JP XVII
potassium bromide disk method under Infrared Spectropho-
tometry <2.25>, and compare the spectrum with the Refer-
ence Spectrum: both spectra exhibit similar intensities of ab-
sorption at the same wave numbers.
(3) A solution of Ketamine Hydrochloride (1 in 10)
responds to the Qualitative Tests <1.09> (2) for chloride.
Absorbance <2.24> E 11zcm (269 nm): 22.0 – 24.5 (after dry-
ing, 30 mg, 0.1 mol/L hydrochloric acid TS, 100 mL).
pH <2.54> Dissolve 1.0 g of Ketamine Hydrochloride in 10
mL of freshly boiled and cooled water: the pH of the solu-
tion is between 3.5 and 4.5.
Purity (1) Clarity and color of solution—Dissolve 1.0 g of
Ketamine Hydrochloride in 5 mL of water: the solution is
clear and colorless.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Keta-
mine Hydrochloride according to Method 1, and perform
the test. Prepare the control solution with 2.0 mL of Stand-
ard Lead Solution (not more than 20 ppm).
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g
of Ketamine Hydrochloride, according to Method 1, and
perform the test (not more than 2 ppm).
(4) Related substances—Dissolve 0.5 g of Ketamine Hy-
drochloride in 10 mL of methanol and use this solution as
the sample solution. Pipet 1 mL of the sample solution, add
methanol to make exactly 200 mL, and use this solution as
the standard solution. Perform the test with these solutions
as directed under Thin-layer Chromatography <2.03>. Spot 2
mL each of the sample solution and standard solution on a
plate of silica gel for thin-layer chromatography. Develop
the plate with a mixture of cyclohexane and isopropylamine
(49:1) to a distance of about 10 cm, and air-dry the plate.
Spray evenly Dragendorff's TS for spraying on the plate, dry
the plate, and then spray evenly hydrogen peroxide TS: the
spots other than the principal spot from the sample solution
is not more intense than the spot from the standard solution.
Loss on drying <2.41> Not more then 0.5z (1 g, 1059C,
3 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 0.5 g of Ketamine Hydro-
chloride, previously dried, dissolve in 1 mL of formic acid,
add 70 mL of a mixture of acetic anhydride and acetic acid
(100) (6:1), and titrate <2.50> with 0.1 mol/L perchloric acid
VS (potentiometric titration). Perform a blank determina-
tion, and make any necessary correction.
Each mL of 0.1 mol/L perchloric acid VS
＝ 27.42 mg of C13H16ClNO.HCl
Containers and storage Containers—Tight containers.
Official Monographs / Ketoconazole 1121
Ketoconazole
ケトコナゾール
C26H28Cl2N4O4: 531.43
1-Acetyl-4-(4-{[(2RS,4SR )-2-(2,4-dichlorophenyl)-
2-(1H-imidazol-1-ylmethyl)-1,3-dioxolan-
4-yl]methoxy}phenyl)piperazine
[65277-42-1]
Ketoconazole, when dried, contains not less than
99.0z and not more than 101.0z of ketoconazole
(C26H28Cl2N4O4).
Description Ketoconazole occurs as a white to light yellow-
ish white powder.
It is soluble in methanol, sparingly soluble in ethanol
(99.5), and practically insoluble in water.
A solution of Ketoconazole in methanol (1 in 20) shows no
optical rotation.
Identification (1) Determine the absorption spectrum of a
solution of Ketoconazole in methanol (3 in 100,000) as di-
rected under Ultraviolet-visible Spectrophotometry <2.24>,
and compare the spectrum with the Reference Spectrum:
both spectra exhibit similar intensities of absorption at the
same wavelengths.
(2) Determine the infrared absorption spectrum of
Ketoconazole as directed in the potassium bromide disk
method under Infrared Spectrophotometry <2.25>, and com-
pare the spectrum with the Reference Spectrum: both spectra
exhibit similar intensities of absorption at the same wave
numbers.
(3) Perform the test with Ketoconazole as directed under
Flame Coloration Test <1.04> (2): a green color appears.
Melting point <2.60> 148 – 1529C
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
Ketoconazole according to Method 2, and perform the test.
Prepare the control solution with 1.0 mL of Standard Lead
Solution (not more than 10 ppm).
(2) Related Substances—Dissolve 0.10 g of Ketoconazole
in 10 mL of methanol, and use this solution as the sample so-
lution. Pipet 5 mL of the sample solution, and add methanol
to make exactly 100 mL. Pipet 1 mL of this solution, add
methanol to make exactly 10 mL, and use this solution as the
standard solution. Perform the test with exactly 10 mL each
of the sample solution and standard solution as directed
under Liquid Chromatography <2.01> according to the fol-
lowing conditions. Determine each peak area of both solu-
tions by the automatic integration method: the area of the
peak other than ketoconazole obtained from the sample so-
lution is not larger than 2/5 times the peak area of ketocona-
zole obtained from the standard solution, and the total area
of the peaks other than ketoconazole from the sample solu-
tion is not larger than the peak area of ketoconazole from
the standard solution.
1122 Ketoconazole Cream / Official Monographs
Operating conditions—
Detector: An ultraviolet absorption photometer (wave-
length: 220 nm).
