Stability of cyclopropane and conjugated linoleic acids during fatty acid quantification in lactic acid bacteria

Seven methods commonly used for fatty acid analysis of microgrganisms and foods were compared to establish the best for the analysis of lyophilized lactic acid bacteria. One of these methods involves fat extraction followed by methylation of fatty acids, while the other methods use a direct methylation of the samples, under different operating conditions (e.g., reaction temperature and time, reagents, and pH). Fatty acid methyl esters were identified by gas chromatography-mass spectrometry and quantified by on-column capillary gas chromatography. Two reliable methods for the analysis of fatty acids in bacteria were selected and further improved. They guarantee high recovery of classes of fragile fatty acids, such as cyclopropane and conjugated acids, and a high degree of methylation for all types of fatty acid esters. These two direct methylation methods have already been successfully applied to the analysis of fatty acids in foods. They represent a rapid and highly reliable alternative to classical time-and solvent-consuming methods and they give the fatty acid profile and the amount of each fatty acid. Using these methods, conjugated linoleic acids were identified and quantified in lactic acid bacteria.

[1]  John L. Ingraham,et al.  EFFECT OF TEMPERATURE ON THE COMPOSITION OF FATTY ACIDS IN ESCHERICHIA COLI , 1962, Journal of bacteriology.

[2]  B. Hennig,et al.  Comparison of methylation methods for the quantitation of conjugated linoleic acid isomers , 1993 .

[3]  I. Mononen,et al.  Gas chromatography analysis of cellular fatty acids and neutral monosaccharides in the identification of lactobacilli , 1987, Applied and environmental microbiology.

[4]  O. Hänninen,et al.  An uncooked vegan diet shifts the profile of human fecal microflora: computerized analysis of direct stool sample gas-liquid chromatography profiles of bacterial cellular fatty acids , 1992, Applied and environmental microbiology.

[5]  F. Ulberth,et al.  One-step extraction/methylation method for determining the fatty acid composition of processed foods , 1992 .

[6]  T. Ross,et al.  Acid habituation of Escherichia coli and the potential role of cyclopropane fatty acids in low pH tolerance. , 1997, International journal of food microbiology.

[7]  W. Colby,et al.  Identification of staphylococci with a self-educating system using fatty acid analysis and biochemical tests , 1996, Journal of clinical microbiology.

[8]  K. Demnerova,et al.  Cellular fatty acids analysis in the identification of lactic acid bacteria. , 1994, International journal of food microbiology.

[9]  K. Grob,et al.  Determination of fat content and fatty acid composition through 1-min transesterification in the food sample : II. Solubilization of the fat, results , 1997 .

[10]  J. R. Lane,et al.  Sequential alkaline saponification/acid hydrolysis/esterification: a one-tube method with enhanced recovery of both cyclopropane and hydroxylated fatty acids , 1993 .

[11]  P. Galzy,et al.  Fatty acid and carotenoid composition of Rhodotorula strains. , 1995, Archives of microbiology.

[12]  E. Eerola,et al.  Computerised gas-liquid chromatography of bacterial cellular fatty acids in analysis of bacterial mixtures , 1996 .

[13]  T. Johnsson,et al.  Cellular Fatty Acid profiles of lactobacillus and lactococcus strains in relation to the oleic Acid content of the cultivation medium , 1995, Applied and environmental microbiology.

[14]  J. Wilson,et al.  Fast atom bombardment-mass spectrometry for bacterial chemotaxonomy: influence of culture age, growth temperature, gaseous environment and extraction technique. , 1992, The Journal of applied bacteriology.

[15]  C. W. Moss,et al.  Comparison of the effects of acid and base hydrolyses on hydroxy and cyclopropane fatty acids in bacteria , 1983, Journal of clinical microbiology.

[16]  M. Delmée,et al.  A Rapid Procedure for the Identification of Lactic Acid Bacteria Based on the Gas Chromatographic Analysis of the Cellular Fatty Acids. , 1991, Journal of food protection.

[17]  N. Rozès,et al.  A rapid method for the determination of bacterial fatty acid composition , 1993 .

[18]  M. Pariza,et al.  Conjugated linoleic acid (9,11- and 10,12-octadecadienoic acid) is produced in conventional but not germ-free rats fed linoleic acid. , 1994, The Journal of nutrition.

[19]  G. Goma,et al.  Efficiency of fatty acid synthesis by oleaginous yeasts: Prediction of yield and fatty acid cell content from consumed C/N ratio by a simple method , 1993, Biotechnology and bioengineering.

[20]  J. Lanser,et al.  Problems associated with identification of Legionella species from the environment and isolation of six possible new species , 1990, Applied and environmental microbiology.

[21]  J. Decallonne,et al.  Gas Chromatographic Analysis of Cellular Fatty Acids in the Identification of Foodborne Bacteria. , 1995, Journal of food protection.

[22]  J. Chardigny,et al.  Fatty acid composition of French infant formulas with emphasis on the content and detailed profile oftrans fatty acids , 1996 .

[23]  S. Laakso,et al.  Temperature adaptation in Lactobacillus fermentum: interconversions of oleic, vaccenic and dihydrosterulic acids. , 1992, Journal of general microbiology.

[24]  K. Liukkonen,et al.  Temperature adaptation in yeasts: the role of fatty acids. , 1990, Journal of general microbiology.

[25]  W. Christie.  PREPARATION OF ESTER DERIVATIVES OF FATTY ACIDS FOR CHROMATOGRAPHIC ANALYSIS , 1993 .

[26]  P. Galzy,et al.  Study of lipids in Lipomyces and Waltomyces , 1994 .

[27]  G. Blank,et al.  Eugenol induced changes in the fatty acid content two Lactobacillus species , 1991 .

[28]  M. Salkinoja-Salonen,et al.  Use of Phage Typing and Fatty Acid Analysis for the Identification of Bacilli Isolated from Food Packaging Paper and Board Machines , 1989 .

[29]  T. E. Cloete,et al.  Taxonomy of lactic acid bacteria associated with vacuum-packaged processed meat spoilage by multivariate analysis of cellular fatty acids. , 1995, International journal of food microbiology.

[30]  J. Folch,et al.  A simple method for the isolation and purification of total lipides from animal tissues. , 1957, The Journal of biological chemistry.

[31]  D. Welch,et al.  Applications of cellular fatty acid analysis , 1991, Clinical Microbiology Reviews.

[32]  J. German,et al.  Isolation and fatty acid analysis of neutral and polar lipids of the food bacterium Listeria monocytogenes , 1996 .

[33]  L. Fay,et al.  Location of double bonds in polyunsaturated fatty acids by gas chromatography-mass spectrometry after 4,4-dimethyloxazoline derivatization , 1991 .

[34]  K. Grob,et al.  Determination of fat content and fatty acid composition through 1-min transesterification in the food sample; principles , 1997 .

[35]  M. Termonia,et al.  Automatic Identification of Microorganisms By Cgc Analysis of Cellular Fatty-acids , 1989 .

[36]  S. Gaskell,et al.  Phospholipids of Lactobacillus spp , 1995, Journal of bacteriology.

[37]  I. Helander,et al.  Cellular fatty acyl and alkenyl residues in Megasphaera and Pectinatus species: contrasting profiles and detection of beer spoilage. , 1995, Microbiology.