We are trying to prepare fatty acid methyl esters of rhamnolipids for GC-MS analysis. We have used one method from a research article, but we faced difficulties with the method as high temperature for 2-3 hours is used in the process. Does anyone have a method where the sample can be easily prepared without loss or damage to the sample?
Hey Rajeev, It's quite common question/problem facing all researchers regarding the preparation and processing of FAMEs sample for GCMS. You can refer my research article for this. I do have optimized the conditions for preparation and processing of FAMEs sample for GCMS.
You can find my My research article in my profile.
The core protocol were taken from: "Carreau J. P., Dubacq J. P., 1978. Adaptation of a macro-scale method to the micro-scale for fatty acid methyl trans-esterification of biological extracts. J. Chromatogr. 151, 384-390".
Article Fatty acids as biomarkers of microalgae
Methylation with diazomethane is done on room temperature and shouldn't produce significant amounts of side products. Just put a bit of your sample in a GC-MS vial and add few drops of diazomethane solution (until the evolution of N2 stops).
BTW, I'm not sure if rhamnolipids can be detected with GC-MS with free hydroxy groups. You should probably try to protect them with TMSCl or something similar first.
read this paper: Journal of Proteome Res, 2013, 15:3755-3768, esp, pp3758. It works wonders and is nicely simple.....
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