 1.67, 2.00, 2.40, and 2.88 mL of this solution, add 5.0 mL of 0.01 N hydrochloric acid to each flask, dilute with Buffer No. 3 to volume, and mix to obtain solutions having concentrations of 0.69, 0.83, 1.0, 1.2, and 1.44 µg of neomycin per mL. Use these solutions to prepare the standard response line.
For nystatin, further dilute the stock solution with dimethylformamide to give concentrations of 256, 320, 400, 500, and 624 Units per mL prior to making the test dilutions. Prepare the standard response line solutions simultaneously with dilutions of the sample to be tested. The Test Dilution of the sample should contain the same amount of dimethylformamide as the test dilutions of the Standard. Use red low-actinic glassware.
For Polymyxin B, prepare the stock solution by adding 2 mL of water for each 5 mg of the weighed USP Reference Standard material.
|
Antibiotic | Test Organism | ATCC * Number |
Amikacin | Staphylococcus aureus | 29737 |
Amphotericin B | Saccharomyces cerevisiae | 9763 |
Bacitracin | Micrococcus luteus | 10240 |
Bleomycin | Mycobacterium smegmatis | 607 |
Candicidin | Saccharomyces cerevisiae | 9763 |
Capreomycin | Klebsiella pneumoniae | 10031 |
Carbenicillin | Pseudomonas aeruginosa | 25619 |
Cephalothin | Staphylococcus aureus | 29737 |
Cephapirin | Staphylococcus aureus | 29737 |
Chlorampheni- col |
Escherichia coli | 10536 |
Chlortetracy- cline |
Staphylococcus aureus | 29737 |
Cloxacillin | Staphylococcus aureus | 29737 |
Colistimethate Sodium |
Bordetella bronchiseptica | 4617 |
Colistin | Bordetella bronchiseptica | 4617 |
Cycloserine | Staphylococcus aureus | 29737 |
Demeclocycline | Staphylococcus aureus | 29737 |
Dihydrostrepto- mycin (CP) |
Bacillus subtilis | 6633 |
Dihydrostrepto- mycin (T) |
Klebsiella pneumoniae | 10031 |
Doxycycline | Staphylococcus aureus | 29737 |
Erythromycin | Micrococcus luteus | 9341 |
Gentamicin | Staphylococcus epidermidis | 12228 |
Gramicidin | Enterococcus hirae | 10541 |
Kanamycin | Staphylococcus aureus | 29737 |
Methacycline | Staphylococcus aureus | 29737 |
Nafcillin | Staphylococcus aureus | 29737 |
Neomycin (CP) | Staphylococcus epidermidis | 12228 |
Neomycin (T) | Klebsiella pneumoniae | 10031 |
Netilmicin | Staphylococcus epidermidis | 12228 |
Novobiocin | Staphylococcus epidermidis | 12228 |
Nystatin | Saccharomyces cerevisiae | 2601 |
Oxytetracycline | Staphylococcus aureus | 29737 |
Paromomycin | Staphylococcus epidermidis | 12228 |
Penicillin G | Staphylococcus aureus | 29737 |
Polymyxin B | Bordetella bronchiseptica | 4617 |
Rolitetracycline | Staphylococcus aureus | 29737 |
Sisomicin | Staphylococcus epidermidis | 12228 |
Spectinomycin | Escherichia coli | 10536 |
Streptomycin (T) |
Klebsiella pneumoniae | 10031 |
Tetracycline | Staphylococcus aureus | 29737 |
Thiostrepton (T) | Enterococcus hirae | 10541 |
Tobramycin | Staphylococcus aureus | 29737 |
Troleandomycin | Klebsiella pneumoniae | 10031 |
Tylosin | Staphylococcus aureus | 9144 |
Vancomycin | Bacillus subtilis | 6633 |
*
American Type Culture Collection, 10801 University Boulevard, Manassas VA 20110-2209. (http://www.atcc.org).
|
Incubation Conditions | Suggested Inoculum Composition | |||||
Test Organism & (ATCC No.) |
Medium | Temp. () | Time | Medium | Amount (mL per 100 mL) | Antibiotics Assayed |
Bacillus subtilis
(6633) |
32 | 32 to 35 | 5 days | 5 | As required | Dihydrostreptomycin |
8 | As required | Vancomycin | ||||
Bordetella bronchiseptica
(4617) |
1 | 32 to 35 | 24 hr. | 10 | 0.1 | Colistimethate Sodium, Colistin, Polymyxin B |
Escherichia coli | 1 | 32 to 35 | 24 hr. | 3 | 0.7 | Chloramphenicol |
(10536) | ||||||
Klebsiella pneumoniae
(10031) |
1 | 36 to 37.5 | 16 to 24 hr. | 3 | 0.05 | Capreomycin |
0.1 | Streptomycin, Troleandomycin, Dihydrostreptomycin |
|||||
39 | 2 | Neomycin | ||||
Micrococcus luteus | 1 | 32 to 35 | 24 hr. | 11 | 1.5 | Erythromycin |
(9341) | ||||||
Micrococcus luteus
(10240) |
1 | 32 to 35 | 24 hr. | 1 | 0.3 | Bacitracin |
Mycobacterium smegmatis (607) |
36 | 36 to 37.5 | 48 hr. | 35 | 1.0 | Bleomycin |
Pseudomonas aeruginosa
(25619) |
1 | 36 to 37.5 | 24 hr. | 10 | 0.5 | Carbenicillin |
Saccharomyces cerevisiae
(9763) |
19 | 29 to 31 | 48 hr. | 13 | 0.2 | Candicidin |
19 | 1.0 | Amphotericin B | ||||
Saccharomyces cerevisiae
(2601) |
19 | 29 to 31 | 48 hr. | 19 | 1.0 | Nystatin |
Staphylococcus aureus
(9144) |
3 | 35 to 39 | 16 to 18 hr. | 39 | 2-3 | Tylosin |
Staphylococcus aureus | 1 | 32 to 35 | 24 hr. | 1 | 0.1 | Cephalothin, Cephapirin, Cloxacillin |
(29737) | ||||||
1 | 0.3 | Nafcillin | ||||
1 | 1.0 | Penicillin G | ||||
3 | 0.1 | Amikacin, Chlortetracycline, Demeclocycline, Doxycycline, Methacycline, Oxytetracycline, Rolitetracycline, Tetracycline |
||||
3 | 0.2 | Kanamycin | ||||
3 | 0.4 | Cycloserine | ||||
3 | 0.15 | Tobramycin | ||||
Staphylococcus
epidermidis |
1 | 32 to 35 | 24 hr. | 11 | 0.25 | Netilmicin |
(12228) | 1 | 4.0 | Novobiocin | |||
11 | 0.03 | Gentamicin, Sisomicin | ||||
11 | 0.4 | Neomycin | ||||
11 | 2.0 | Paromomycin | ||||
Enterococcus hirae
(10541) |
3 | 36 to 37.5 | 16 to 18 hr. | 3 | 1.0 | Gramicidin |
40 | 36 to 37.5 | 18 to 24 hr. | 41 | 0.2 | Thiostrepton | |
NOTEFor Pseudomonas aeruginosa (ATCC 25619) in the assay of Carbenicillin, use 0.5 mL of a 1:25 dilution of the stock suspension per 100 mL of Medium 10.
|