912 autoanalyser

Anthropometric measurements including height, weight, and waist circumference, were obtained using standardized techniques. The body mass index (BMI) was calculated as the weight in kilograms divided by the square of height in meters. Fasting plasma glucose (FPG) (glucose oxidase-peroxidase method), serum cholesterol (cholesterol oxidase-peroxidase- amidopyrine method), serum triglycerides (glycerol phosphate oxidase-peroxidase-amidopyrine method), high density lipoprotein cholesterol (HDL-C) (direct method-polyethylene glycol-pretreated enzymes), and creatinine (Jaffe’s method) were measured using a Hitachi-912 Autoanalyser (Hitachi, Mannheim, Germany). The intra- and inter assay coefficient of variation for the biochemical assays ranged between 3.1% and 5.6%. Glycated hemoglobin (HbA1c) was estimated by high pressure liquid chromatography using a variant machine (Bio-Rad, Hercules, CA). The intra- and inter-assay coefficient of variation of HbA1c was less than 5%. The plasma concentrations of high-sensitivity C-reactive protein (hsCRP) were measured by high sensitive nephelometric assay. The intra- and the inter-assay coefficients of variation for hsCRP were 4.2% and 6.8%, respectively, and the detection limit was 0.15 mg/L. Both the intra and inter assay variations were determined in our biochemical lab.

Fasting plasma glucose (glucose oxidase-peroxidase method), serum cholesterol (cholesterol oxidase-peroxidase-amidopyrine method), serum triglycerides (glycerol phosphate oxidase-peroxidase-amidopyrine method) and HDL cholesterol (direct method-polyethylene glycol-pretreated enzymes) were measured using Hitachi-912 Autoanalyser (Hitachi, Mannheim, Germany). The intra and inter assay co-efficient of variation for the biochemical assays were <5%. Low-density lipoprotein (LDL) cholesterol was calculated using Friedewald formula. Glycated hemoglobin (HbAlc) was estimated by high-pressure liquid chromatography using the variant analyzer (Bio-Rad, Hercules, Calif., USA). Serum Insulin was estimated using enzyme-linked immunosorbent assay (Calbiotech, CA). The intra-assay and the inter-assay coefficients of variation for insulin assay was <10%. Insulin resistance was calculated using the homeostasis assessment model (HOMA-IR) using the formula: {fasting insulin (μIU/mL) x fasting glucose (mmol/L)} / 22.5.

Fasting plasma glucose (glucose oxidase-peroxidase method), serum cholesterol (cholesterol oxidase-peroxidase-amidopyrine method), serum triglycerides (glycerol phosphate oxidase-peroxidase-amidopyrine method) and HDL cholesterol (direct method-polyethylene glycol-pretreated enzymes) were measured using Hitachi-912 Autoanalyser (Hitachi, Mannheim, Germany). The intra- and inter-assay co-efficient of variation for the biochemical assays were <5%. Low-density lipoprotein (LDL) cholesterol was calculated using the Friedewald formula [16 (link)]. Glycated hemoglobin (HbAlc) was estimated by high-pressure liquid chromatography using the variant analyzer (Bio-Rad, Hercules, Calif., USA). Serum insulin was estimated using enzyme-linked immunosorbent assay (Calbiotech, CA). The intra-assay and the inter-assay coefficients of variation for insulin assay was <10%. Insulin resistance was calculated using the homeostasis assessment model (HOMA-IR) using the formula: fasting insulin (μIU/mL)*fasting glucose (mmol/L) / 22.5.

Anthropometric measurements including weight, height, and waist circumference were measured using standardized techniques. The body mass index (BMI) was calculated using the formula, weight (kg)/height (m²), and obesity was classified as BMI ≥ 25 according to WHO Asia Pacific Guidelines for Asians (The Asia Pacific perspective 2000). Fasting plasma glucose (glucose oxidase–peroxidase method) was measured using Hitachi-912 Autoanalyser (Hitachi, Mannheim, Germany). Glycated hemoglobin (HbA1c) was estimated by high-performance liquid chromatography using a Variant™ machine (Bio-Rad, Hercules, CA, USA). Serum insulin, serum vitamin B₁₂, and folic acid concentration were estimated using the electrochemiluminescence using a Roche e601Cobas immunoassay analyzer (Roche Diagnostics, Indianapolis, IN, USA). The intra- and inter-assay coefficients of variation for vitamin B₁₂ assay were 0.95% and 4.08%. Serum homocysteine was measured using enzymatic assay using the Beckman Coulter AU2700 (Fullerton, CA, USA) Biochemistry analyzer.

Blood samples were collected after 10–12 h fasting and serum and plasma were separated. All of the samples were stored at −80 °C for later biochemical analysis. Fasting serum levels of glucose and lipid profiles, including total cholesterol (TC), triglyceride (TG) and high density lipoprotein (HDL-C) were measured by a Hitachi 912-autoanalyser (Hitachi, Mannheim, Germany) using commercial kits. Low density lipoprotein (LDL) was calculated by friedewald equation. Insulin was measured by radioimmunoassay. Serum irisin concentration was measured using the enzyme-linked immunosorbent assay (ELISA) kits (Biovendor Laboratory Medicine, Modrice, Czech Republic).