Abstract
Oleic acid has a partition coefficient, upper phase/lower phase, of 1.9 (22 degrees C) in the liquid-liquid partition system described herein. Tri-, di-, and monoolein are found almost exclusively in the lower (organic) phase. Oleic acid can be quantitatively removed from mixtures of triglyceride and partial glycerides by means of this partition system under conditions resembling those in a lipase assay.
Highlights
If chloroform is included in the organic phase, two improvements over Borgstrom’s system can be brought about: (u) free fatty acids are separated from partial glyceride products of lipolysis as well as from the triglyceride substrate, and (6) the free fatty acids are found in the upper phase so that sampling for radioassay is convenient and rapid
The percentage values were determined from an aliquot of the upper phase as described in the text
From a practical point of view, it is important to know whether the distribution of the oleic acid remains constant at different temperatures and in the presence of
Summary
LIPASEACTIVITY has been most commonly assayed by titration of fatty acids produced from hydrolysis of a triglyceride emulsion, The use of radioactive triglycerides greatly increases the sensitivity of the assay but isolation of the fatty acids from the glycerides is timeconsuming. When we desired to express lipase activity in absolute terms, the total amount of radioactive fatty acid could be calculated from the corresponding upper phase value and the partition coefficient of the fatty acid. The latter had to be determined in a separate experiment with labeled fatty acid and varies to some extent as discussed below. The solvent mixtures were added immediately, except when otherwise indicated, and the procedure described above was followed
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