464 Amoxicillin Capsules / Official Monographs JP XVIII

Operating conditions— according to the following conditions. Determine each peak

Detector: An ultraviolet absorption photometer (wave- area of both solutions by the automatic integration method:
length: 230 nm). the area of each peak other than amoxicillin obtained from
Column: A stainless steel column 4.6 mm in inside diame- the sample solution is not larger than the peak area of
ter and 15 cm in length, packed with octadecylsilanized silica amoxicillin from the standard solution.
gel for liquid chromatography (5 mm in particle diameter). Operating conditions—
Column temperature: A constant temperature of about Proceed as directed in the operating conditions in the
259 C. Purity (3) under Amoxicillin Hydrate.
Mobile phase: Dissolve 1.361 g of sodium acetate trihy- System suitability—
drate in 750 mL of water, adjust to pH 4.5 with acetic acid Test for required detectability and system repeatability:
(31), and add water to make 1000 mL. To 950 mL of this so- Proceed as directed in the system suitability in the Purity (3)
lution add 50 mL of methanol. under Amoxicillin Hydrate.
Flow rate: Adjust so that the retention time of amoxicillin System performance: When the procedure is run with 10
is about 8 minutes. mL of the standard solution under the above operating con-
System suitability— ditions, the number of theoretical plates and the symmetry
System performance: When the procedure is run with factor of the peak of amoxicillin is not less than 2500 and
10 mL of the standard solution under the above operating not more than 1.5, respectively.
conditions, the number of theoretical plates of the peak of
Water <2.48> Not more than 15.0z (0.1 g, volumetric
amoxicillin is not less than 2500.
titration, direct titration).
System repeatability: When the test is repeated 6 times
with 10 mL of the standard solution under the above operat- Uniformity of dosage units <6.02> It meets the requirement
ing conditions, the relative standard deviation of the peak of the Mass variation test.
area of amoxicillin is not more than 1.0z.
Dissolution <6.10> When the test is performed at 100 revo-
Containers and storage Containers—Tight containers. lutions per minute according to the Paddle method using the
sinker, using 900 mL of water as the dissolution medium, the
dissolution rate in 60 minutes of Amoxicillin Capsules is not
Amoxicillin Capsules less than 75z.
Start the test with 1 capsule of Amoxicillin Capsules, with-
アモキシシリンカプセル draw not less than 20 mL of the medium at the specified
minute after starting the test, and filter through a membrane
filter with a pore size not exceeding 0.45 mm. Discard not less
Amoxicillin Capsules contain not less than 92.0z
than 10 mL of the first filtrate, pipet V mL of the subsequent
and not more than 105.0z of the labeled potency of
filtrate, add water to make exactly V? mL so that each mL
Amoxicillin (C16H19N3O5S: 365.40).
contains about 56 mg (potency) of Amoxicillin Hydrate, and
Method of preparation Prepare as directed under Cap- use this solution as the sample solution. Separately, weigh
sules, with Amoxicillin Hydrate. accurately an amount equivalent to about 28 mg (potency) of
Amoxicillin RS, dissolve in water to make exactly 100 mL.
Identification Take out the contents of Amoxicillin Cap-
Pipet 5 mL of this solution, add water to make exactly 25
sules, to a quantity of the contents, equivalent to 8 mg
mL, and use this solution as the standard solution. Perform
(potency) of Amoxicillin Hydrate, add 2 mL of 0.01 mol/L
the test with exactly 50 mL each of the sample solution and
hydrochloric acid TS, shake for 30 minutes, filter, and use
standard solution as directed under Liquid Chromatography
the filtrate as the sample solution. Separately, dissolve an
<2.01> according to the following conditions, and determine
amount equivalent to 8 mg (potency) of Amoxicillin RS in 2
the peak areas, AT and AS, of amoxicillin in each solution.
