You first need to figure out the molecular weight of your target and run the appropriate calibration standards for the molecular mass range. POPC cannot be analyzed via GC so I'm a little confused as to what you're referring to.

You also should double check that your library/database has the fragmentation patterns for Phospholipase B and, if not, you need to update your frag lib or add them manually. You should be running an internal standard if you're trying to do any quantitative analysis, unless you just need retention time and fragmentation analysis. Not sure what positional specificity is referring to?

In Case of phospholipase b you should end up with free palmitic acid and oleic acid. There are a lot Informationen about analysis of free Fatty acids e.g. mstfa. Control: minus Enzyme.

Then you could See if the reaction Works. But in my opinion it is more easy to use lcms, where you can monitor educt and Products as well as side products without fear to measure degradation products arison within gc.

Thank You for reponse

Your kidding. Phospholipase B is an enzyme and NOT soluble in common GC solvents like Hexane. Try HPLC instead!

A novel phospholipase B from Streptomyces sp. NA684 – purification, characterization, gene cloning, extracellular production and prediction of the catalytic residues

Yusaku Matsumoto1, Shingo Mineta1, Kazutaka Murayama2 and Daisuke Sugimori1

1 Department of Symbiotic Systems Science and Technology, Graduate School of Symbiotic Systems Science and Technology, Fukushima

University, Japan

2 Division of Biomedical Measurements and Diagnostics, Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan

Please see this refrence Sir Bruce Neagle