Rapid LC-MS-MS Analysis of Biomarkers of Drug-Induced Phospholipidosis in Rats Treated with Amiodarone and Gentamicin

摘要

Cationic amphiphilic drug-induced phospholipidosis (PL) is characterized by accumulation of phospholipids in tissue resulting in lamellar phospholipid inclusions inside cells. Buildup of phospholipids may be reversed when drug treatment is removed. In two recent reports (1, 2) bis mono-phosphatidic acids (BMP's) were observed at elevated levels in plasma or urine of animals treated with PL-inducing drugs. Specifically, 16:0/18:1 BMP in serum (2) and 22:6 BMP in urine (3) were identified as potential biomarkers. These methods employed long chromatographic separations suitable for biomarker identification but lacked the throughput needed for a clinical assay. Two rapid (4 and 10min) LC-MS-MS assays were developed using two alternate chromatographic approaches: 1) the use of sub-2(mu)m particles and high pressure (UPLC) to resolve BMP's from isomeric phosphatidylglycerols (PG) and 2) 2-3(mu)m particles and conventional HPLC to provide a higher throughput method. After chromatographic separation, the potential biomarkers were detected via negative ion ESI and tandem mass spectrometry (LC-MS-MS).