D 3 hydroxybutyrate kit

We performed all metabolite assays at the Laboratorio Clínico Veterinario (Universidad Austral de Chile) using an auto-analyzer (Metrolab 2300, Wiener Lab). Briefly, total triglycerides (triglycerides plus free glycerol) was measured by enzymatic colorimetric assay (Triglycerides liquicolormono GPO-PAP kit, HUMAN GmbH) adapted to small sample volumes in 300 μL flat-bottom microplates. Glycerol concentration was determined by a coupled enzyme assay (Sigma-Aldrich MAK117). Concentration of uric acid was determined using the enzymatic colorimetric test with lipid clearing factor (uric acid liquicolor PAP kit, HUMAN GmbH). β-hydroxybutyrate was measured using colorimetric enzymatic reaction (D-3-hydroxybutyrate kit; Ranbut, Randox Laboratories). Triglyceride levels were calculated by subtracting free glycerol from total triglyceride levels^{23 (link),39 (link)}.

Blood glucose, blood sodium, blood potassium, C-reactive protein (CRP), procalcitonin (PCT), activated partial thromboplastin time (APTT), prothrombin activity (PTA), serum creatinine (SCR), cystatin C, creatine kinase, pH, HCO3-, base excess (BE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum albumin, blood amylase, triglyceride (TG), total cholesterol (TC) and low density lipoprotein (LDL) were measured using an automatic biochemical analyser (AU5821, Beckman Coulter, USA). β-hydroxybutyrate was measured using a colorimetric enzymatic reaction (D-3-hydroxybutyrate kit; Ranbut, Randox Laboratories). White blood cells, neutrophils, lymphocytes, and platelets were determined by an automatic blood cell analyser (BC-6800, Mindray, Shenzhen, China). Troponin I was measured by chemiluminescence immunoassay using a DXI800 (Beckman Coulter, USA). Haemoglobin A1c was measured by high-performance liquid chromatography (BC-6800, Mindray, Shenzhen, China).