I am working on total lipid quantification of microalgal system via in situ FAME derivatization but having difficulties in generating the calibration equations of the standards.
Hi dear searcher, when you determinate the FAME you are GC with FID detector this detector give the amount as area percent but if you want to quantify (mg or mmol) you can you Internal standard calibration and the used logicial give you the amount automatically by logitial calibration
I do not think that FA quantification with C17:0 ME as IS is a reliable way to quantify lipids. First, you are assuming that all FA from the different lipid fractions (phospholipids and acylglycerides) will be fully methylated and that they will be extracted as efficiently as C17:0 ME. And second, you will be disregarding any non-saponifiable lipids.
I was talking in light of some of the published protocols. Anyway, if not, then what should be the best possible way for determining the lipid content in microalgae, is it the solvent extraction method or fluorescence microscopy(which I think is not adequately precise) or anything else #Jorge Ruiz Carrascal.
Extraction methods based on mixtures of solvents (i.g. Folch) are working pretty fine, but of course it depends on the amount of sample and the type of lipids (I'm not familiar with algae).
Well, that is fine Jorge. But again,to quantify relative concentrations of individual fatty acids in the solvent extracted lipids, one has to go either for TLC or GC. TLC is again a not so precise process. Then how should one go if he/she has to quantify proportions of different FAs in lipid sample of the algae (in my case it is Dunaliella sp)?
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