Hi there,
My project aims to measure variety of oxidised phospholipid species of PC,PE and PS classes using targeted approaches. For relative quantification, I aim to normalise my data by measuring total phospholipid content in biological samples. Can I please request for a protocol that has been used to measure total phospholipid content in biological samples.
You have the answer in this reference :
CUVELIER I. et al
Clin Chem 1985; 31/5 : 763-766
Ini general terms, you have to homogenize your sample, then perform an organic extraction (CHCl3/MeOH/H2O, Bligh and Dyer or a variant to that protocol, and perform a Pi determination after destruction of the PLs as per Rouser 1970.
This all is pretty well described in various publications I am sure (I do not have them at hand here): You may want to stick to a protocol where the biological sample under study resembles your own samples best, and you may want to consider pre-treatment with proteases/DNAses/cell wall degradigg enzymes (all depending on what you are working with) to facilitate lipid extraction an homogenisation.
Good luck, Matthijs
Hi,
In our Lab we are using Bligh and Dyer too. Here is the protocol:
Good Luck!
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