Dear researchers, We are starting a new experiment involving short chain fatty acids and we wanted to measure them in serum samples using UPLC-MS/MS.
For the protocol set up, we found some preliminary information, but we still need some details of the procedure, such as the proper gradient and column. Therefore, I was wondering if someone could shed some light about the technique and share with us the information about experimental procedure. I would really appreciate any further detail about it. Thank you in advance for your time. Javier Moleón Moya
Hi Javier,
We have a method set up for SCFA and we use it for faeces samples using HPLC MS/MS.
First SCFA are derivatized using 200mM 3NPH in methanol:water (1:1, v/v) 30 min at 40C, cooled down on ice for 1min and reaction stopped by diluting 1:10 with 10% MeOH.
Then analysed by MS using a C18 column. MPa: 0.01%FA (aq) MPb: 0.01%FA MeOH. 15min run standard gradient starting 20%B up to 100%B.
I hope this helps but let me know if you have any question.
Thanks,
Check the internet, especially the Waters, Agilent, and Phenomenex websites. You will have to homogenize your sample (and break the cell membranes) before analysis.
Several analytical techniques have been used to analyze SCFAs in biological samples using Gas Chromatography (GC) and Liquid Chromatography (LC) coupled with various kinds of detectors, such as Ultraviolet (UV), Flame Ionization Detector (FID), and Mass Spectrometry (MS).
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