mL of 0.01 mol/L hydrochloric acid TS, and use this solu-
tion as the standard solution. Perform the test with these so- Dissolution rate (z) with respect to the labeled amount
lutions as directed under Thin-layer Chromatography <2.03>. of amoxicillin (C16H19N3O5S)
Spot 5 mL each of the sample solution and standard solution ＝ MS × AT/AS × V?/V × 1/C × 180
on a plate of silica gel for thin-layer chromatography. De-
MS: Amount [mg (potency)] of Amoxicillin RS taken
velop the plate with a mixture of tetrahydrofuran, water and
C: Labeled amount [mg (potency)] of amoxicillin
formic acid (50:5:2) to a distance of about 10 cm, and air-
(C16H19N3O5S) in 1 capsule
dry the plate. Spray evenly a solution of ninhydrin in ethanol
(95) (1 in 20) on the plate, and heat the plate at 1109
C for 15 Operating conditions—
minutes: the principal spot obtained from the sample solu- Proceed as directed in the operating conditions in the
tion and the spot from the standard solution show a red- Assay under Amoxicillin Hydrate.
purple color and the same R f value. System suitability—
System performance: When the procedure is run with 50
Purity Related substances—Take out the contents of
mL of the standard solution under the above operating con-
Amoxicillin Capsules, to a quantity of the contents, equiva-
ditions, the number of theoretical plates and the symmetry
lent to 0.1 g (potency) of Amoxicillin Hydrate, add 30 mL of
factor of the peak of amoxicillin are not less than 2500 and
a solution of boric acid (1 in 200), shake for 15 minutes, and
not more than 2.0, respectively.
add a solution of boric acid (1 in 200) to make 50 mL. Cen-
System repeatability: When the test is repeated 6 times
trifuge this solution, and use the supernatant liquid as the
with 50 mL of the standard solution under the above operat-
sample solution. Pipet 1 mL of the sample solution, add a
ing conditions, the relative standard deviation of the peak
solution of boric acid (1 in 200) to make exactly 100 mL, and
area of amoxicillin is not more than 1.5z.
use this solution as the standard solution. Perform the test
with exactly 10 mL each of the sample solution and standard Assay Weigh accurately the mass of not less than 10
solution as directed under Liquid Chromatography <2.01> Amoxicillin Capsules, take out the contents, and weigh accu-

The JP Drugs are to be tested according to the provisions given in the pertinent monographs, General Notices, General Rules for Crude Drugs,
General Rules for Preparations, and General Tests for their conformity to the Japanese Pharmacopoeia. (See the General Notices 5.)
JP XVIII Official Monographs / Amphotericin B 465

rately the mass of the emptied shells. Weigh accurately an It contains not less than 840 mg (potency) per mg,
amount equivalent to about 0.1 g (potency) of Amoxicillin calculated on the dried basis. The potency of Am-
Hydrate, add 70 mL of water, shake for 15 minutes, and add photericin B is expressed as mass (potency) of am-
water to make exactly 100 mL. Centrifuge this solution, and photericin B (C47H73NO17).
use the supernatant liquid as the sample solution. Separately,
Description Amphotericin B occurs as a yellow to orange
weigh accurately an amount equivalent to about 20 mg (po-
powder.
tency) of Amoxicillin RS, dissolve in water to make exactly
It is freely soluble in dimethylsulfoxide and practically
20 mL, and use this solution as the standard solution. Per-
insoluble in water and in ethanol (95).
form the test with exactly 10 mL each of the sample solution
and standard solution as directed under Liquid Chromatog- Identification (1) Dissolve 5 mg of Amphotericin B in 10
raphy <2.01> according to the following conditions, and mL of dimethylsulfoxide. To 1 mL of this solution add 5 mL
determine the peak areas, AT and AS, of amoxicillin in each of phosphoric acid: a blue color develops between the two
solution. layers, and the solution becomes blue by shaking. After ad-
dition of 15 mL of water it becomes yellow to light yellow-
Amount [mg (potency)] of amoxicillin (C16H19N3O5S)
brown by shaking.
＝ M S × AT / AS × 5
(2) Dissolve 25 mg of Amphotericin B in 5 mL of
MS: Amount [mg (potency)] of Amoxicillin RS taken dimethylsulfoxide, and add methanol to make 50 mL. To 1
mL of this solution add methanol to make 50 mL. Determine
Operating conditions—
the absorption spectrum of this solution as directed under
Column temperature, mobile phase, and flow rate: Pro-
Ultraviolet-visible Spectrophotometry <2.24>, and compare
ceed as directed in the operating conditions in the Assay
the spectrum with the Reference Spectrum or the spectrum
under Amoxicillin Hydrate.
of a solution of Amphotericin B RS prepared in the same
Detector: An ultraviolet absorption photometer (wave-
manner as the sample solution: both spectra exhibit similar
length: 254 nm).
intensities of absorption at the same wavelengths.
