Analysis by High Performance Liquid Chromatography of The Water-Soluble Precursors of Choline and Ethanolamine Phosphoglycerides in the Rat Brain

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Abstract
The simultaneous quantitation of PC, PE, CDP-cho-line and CDP-ethanolamine has been performed by high performance liquid chromatography of purified rat brain extracts. The water-soluble intermediates of the pho-sphoglyceride biosynthesis were extracted from the rat brain by homogenization in 1 M PCA and purified by chro-matography on neutral alumina and Dowex 1. This procedure removes almost all of the UV-contaminating substances and inorganic phosphate, which interfere in the a-nalytical chromatographic process. The high performance liquid chromatography was carried out using the styre-ne-type Aminex A-14 resin. The elution was performed for 5 min with 0.1 M 2-amino-2-methyl-1-propanol, 0.02 M NaCl, pH 10.3 buffer, and then with 0.1 M 2-amino-2-me-thyl-1-propanol, 0.17 M NaCl, pH 11.0 solution, until the end of the run. Satisfactory separations of PC, PE, CDP-choline and CDP-ethanolamine were achieved within 30 min. 5′-CMP can also be dosed. The cytidine-containing compounds have been detected and quantified by UV absorption at 280 nm, while PC and PE by phosphorus determination in the chromatographic fractions. The method allows the quantitative determination in subgram amounts of wet tissue.

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