Methodology for simultaneous estimation of vitamins A and E in animal feeds using high performance liquid chromatogrphy.
2004
Kaur, H. | Ramani, N. K. | Garg, M. R. | Kumar, P., National Dairy Research Institute, Karnal (India)
Validation of International Organisation for Standardisation (ISO) method for vitamin A analysis in feeds was carried out. The method was extended for simultaneous estimation of vitamins A and E in animal feeds. The separation of vitamins A and E was achieved at 1.95 and 6.95 min, respectively, with solvent system consisting of methanol: water in the ratio of 95:5, as used in ISO mehtod for the analysis of vitamins A alone. Various solvent systems comprising tetrahydrofuran, acetonitrile and water were tried at different flow rates to maximize the separation of 2 vitamins in shortest time so that the method can be easily adopted by feed industry of large scale analysis of feed samples. Solvent system consisting of acetonitrile, tetrahydrofuran and HPLC grade water in the ratio of 47:42:11(v/v/v) resulted in the separation of retinol and ¦-tocopherol at retention time of 1.72 and 3.53 min at 325 and 290 nm wavelengths, respectively, at flow rate of 1.5 ml/min. The developed method can be routinely used for the analysis of vitamins A and E in feed samples.
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