Determination of alpha-, gamma(+beta)-, and delta-tocopherols in a variety of liver tissues by reverse-phase high pressure liquid chromatography

A modified version of a previously published extraction technique using acetone as the extracting medium and separation on an HPLC equipped with a Spherisorb C-18 ODS2 column packed with 3 mu m particles along with a suitable flow rate and mobile phase significantly improves the resolution of the chromatographic output (fluorescence detector), allowing for accurate measurement of alpha-, delta- and gamma(+beta)-tocopherols. It was found that two extractions of the same sample yielded approximately 97% of the total extractable alpha-tocopherol. Vitamin C was not needed as an anti-oxidant to protect alpha-tocopherol during extraction from chicken liver but was needed when using fish liver. alpha-tocopherol was found not to be evenly distributed in beef, chicken or fish liver.