I am working with Nitrogen fixating organisms especially cyanobacteria and Heterotrophic bacteria. For quantifying nitrogen fixation rates by these micro organisms ARA used. I need to know the about the Gas chromatography condition separate acetylene and ethylene (I am using Elite Capillary Alumina KCL column (0.53*50m)). I am waiting for your valuable comments.
Please refer to the publication below. It is an open access publication and has the conditions for GC. all the best.
Article Improvement of plant growth and seed yield in Jatropha curca...
Publications using this test including ethane determination (for alternative nitrogenase):
Tsygankov A.A., Serebryakova L., Sveshnikov D., Rao K., Gogotov I., Hall D.Hydrogen production by three different nitrogenases in whole cells of Anabaena variabilis and the dependence on pH..Int. J. of Hydrogen Energy. 1997. V.22, P.859-867
Tsygankov A.A., Serebryakova L.T., Rao K.K., Hall D.O.Acetylene reduction and hydrogen photoproduction by wild type and mutant of Anabaena at different CO2 and O2 concentrations.FEMS Microbiol. Lett. 1998, 167, 13-17.
You don´t say what chromatograph you are using.
I used a gas chromatograph with Flame Ionization Detector (FID) with a HP-PLOT/AL203 column.
If you are using FID, methods in the following paper should also work for you. You might have to vary a bit the injector and detector temperatures. By varying the rate of temperature increase you will be able to separate better the curves of ethilene and acetylene.
Article Nitrogen Fixation in Preserved, Reforested, Naturally Regene...
I am giving you a recent and a good alternative of what you are asking
Microbial assay of N2 fixation rate, a simple alternate for acetylene reduction assay
lSubhajitDasTarun KumarDe
https://doi.org/10.1016/j.mex.2017.11.010
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