ORIGINAL PAPER
 
KEYWORDS
TOPICS
ABSTRACT
The objectives of our study were to enhance and evaluate an improved saponification method for the quantification of fatty acids (FA), with a specific focus on highly-unsaturated long-chain polyunsaturated FA (LPUFA) that are easily peroxidised in pig erythrocytes. These erythrocytes serve as a valuable large animal model for human studies. We implemented a modified saponification procedure, involving a shorter initial saponification at 95 °C for 2 min, followed by overnight saponification at 22–25 °C, and subsequent gentle base and acid-catalysed methylation of FA. These analytical procedures proved to be suitable for gas-chromatographic analysis of highly-unsaturated LPUFA, which are prone to peroxidation in processed piglet erythrocytes. The modified initial saponification at 95 °C, followed by overnight saponification at 22–25 °C, and subsequent methylation, were used to assess the suitability of erythrocytes for the determination of FA, tocopherol and malondialdehyde (MDA) concentrations in monogastric mammalian tissues. The content of docosahexaenoic acid (DHA) in erythrocytes can serve as a marker for DHA bioaccumulation in animal tissues and dietary contents of n-3LPUFA (especially DHA). The concentrations of δ-, γ-, α-tocopherols, total cholesterol and MDA increased in piglet erythrocytes during 16 days of exposure to dietary fat. The levels of FA and tocopherols can therefore be used in erythrocytes as indicators of the bioaccumulation yield of these compounds in animal tissues. Additionally, we propose that FA and tocopherol profiles in erythrocytes may be used as indicators of the concentrations of these components in diets and the effectiveness of their bioaccumulation in animal tissues.
ACKNOWLEDGEMENTS
The authors would like to express their gratitude to Dr. Janine Donaldson for her valuable contributions and feedback during the preparation of the manuscript. Her insights and expertise greatly enhanced the quality of this research.
FUNDING
We would like to acknowledge the financial support provided by the National Science Centre, Grant No. 2020/04/X/NZ9/00358 (MINIATURA 4), as well as the support from the statutory funds of The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Jabłonna, Poland.
CONFLICT OF INTEREST
The Authors declare that there is no conflict of interest.
 
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