Analysis of Cellular Fatty Acids of Bacteria by Gas-Liquid Chromatography 



SUMMARY OF GLC ANALYSIS OF BACTERIAL CELLULAR FATTY ACIDS 



A. Saponification - 4 ml of 5% NaOH in 50% methanol is added to cells from a slant culture 

 and heated for 30 min at 100°C. The sample is cooled. 



B. Methylation - The pH is adjusted to 2.0 with 6 N HCl; 4 ml of 10% boron tri- 

 chloride-methanol reagent is added, and tlie mixture is heated at 80" C for 5 min. Sample is 

 cooled. 



C. Extraction Methyl esters extracted from mixture with two 10-ml aliquots of a 1:1 

 mixture of diethyl ether;hexane. Solvent containing methyl esters is reduced to about 0.1 ml. 



D. GLC 1-3 ij\ of methyl ester sample is injected into non-polar GLC column and analyzed. 

 Retention times of peaks are recorded and compared to reference standards. Quantitative data 

 are obtained, and identities of fatty acids are confirmed by additional procedures (hydrogena- 

 tion, acetylation, analysis on polar column, mass spectrometry). 



REFERENCES 



1. Lechevalier, M. P. 1977. Lipids in bacterial taxonomy A taxonomist's view. pp. 109-210. //; A. I. Laskin 

 and H. Lechavalier (eds.). Critical Review of Microbiology, Vol. 5. CRC Press, Cleveland, Ohio. 



2. Moss, C.W. 1978. New methodology for identification of nonfermenters: Gas-liquid chromatographic chemo- 

 taxonomy. pp. 171-201. //; G. L. Gilardi (ed.). Glucose Nonfeimentating Gram-Negative Bacteria in Clinical 

 Microbiology. CRC Press, West Palm Beach. Florida. 



3. Moss, C. W., and W. B. Cherry. 1978. Characterization of the C-15 branched-chain fatty acids of Conv;e- 

 bactehum acnes by gas chromatography. J. Bacteriol. 95:241-243. 



4. Moss, C.W., M. A. Lambert, and W. H. Mervin. 1974. Comparison of rapid methods for analysis of bacterial 

 fatty acids. Appl. Microbiol. 28:80-86. 



5. Moss, C. W., R. E. Weaver, S. B. Dees, and W. B. Cherry. 1977. Cellular fatty acid composition of isolates 

 from Legionnaires' disease. J. Clin. Microbiol. 6: 140-143. 



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