(floo sye' toe seen).
» Flucytosine contains not less than 98.5 percent and not more than 101.0 percent of C4H4FN3O, calculated on the dried basis.
Packaging and storage Preserve in tight, light-resistant containers.
USP Reference standards 11
Solution: 8 µg per mL.
Medium: dilute hydrochloric acid (1 in 100).
Absorptivities at 285 nm, calculated on the dried basis, do not differ by more than 2.0%.
B: The RF value of the principal spot in the specimen chromatogram in the test for Fluorouracil corresponds to that obtained with the solution of USP Flucytosine RS.
Loss on drying 731 Dry it at 105 for 4 hours: it loses not more than 1.5% of its weight.
Residue on ignition 281: not more than 0.1%.
Heavy metals, Method II 231: 0.002%.
Fluoride ions [noteAll glassware and/or plasticware used in this test should be scrupulously clean and even free from trace amounts of fluoride. The use of plasticware to contain the solutions while the potential is measured is recommended. ]
Buffer solution To 110 g of sodium chloride in a 2-L volumetric flask add 1 g of sodium citrate and 700 mL of water, and dissolve with shaking. Carefully add 150 g of sodium hydroxide, and dissolve with shaking. Cool to room temperature, and while stirring, cautiously add 450 mL of glacial acetic acid. Cool to room temperature, add 600 mL of isopropyl alcohol, dilute with water to volume, and mix. The pH of this solution is between 5.0 and 5.5.
Standard stock solution Accurately weigh 2.211 g of sodium fluoride, previously dried at 150 for 4 hours, into a 1-L volumetric flask, and dissolve in about 200 mL of water. Add 1.0 mL of sodium hydroxide solution (1 in 250), dilute with water to volume, and mix. Each mL of this solution contains 1 mg of fluoride ion. Store the solution in a closed plastic container.
Standard preparations Dilute a portion of Standard stock solution quantitatively and stepwise with Buffer solution to obtain a Standard preparation having a fluoride concentration of 1 µg per mL. Prepare the final dilution in a 100-mL volumetric flask. In the same manner, prepare additional Standard preparations having fluoride concentrations of 3 µg per mL, 5 µg per mL, and 10 µg per mL, respectively.
Test preparation Place 1 g of Flucytosine, accurately weighed, in a 100-mL volumetric flask, and dissolve in and dilute with Buffer solution to volume.
Procedure Concomitantly measure the potential (see Titrimetry 541), in mV, of the Standard preparations and the Test preparation, with a suitable pH meter equipped with a fluoride-specific ion electrode and a glass-sleeved calomel reference electrode that has been modified in the following manner. Mix 70 mL of freshly prepared saturated potassium chloride solution with 30 mL of isopropyl alcohol, fill the electrode with the clear supernatant, and allow the electrode to remain in the mixture for not less than 2 hours prior to use, or preferably overnight.
When taking the measurements, transfer the solution to a 150-mL beaker, and immerse the electrodes. Insert a polytef-coated stirring bar into the beaker, place the beaker on a magnetic stirrer having an insulated top, and allow to stir until equilibrium is attained (about 1 to 2 minutes). Rinse and dry the electrodes between measurements, taking care not to scratch the crystal in the specific ion electrode.
Measure the potential of each Standard preparation, and plot the fluoride concentration, in mg per 100 mL, versus the potential, in mV, on semilogarithmic paper. Measure the potential of the Test preparation, and determine from the standard curve the fluoride concentration, in mg per 100 mL. Calculate the percentage of fluoride in the portion of Flucytosine taken by the formula:
C / 10
in which C is the fluoride concentration, in mg per 100 mL, from the standard curve: not more than 0.05% of fluoride is found.
Fluorouracil Dissolve 250 mg in 10 mL of a mixture of glacial acetic acid and water (4:1). Apply 20 µL of this solution to a thin-layer chromatographic plate (see Chromatography 621) coated with a 0.5-mm layer of chromatographic silica gel mixture. To the same plate apply 20 µL, in 10-µL increments, of a 0.025 mg per mL solution of USP Fluorouracil RS in a mixture of glacial acetic acid and water (4:1). Develop the chromatogram in a mixture of chloroform and glacial acetic acid (13:7) until the solvent front has moved not less than 14 cm from the origin. Remove the plate from the developing chamber, and allow the solvent to evaporate. Locate the spots on the plate by observing under short-wavelength UV radiation: any spot from the solution under test is not greater in size and intensity than the spot at the respective RF produced by the Standard solution, corresponding to not more than 0.1% of fluorouracil.
Assay Place about 400 mg of Flucytosine, accurately weighed, in a 250-mL beaker, add 150 mL of a mixture of 2 volumes of glacial acetic acid and 1 volume of acetic anhydride, and dissolve, warming gently if necessary. Titrate potentiometrically with 0.1 N perchloric acid VS, using a calomel-glass electrode system. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 12.91 mg of C4H4FN3O.
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USP35NF30 Page 3209Pharmacopeial Forum: Volume No. 30(5) Page 1621