I would like to characterize and quantify different classes of lipids of different samples of pollen. I was thinking to use HPLC or GC-MS. Someone know if these techniques are good for that? Someone has a suggestion of protocol to follow?
http://hortsci.ashspublications.org/content/28/12/1191.full.pdf
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Hey Priscila,
A technique that you have not mentioned is the best one to use: LC-MS/MS! : )
1. With GC-MS you can analyze and quantify free fatty acids, fatty acid methyl esters (FAMEs) and sterols etc. You need not buy standards etc.
2. With HPLC you need to buy standards to be able to do anything for retention time matching. Also, need to collect the HPLC-separated fractions and do again LC-MS/MS for structural validation using mass-spectrometry confirmation etc.
The best way to go would be (UP)LC-ESI-MS/MS which is available at most core facilities, you may or may not buy standards and in a established laboratory you can identify and quantify 300-800 lipid species using typical metabolomics/ lipidomics workflows. Given that pollens would be in limited amounts nothing better than LC-MS's sensitivity and for MS/MS experiments.
A lot of literature is available in Google on lipidomics using LC-MS/MS and you can follow papers that have been cited 1000s of times to get a robust method for lipid extraction (sample preparation) and LC-MS.
Good luck.
Biswa
Hello Priscila ,
Could you please clarify what classes of lipids you want to differentiate? I mean with examples of lipid class.
Thank you everybody for the answers.
I want to differentiate fatty acids and sterols, for example.
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