Abstract
AbstractThe fractionation and analysis of the lipid classes and fluorescent substances of animal tissues by high performance liquid chromatography (HPLC) using a combination of fluorescence and flame ionization detectors is described. The lipid classes and fluorescent substances are extracted from rat kidney and liver tissue by a new method that involves preextraction of nonlipid and aqueous‐soluble fluorescent substances with hot dilute (0.05 N) acetic acid. The lipid classes and organic‐soluble fluroscent substances are extracted from the residual tissue in three extractions: the first with chloroform/methanol, 1:1, v/v; the second with chloroform/methanol, 1:2, v/v; and the third with methanol. The fractionation of these compounds by HPLC is carried out with a column 0.2×45 cm, packed with a special adsorbent prepared by reacting silicic acid with ammonium hydroxide. The eluent is passed through a fluorescence detector, which provides a profile of the fluorescent compounds, and then to a flame ionization detector for analysis of the lipid classes. The method is demonstrated on rat blood serum, liver and kidney tissue.
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