Cobas pentra centrifugal spectrophotometer

Blood was collected from the antecubital vein for analyses of several blood variables. Plasma concentrations of free fatty acids (NEFA-HR set; Wako Chemicals), free glycerol (Glycerol kit; R-Biopharm), total glycerol (ABX Triglyceriden CP; Horiba ABX), and glucose (ABX Glucose HK CP; Horiba ABX) were measured on a COBAS PENTRA centrifugal spectrophotometer (Horiba ABX). Plasma triglyceride concentrations were calculated by subtracting free glycerol from the total glycerol concentrations, and serum insulin was analyzed on a Gamma Counter (2470 Automatic Gamma Counter Wizard2 Wallac; Perkin-Elmer) with a Human Insulin specific RIA kit (Millipore). Serum TSH was measured by an Electrochemiluminescence Immunoassay Kit on a COBAS 6000 system (Roche Diagnostica), and free thyroxine (FT4) was analyzed by a solid-phase time-resolved fluoroimmunoassay kit on an AutoDELFIA system (PerkinElmer). Plasma inflammatory marker C-reactive protein (CRP) was measured with a particle-enhanced immunoturbidimetric assay on a COBAS PENTRA system (Horiba ABX). BA concentrations were determined in plasma by phase liquid chromatography associated to tandem mass spectrometry (LC-MS/MS, in Multiple Reaction Monitoring [MRM] mode) after extraction by protein precipitation. The use of internal deutered standards allowed the quantification.