1. I'm using DAD detector with MeCN/MeOH as a mobile phase
2. Unable to get PI peak which I guess is overlapping with oil peak
3. Also there is a question related to storage of working standards
It sounds like you are trying operate an HPLC system for the first time. Many years of training and experience are needed to acquire the basic skills and knowledge needed. Please research the topic as there are thousands of papers and articles on this topic freely available. Perhaps someone local to you with experience in using HPLC can assist you? Note: Phospholipids do not absorb in the UV/VIS range so you will need to derivatize them or use a different type of detector (ELSD, CAD or MS work well). Using low UV wavelengths coupled to chloroform in your sample or mobile phase will also obscure all signals.
Mr. Tribhuwan,
For lipidomics you need MS. The quantification can be carried out exactly by means of the following method:
1. Article Stochastic dynamic electrospray ionization mass spectrometri...
(Consider as well as the reference section therein.)
The equation appears universal law, thus you can use a broad spectrum of MS methods.
Currently under consideration is our new law govening temperature dependence on the MS intensity. When the paper appears, I shall update my answer, herein, because of it is applicable to a broad range of temperatures. Thus, it is, in particular, very useful to lipidomics.
You should bear in mind that the reference above provides exact model, thus the mass spectrometry appears exact method for analysis, in general.
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