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A reliable and sensitive method for the determina- tion of ethanolamine and serine, the bases present in kephalin, is necessary in the study of phospho- lipid metabolism. The procedure described by Artom (1945) is reliable but not sensitive (range 10-50 1tmoles), whereas that described by Levine & Chargaff (1951) is sensitive (range 0 1-0 3panole) but unreliable. The method devised by Axelrod, Reichenthal & Brodie (1953) involves the hydrolysis of the fat, the reaction of the free bases with 1-fluoro-2:4-dinitrobenzene (FDNB), the separation of the N-dinitrophenyl (DNP) bases and their quiantitative estimation by their light absorption at a wavelength at which FDNB does not inter- fere. The present paper describes a method which involves the reaction of the lipids with FDNB, the complete removal of excess of reagents, the hydrolysis of the DNP-lipids and the determination of DNP-ethanolamine and DNP-serine by a differential colour reaction which does not involve their separation.
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Collins et al. (1958) studied this question.
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