Kanamycin monosulfate EUROPEAN PHARMACOPOEIA 11.
07/2021:0032 System suitability : reference solution (b) :
– the chromatogram shows 3 clearly separated spots.
Results : the principal spot in the chromatogram obtained
with the test solution is similar in position, colour and size
to the principal spot in the chromatogram obtained with
KANAMYCIN MONOSULFATE reference solution (a).
B. Dissolve 0.5 g in 10 mL of water R. Add 10 mL of picric acid
solution R. Initiate crystallisation if necessary by scratching
Kanamycini monosulfas the wall of the tube with a glass rod and allow to stand.
Collect the crystals, wash with 20 mL of water R and filter.
Dry at 100 °C. The crystals melt (2.2.14) at about 235 °C,
with decomposition.
C. Dissolve about 50 mg in 2 mL of water R. Add 1 mL of a
10 g/L solution of ninhydrin R and heat for a few minutes
on a water-bath. A violet colour develops.
D. It gives the reactions of sulfates (2.3.1).
TESTS
Solution S. Dissolve 0.20 g in carbon dioxide-free water R and
dilute to 20.0 mL with the same solvent.
C18H38N4O15S,H2O Mr 601
[5965-95-7] pH (2.2.3): 6.5 to 8.5 for solution S.
Specific optical rotation (2.2.7) : + 112 to + 123 (dried
DEFINITION substance), determined on solution S.
6-O-(3-Amino-3-deoxy-α-D-glucopyranosyl)-4-O-(6-amino-
Kanamycin B. Thin-layer chromatography (2.2.27) as
6-deoxy-α-D-glucopyranosyl)-2-deoxy-D-streptamine sulfate
described under Identification A with the following
monohydrate.
modifications.
Antimicrobial substance produced by the growth of certain Test solution. Dissolve 0.1 g of the substance to be examined
strains of Streptomyces kanamyceticus. in water R and dilute to 20 mL with the same solvent.
Content : minimum 750 IU/mg (dried substance). Reference solution. Dissolve 4 mg of kanamycin B sulfate CRS
PRODUCTION in water R and dilute to 20 mL with the same solvent.
It is produced by methods of manufacture designed to Application : 4 μL.
eliminate or minimise substances lowering blood pressure. Detection : spray with ninhydrin and stannous chloride
reagent R. Heat at 110 °C for 15 min.
CHARACTERS Limit :
Appearance : white or almost white, crystalline powder. – kanamycin B : any spot corresponding to kanamycin B in
Solubility : freely soluble in water, practically insoluble in the chromatogram obtained with the test solution is not
acetone and in ethanol (96 per cent). more intense than the spot in the chromatogram obtained
with the reference solution (4.0 per cent).
IDENTIFICATION
Loss on drying (2.2.32) : maximum 1.5 per cent, determined
A. Thin-layer chromatography (2.2.27). on 1.000 g by drying at 60 °C at a pressure not exceeding
Test solution. Dissolve 10 mg of the substance to be 670 Pa for 3 h.
examined in water R and dilute to 10 mL with the same Sulfated ash (2.4.14): maximum 0.5 per cent, determined on
solvent. 1.0 g.
Reference solution (a). Dissolve 10 mg of kanamycin
Sulfate : 15.0 per cent to 17.0 per cent of sulfate (dried
monosulfate CRS in water R and dilute to 10 mL with the
substance).
same solvent.
Dissolve 0.250 g in 100 mL of water R and adjust the solution
Reference solution (b). Dissolve 10 mg of kanamycin to pH 11 with concentrated ammonia R. Add 10.0 mL of 0.1 M
monosulfate CRS, 10 mg of neomycin sulfate CRS and 10 mg
barium chloride and about 0.5 mg of phthalein purple R.
of streptomycin sulfate for identification CRS in water R and Titrate with 0.1 M sodium edetate adding 50 mL of ethanol
dilute to 10 mL with the same solvent. (96 per cent) R when the colour of the solution begins to
Plate : suitable plate coated with a 0.75 mm layer of a change and continue the titration until the violet-blue colour
mixture prepared as follows : mix 0.3 g of carbomer R with disappears.
240 mL of water R and allow to stand, with moderate 1 mL of 0.1 M barium chloride is equivalent to 9.606 mg of
shaking, for 1 h ; adjust to pH 7 by the gradual addition, SO4.
with continuous shaking, of dilute sodium hydroxide
solution R and add 30 g of silica gel H R. Pyrogens (2.6.8). If intended for use in the manufacture
of parenteral preparations without a further appropriate
Pretreatment : heat the plate at 110 °C for 1 h, allow to cool
procedure for the removal of pyrogens, it complies with the
and use immediately.
test for pyrogens. Inject per kilogram of the rabbit’s mass 1 mL
Mobile phase : 70 g/L solution of potassium dihydrogen of a 10 mg/mL solution of the substance to be examined in
phosphate R. water for injections R.
Application : 10 μL.
ASSAY
Development : over a path of 12 cm.
Carry out the microbiological assay of antibiotics (2.7.2). Use
Drying : in a current of warm air. kanamycin monosulfate CRS as the reference substance.
Detection : spray with a mixture of equal volumes of a 2 g/L
solution of 1,3-dihydroxynaphthalene R in ethanol (96 per STORAGE
cent) R and a 460 g/L solution of sulfuric acid R. Heat at If the substance is sterile, store in a sterile, tamper-evident
150 °C for 5 min to 10 min. container.
3170 See the information section on general monographs (cover pages)
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