Phytonadione Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of phytonadione (C31H46O2).
• A. Ultraviolet Absorption
Standard solution: 0.01 mg/mL of USP Phytonadione RS in dehydrated alcohol
Sample solution: A portion of finely powdered Tablets, equivalent to 0.01 mg/mL of phytonadione in dehydrated alcohol. Shake vigorously, and filter. Use the filtrate.
Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of the Standard solution concomitantly measured.
• B. HPLC Identification Test
Analysis: Proceed as directed in the Assay.
Acceptance criteria: The retention time of the major peak of the Sample solution corresponds to that of the Standard solution.
[NoteUse low-actinic glassware throughout the Assay, and otherwise protect the solutions from light. ]
Mobile phase: Dehydrated alcohol and water (95:5)
Standard solution: 0.10 mg/mL of USP Phytonadione RS in dehydrated alcohol
Sample solution: 0.1 mg/mL of phytodione in dehydrated alcohol prepared as follows. Mix a portion of finely powdered Tablets (NLT 20) with dehydrated alcohol to obtain a nominal concentraion of 0.4 mg/mL of phytonadione. Shake by mechanical means for 15 min, dilute with dehydrated alcohol to 0.10 mg/mL, and filter.
Detector: UV 254 nm
Column: 4-mm × 30-cm; packing L1
Flow rate: 1.5 mL/min
Injection size: 10 µL
Sample: Standard solution (three replicate injections)
Column efficiency: NLT 915 theoretical plates, determined from the analyte peak
Tailing factor: NMT 2.0
Relative standard deviation: NMT 2.0%
[NoteZ- and E-isomers coelute in this chromatographic system. ]
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of phytonadione (C31H46O2) in the portion of Tablets taken:
Result = (rU/rS) × (CS/CU) × 100
Acceptance criteria: 90.0%110.0%
• Disintegration 701
Time: 30 min
• Uniformity of Dosage Units 905: Meet the requirements
• Packaging and Storage: Preserve in well-closed, light-resistant containers.
• USP Reference Standards 11
Auxiliary Information Please check for your question in the FAQs before contacting USP.
USP35NF30 Page 4319
Chromatographic columns text is not derived from, and not part of, USP 35 or NF 30.