Determination of α-tocopherol by reversed-phase HPLC in feed and animal-derived foods without saponification

J Sci Food Agric. 2016 Jan 30;96(2):522-9. doi: 10.1002/jsfa.7119. Epub 2015 Feb 27.

Abstract

Background: The analysis of α-tocopherol in feed and animal-derived foods usually involves a saponification step. However, since saponification often leads to losses of α-tocopherol, a method for the determination of α-tocopherol in feed and in animal-derived foods was developed without a saponification step.

Results: In this method, α-tocopherol is extracted with hot ethanol and the co-extracted fat is removed by centrifugation. Removal of the fat fraction is made possible by the addition of water, to achieve an ethanol:water ratio of 40:7, followed by cooling on ice before centrifugation. This procedure allows removal of the fat fraction, while α-tocopherol is retained. Matrices differing in gross composition and α-tocopherol content were analyzed: fresh pork, cooked ham, subcutaneous fat, liver, egg yolk, milk and a compound pig feed. Higher α-tocopherol concentrations were found for this novel method compared to a conventional method with saponification, particularly for subcutaneous fat (P < 0.05). Recoveries were higher (P < 0.05) for the novel method (82-103%), compared to the saponification method (66-90%; for subcutaneous fat < 25%).

Conclusion: Determining α-tocopherol in feed and animal-derived foods using pure ethanol without saponification results in higher extraction yields and recoveries compared to the saponification method.

Keywords: egg; fat; feed; meat; milk; vitamin E.

MeSH terms

  • Animal Feed / analysis*
  • Animals
  • Cattle
  • Chromatography, High Pressure Liquid / methods*
  • Egg Yolk / chemistry*
  • Food Analysis / methods
  • Liver / chemistry
  • Meat / analysis*
  • Milk / chemistry*
  • Subcutaneous Fat / chemistry
  • Swine
  • alpha-Tocopherol / chemistry*

Substances

  • alpha-Tocopherol