Column: A stainless steel column 4.6 mm in inside diame-
ter and 10 cm in length, packed with octadecylsilanized silica
gel for liquid chromatography (3 mm in particle diameter).
Column temperature: A constant temperature of about
259 C.
Mobile phase A: Acetonitrile for liquid chromatography.
Mobile phase B: A solution of tetrabutylammonium
hydrogensulfate (17 in 5000).
Flowing of mobile phase: Control the gradient by mixing
the mobile phases A and B as directed in the following table.
Time after injection Mobile phase A Mobile phase B
of sample (min) (volz) (volz)
0 – 10 5 → 50 95 → 50
10 – 15 50 50
Flow rate: 2.0 mL per minute.
Time span of measurement: For 15 minutes after injec-
tion, beginning after the solvent peak.
System suitability—
Test for required detectability: Pipet 2 mL of the standard
solution, and add methanol to make exactly 20 mL. Confirm
that the peak area of ketoconazole obtained from 10 mL of
this solution is equivalent to 7 to 13z of that of ketocona-
zole obtained from 10 mL of the standard solution.
System performance: When the procedure is run with 10
mL of the standard solution under the above operating con-
ditions, the number of theoretical plates and the symmetry
factor of the peak of ketoconazole are not less than 40,000
and not more than 1.5, respectively.
System repeatability: When the test is repeated 6 times
with 10 mL of the standard solution under the above operat-
ing conditions, the relative standard deviation of the peak
area of ketoconazole is not more than 2.5z.
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
4 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 0.2 g of Ketoconazole, pre-
viously dried, dissolve in 70 mL of a mixture of 2-butanone
and acetic acid (100) (7:1), and titrate <2.50> with 0.1 mol/L
perchloric acid VS (potentiometric titration). Perform a
blank determination in the same manner, and make any nec-
essary correction.
Each mL of 0.1 mol/L perchloric acid VS
＝ 26.57 mg of C26H28Cl2N4O4
Containers and storage Containers—Tight containers.
Storage—Light-resistant.
Ketoconazole Cream
ケトコナゾールクリーム
Ketoconazole Cream contains not less than 95.0z
and not more than 105.0z of the labeled amount of
ketoconazole (C26H28Cl2N4O4: 531.43).
Method of preparation Prepare as directed under Creams,
with Ketoconazole.
JP XVII
Identification To a quantity of Ketoconazole Cream,
equivalent to 0.1 g of Ketoconazole, add 20 mL of 2-
propanol, shake for 20 minutes, centrifuge, and use the su-
pernatant liquid as the sample solution. Separately, dissolve
25 mg of ketoconazole in 5 mL of 2-propanol, and use this
solution as the standard solution. Perform the test with these
solutions as directed under Thin-layer Chromatography
<2.03>. Spot 5 mL each of the sample solution and standard
solution on a plate of silica gel with fluorescent indicator for
thin-layer chromatography. Develop the plate with a mixture
of ethyl acetate, hexane, methanol, water and ammonia so-
lution (28) (40:40:25:2:1) to a distance of about 12 cm, and
air-dry the plate. Examine under ultraviolet light (main
wavelength: 254 nm): the principal spot obtained from the
sample solution has the same R f value as the spot obtained
from the standard solution.
Assay Weigh accurately an amount of Ketoconazole
Cream, equivalent to about 25 mg of ketoconazole
(C26H28Cl2N4O4), dissolve in methanol to make exactly 100
mL. Pipet 10 mL of this solution, add exactly 4 mL of the
internal standard solution, add methanol to make 50 mL,
and use this solution as the sample solution. Separately,
weigh accurately about 25 mg of ketoconazole for assay,
previously dried at 1059C for 4 hours, and dissolve in metha-
nol to make exactly 50 mL. Pipet 5 mL of this solution, add
exactly 4 mL of the internal standard solution, add methanol
to make 50 mL, and use this solution as the standard solu-
tion. Perform the test with 10 mL each of the sample solution
and standard solution as directed under Liquid Chromatog-
raphy <2.01> according to the following conditions, and cal-
culate the ratios, QT and QS, of the peak area of ketocona-
zole to that of the internal standard.
Amount (mg) of ketoconazole (C26H28Cl2N4O4)
＝ MS × QT/QS
MS: Amount (mg) of ketoconazole for assay taken
Internal standard solution—A solution of xanthone in meth-
anol (1 in 10,000).
Operating conditions—
Detector: An ultraviolet absorption photometer (wave-
length: 230 nm).
Column: A stainless steel column 4.6 mm in inside diame-
ter and 15 cm in length, packed with octadecylsilanized silica
gel for liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about
409C.
Mobile phase: To ammonium acetate solution (1 in 200)
add acetic acid (100) to adjust the pH to 5.0. To 250 mL of
this solution add 750 mL of methanol.
Flow rate: Adjust so that the retention time of ketocona-
zole is about 8 minutes.
System suitability—
System performance: When the procedure is run with 10
mL of the standard solution under the above operating con-
ditions, the internal standard and ketoconazole are eluted in
this order with the resolution between these peaks being not
less than 5.
System repeatability: When the test is repeated 6 times
with 10 mL of the standard solution under the above operat-
ing conditions, the relative standard deviation of the ratio of
the peak area of ketoconazole to that of the internal stand-
ard is not more than 1.0z.
Containers and storage Containers—Tight containers.