Advia 2400 chemistry analyzer

A volume of 5 ml of blood was withdrawn from the antecubital vein after fasting for a minimum of 8 h. Blood samples were analyzed by Gribbles Pathology Laboratory (Selangor, Malaysia). Serum TG, HDL cholesterol, and TC were measured by using enzymatic methods (Advia 2400 Chemistry Analyzer, Siemens, Tokyo, Japan). The blood glucose was measured by using the enzymatic method using hexokinase and glucose-6-phosphate dehydrogenase enzymes (Advia 2400 Chemistry Analyzer, Siemens, Tokyo, Japan). High-sensitivity C-reactive protein (hs-CRP) was measured via particle-enhanced turbidimetric assay. This laboratory obtained the approval of International Organization of Standardization (ISO: MS ISO 15189) in compliance with the standard quality.

Following the 14-day treatment, the animals were anesthetized and blood was collected via cardiac puncture (~5 mL) into plain tubes. To separate the sera, the samples were allowed to clot for 30 min and were centrifuged at 2500 rpm for 15 min. Separated into two portions, the sera were stored at −20 °C until further use. Lipid profile and kidney function parameters were determined at a local reference laboratory, Gribbles Pathology. Serum lipid profile was determined using ADVIA 2400 Chemistry Analyzer (Siemens, Erlangen, Germany). Kidney function parameters were analyzed by using Olympus AU640 Chemistry Immuno-Analyzer (Olympus, Tokyo, Japan). The concentration of serum insulin was determined by using an ultra-sensitive rat insulin ELISA kit (Crystal Chem Inc., Illinois, IL, USA) with inter- and intra-assay precision of CV ≤10 as per the manufacturer’s handbook.

Serum total cholesterol, HDL cholesterol, triglycerides, sodium, potassium, and glucose assays were measured using an enzymatic method (SIEMENS Advia 2400 Chemistry Analyzer). LDL cholesterol was calculated according to the Friedewald formula (LDL = total cholesterol–HDL cholesterol–triglycerides/5; mg/dL). Lp(a), Apo B100, and Apo A1 were determined by an immunoturbidimetry procedure (Abbott ARCHITECT cSystem). Serum total cortisol was measured by electrochemiluminescent assay (Cobas e411, ROCHE Diagnostic, Indianapolis, IN, USA), serum aldosterone by chemiluminescent immunoassay technology (LIAISON XL USA), PRA by ELISA competitive assay (IBL company, Minneapolis, MN, USA), and plasma ACTH by chemiluminescent assay (IMMULITE 2000XP SIEMENS Diagnostics, Manchester, UK).