Vincaleukoblastine, 22-oxo-, sulfate (1:1) (salt).
Leurocristine sulfate (1:1) (salt) [2068-78-2].
» Vincristine Sulfate contains not less than 95.0 percent and not more than 105.0 percent of C46H56N4O10·H2SO4, corrections being applied for loss in weight.
CautionHandle Vincristine Sulfate with great care since it is a potent cytotoxic agent.
Packaging and storage Preserve in tight, light-resistant containers, and store in a freezer.
USP Reference standards 11
USP Vincristine Sulfate RS.
USP Vinblastine Sulfate RS. [noteNo Loss on drying determination is needed for
USP Vinblastine Sulfate RS.]
B: A solution (1 in 10) responds to the tests for Sulfate 191.
pH 791: between 3.5 and 4.5, in a solution (1 in 1000).
Loss on drying (see Thermal Analysis 891) [noteIn this procedure, perform weighings rapidly with minimum exposure of the substances to air.] Determine the percentage of volatile substances by thermogravimetric analysis on an appropriately calibrated instrument, using about 10 mg of Vincristine Sulfate, accurately weighed. Heat the specimen at the rate of 5 per minute between ambient temperature and 200 in an atmosphere of nitrogen at a flow rate of 40 mL per minute. From the thermogram, determine the accumulated loss in weight between ambient temperature and a point on the plateau before decomposition is indicated (at about 160):it loses not more than 12.0% of its weight.
Solvent A Prepare a filtered and degassed mixture of water and diethylamine (985:15), adjusted with phosphoric acid to a pH of 7.5.
Solvent B Use methanol.
High load test preparation Prepare as directed for Assay preparation in the Assay.
Low load test preparation Pipet 1 mL of High load test preparation into a 25-mL volumetric flask, dilute with water to volume, and mix.
Chromatographic system (see Chromatography 621)Use the liquid chromatograph equipped as directed in the Assay. The mobile phase is maintained at a flow rate of about 2 mL per minute, with an initial gradient of 62% of Solvent B and 38% of Solvent A for 12 minutes, then changed to increase Solvent B at a rate of 2% per minute, so that after 15 minutes it will comprise 92% of the mixture, then changed to decrease Solvent B at a rate of 15% per minute, so that after 2 minutes it will again comprise 62% of the mixture, then maintained at this ratio for 5 minutes.
Procedure Separately inject equal volumes (about 200 µL) of the Low load test preparation and of the High load test preparation into the chromatograph, record the chromatograms, and measure the peak responses, ri, of any related substances appearing after the solvent peak in the chromatogram of the High load test preparation. Calculate the total percentage of responses due to related substances taken by the formula:
100rt / (rt + 25rv)in which rt is the sum of the ri responses, and rv is the vincristine peak response in the chromatogram of the Low load test preparation. Not more than 4.0% is found. Calculate the percentage response of each related substance taken by the formula:
100ri / (rt + 25rv).Not more than 1.0% of response due to any individual related substance is found.
Diethylamine solution Mix 5 mL of diethylamine with 295 mL of water, and adjust with phosphoric acid to a pH of 7.5.
Mobile phase Prepare a filtered and degassed mixture of methanol and Diethylamine solution (70:30). Make adjustments if necessary (see System Suitability under Chromatography 621).
Standard preparation Dissolve an accurately weighed quantity of USP Vincristine Sulfate RS in water to obtain a solution having a known concentration of about 1 mg per mL.
Assay preparation Equilibrate a portion of Vincristine Sulfate for 30 minutes with the ambient humidity, transfer about 10 mg, accurately weighed, to a 10-mL volumetric flask, dissolve in and dilute with water to volume, and mix. Using another portion of the equilibrated specimen, determine the loss in weight as directed for USP Vincristine Sulfate RS.
System suitability preparation Transfer 5 mg of USP Vincristine Sulfate RS and 5 mg of Vinblastine Sulfate RS, each accurately weighed, to a 5-mL volumetric flask, dissolve in and dilute with water to volume, and mix.
Chromatographic system (see Chromatography 621)The liquid chromatograph is equipped with a 297-nm detector, a precolumn packed with porous silica gel, a 2- to 5-cm guard column containing packing L1, and a 4.6-mm × 25-cm analytical column that contains packing L7. The flow rate is about 1.5 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the relative standard deviation for replicate injections is not more than 2.0%. Similarly chromatograph 10 µL of the System suitability preparation, and record the peak responses: the resolution, between vincristine sulfate and vinblastine sulfate is not less than 4.0. [noteFor a particular column, the resolution may be increased by increasing the proportion of water in the Mobile phase.]
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 responses for the major peaks. Calculate the quantity, in mg, of C46H56N4O10·H2SO4 in the portion of Vincristine Sulfate taken, corrected for loss in weight in the Assay preparation, by the formula:
10C(rU / rS)in which C is the concentration, in mg per mL, of USP Vincristine Sulfate RS, corrected for loss in weight in the Standard preparation; and rU and rS are the peak 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 3861Pharmacopeial Forum: Volume No. 32(5) Page 1470
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.