Abstract
Vitamin K1 isomers and related compounds are separated using a silica HPLC column and a normal phase solvent system. The correct silica type and mobile phase equilibration are important considerations in order to achieve good resolution and retention time reproducibility.
Introduction
Vitamin K is believed to be an important factor in preventing a number of disease states and is essential in the synthesis of blood clotting agents. Consequently, there is obvious interest in monitoring this compound in foods, supplements, and humans. Vitamin K1, also known as phylloquinone or phytomenadione, is the dietary form of vitamin K and is an indicator of overall vitamin K status. The vitamin itself is consumed in the form of green leafy foods such as cabbages, onions, and broccoli. Where subjects are deficient in this vitamin, cholestasis will result and a synthetic oral supplement can be administered. The British Pharmacopoeia (BP) and European Pharmacopoeia (EP) monographs include a normal-phase HPLC method for the determination of the cis and trans isomers and the trans-epoxy modification of vitamin K1. The method in this application note is derived from this methodology. This application note demonstrates the separation of these compounds using a normal phase approach and demonstrates that different silicas can be chromatographically very different.
Experimental Details
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