By tomtom on Monday, March 31, 2003 - 11:03 pm:
Dear Chromatographers!
I have to quantify biomass of Methanogens using Coenzyme M (sulfonic acid) and an Agilent 1100 with a PDA-Detector.
Does anybody know a simple procedure for derivatisation of sulfonic acids for subsequent anaylsis by HPLC/PDA?
What will be the setup for LC: column, eluent..?
Thank you in advance
tomtom
By Anonymous on Thursday, April 10, 2003 - 02:12 am:
I am not familiar with your special problem and your question gives only low information.
Therefore only some general remarks.
First: All additional steps will complicate your system - is derivatization really a must?
Generally you have two possibilities (pre- and post-column deriv.).In the first case you add the problem of separating the excess of the reagent.With post-column derivat. you
need a reagent pump, pulse damper, rection coil, oven... Your reagent has to be invisible at your wavelength and the derivatization reagent acts in the presence of the eluent.
Reagents that work with the acid don’t always react with the salt-form.
You will find some literature:
-Knapp, Handbook of Analytical Derivatization Reactions (Wiley&Sons)
-Blau,King, Handbook of Derivatives for Chromatography (Heyden)
- Journal of Chrom. Science, Vol 17 (1979) -- two reviews: Silylation and UV-VIS-Derivatization.
Trimethyl-silylation (of thiol and acid) e.g. with MSTFA of a dry sample and following GC might be a different approach.
PETER