Development of a method for the application of solid-phase microextraction to monitor biodegradation of volatile hydrocarbons during bacterial growth on crude oil |
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Authors: | J D Van Hamme O P Ward |
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Institution: | (1) Microbial Biotechnology Laboratory, Department of Biology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1, CA |
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Abstract: | A quantitative solid-phase microextraction, gas chromatography, flame ionization detector (SPME-GC-FID) method for low-molecular-weight
hydrocarbons from crude oil was developed and applied to live biodegradation samples. Repeated sampling was achieved through
headspace extractions at 30°C for 45 min from flasks sealed with Teflon Mininert. Quantification without detailed knowledge
of oil–water–air partition coefficients required the preparation of standard curves. An inverse relationship between retention
time and mass accumulated on the SPME fibre was noted. Hydrocarbons from C5 to C16 were dated and those up to C11 were quantified. Total volatiles were quantified using six calibration curves. Biodegradation of volatile hydrocarbons during
growth on crude oil was faster and more complete with a mixed culture than pure isolates derived therefrom. The mixed culture
degraded 55% of the compounds by weight in 4 days versus 30–35% by pure cultures of Pseudomonas aeruginosa, Rhodococcus globerulus or a co-culture of the two. The initial degradation rate was threefold higher for the mixed culture, reaching 45% degradation
after 48 h. For the mixed culture, the degradation rate of individual alkanes was proportional to the initial concentration,
decreasing from hexane to undecane. P. fluorescens was unable to degrade any of the low-molecular-weight hydrocarbons and methylcyclohexane was recalcitrant in all cases. Overall,
the method was found to be reliable and cost-effective. Journal of Industrial Microbiology & Biotechnology (2000) 25, 155–162.
Received 04 March 2000/ Accepted in revised form 25 June 2000 |
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Keywords: | : biodegradation crude oil SPME volatile hydrocarbons |
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