Supplement I to the Japanese Pharmacopoeia Fourteenth Edition

Supplement I, JP XIV
tem suitability test.
System performance: Dissolve 5 mg of josamycin propionate
and 2 mg of josamycin in 50 mL of the mobile
phase. When the procedure is run with 10 mL of this solution
under the above operating conditions, josamycin and
josamycin propionate are eluted in this order with the resolution
between these peaks being not less than 25.
System repeatability: When the test is repeated 6 times
with 10 mL of the solution for system suitability test under
the above operating conditions, the relative standard deviation
of the peak area of josamycin propionate is not more
than 1.5z.
Loss on drying Not more than 1.0z (1 g, in vacuum,
phosphorus (V) oxide, 609C, 3 hours).
O‹cial Monographs for Part I
Add the following:
Kanamycin Monosulfate
一 硫 酸 カナマイシン
1481
Residue on ignition
Not more than 0.10z (1 g).
Assay Perform the test according to the Cylinder-plate
method as directed under the Microbial Assay for Antibiotics
according to the following conditions.
(1) Test organism—Bacillus subtilis ATCC 6633
(2) Culture medium—Use the medium ii in 3) Medium
for other organisms under (1) Agar media for seed and base
layer. Adjust the pH of the medium so that it will be 7.9 to
8.1 after sterilization.
(3) Standard solutions—Weigh accurately an amount of
Josamycin Propionate Reference Standard, equivalent to
about 20 mg (potency), dissolve in 10 mL of methanol, add
1/15 mol/L phosphate buŠer solution, pH 5.6 to make exactly
50 mL, and use this solution as the standard stock solution.
Keep the standard stock solution at 59C orbelow,and
use within 3 days. Take exactly a suitable amount of the
standard stock solution before use, add 1/15 mol/L phosphate
buŠer solution, pH 5.6 to make solutions so that each
mL contains 80 mg (potency) and 20 mg (potency), and use
these solutions as the high concentration standard solution
and the low concentration standard solution, respectively.
(4) Sample solutions—Weigh accurately an amount of
Josamycin Propionate, equivalent to about 20 mg (potency),
dissolve in 10 mL of methanol, add 1/15 mol/L phosphate
buŠer solution, pH 5.6 to make exactly 50 mL. Take exactly
a suitable amount of this solution, add 1/15 mol/L phosphate
buŠer solution, pH 5.6 to make solutions so that each
mL contains 80 mg (potency) and 20 mg (potency), and use
these solutions as the high concentration sample solution
and the low concentration sample solution, respectively.
Containers and storage Containers—Tight containers.
Storage—Light-resistant.
C 18 H 36 N 4 O 11 .H 2 SO 4 : 582.58
O-3-Amino-3-deoxy-a-D-glucopyranosyl-(1→6)-O-[6-
amino-6-deoxy-a-D-glucopyranosyl-(1→4)]-2-deoxy-
D-streptamine monosulfate [25389-94-0 ]
Kanamycin Monosulfate contains not less than 750
mg (potency) per mg, calculated on the dried basis. The
potency of Kanamycin Monosulfate is expressed as
mass (potency) of kanamycin (C 18 H 36 N 4 O 11 : 484.50).
Description Kanamycin Monosulfate occurs as a white
crystalline powder.
It is freely soluble in water, and practically insoluble in
ethanol (99.5).
Identiˆcation (1) Dissolve 50 mg of Kanamycin
Monosulfate in 3 mL of water, and add 6 mL of anthrone
TS: a blue-purple color develops.
(2) Dissolve 20 mg each of Kanamycin Monosulfate and
Kanamycin Monosulfate Reference Standard in 1 mL of
water, and use these solutions as the sample solution and the
standard solution. Perform the test with these solutions as
directed under the Thin-layer Chromatography. Spot 5 mL
each of the sample solution and the standard solution on a
plate of silica gel for thin-layer chromatography. Develop
the plate with the supernatant layer of a mixture of chloroform,
ammonia solution (28) and methanol (2:1:1) to a
distance of about 10 cm, and air-dry the plate. Spray evenly
asolutionof0.2z ninhydrin-water saturated 1-butanol TS
on the plate, and heat at 1009C for 10 minutes: the principal
spots obtained from the sample solution and the standard
solution show a purple-brown color and the same Rf value.
(3) To a solution of Kanamycin Monosulfate (1 in 5) add
1 drop of barium chloride TS: a white precipitate is formed.
Optical rotation [a] 20
D : +112 – +1239(0.2 g calculated on
the dried basis, water, 20 mL, 100 mm).
Sulfuric acid Weigh accurately about 0.25 g of Kanamycin
Monosulfate, dissolve in 100 mL of water, adjust the pH to
11.0 with ammonia solution (28), add exactly 10 mL of
0.1 mol/L barium chloride VS, and titrate with 0.1 mol/L