A method for simultaneous determination of retinol and all 4 tocopherols was developed using HPLC equipped with a reversed-phase column. Using this method, β-, γ-tocopherol and retinol, which had been difficult to separate by HPLC equipped with a reversed-phase column, can be separately determined. HPLC conditions were as follows ; column : YMC-PACK A 302 S-5 120A ODS, detector ; fluorophotometer (excitation wave length 340 nm and emission wave length 460 nm for retinol, excitation wave length 298 nm and emission wave length 325 nm for tocopherols), mobile phase : 65% isopropanol, sample solvent : 99.5% methanol, temperature : 30℃. Retinol and tocopherols in umbilical cord plasma, adult plasma, and liver of rats were determined using this method. This method can simultaneously determine retinol and each tocopherol homologue in biological samples.
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