I have done fermentation by microbes and i wanted to do qualitative and quantitative assays with the help of Gas chromatography mass spectra, Please kindly help me with the method.
Thanks.
It is very hard to separate ethanol from aquas broth and for proper GC/MS analysis you need water free samples. Therefore conventional GC/MS is not advised for ethanol estimation for reliable/error free result as well as for good health of your machine. Although headspace GC/MS could be perform where headspace gas over the fermentation broth is analyzed. As ethanol is volatile in nature some trace could be found in headspace air. HPLC or LC/MS would be better option.
Although some specialized GC/MS with some interesting specification is there which use water vapor filter which can be used. For that you can find out protocol from following paper.
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Apart from that in my work in bio-ethanol production I have use HPLC. If you want u can have protocol of HPLC from there.
Thank you,
Hi,
You can use SPAM with GC/MS . GC/MS is not analysis the sample with water such as media growth.
I used the following protocol in my work, you can find in my research papers on bioethanol production from bacterial fermentation.
C18 column (4.6 × 250 × 5 μm) using a mobile phase of 0.05 M H2SO4, flow rate of 0.5 ml min−1 and an injection volume of 20 μl. RID technique was used for detection.
Hello Ratan Chowdhury
;What the colleagues have said in their previous answers is valid in the case that you want to inject your sample directly to the GC. However, it is a common practice for aqueous fermentation broths to perform a solvent extraction to a non polar organic solvent with low boiling point, and then carry out the GC analysis of the organic compound you want to detect and quantify. In your case, as you are interested in ethanol, you can take advantage of the high miscibility range of this compound in non-polar solvents, as is the case of benzene and toluene. So if you have a GC-MS (that not everyone has) I suggest to perform an extraction with toluene, and then inject the organic phase to the GC. This aproach will also help you to avoid the mess for treating the aqueous fermentation broth previous to HPLC analysis.
Hope this helps and good luck.
Regards,
Rogelio.
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