KP X 1115
ratios, QT and QS, of the peak area of sulbactam to that um for Injection contains Sulbactam Sodium and Am-
of the internal standard. picillin Sodium in the ratio of 1 : 2 (potency).
Amount [μg (potency)] of sulbactam (C8H11NO5S) Description Sulbactam Sodium･Ampicillin Sodium
Q for Injection appears as white to grayish white crystal-
= Amount [μg (potency)] of Sulbactam RS × T line powder.
QS
Identification (1) Sulbactam sodium⎯The retention
Internal standard solution—A solution of ethyl
time of the principal peak of sulbactam from the test
parahydroxybenzoate in the mobile phase (7 in 1000)
solution corresponds to that of the principal peak of
sulbactam from the standard solution, as obtained in
Operating conditions
the Assay (1).
Detector: An ultraviolet absorption photometer
(wavelength: 220 nm) (2) Ampicillin sodium⎯The retention time of the
Column: A stainless steel column, about 3.9 mm in principal peak of ampicillin from the test solution cor-
internal diameter and about 30 cm in length, packed responds to that of the principal peak of ampicillin
with octadecylsilanized silica gel for liquid chromatog- from the standard solution, as obtained in the Assay (2).
raphy (10 µm in particle diameter)
Column temperature: A constant temperature of pH The pH of a solution prepared by dissolving an
about 35 °C amount of Sulbactam Sodium･Ampicillin Sodium for
Mobile phase: To 250 mL of acetonitrile add 0.005 Injection, equivalent to 1.0 g (potency) of ampicillin
mol/L tetrabutylammonium hydroxide TS to make ex- sodium, in 100 mL of water is between 8.0 and 10.0.
actly 1000 mL
Flow rate: Adjust the flow rate so that the retention Water Not more than 2.0 % (0.5 g, volumetric titra-
time of sulbactam is about 6 minutes. tion, direct titration)
System suitability
System performance: When the procedure is run Sterility Test It meets the requirement.
with 10 μL of the standard solution under the above
operating conditions, sulbactam and the internal stand- Bacterial Endotoxins Less than 0.20 EU/mg (poten-
ard are eluted in this order with the resolution between cy) of ampicillin.
these peaks being not less than 1.5.
System repeatability: When the test is repeated 6 Foreign Insoluble Matter Test It meets the re-
times with 10 μL each of the standard solution under quirement.
the above operating conditions, the relative standard
deviation of the peak areas of sulbactam is not more Insoluble Particulate Matter Test for Injections It
than 1.0 %. meets the requirement.
0.005 mol/L Tetrabutylammonium hydroxide TS⎯To Uniformity of Dosage Units It meets the require-
10 mL of tetrabutylammonium hydroxide TS add 700 ment.
mL of water, adjust the pH to 4.0 with diluted phos-
phoric acid (1 in 10), and add water to make 1000 mL. Assay Dissolve Sulbactam Sodium･Ampicillin Sodi-
um for Injection in water according to the labeled
Containers and Storage Containers—Tight con- amount. Pipet a suitable amount of this solution, add
tainers. the mobile phase so that each mL contains 0.25 mg
(potency) of sulbactam and 0.5 mg (potency) of ampi-
cillin, and use this solution as the test solution. Sepa-
rately, weigh accurately a suitable amount each of
Sulbactam Sodium·Ampicillin Ampicillin RS and Sulbactam RS, dissolve in the mo-
Sodium for Injection bile phase so that each mL contains 0.25 mg (potency)
of sulbactam and 0.5 mg (potency) of ampicillin, and
use this solution as the standard solution. Perform the
Sulbactam Sodium･Ampicillin Sodium for Injection is
test with 10 μL each of the test solution and standard
a preparation for injection, which is dissolved before solution as directed under Liquid Chromatography ac-
use. cording to the following conditions, and determine the
Sulbactam Sodium･Ampicillin Sodium for Injection peak areas, AT1, AS1, AT2, and AS2, of sulbactam and
contains not less than 90.0 % and not more than 120.0 % ampicillin in the test solution and standard solution.
of the labeled amount of sulbactam (C8H11NO5S:
233.24) and ampicillin (C16H19N3O4S: 349.41). Amount [μg (potency)] of sulbactam (C8H11NO5S)
Method of Preparation Prepare as directed under
Injections so that Sulbactam Sodium･Ampicillin Sodi-
1116 Monographs, Part I
= Concentration of sulbactam in the standard solution containers.
