Phytonadione Tablets
DEFINITION
Phytonadione Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of phytonadione (C31H46O2).
IDENTIFICATION
•  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.
ASSAY
•  Procedure
[Note—Use 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.
Chromatographic system 
Mode:  LC
Detector:  UV 254 nm
Column:  4-mm × 30-cm; packing L1
Flow rate:  1.5 mL/min
Injection size:  10 µL
System suitability 
Sample:  Standard solution (three replicate injections)
Suitability requirements 
Column efficiency:  NLT 915 theoretical plates, determined from the analyte peak
Tailing factor:  NMT 2.0
Relative standard deviation:  NMT 2.0%
[Note—Z- and E-isomers coelute in this chromatographic system. ]
Analysis 
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
rU== peak area from the Sample solution
rS== peak area from the Standard solution
CS== concentration of USP Phytonadione RS in the Standard solution (mg/mL)
CU== nominal concentration of phytonadione in the Sample solution (mg/mL)
Acceptance criteria:  90.0%–110.0%
PERFORMANCE TESTS
•  Disintegration 701
Time:  30 min
•  Uniformity of Dosage Units 905: Meet the requirements
ADDITIONAL REQUIREMENTS
•  Packaging and Storage: Preserve in well-closed, light-resistant containers.
•  USP Reference Standards 11
USP Phytonadione RS Click to View Structure
Auxiliary Information— Please check for your question in the FAQs before contacting USP.
Topic/Question Contact Expert Committee
Monograph Natalia Davydova
Scientific Liaison
1-301-816-8328
(DS2010) Monographs - Dietary Supplements
701 Margareth R.C. Marques, Ph.D.
Senior Scientific Liaison
1-301-816-8106
(GCDF2010) General Chapters - Dosage Forms
Reference Standards RS Technical Services
1-301-816-8129
rstech@usp.org
USP35–NF30 Page 4319