How do you monitor or quantify residual glycerol concentrations in fermentations? HPLC, enzymatic kits or other? What are we looking for primarily (accuracy, fast, cheap)?
We use HPLC with an Aminex HPX-87H column with RI detection. Mobile phase is Water with 0.005 M H2SO4 (cheap). Total run is 23 minutes. The advantage is that ethanol, glucose, maltose, methanol and somes organic acids (e.g acetic acid) can be detected in the same run. Due to the temperature and the acidic conditions sucrose can't be quantified by this method. Good work
Thanks Sébastien. We use a similar procedure. We consider a quite slow method when willing to have (close to) real time monitoring and/or processing many samples.
Enzymatic kits have some interferences with media prodcuts or metabolites.
We are developping alternative methods.
I am using HPLC with an Aminex HPX-87H column with RI detector. The mobile phase is 5mM with flow rate 0.6 ml /min. The column should maintained at 60C and the rt time for glycerol is 14.4 min. If you are using the auto-sample for HPLC i think it is best method to detect glycerol con up to 10ppm in the fermented samples
We just had a publication accepted in Talanta comparing mtehods HPLC (as you described), enzymatic kit and a new method using Dotblot+image analysis. It enables a 1 minute quantification to monitor your fermentation or reaction.
Merry Christmas everyone
The new method for glycerol monitoring was published
http://www.sciencedirect.com/science/article/pii/S0039914013010047
suitable to fermentation media as well as other applications
Dear Dr. Xavier Turon,
Thank you very much for your exciting work. My lab is wasting lot of HPLC grade water and running HPLC for routine experiments is tedious. How to implement your technique? Is it suitable for beginners in MATLAB? Any codings required? Other than MATLAB, what can be used?
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