[μg (potency)/mL]
A
× T 1 × Test solution dilution factor
AS 1 Sulbenicillin Sodium
NaO O
Amount [μg (potency)] of ampicillin (C16H19N3O4)
= Concentration of ampicillin in the standard solution O
[μg (potency)/mL] SO3Na N CH3
AT 2 H
× × Test solution dilution factor N CH3
AS 2 S
H H
O
Operating conditions
Detector: An ultraviolet absorption photometer
(wavelength: 230 nm). C16H16N2Na2O7S2: 458.42
Column: A stainless steel column about 4 mm in in-
ternal diameter and about 30 cm in length, packed with Disodium (3S)-2,2-dimethyl-6b-[(2R)-2-phenyl-2-
octadecylsilanized silica gel for liquid chromatography sulfonatoacetamido]penam-3-carboxylate [28002-18-8]
(1.5 to 10 μm in particle diameter).
Mobile phase: a mixture of 0.005 mol/L Sulbenicillin Sodium contains not less than 900 μg
tetrabutylammonium hydroxide TS and acetonitrile (potency) and not more than 970 μg (potency) per mg
(1650 : 350). of sulbenicillin (C16H18N2O7S2: 414.45), calculated on
Flow rate: 2.0 mL/minute. the anhydrous basis.
0.005 mol/L Tetrabutylammonium hydroxide TS⎯To Description Sulbenicillin Sodium appears as white to
6.6 mL of a 40 % solution of tetrabutylammonium hy- pale yellowish white powder.
droxide add water to make 1800 mL, adjust the pH to Sulbenicillin Sodium is very soluble in water, freely
5.0 ± 1 with 1 mol/L phosphoric acid, and add water to soluble in methanol, and slightly soluble in ethanol
make 2000 mL. (99.5).
Sulbenicillin Sodium is hygroscopic.
System suitability
System performance: When the procedure is run Identification (1) Determine the infrared spectra of
with 10 μL of the system suitability solution under the Sulbenicillin Sodium and Sulbenicillin Sodium RS as
above operating conditions, the relative retention time directed in the potassium bromide disk method under
of ampicillin with respect to sulbactam alkaline Infrared Spectrophotometry: both spectra exhibit simi-
degredation product is about 0.7, with the resolution lar intensities of absorption at the same wave numbers.
between these peaks being not less than 4.0. When the (2) Sulbenicillin Sodium responds to the Qualitative
procedure is run with 10 μL of the standard solution Tests (1) for sodium salt.
under the above operating conditions, the relative re-
tention time of ampicillin with respect to sulbactam is Specific Optical Rotation [α ] D : +167 ~ +182° (1
20
about 0.35. The number of theoretical plates and sym-
g calculated on the anhydrous basis, water, 20 mL, 100
metry factor of sulbactam are not less than 3500 and
mm)
not more than 1.5, respectively.
System repeatability: When the test is repeated 5
pH The pH of a solution obtained by dissolving 0.20
times with 10 μL each of the standard solution under
g of Sulbenicillin Sodium in 10 mL of water is between
the above operating conditions, the relative standard
4.5 and 7.0.
deviation of the peak areas of sulbactam is not more
than 2.0 %.
Purity (1) Clarity and color of solution⎯Dissolve
2.5 g of Sulbenicillin Sodium in 5 mL of water: the
System suitability solution⎯Dissolve about 30 mg
solution is clear and colorless to pale yellow.
of Sulbactam RS in 0.01 mol/L sodium hydroxide solu-
(2) Heavy metals⎯Proceed with 1.0 g of
tion to make 100 mL, allow to stand for 30 minutes,
Sulbenicillin Sodium according to Method 1, and per-
and adjust the pH to 5.0 ± 1 with 1 mol/L phosphoric
form the test. Prepare the control solution with 2.0 mL
acid. To 5 mL of this solution add 4.25 mL of acetoni-
of standard lead solution (not more than 20 ppm).
trile and 0.005 mol/L tetrabutylammonium hydroxide
TS to make 25 mL. To 1 mL of this solution add 15 mg (3) Arsenic⎯Proceed with 1.0 g of Sulbenicillin
of Ampicillin RS and add the mobile phase to make 25 Sodium according to Method 1, and perform the test
mL. (not more than 2 ppm).
(4) Related substances⎯Dissolve 0.1 g of
Containers and Storage Containers—Hermetic Sulbenicillin Sodium in 15 mL of the mobile phase,
