Cefprozil (E)-isomer ratio
Calculate the ratio of the cefprozil (
E)-isomer to the total cefprozil taken by the formula:
E / (E + Z),
in which
E is the content of cefprozil (
E)-isomer, in µg per mg, as determined in the
Assay, and
Z is the content of cefprozil (
Z)-isomer, in µg per mg, as determined in the
Assay; the ratio is between 0.06 and 0.11.
Assay
Mobile phase
Dissolve 20.7 g of monobasic ammonium phosphate in 1800 mL of water, and adjust, if necessary, with phosphoric acid to a pH of 4.4. Add 200 mL of acetonitrile, and mix. Filter this solution through a filter having a porosity of 0.5 µm or finer, and degas. Make adjustments if necessary (see
System Suitability under
Chromatography 621.
[NOTEDecreasing the proportion of acetonitrile increases retention times and improves the separation of the cefprozil isomer peaks.
]
Standard cefprozil
(
Z)
-isomer preparationDissolve an accurately weighed quantity of
USP Cefprozil (Z)-Isomer RS. quantitatively in water to obtain a solution having a known concentration of about 0.25 mg per mL.
[NOTEUse this solution within 6 hours.
]
Standard cefprozil
(
E)
-isomer preparationDissolve an accurately weighed quantity of
USP Cefprozil (E)-Isomer RS. quantitatively in water to obtain a stock solution having a known concentration of about 0.25 mg per mL. Transfer 5.0 mL of this stock solution to a 50-mL volumetric flask, dilute with water to volume, and mix.
[NOTEUse this solution within 6 hours.
]
Resolution solution
Prepare a mixture of equal volumes of the Cefprozil (Z)-isomer standard preparation and of the stock solution used to prepare the Cefprozil (E)-isomer standard preparation. [NOTEUse this solution within 6 hours.]
Assay preparation
Transfer about 15 mg of Cefprozil, accurately weighed, to a 50-mL volumetric flask, dilute with water to volume, and shake to assure dissolution. [NOTEUse this solution within 6 hours.]
Chromatographic system
(see
Chromatography 621)The liquid chromatograph is equipped with a 280-nm detector and a 3.9-mm × 25-cm column that contains 5-µm packing L1. The flow rate is about 1 mL per minute. Chromatograph the
Resolution solution, and record the responses as directed under
Procedure: the relative retention times are about 0.7 for cefprozil (
Z)-isomer and 1.0 for cefprozil (
E)-isomer, and the resolution,
R, between the cefprozil (
Z)-isomer peak and the cefprozil (
E)-isomer peak is not less than 2.5. Chromatograph the
Cefprozil (
Z)-
isomer standard preparation, and record the responses as directed under
Procedure: the column efficiency, determined from the cefprozil (
Z)-isomer peak, is not less than 2500 theoretical plates when calculated by the formula:
5.545(tr / Wh / 2)2,
the tailing factor, determined from the cefprozil (
Z)-isomer peak, is not less than 0.9 and not more than 1.1, when calculated by the formula:
W0.1 / 2f,
in which
W0.1 is the width of the peak at 10% height, and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure
Separately inject equal volumes (about 10 µL) of the
Cefprozil (
Z)-
isomer standard preparation, the
Cefprozil (
E)
-isomer 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 cefprozil (
Z)-isomer and cefprozil (
E)-isomer in each mg of the Cefprozil taken by the formula:
50(CP / M)(rU / rS),
in which
C is the concentration, in mg per mL, of USP Cefprozil (
Z)-Isomer RS in the
Cefprozil (
Z)
-isomer standard preparation or of the USP Cefprozil (
E)-Isomer RS in the
Cefprozil (
E)
-isomer standard preparation, as appropriate;
P is the assigned potency, in µg per mg, of the appropriate USP Reference Standard;
M is the quantity, in mg, of Cefprozil taken to prepare the
Assay preparation; and
rU and
rS are the peak responses of the cefprozil (
Z)-isomer or the cefprozil (
E)-isomer, as appropriate, obtained from the
Assay preparation and the relevant
Standard preparation, respectively. Calculate the quantity, in µg, of cefprozil (C
18H
19N
3O
5S) in each mg of the Cefprozil taken by adding the values, in µg per mg, obtained for the cefprozil (
Z)-isomer and for cefprozil (
E)-isomer.