I want to prepare Volatile fatty acids standard mixture from individual acids bought from Sigma Aldrich. I have seen different volume mixture contradicting. I need a standard procedure/ volume mixture that will help to create a good calibration curve for quantifying the unknown VFA in rumen fluid using GC
Kindly check https://onlinelibrary.wiley.com/doi/full/10.1002/elsc.201600095
Also check https://academic.oup.com/chromsci/article/56/1/1/4561566
The curve you need will depend on your sample dilution, initial concentration (of course) and your LOQ of your machine/column/protocol, so keep that in mind. Recently did some work with rumen fluid, will private message you more detailed answer but our standard curve was as below (in case someone else needs to see this in the future). Will link the paper once it's out (currently pre-print).
Master solution = 25.33 mM acetic, 24.99 mM propionic, and 10.0 mM each of butyric, iso-butyric, valeric and iso-valeric acids. Calculate the amount needed to make those mM concentrations from the molecular weight of each acid (can easily find this on Sigma's site).
Curve = 0.001, 0.05, 0.1, 0.2, 0.4, 0.6, 0.8 and 1.0-fold of the master solution (so minimum acetic acid = 0.025 mM, maximum = 25.33 mM, and so on). Also of course ran many blanks to check for any baseline issues.
We found that adding formic acid to our internal standard mixture (added to all samples and standards) helped further reduce ghost peaks in subsequent injections. This may not be as much an issue with a different column or protocol but we found it helpful and easy to do with our sample prep.
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