On-line monitoring of bioreactions of Bacillus polymyxa and Klebsiella oxytoca by membrane introduction tandem mass spectrometry with flow injection analysis sampling

Abstract

Membrane Introduction mass spectrometry with flow Injection analysis sampling has been utilized for on-line monitoring of the major products and the volatile metabolites of fermentation by the Bacillus polymyxa and Klebsiella oxytoca organisms. A flow Injection sampling system was used to rapidly deliver fermentation broth or an external standard to the mass spectrometer. Analyte Introduction occurred via a direct Insertion membrane probe In which the aqueous solutions flowed past a membrane located within the Ion source of the mass spectrometer. For both organisms, concentrations of the liquid-phase products, acetic acid, acetoln, 2,3-butanediol, and ethanol, were monitored as a function of time after permeation through the membrane and ionization by chemical Ionization. Tandem mass spectrometry confirmed that these measurements were made without Interference. Off-line gas chromatography was utilized to test the accuracy of these measurements, and excellent agreement was found. Dissolved oxygen In the fermentation broth was also monitored, and carbon dioxide and oxygen were followed In the offgases. The 02 and C02 measurements were compared with other common measurement techniques (galvanic 02 electrode, Infrared absorption, and paramagnetic 02 analysis), and very good agreement was found. The use of tandem mass spectrometry has allowed the detection of additional compounds that were previously not known to be present In measurable amounts.

On‐line monitoring of bioreductions via membrane introduction mass spectrometry

“…The VOC are adsorbed from the solution onto the membrane, diffuse through the membrane, and evaporate from the membrane surface directly into the high vacuum ion-source region of the mass spectrometer. The ability to perform rapid and direct (without extraction or sample pretreatment) and quantitative analysis of VOC in a variety of matrixes is, therefore, one of the most attractive features of MIMS Hayward et al, 1990). The main disadvantage is the inability to readily distinguish and quantify stereoisomers.…”

“…Membrane introduction mass spectrometry has also been shown to be very suitable for real-time and on-line monitoring of a number of chemical processes such as (a) the major products and the volatile metabolites from fermentation by microorganisms (Hayward et al, 1990); (b) the conversion of chlorine to chloramines, an environmentally relevant reaction (Kotiaho et al, 1991); (c) the chlorination of phenol and related compounds, models of humic substances, by sodium hypochloride (Rios et al, 2000); (d) the oxidation of benzene derivatives by Fenton's reagent (Augusti et al, 1998); (e) the photolysis of aryl methyl ether in aqueous and aqueous-methanolic solutions (Wong et al, 1996); (f) the hydrolysis of epichlorohydrin, a significant compound in the polymer industry (Johnson et al, 1999); (g) the catalytic hydrodechlorination of aromatic chlorides, an important process to treat organochloro compounds ; and to the study of the stability constants of small organic guest molecules into cyclodextrin hosts in aqueous medium (Burgos et al, 2003).…”

On‐line monitoring of CO2 production in Lactococcus lactis during physiological pH decrease using membrane inlet mass spectrometry with dynamic pH calibration

“…Membrane inlet mass spectrometry (MIMS) is a wellknown technique for the monitoring of dissolved gasses in fermentation (Hayward et al, 1990;Lloyd et al, 2002), but to our knowledge it has hitherto not been used for quantitative carbon dioxide detection in processes where pH varies considerably. This is because carbon dioxide in aqueous solution exists as a mixture of CO 2(aq) and H 2 CO 3 , which is a weak acid.…”

On‐line biodegradation monitoring of nitrogen‐containing compounds by membrane inlet mass spectrometry

“…MIMS processes have been reported by several groups for monitoring biodegradation or fermentation of volatile organic compounds (VOCs) and by-products. 1 -7 The on-line monitoring of the major products (acetic acid, acetoin, 2,3-butanediol and ethanol) of fermentation by the Bacillus polymyxa and Klebsiella oxytoca by MIMS has been reported by Hayward et al 1 Hansen and colleagues carried out on-line monitoring of phenoxyacetic acid, a precursor of penicillin V, during Penicillium chrysogenum fermentation 2,3 and Johnson et al monitored on-line fermentation broths in a pilot plant by MIMS, using a 9000 dm 3 fermentation reactor in which samples were continuously taken by splitting flow from the fermentation broth and analysed for ethanol content by flow injection. Minor components such as acetic acid and lactic acid were also identified.…”