Identification of dissolved volatile metabolites in microbial cultures by membrane inlet tandem mass spectrometry

A membrane inlet tandem mass spectrometer is used for easy identification of volatile metabolites in microbial cultures. Filtrates of the cultures are without further purification placed into a measuring cell and introduced via a membrane inlet into a triple-quadrupole mass spectrometer. The samples are analysed with electron impact and chemical ionization mass spectrometry and tandem mass spectrometry. An extra dimension of separation is obtained by utilizing differences in response time due to differences in transport rates through the membrane. The following examples are used to demonstrate the technique. (i) Filtrates from cultures of three parasitic protozoa (trichomonads); two different strains of Trichomonas vaginalis isolated from humans and from Tritrichomonas foetus isolated from cattle were analysed. Indole was found in all samples; dimethyl disulphide was also present in cultures of the two organisms isolated from humans. (ii) The yeast Brettanomyces was grown in the presence of coumaric acid, ferulic acid, vanillic acid or syringic acid and the filtrate analysed for volatile products. In all cases ethyl acetate was found; coumaric acid biotransformation also gave 4-ethylphenol, whereas ferulic acid also gave 4-ethyl-2-methoxyphenol.

[1]  P. Nicholson,et al.  Determination of volatile organic compounds in aqueous systems by membrane inlet mass spectrometry , 1987 .

[2]  M. Klapper,et al.  Permeable membrane-mass spectrometric measurement of reaction kinetics , 1981 .

[3]  J. Weaver,et al.  Use of variable p(H) interface to a mass spectrometer for the measurement of dissolved volatile compounds. , 1979, The Review of scientific instruments.

[4]  Elmar Heinzle,et al.  Mass spectrometry in biotechnological process analysis and control. , 1987 .

[5]  David Lloyd,et al.  Quadrupole mass spectrometry in the monitoring and control of fermentations , 1985 .

[6]  Ramani Narayan,et al.  On-line monitoring of bioreactions of Bacillus polymyxa and Klebsiella oxytoca by membrane introduction tandem mass spectrometry with flow injection analysis sampling , 1990 .

[7]  F. R. Lauritsen.  A new membrane inlet for on-line monitoring of dissolved, volatile organic compounds with mass spectrometry , 1990 .

[8]  R. Cooks,et al.  Membrane interface for selective introduction of volatile compounds directly into the ionization chamber of a mass spectrometer , 1987 .

[9]  H. Degn,et al.  Stopped flow mass spectrometry: applications to the carbonic anhydrase reaction. , 1986, Journal of biochemical and biophysical methods.

[10]  H. Degn,et al.  Mass spectrometric measurements of steady-state kinetics of cyanobacterial nitrogen fixation by monitoring dissolved N2 in an open system , 1981 .

[11]  J W Brantigan,et al.  A clinical catheter for continuous blood gas measurement by mass spectrometry. , 1976, Journal of applied physiology.

[12]  D. Lloyd,et al.  The influence of oxygen and organic hydrogen acceotors on glycolytic dioxide production in Brettanomyces anomalus , 1988 .

[13]  Christie G. Enke,et al.  Triple Quadrupole Mass Spectrometry , 1979 .

[14]  J. C. Tou,et al.  Novel mass spectrometric sampling device. Hollow fiber probe , 1974 .

[15]  D. Lloyd,et al.  Continuous measurement of dissolved gases in biochemical systems with the quadrupole mass spectrometer. , 1985, Methods of biochemical analysis.

[16]  G. Hoch,et al.  A mass spectrometer inlet system for sampling gases dissolved in liquid phases. , 1963, Archives of biochemistry and biophysics.

[17]  H. Degn,et al.  Gas-exchange rates in the Belousov-Zhabotinskii reaction determined with membrane inlet mass spectrometry , 1989 .

[18]  I. Berecz,et al.  Gas concentration determination in fermentors with quadrupole mass spectrometer , 1983 .

[19]  L. Szigeti,et al.  Mass spectrometric monitoring of 2-oxoglutaric acid in fermentation broth , 1984 .

[20]  D. Lloyd,et al.  Direct measurement of dissolved gases in microbiological systems using membrane inlet mass spectrometry , 1983 .

[21]  B. B. Jensen,et al.  [48] Measurement of hydrogen exchange and nitrogen uptake by mass spectrometry , 1988 .

[22]  G. Jeffery.  Extended low-temperature preservation of human malaria parasites. , 1957, The Journal of parasitology.

[23]  R. Cooks,et al.  Direct insertion membrane probe for selective introduction of organic compounds into a mass spectrometer , 1990 .

[24]  R. Cooks,et al.  An Exceedingly Simple Mass Spectrometer Interface with Application to Reaction Monitoring and Environmental Analysis , 1985 .

[25]  J. Lundsgaard,et al.  Fast-responding flow-independent blood gas catheter for oxygen measurement. , 1980, Journal of applied physiology: respiratory, environmental and exercise physiology.

[26]  L. Joergensen.  The methane mono-oxygenase reaction system studied in vivo by membrane-inlet mass spectrometry. , 1985, The Biochemical journal.

[27]  H. Degn,et al.  A membrane‐inlet tandem mass spectrometer for continuous monitoring of volatile organic compounds , 1990 .

[28]  H. Busse,et al.  Oscillatory CO2 evolution in glycolysing yeast extracts , 1990, Yeast.

[29]  I. J. Dunn,et al.  On-line mass spectrometry in fermentation , 1984 .

[30]  L. S. Diamond.  The establishment of various trichomonads of animals and man in axenic cultures. , 1957, The Journal of parasitology.