Vincristine Sulfate for Injection
Vincristine Sulfate for Injection is a sterile mixture of Vincristine Sulfate with suitable diluents. It contains NLT 90.0% and NMT 110.0% of the labeled amount of vincristine sulfate (C46H56N4O10·H2SO4).
[CautionHandle Vincristine Sulfate for Injection with great care because it is a potent cytotoxic agent. ]
Change to read:• A. Thin-Layer Chromatographic Identification Test 201
Standard solution: 10 mg/mL of USP Vincristine Sulfate RS or USP Vincristine Sulfate (Assay) RS(RB 1-Jul-2011) in dichloromethane and methanol (3:1)
Sample stock solution: 25 mg/mL of vincristine sulfate from Vincristine Sulfate for Injection in water
Sample solution: 10 mg/mL of vincristine sulfate from Sample stock solution in methanol
Adsorbent: 0.25-mm layer of chromatographic silica gel mixture
Application volume: 20 µL
Developing solvent system: Fresh ether, methanol, and methylamine solution (2 in 5) (19:2:1)
Spray reagent: Dissolve 2.0 g of ceric ammonium sulfate in 100 mL of water with heating and stirring, and slowly add 100 mL of phosphoric acid. Filter if necessary.
Analysis: Develop the chromatographic plate in a methanol prewash tank; for maximum sensitivity, dry it NMT 2 h before use. Score it about 15 cm above the points of application. Apply the Sample solution and the Standard solution at points about 2.5 cm from the lower edge of the plate, and dry thoroughly (a current of cool air may be used to help dry the spots). Place the plate in the nonequilibrated developing chamber that contains a paper liner around the back and sides and Developing solvent system to a depth of about 2 cm. Remove the plate when the solvent moves to the scored line (about 80 min), and discard the solvent system. Dry the plate in a fume hood at room temperature, heat on a metal plate on a steam bath for about 15 min, and spray the plate while still hot with Spray reagent. Continue heating the plate for 15 min to stabilize the spots.
Acceptance criteria: The RF value and the color of the principal spot from the Sample solution correspond to those from the Standard solution.
• B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
Change to read:• Procedure
Solution A: Diethylamine and water (1:59). Adjust with phosphoric acid to a pH of 7.5.
Mobile phase: Methanol and Solution A (70:30)
Standard solution: 1.2 mg/mL(RB 1-Jul-2011) of USP Vincristine Sulfate RS or USP Vincristine Sulfate (Assay) RS(RB 1-Jul-2011) in water
System suitability solution: 1 mg/mL of (RB 1-Jul-2011) USP Vinblastine Sulfate RS in the Standard solution(RB 1-Jul-2011)
Sample solution: 1.2 mg/mL(RB 1-Jul-2011) of vincristine sulfate from Vincristine Sulfate for Injection in water. [NoteShake to mix. ]
Detector: UV 297 nm
Precolumn: Porous silica gel packing
Guard: 2- to 5-cm; packing L1
Analytical: 4.6-mm × 25-cm; packing L7
Flow rate: 1.5 mL/min
Injection size: 10 µL
Samples: Standard solution and System suitability solution
Resolution: NLT 4.0 between vincristine sulfate and vinblastine sulfate, System suitability solution. [NoteFor a particular column, the resolution may be increased by increasing the proportion of water in the Mobile phase. ]
Relative standard deviation: NMT 2.0%, Standard solution
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of vincristine sulfate (C46H56N4O10·H2SO4) in the portion of Vincristine Sulfate for Injection taken:
Result = (rU/rS) × (CS/CU) × 100
Acceptance criteria: 90.0%110.0%
Change to read:• Uniformity of Dosage Units 905
Procedure for content uniformity
Buffer solution: Dissolve 6.3 g of ammonium formate in about 900 mL of water. Adjust with formic acid to a pH of 5.0 while stirring, and dilute with water to 1000 mL.
Standard solution: 4050(RB 1-Jul-2011) µg/mL of USP Vincristine Sulfate RS or USP Vincristine Sulfate (Assay) RS(RB 1-Jul-2011) in Buffer solution
Sample solution: Dissolve the contents of one container of Vincristine Sulfate for Injection in a suitable volume of Buffer solution to obtain a solution containing 4050 µg/mL of vincristine sulfate.
Cell: 1 cm
Analytical wavelength: 262 nm
Blank: Buffer solution
Samples: Standard solution and Sample solution
Calculate the quantity, in mg, of vincristine sulfate (C46H56N4O10·H2SO4) in the container of Vincristine Sulfate for Injection taken:
Result = (AU/AS) × CS × V × F
Acceptance criteria: Meets the requirements for solids
• Organic Impurities
Solution A: Diethylamine and water (3:197). Adjust with phosphoric acid to a pH of 7.5.
Solution B: Methanol
Mobile phase: See Table 1.
Standard solution, System suitability solution, and System suitability: Proceed as directed in the Assay.
Sample solution A: 1 mg/mL of vincristine sulfate from Vincristine Sulfate for Injection in water
Sample solution B: 0.04 mg/mL of vincristine sulfate in water from Sample solution A
Chromatographic system: Proceed as directed in the Assay, except use a Flow rate of 2 mL/min and an Injection size of 200 µL.
Samples: Sample solution A and Sample solution B
Calculate the percentage of each impurity in the portion of Vincristine Sulfate for Injection taken:
Result = [rUA/(SrUA + 25rUB)] × 100
Calculate the percentage of total impurities in the portion of Vincristine Sulfate for Injection taken:
Result = [SrUA/(SrUA + 25rUB)] × 100
Individual impurities: NMT 2.0%
Total impurities: NMT 5.0%
• Sterility Tests 71: Meets the requirements
• Bacterial Endotoxins Test 85: It contains NMT 100.0 USP Endotoxin Units/mg of vincristine sulfate.
• Constituted Solution: At the time of use, it meets the requirements for Injections 1, Constituted Solutions.
• Other Requirements: It meets the requirements in Injections 1, Labeling.
• Packaging and Storage: Preserve as described in Injections 1, Containers for Sterile Solids, and store in a refrigerator.
• Labeling: The label states, For Intravenous Use OnlyFatal If Given By Other Routes.
Where labeled as containing more than 2 mg, it must also be labeled as a Pharmacy Bulk Package (see Injections 1). The labeling directs that the drug be dispensed only in containers enclosed in an overwrap labeled as directed below. When packaged in a Pharmacy Bulk Package, it is exempt from the requirement in Injections 1 that the closure be penetrated only one time after constitution with a suitable sterile transfer device or dispensing set, when it contains a suitable substance or mixture of substances to prevent the growth of microorganisms.
When dispensed, the container or syringe (holding the individual dose prepared for administration to the patient) must be enclosed in an overwrap bearing the statement, Do Not Remove Covering Until Moment of Injection. For Intravenous Use OnlyFatal If Given By Other Routes.
Change to read:• USP Reference Standards 11
USP Endotoxin RS
USP Vinblastine Sulfate RS
[NoteNo loss on drying determination is needed for USP Vinblastine Sulfate RS. ]
USP Vincristine Sulfate (Assay) RS(RB 1-Jul-2011)
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USP35NF30 Page 5025Pharmacopeial Forum: Volume No. 37(1)