Packaging and storage—
Preserve in tight containers.
Labeling—
Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms.
Identification—
A:
Dissolve about 10 mg in 1 mL of water, add 1 mL of a solution of ninhydrin in butyl alcohol (1 in 500), and add 0.5 mL of pyridine. Heat in a steam bath for 5 minutes, and add 10 mL of water: a deep purple color is produced.
B:
It meets the requirements of the tests for
Sulfate 
191
.
C:
The retention time of the peak for kanamycin in the chromatogram of the
Assay preparation corresponds to that in the chromatogram of the
Standard preparation, as obtained in the
Assay.
pH 791:
between 6.5 and 8.5, in a solution (1 in 100).
Loss on drying 731—
Dry about 100 mg, accurately weighed, in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60
for 3 hours: it loses not more than 4.0% of its weight.
Residue on ignition 281:
not more than 1.0%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid.
Chromatographic purity—
Dissolve a quantity of Kanamycin Sulfate in water to obtain a test solution having a concentration of 30 mg per mL. Dissolve a suitable quantity of
USP Kanamycin Sulfate RS in water to obtain a Standard solution having a known concentration of 30 mg per mL. Dilute a portion of this solution quantitatively with water to obtain a
Diluted standard solution having a concentration of 0.90 mg per mL. Apply separate 1-µL portions of the three solutions to the starting line of a suitable thin-layer chromatographic plate (see
Chromatography 621) coated with a 0.25-mm layer of chromatographic silica gel and heated at 110
for 1 hour and cooled immediately before use. Allow the spots to dry, and develop the chromatogram in a suitable chamber, previously equilibrated for 90 minutes with a developing solvent of monobasic potassium phosphate solution (7.5 in 100), until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, and air-dry. Spray the plate with a solution of ninhydrin in butyl alcohol (1 in 100). Dry the plate at 110
for 10 minutes, and examine the chromatograms: the chromatograms show principal spots at about the same
RF value, and no secondary spot, if present in the chromatogram from the test solution, is more intense than the principal spot obtained from the
Diluted standard solution.
Assay—
Mobile phase—
Use a 0.115 N sodium hydroxide solution. Make adjustments if necessary (see
System Suitability under
Chromatography 621).
Standard preparation—
Quantitatively dissolve an accurately weighed quantity of
USP Kanamycin Sulfate RS in water to obtain a solution having a known concentration of about 0.008 mg per mL.
Assay preparation—
Transfer about 40 mg of Kanamycin Sulfate, accurately weighed, to a 250-mL volumetric flask, dilute with water to volume, and mix. Transfer 5.0 mL of this solution to a 100-mL volumetric flask, dilute with water to volume, and mix.
Chromatographic system (see Chromatography 621)—
The liquid chromatograph is equipped with an electrochemical detector, a gold working electrode, a pH silver–silver chloride reference electrode, a guard column that contains packing L47, and a 4-mm × 25-cm analytical column that contains packing L47. The electrochemical detector is used in the integrated amperometric mode with a range of 300 nC and an output of 1 V full-scale. The potential is programmed as follows.
| Time (seconds) |
Potential (V) |
Integration |
| 0.00 |
+0.04 |
|
| 0.30 |
+0.04 |
begins |
| 0.50 |
+0.04 |
ends |
| 0.51 |
+0.80 |
|
| 0.70 |
+0.80 |
|
| 0.71 |
–0.80 |
|
| 0.90 |
–0.80 |
|
The flow rate is about 0.5 mL per minute. Chromatograph the
Resolution solution, and record the peak responses as directed for
Procedure: the relative retention times are about 1.0 for kanamycin and 1.3 for amikacin; and the resolution,
R, between kanamycin and amikacin is not less than 3. Chromatograph the
Standard preparation, and record the peak responses as directed for
Procedure: the tailing factor is not more than 2; and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure—
Separately inject equal volumes (about 20 µL) of the
Standard preparation and the
Assay preparation into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in µg, of kanamycin (C
18H
36N
4O
11) in each mg of Kanamycin Sulfate taken by the formula:
5000(CP/W)(rU / rS),
in which
C is the concentration, in mg per mL, of
USP Kanamycin Sulfate RS in the
Standard preparation; P is the designated content, in µg per mg, of kanamycin in
USP Kanamycin Sulfate RS;
W is the weight, in mg, of Kanamycin Sulfate taken to prepare the
Assay preparation; and
rU and
rS are the kanamycin peak areas obtained from the
Assay preparation and the
Standard preparation, respectively.
Auxiliary Information—
Staff Liaison :
Brian D. Gilbert, Ph.D., Scientist
Expert Committee : (MDANT05) Monograph Development-Antibiotics
USP29–NF24 Page 1212
Pharmacopeial Forum : Volume No. 27(6) Page 3312
Phone Number : 1-301-816-8223
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