A rapid and sensitive LC–MS/MS method for quantitative analysis of cardiolipin (18:2)4 in human leukocytes and mouse skeletal muscles

Publication date: 5 September 2018Source: Journal of Pharmaceutical and Biomedical Analysis, Volume 158Author(s): Gang Xu, Xiao Liu, Yachun Shu, Jagan A. Pillai, Yan XuAbstractA rapid and sensitive LC–MS/MS method has been developed for quantitative analysis of cardiolipin (18:2)4 or CL (18:2)4 in human leukocytes and mouse mitochondria. The structural analog CL (14:0)4 was used as the internal standard. Both CL (18:2)4 and the IS were extracted using a modified Folch method, and separated on a Waters XBridge® BEH C18 XP column using a mobile phase of 0.1% ammonium hydroxide in acetonitrile/water (90:10, v/v) pumped at a flow rate of 0.4 mL/min. Quantitation was achieved by negative ESI-MS/MS in MRM mode. The total run time was 2.00 min with retention times of 0.74 min for the IS and 0.84 min for CL (18:2)4, respectively. The method was validated according to the US-FDA guidance for bioanalytical method validation using human leukemia and lymphoma cell lines, which had a calibration range of 0.120–60.2 nM with a correlation coefficient>0.999. The intra- and inter-assay accuracy and precision were ≤±5% and ≤8%. The IS normalized matrix factors of CL (18:2)4 and the IS normalized recoveries of CL (18:2)4 ranged 0.92–1.04, and 95–101%, respectively. The stability studies showed that CL (18:2)4 was stable under various test conditions. The developed method was successfully applied to the measurement of CL (18:2)4 in various biological samples including K56...
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research