Reducing glycerophosphocholine lipid matrix interference effects in biological fluid assays by using high-turbulence liquid chromatography.
Matrix interferences can severely affect quantitative assays of biological samples when electrospray ionization (ESI) is employed with liquid chromatography/tandem mass spectrometry (LC/MS/MS). A major source of matrix interferences for plasma sample analyses is the presence of glycerophosphocholine (GPCho) lipids. The efficiency of online high-turbulence liquid chromatography (HTLC) extraction for eliminating these lipids is evaluated and the interfering effects of endogenous lipids on human plasma assays are measured for pharmaceutical compounds having a wide variety of chemical properties. It is found that GPCho lipids, represented by 16:0, 18:1 and 18:0 LPC (lysophosphatidylcholine) and 16:0-18:2 PC, cause variations for hydrophobic compound analyses even when optimal online HTLC extraction conditions are employed. The efficiency for lipid removal depends on the organic content of the transfer solvent, but turbulent flow loading has no significant effect.
Department of Clinical PK/PD, Merck Research Laboratories, Merck & Co. Inc., West Point, PA 19486-0004, USA. Lihong_Du@Merck.com
Lihong
L
Du
Robert L
RL
White
D001681:Biological Assay; D002853:Chromatography, Liquid / Q000379:methods*; D005997:Glycerylphosphorylcholine / Q000097:blood* / Q000737:chemistry; D006801:Humans; D008055:Lipids / Q000097:blood* / Q000737:chemistry; D015394:Molecular Structure; D009862:Online Systems; D012015:Reference Standards; D012996:Solutions / Q000737:chemistry
D008055:Lipids; D012996:Solutions; D005997:Glycerylphosphorylcholine