How could I analyze phospholipids extracted from bacterial cell wall?For analysis of phospholipids extracted from bacterial cell wall, can any one suggest a method rather than using molybdenum spray in 2 D TLC ?
Could I use LC-MS, and what is the suitable protocol for separation and identification without using standards?
One can try with GC-MS. The peaks can be identified from NIST Library and there is no need for standards. Can be tried.
thanks Dr. Bimal but I read that phospholipids not volatile so it ia difficult to use GC-Ms
Hi Noha,
yes you are right, phospholipids can be profiled (and identified even without stds) using LCMS. If you browse on internet you will find many articles that talk about this approach.
To start with, I can suggest you this interesting article:
http://www.ncbi.nlm.nih.gov/pubmed/24548922
and then I can say that the LC method is working (for all the lipids classes), and that for the phospholipids for example, I see them in ESi+ as sodium adducts or simply as [M-H]+ and not as NH3 adducts.
Don`t worry for the machine, they use a qtof mass spectrometer but you can do also with a MS iontrap and a triple quadrupole to perform MS/MS studies
Try to have a look to articles that talk about neutral loss for identification of phospholipids, maybe this approach will speed up your identification.
Good luck for this!!
Best regards,
Paolo
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