Abstract: In order to discuss the distribution and function of lipids in thylakoid membranes and its protein complex,lipids were extracted from photosystem I separated by DOC-PAGE from thylakoid membranes in the leaves of Pisum sativum L.An RP-HPLC-UV method with gradient elution was established for the separation of lipids obtained from thylakoid membranes,and baseline separation of the sample components could be realized in 25 min with this method.From the UV spectrum extracted from the DAD 3D graph,the main components separated in HPLC can be characterized preliminarily as lipids.