Pyrogen—
Where the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements of the
Pyrogen Test 
151
, the test dose being 1.0 mL per kg of a solution in pyrogen-free sodium carbonate solution (prepared by dissolving 25.6 g of sodium carbonate, previously heated at 170
for not less than 4 hours, in 1000 mL of Sterile Water for Injection) containing 40 mg per mL.
Water, Method I 921:
not more than 7.0%, the
Test Preparation being prepared as directed for a hygroscopic specimen, except to use a mixture of 20 mL of formamide (previously dried over anhydrous sodium sulfate for 24 hours) and methanol (2:1), instead of methanol, to dissolve the specimen, and to determine the water content of the formamide and methanol mixture.
Assay—
Mobile phase—
Dissolve 13.1 g of ammonium sulfate in 850 mL of water, adjust with 2 N ammonium hydroxide to a pH of 6.5 ± 0.1, add 150 mL of acetonitrile, and mix. Filter through a suitable filter of 0.5 µm or finer porosity, and degas. Make adjustments if necessary (see
System Suitability under
Chromatography 621).
Standard preparation—
Dissolve an accurately weighed quantity of
USP Cefotiam Hydrochloride RS, quantitatively in water to obtain a solution having a known concentration of about 1000 µg of cefotiam (C
18H
23N
9O
4S
3) per mL. Transfer 5.0 mL of this solution to a 100-mL volumetric flask, dilute with
Mobile phase to volume, and mix. This solution contains the equivalent of about 50 µg of cefotiam (C
18H
23N
9O
4S
3) per mL. Use this solution without delay.
Assay preparation—
Transfer about 60 mg of Cefotiam Hydrochloride, accurately weighed, to a 50-mL volumetric flask, add water to volume, and mix. Transfer 5.0 mL of this solution to a 100-mL volumetric flask, dilute with Mobile phase to volume, and mix. Use this solution without delay.
System suitability solution—
Prepare a solution of
USP Cefotiam Hydrochloride RS in water containing about 1 mg per mL. Heat this solution at 95
for 3 minutes, and cool. Transfer 1 mL of this solution to a 100-mL volumetric flask, dilute with
Mobile phase to volume, and mix.
Chromatographic system
(see
Chromatography 621)—The liquid chromatograph is equipped with a 254-nm detector and a 4-mm × 25-cm column that contains packing L1. The flow rate is about 1.5 mL per minute. Chromatograph the
Standard preparation, and record the peak responses as directed for
Procedure: the column efficiency, determined from the cefotiam peak, is not less than 1985 theoretical plates when calculated by the formula:
5.545(tr / Wh / 2)2,
the tailing factor for the cefotiam peak is not more than 1.8, and the relative standard deviation for replicate injections is not more than 1.0%. Chromatograph the
System suitability solution, and record the peak responses as directed for
Procedure: the relative retention times are about 0.6 for de-tetrazol-cefotiam and 1.0 for cefotiam; and the resolution,
R, between the de-tetrazol-cefotiam peak and the cefotiam peak is not less than 4.0.
Procedure—
Separately inject equal volumes (about 10 µL) of the
Standard preparation and the
Assay preparation into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in µg, of cefotiam (C
18H
23N
9O
4S
3) in each mg of the Cefotiam Hydrochloride taken by the formula:
1000(C/W)(rU / rS),
in which
C is the concentration, in µg per mL, of cefotiam (C
18H
23N
9O
4S
3) in the
Standard preparation, based on the quantity of
USP Cefotiam Hydrochloride RS taken to prepare the
Standard preparation, the designated cefotiam (C
18H
23N
9O
4S
3) content, in µg per mg, of
USP Cefotiam Hydrochloride RS, and the extent of dilution;
W is the weight, in mg, of Cefotiam Hydrochloride taken to prepare the
Assay preparation; and
rU and
rS are the cefotiam peak responses obtained from the
Assay preparation and the
Standard preparation, respectively.
;)
