(kan'' a mye' sin sul' fate).
d-Streptamine, O-3-amino-3-deoxy--d-glucopyranosyl-(1®6)-O-[6-amino-6-deoxy--d-glucopyranosyl(1®4)]-2-deoxy-, sulfate (1:1) (salt).
Kanamycin sulfate (1:1) (salt)
» Kanamycin Sulfate has a potency equivalent to not less than 750 µg of kanamycin (C18H36N4O11) per mg, calculated on the dried basis.
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.
USP Reference standards 11
USP Endotoxin RS
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.
Crystallinity 695: meets the requirements.
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.
Other requirements Where the label states that Kanamycin Sulfate is sterile, it meets the requirements for Sterility and Bacterial endotoxins under Kanamycin Injection. Where the label states that Kanamycin Sulfate must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements for Bacterial endotoxins under Kanamycin Injection.
Mobile phase Use a 0.115 N sodium hydroxide solution. Make adjustments if necessary (see System Suitability under Chromatography 621).
Resolution solution Prepare a solution in water containing about 0.02 mg of USP Amikacin RS per mL and 0.008 mg of USP Kanamycin Sulfate RS per mL.
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 silversilver 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.
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 (C18H36N4O11) 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.
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USP35NF30 Page 3616Pharmacopeial Forum: Volume No. 27(6) Page 3312