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1 Ars Separatoria Acta

1 2002

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Determination of conjugated fatty acid in ovine milk, meat, fat and intestinal digesta

Czauderna M., Kowalczyk J., Potkański A., Szumacher-Strabel N.

Ars Separatoria Acta

2002

No. 1

104-110

Positional and geometric isomers of geometric isomers of linoleic acid (CLA) were separated from interfering species on commercially available two reversed-phase C18-columns (Nova Pak, Waters) in gradient systems composed of acetonitrile and water, utilizing photodiode array detection. The biological samples were hydrolyzed with 2 M NaOH for 35 min at 85°C. After cooling, the hydrolysates were acidified with 4 M HCl and the free fatty acids were extracted with dichloromethane. The CLA isomers were determined directly using UV detection at 234.5 nm or after pre -column derivatization with 2,4’-dibromoacetophenone in the presence of triethylamine and UV detection at 256 and 235 nm. HPLC system with pre-column derivatization enables more efficient fractionation of the CLA isomers than the direct HPLC system. On the other hand, elimination of derivatization procedure provides a less expensive, more specific and simpler analytical tool for determination of CLA than HPLC method with precolumn derivatization. The presented HPLC methods provide analytical tools for simple quantification of CLA in ovine meat, milk, fat and intestinal digesta samples.