We are provided a Phospholipid (such as Phosphatidylcholine) and we are now required to determine the number of phosphate moieties present in the single phospholipid molecule. How can we experimentally determine/ prove this?
There is, by definition based on chemical structure, only one phosphate moiety in every single phospholipid molecule. You can find that evidence in textbooks, see e.g. Lehninger's Principles of Biochemistry, etc.
Best: Karoly
Perhaps Vaibhav means that they have a complex mixture of PLs in a PL extract, so that some molecules might actually have two P groups (cardiolipins), and some might have lost the P group due to the activity of specific phospholipases (PLase C). If this is the case, mass spectrometry appears to be the state of the art technique for PL structural identification.
The Condition here is we don't know how many phosphates are there in the given molecule. What we know is that it is a phospholipid. So how do we determine experimentally that how many phosphates are there in the given phospholipid?
Hi Vaibhav,
I agree Jorge that MS (Lipidomics) would be the state of the art technique, though it needs special knowledge and expensive equipment. A simplistic approach would be to separate your phospholipid samples on a TLC plate. You can find many examples on what mobil phases to use in the literature. Then, you could stain your sample and appropriate control lipids on the TLC plate with primulin, which is sensitive to the total lipid content, and consecutively with molibdenum blue, which reacts only with phosphate groups. You can purchase these reagents from Sigma-Aldrich.
I hope this helps. Best,
Karoly
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