Phytonadione and Phytonadione, Vitamin K Isomers Analyzed by HPLC - AppNote
May 8, 2012
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Phytonadione Separation by Shape Selectivity
In this Method, we separate the two E and Z isomers of Phytonadione on the basis of shape selectivity. The below chromatogram is a five injection overlay with a resolution value of 1.5.
Peaks: 1. Impurity, 2. Phytonadione (E isomer), 3. Phytonadione (Z isomer),
Method Conditions
Column:
Cogent UDC-Cholesterol™, 4 μm, 100 Å
Catalog No.
:
69069-15P
Dimensions:
4.6 x 150 mm
- A: 50% DI Water / 50% MeOH / 0.1% Formic Acid
- B: 97% Acetonitrile / 3% DI Water / 0.1% Formic Acid
| Time (minutes) | %B |
|---|---|
| 0 | 80 |
| 15 | 92 |
| 16 | 80 |
Temperature: 12˚C
Post time: 2 minutes
Flow rate: 1.5 mL / minute
Detection: UV @ 254 nm
Sample Preparation:
- Stock Solution: 10 μL / mL Phytonadione in Acetonitrile diluent. (The solution was vortexed for 10 minutes.)
- Working Solution: Stock solution was diluted 1:10 with Acetonitrile.
Note: Phytonadione (a.k.a. Phylloquinone, Vitamin K1) is a lipophilic vitamin that can be obtained in the diet from leafy green vegetables. It plays an essential role in blood clotting by acting as a cofactor for formation of coagulation factors II, VII, IX, and X. The letter designation for Vitamin K was based on the first letter of “Koagulationsvitamin” (coagulation vitamin), which is from the German journal that first published its identification by Danish biochemist Henrik Dam.
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