Metabolic Biomarker Assessment Protocol

Blood samples were taken in the fasting state between 08:00 and 10:00 hours and transported to the laboratory facility at room temperature or 4°C, depending on the sample requirements. On the same day, HbA_1c (EDTA-anticoagulated) was analysed using an NGSP-certified turbidimetric inhibition immunoassay on a Cobas Integra 800 CTS analyser (Roche Diagnostics Nederland, Almere, the Netherlands). Serum creatinine was measured on a Roche Modular P chemistry analyser (Roche, Basel, Switzerland) and renal function was calculated as estimated (e)GFR with the formula developed by the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) [31 (link)]. Total cholesterol and HDL-cholesterol were measured using an enzymatic colorimetric method, triacylglycerol using a colorimetric UV method, and LDL-cholesterol using an enzymatic method, on a Roche Modular P chemistry analyser (Roche). Fasting blood glucose was measured using a hexokinase method.