Evaluation of HPLC method for the rapid and simple determination of α-tocopherol acetate in feed premixes
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Research Institute of Animal Production, Department of Feed Sciences, 32-083 Balice, Poland
Research Institute of Animal Production, Central Laboratory 32-084 Morawica, Poland
Publication date: 2002-08-02
J. Anim. Feed Sci. 2002;11(3):527–536
Alpha-tocopherol was determined in feeds for chickens and pigs. Samples for chromatographic (HPLC) analysis were prepared using a method encompassing one-step extraction with a 30:70 (v/v) mixture of acetonexhloroform. The samples were eluted isocratically from a LiChroCART™ 250-4 column with UV detection using a UV-VIS detector at a wavelength of 290 nm. The eluent was methanol at a flow rate of 1 ml/min. 20 µI samples were injected into the column using an autosampler. The retention time of the a-tocopherol acetate peak was 13.41 min with a coefficient of variation of 2.78%. The repeatability of the area under the curve, expressed as the coefficient of variation, was 22.7%) for α-tocopherol concentrations ranging from 0.5 to 1 µg/ml, 8.3%o for a range from 1-11 µg/ml, and 1.8% for a range from 11 to 3000 µg/ml. The coefficient of variation for the full assay of a-tocopherol acetate in samples encompassing all of its stages, starting from the weighing of a sample to its chromatographic analysis, did not exceed 7.3%. The recovery of tocopherol measured against an external standard was 92.4%, and against a standard processed through the sample preparation steps 93.2%. Alpha-tocopherol acetate standard stored at 4°C and protected from the light was found to be very stable. Two of the advantages of the developed method are its speed and ease of performance.
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