Hi,
I ' ve already tried twice to do a Phospholipid-Assay (Stewart), i.e. to establish a standard curve for Phosphatidylcholin, but it hasnt worked yet. That means macroscopically it looks fine, because the lower chloroform layer gets the more red the higher is the content of PC. But I can't confirm this result by measuring it at 488 nm. Do someone know what could be the reason for that?
These are my values (without a background value measured). For each content I made up two approaches with the same µg PC.
Content of PC per tube: Extinction approach 1 // Extinction approach 2
0 µg: -2,912 // -2,41
10 µg: -3,22 //-1,9
25 µg: -3,01 // -1,8
50 µg: -2,9 // -0,93
75µg: -2,74 // -2,95
100µg: -2,89 // -2,79
I'd be very glad about an answer!