Column: A stainless steel column 4 mm in inside diameter
and 30 cm in length, packed with octadecylsilanized silica gel Purity Amphotericin A—Weigh accurately about 50 mg
for liquid chromatography (10 mm in particle diameter). each of Amphotericin B and Amphotericin B RS, add ex-
System suitability— actly 10 mL each of dimethylsulfoxide to dissolve, and add
System performance: When the procedure is run with 10 methanol to make exactly 50 mL. Pipet 4 mL each of these
mL of the standard solution under the above operating con- solutions, add methanol to make exactly 50 mL, and use
ditions, the number of theoretical plates and the symmetry these solutions as the sample solution and standard solution
factor of the peak of amoxicillin are not less than 2500 and (1), respectively. Separately, weigh accurately about 20 mg
not more than 1.5, respectively. of Nystatin RS, add exactly 40 mL of dimethylsulfoxide to
System repeatability: When the test is repeated 6 times dissolve, then add methanol to make exactly 200 mL. Pipet 4
with 10 mL of the standard solution under the above operat- mL of this solution, add methanol to make exactly 50 mL,
ing conditions, the relative standard deviation of peak areas and use this solution as the standard solution (2). Perform
of amoxicillin is not more than 1.0z. the test with these solutions as directed under Ultraviolet-
visible Spectrophotometry <2.24> using a solution obtained
Containers and storage Containers—Tight containers.
in the same manner as the sample solution as the blank, and
determine the absorbances at 282 nm and at 304 nm. Calcu-
late the amount of amphotericin A by the following equa-
Amphotericin B tion: not more than 5z for Amphotericin B used for injec-
tions, and not more than 15z for Amphotericin B not used
アムホテリシン B
for injections.
Amount (z) of amphotericin A
M × {(ASa1 × AT2) － (ASa2 × AT1)} × 25
＝ S
MT × {(ASa1 × ASb2) － (ASa2 × ASb1)}
MS: Amount (mg) of Nystatin RS taken
MT: Amount (mg) of Amphotericin B taken
ASa1: Absorbance at 282 nm of the standard solution (1)
ASb1: Absorbance at 282 nm of the standard solution (2)
ASa2: Absorbance at 304 nm of the standard solution (1)
ASb2: Absorbance at 304 nm of the standard solution (2)
AT1: Absorbance at 282 nm of the sample solution
AT2: Absorbance at 304 nm of the sample solution
C47H73NO17: 924.08
(1R,3S,5R,6R,9R,11R,15S,16R,17R,18S,19E,21E, Loss on drying <2.41> Not more than 5.0z (0.1 g, in vacu-
23E,25E,27E,29E,31E,33R,35S,36R,37S )-33-(3- um, 609C, 3 hours).
Amino-3,6-dideoxy-b-D-mannopyranosyloxy)-
Assay Perform the test according to the Cylinder-plate
1,3,5,6,9,11,17,37-octahydroxy-15,16,18-trimethyl-13-oxo-
method as directed under Microbial Assay for Antibiotics
14,39-dioxabicyclo[33.3.1]nonatriaconta-
<4.02> according to the following conditions.
19,21,23,25,27,29,31-heptaene-36-carboxylic acid
(i) Test organism—Saccharomyces cerevisiae ATCC
[1397-89-3]
9763
(ii) Culture medium—Use the medium 2) under (1) Agar
Amphotericin B is a polyene macrolide substance
media for seed and base layer.
having antifungal activity produced by the growth of
(iii) Preparation of cylinder-agar plate—Proceed as di-
Streptomyces nodosus.
The JP Drugs are to be tested according to the provisions given in the pertinent monographs, General Notices, General Rules for Crude Drugs,
General Rules for Preparations, and General Tests for their conformity to the Japanese Pharmacopoeia. (See the General Notices 5.)