Abstract
A sensitive method for the determination of human serum trans-phylloquinone levels has been developed. Serum samples were extracted with hexane and subjected to preliminary separation on a silica semipreparative HPLC column with an 80% recovery as determined by the addition of [ 3H]phylloquinone. The portion of the eluate containing trans-phylloquinone was dried and injected into a μBondapak C-18 analytical HPLC column, and the concentration of the vitamin was determined by reductive electrochemical detection utilizing a glassy carbon electrode and and Ag AgCl reference electrode. The presumed vitamin peak was confirmed as trans-phylloquinone by rechromatography at different HPLC conditions, hydrodynamic voltamography, and photodegradation. As little as 300 pg/ml rans-phylloquinone could be detected, and normal human serum concentrations were found to be in the range of 1 ng/ml.
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