New HPLC method for separation of blood plasma phospholipids.
The aim of the present work was to develop a new HPLC method for separation of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and lysophosphatidylcholine (LPC) from small-volume samples of blood plasma. Human plasma glycerophospholipids were separated by liquid-liquid extraction method followed by solid phase extraction (SPE) on aminopropyl columns. Reversed-phase Sephasil C8 column (10 cm x 2.1 mm, I.D. 5 microm) and micropreparative chromatograph "SMART" were used for separation of PC, PE, LPC and PI from SPE phospholipids extract. Binary-step gradient of eluent A: acetonitrile-methanol (130:5, v/v) and B (0.01% trifluoroacetic acid) provided good, fast and reproducible resolution of investigated phospholipids classes in 12 min at 30 degrees C. Eluted phospholipids were detected at wavelengths lambda=235 and 254 nm. This method made it possible to determine quantitatively: 5 microg ml(-1) PC, 1 microg ml(-1) LPC, 4 microg ml(-1) PE and 3 microg ml(-1) PI in blood plasma samples.
Suchocka
Zofia
Z
Department of Biochemistry and Clinical Chemistry, The Warsaw Medical University, 1 Banacha Str., 02-097 Warsaw, Poland. zos@farm.amwaw.edu.pl
Gronostajska
Dorota
D
Suchocki
Piotr
P
Pachecka
Jan
J
D002648:Child; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D010743:Phospholipids / Q000097:blood*; D010949:Plasma; D013678:Technology, Pharmaceutical / Q000379:methods*
D010743:Phospholipids