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. 1981 Apr 1;195(1):301–306. doi: 10.1042/bj1950301

Alkaline O leads to N-transacylation. A new method for the quantitative deacylation of phospholipids.

N G Clarke, R M Dawson
PMCID: PMC1162886  PMID: 7306057

Abstract

1. Quantitative O-deacylation of phospholipids has been achieved by incubation with a reagent containing monomethylamine, methanol and water. The reaction is primarily an O leads to N-transacylation with N-methyl fatty acid amides being formed. 2. The reagent can be removed easily by volatilization and under defined conditions no secondary decomposition of the phosphorus-containing deacylation products occurs. 3. The water-soluble phosphorus compounds derived by deacylation of mammalian tissue O-diacylated phospholipids have been completely separated by a single-dimensional paper ionophoresis with a volatile pH9 buffer. 4. The O-deacylated alkyl and alkenyl phospholipids have been examined by t.l.c. before and after catalytic hydrolysis with Hg2+. 5. A complete analysis of rat brain phospholipids by the above methods agrees closely with that obtained by other procedures.

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Selected References

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  1. ABDEL-LATIF A. A., ABOOD L. G. INCORPORATION OF ORTHO (32P)PHOSPHATE INTO THE SUBCELLULAR FRACTIONS OF DEVELOPING RAT BRAIN. J Neurochem. 1965 Mar;12:157–166. doi: 10.1111/j.1471-4159.1965.tb06752.x. [DOI] [PubMed] [Google Scholar]
  2. ANSELL G. B., SPANNER S. THE ALKALINE HYDROLYSIS OF THE ETHANOLAMINE PLASMALOGEN OF BRAIN TISSUE. J Neurochem. 1963 Dec;10:941–945. doi: 10.1111/j.1471-4159.1963.tb11921.x. [DOI] [PubMed] [Google Scholar]
  3. BROCKERHOFF H. BREAKDOWN OF PHOSPHOLIPIDS IN MILD ALKALINE HYDROLYSIS. J Lipid Res. 1963 Jan;4:96–99. [PubMed] [Google Scholar]
  4. Bjerve K. S., Daae L. N., Bremer J. The selective loss of lysophospholipids in some commonly used lipid-extraction procedures. Anal Biochem. 1974 Mar;58(1):238–245. doi: 10.1016/0003-2697(74)90463-1. [DOI] [PubMed] [Google Scholar]
  5. Brockerhoff H., Yurkowski M. Simplified preparation of L-alpha-glyceryl phosphoryl choline. Can J Biochem. 1965 Oct;43(10):1777–1777. doi: 10.1139/o65-197. [DOI] [PubMed] [Google Scholar]
  6. DAWSON R. M. A hydrolytic procedure for the identification and estimation of individual phospholipids in biological samples. Biochem J. 1960 Apr;75:45–53. doi: 10.1042/bj0750045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DAWSON R. M., HEMINGTON N., DAVENPORT J. B. Improvements in the method of determining individual phospholipids in a complex mixture by successive chemical hydrolyses. Biochem J. 1962 Sep;84:497–501. doi: 10.1042/bj0840497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DAWSON R. M. The measurement of 32P labelling of individual kephalins and lecithin in a small sample of tissue. Biochim Biophys Acta. 1954 Jul;14(3):374–379. doi: 10.1016/0006-3002(54)90195-x. [DOI] [PubMed] [Google Scholar]
  9. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  10. HANAHAN D. J., WATTS R. The isolation of an alpha'-alkoxy-beta-acyl-alpha-glycerophosphorylethanolamine from bovine erythrocytes. J Biol Chem. 1961 Sep;236:PC59–PC60. [PubMed] [Google Scholar]
  11. MARUO B., BENSON A. A. Cyclic glycerophosphate formation from the glycerolphosphatides. J Biol Chem. 1959 Feb;234(2):254–256. [PubMed] [Google Scholar]
  12. NORTON W. T. Reaction of mercuric chloride with plasmalogen. Nature. 1959 Oct 10;184(Suppl 15):1144–1145. doi: 10.1038/1841144a0. [DOI] [PubMed] [Google Scholar]
  13. Vaskovsky V. E., Kostetsky E. Y. Modified spray for the detection of phospholipids on thin-layer chromatograms. J Lipid Res. 1968 May;9(3):396–396. [PubMed] [Google Scholar]
  14. Wells M. A., Dittmer J. C. A comprehensive study of the postnatal changes in the concentration of the lipids of developing rat brain. Biochemistry. 1967 Oct;6(10):3169–3175. doi: 10.1021/bi00862a026. [DOI] [PubMed] [Google Scholar]
  15. Wuthier R. E. Two-dimensional chromatography on silica gel-loaded paper for the microanalysis of polar lipids. J Lipid Res. 1966 Jul;7(4):544–550. [PubMed] [Google Scholar]

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