Determination of alpha-tocopherol in plasma by high performance liquid chromatography with fluorescence detection and stability of alpha-tocopherol under different conditions.
In order to study the clinical effects of the antioxidant alpha-tocopherol (tocofersolan, CAS 30999-06-5) a simple, robust, sensitive and reliable high performance liquid chromatography (HPLC) method using fluorescence detection for the daily measurement of alpha-tocopherol concentration in plasma is described. Several methods of HPLC analysis using fluorescence techniques are published, however, investigations of the stability of alpha-tocopherol during sample preparation under different conditions are relevant in the routine measurements and have to date not been reported. Therefore a modified method, both sensitive and robust, qualified for the day-by-day measurement of alpha-tocopherol in plasma was established and the stability of alpha-tocopherol during sample preparation was analysed under different conditions. This method enables the complete separation of alpha-tocopherol from other compounds within 14 min. In addition, investigations on the stability of alpha-tocopherol during sample preparation showed the necessity of rapid sample preparation without delay. Prepared samples can be stored in liquid nitrogen for at least 56 days.
Rümelin
Andreas
A
Clinic of Anaesthesiology, Johannes Gutenberg University, Mainz, Germany. ruemelin@mail.uni-mainz.de
Humbert
Till
T
Fauth
Ulrich
U
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D015203:Reproducibility of Results; D013048:Specimen Handling; D013050:Spectrometry, Fluorescence; D024502:alpha-Tocopherol / Q000097:blood*
D024502:alpha-Tocopherol