» Ribavirin contains not less than 98.9 percent and not more than 101.5 percent of C8H12N4O5, calculated on the dried basis.
Packaging and storage Preserve in tight containers.
USP Reference standards 11
USP Ribavirin RS.
A: Infrared Absorption 197K.
B: Thin-Layer Chromatographic Identification Test 201
Test solution: 10 mg per mL.
Developing solvent system: a mixture of acetonitrile and 0.1 M ammonium chloride (9:2).
Spray reagent Mix 0.5 mL of anisaldehyde, 0.5 mL of sulfuric acid, 0.1 mL of glacial acetic acid, and 9 mL of alcohol.
Procedure Proceed as directed in the chapter. Allow the plate to air-dry for about 15 minutes, spray with Spray reagent, heat the plate at 110 for 30 minutes, and locate the spots on the plate by examining the plate in daylight.
Specific rotation 781S: between 33.5 and 37.0 (t = 20).
Test solution: 10 mg per mL, in water.
pH 791: between 4.0 and 6.5, in a solution (1 in 50), to each 50 mL of which has been added 0.2 mL of a saturated potassium chloride solution.
Loss on drying 731: Dry it at 105 for 5 hours: it loses not more than 0.5% of its weight.
Residue on ignition 281: not more than 0.25%.
Heavy metals, Method II 231: 0.001%.
Mobile phase, Standard preparation, Test solution, and Chromatographic system Prepare as directed in the Assay.
Procedure Inject about 10 µL of the Test solution into the chromatograph, record the chromatogram, and measure the responses of all the peaks, except that of the solvent peak. Calculate the percentage of each peak, other than that of the solvent peak and the main ribavirin peak, by the formula:
100ri / rtin which ri is the response of the individual peak, and rt is the sum of the responses of all the peaks in the chromatogram: not more than 0.25% of any individual peak is found, and the sum of all such peaks does not exceed 1.0%.
Mobile phase Adjust water with sulfuric acid to a pH of 2.5 ± 0.1. 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 Ribavirin RS quantitatively in Mobile phase to obtain a solution having a known concentration of about 0.025 mg per mL.
Test solution Transfer about 50 mg of Ribavirin, accurately weighed, to a 100-mL volumetric flask, add about 50 mL of Mobile phase, swirl to dissolve, dilute with Mobile phase to volume, and mix.
Assay preparation Transfer 5.0 mL of the Test 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 207-nm detector and a 7.8-mm × 10-cm column that contains packing L17 and is operated at 65 ± 0.5. The flow rate is about 1 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the tailing factor for the ribavirin peak is not less than 0.7 and not more than 1.5, and the relative standard deviation for replicate injections is not more than 0.5%.
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 peak area responses for the major peaks. Calculate the quantity, in mg, of C8H12N4O5 in the portion of Ribavirin taken by the formula:
2000C(rU / rS)in which C is the concentration, in mg per mL, of USP Ribavirin RS in the Standard preparation, and rU and rS are the ribavirin peak area responses obtained from the Assay preparation and the Standard preparation, respectively.
Auxiliary Information Please check for your question in the FAQs before contacting USP.Chromatographic Column
USP32NF27 Page 3496Pharmacopeial Forum: Volume No. 27(3) Page 2577
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.