Enzytec glycerol kit

Fasting blood (EDTA plasma) was drawn on the day of either scan in the case of the DTC patients, and on day 30 of placebo treatment for the subjects at risk of developing type 2 diabetes. TSH, fT4, CRP, glucose, and cholesterol levels were analysed. TSH was measured using an electrochemiluminescent immunometric assay (Cobas 6000, Roche Diagnostics, Mannheim, Germany) with a reference range of 0.4–4.3 mU/l. fT4 was measured using a competitive immunofluorimetric assay (AutoDelfia, Perkin Elmer, Turku, Finland) with a reference range of 8–18 pmol/l. CRP was measured using a turbidimetric assay (Cobas 8000) with a reference range of < 10 mg/l and a detection limit of > 1 mg/l. Glucose was measured using a hexokinase-based assay (Cobas 6000) with a reference range of 3.1–7.8 mmol/l. Cholesterol levels were determined using spectrophotometric analysis (reference ranges total cholesterol < 5.0 mmol/l; LDL < 3.0 mmol/l). Plasma free fatty acids (FFA) and total glycerol were measured using enzymatic assays automated on a Cobas Fara/Mira analyser, using Wako Nefa C test kit (Wako Chemicals, Richmond, VA, USA) and Enzytec glycerol kit (R-biopharm, Darmstadt, Germany) respectively.

Blood samples were collected in EDTA-containing tubes and immediately spun at high speed and frozen in liquid nitrogen and subsequently stored at −80°C until assayed. Plasma free fatty acids, triglycerides, and glucose were measured with enzymatic assays automated on a Cobas Fara/Mira (FA: Wako Nefa C test kit; Wako Chemicals, Neuss, Germany) (glucose: hexokinase method; Roche, Basel, Switzerland) (triglycerides: ABX Pentra CP reagents, Horiba ABX Diagnostics, Montpellier, France) (glycerol: Enzytec™ glycerol kit, R-Biopharm, Germany). The liver enzymes Gamma-GT, ASAT and ALAT were routinely measured during the screening visit and analysed in the clinical chemistry department in the hospital.