Journal of Chromatography A, 1025 (2004) 313–317 Short communication Simultaneous determination of Vitamin E homologs in chicken meat by liquid chromatography with fluorescence detection A.K. Hewavitharana a,∗ , M.C. Lanari a , C. Becu b a CSIRO, Food Science Australia, Cannon Hill, Qld. 4170, Australia b Ecole Nationale Supérieure de Biologie Apliquee a la Nutrition et a la Alimentation (ENSBANA), Dijon, France Received 17 July 2003; received in revised form 7 October 2003; accepted 16 October 2003 Abstract A simple and reliable method for the simultaneous determination of all eight homologs of Vitamin E in chicken meat is described. All analytes, including the internal standard (-tocopherol acetate), were eluted within 35 min and detected using their native fluorescence (295 nm excitation and 330 nm emission). Chromatography using hexane based eluent on a normal phase silica column included an initial column conditioning step to prevent irreversible adsorption of tocopherols and tocotrienols on silica. Lowest detectable levels of -tocopherol, -tocopherol, -tocotrienol, -tocotrienol, -tocotrienol and -tocotrienol were 0.73, 0.86, 1.0, 1.2, 1.7 and 1.3 ng, respectively. © 2003 Elsevier B.V. All rights reserved. Keywords: Meat; Food analysis; Vitamins; Tocopherols; Tocotrienols 1. Introduction Vitamin E plays a fundamental role in the prevention of radical formation in biological systems like plasma, mem- branes and tissues. Vitamin E is the collective name for the eight naturally occurring forms , ,  and  tocopherols and , ,  and  tocotrienols. Historically, -tocopherol was reported to have the highest biological activity [1] thus most methods available for Vitamin E assay were developed exclusively for the determination of -tocopherol. However, many studies have demonstrated that the antioxidant activ- ities of ,  and  tocopherols were also important [2]. Yamaoka et al. [3] showed that -tocotrienol had a higher antioxidant activity than -tocopherol in a phopholipid li- posome solution. Dietary tocotrienols reduced plasma lipid peroxidation in humans [4], and -tocotrienol decreased the susceptibility of low-density lipoptoteins (LDLs) to copper induced oxidation [5]. The use of plant extracts rich in tocopherols and to- cotrienols in the pharmaceutical and food industries is ∗ Corresponding author. Present address: Investigative Chemistry, Queensland Health Scientific Services, P.O. Box 594, Archerfield, Qld 4108, Australia. Fax: +61-7-3274-9123. E-mail address: Amitha Hewavitharana@health.qld.gov.au (A.K. Hewavitharana). becoming increasingly popular, generating a great need for a fast and efficient technique for separating and quanti- fying the individual Vitamin E analogs. Although several methods have been developed to determine other forms of Vitamin E in various matrices [2], no methods were reported for the separation and/or quantification of all eight vitamin analogs in animal muscle The aim of the present study was to develop a technique for measuring the composition of tocopherols and tocotrienols in meat from chickens supplemented with extracts rich in these antioxi- dants. In this study, we selected an extraction method that claimed to exhibit the minimum loss of Vitamin E [6] which did not include saponification, then further modified to minimize oxidative losses. Precautions were taken in order to prevent inaccuracies in quantification due to irre- versible adsorption of analytes on to the silica column, and fluorescence quenching of tocopherol signal at the detection step of chromatographic analysis. The accuracy, precision and robustness of the method were improved by incorpo- rating an internal standard from the extraction step through to the chromatographic analysis. As well, the use of flu- orescence rather than UV as the detection mode provided the sensitivity and the selectivity required for the accurate determination of low levels of these homologs in muscle tissue. 0021-9673/$ – see front matter © 2003 Elsevier B.V. All rights reserved. doi:10.1016/j.chroma.2003.10.052