Blood glucose was measured at the time of the blood collection using a glucometer (Freestyle Disectronic, Vianen, The Netherlands). Blood samples were used to prepare EDTA plasma by centrifugation (10 min, 6000 rpm). In freshly prepared plasma, total triglyceride and cholesterol concentrations were measured using enzymatic assays (triglyceride GPO-PAP and cholesterol CHOD-PAP, Roche Diagnostics, Almere, The Netherlands).
Cholesterol chod pap
Manufactured by Roche
Sourced in Germany, Switzerland
The Cholesterol CHOD-PAP is a laboratory equipment product used for the quantitative determination of cholesterol in serum and plasma. It employs the CHOD-PAP method, which involves the enzymatic colorimetric determination of cholesterol.
Automatically generated - may contain errors
Lab products found in correlation
Triglycerides gpopap, by Roche (3 mentions)
Triglyceride gpo pap, by Roche (2 mentions)
Free cholesterol fs, by DiaSys (2 mentions)
Phospholipids fs, by DiaSys (2 mentions)
Superose 6 pc 3.2 300 column, by GE Healthcare (1 mentions)
Ezrmi 13k, by Merck Group (1 mentions)
Ldl cholesterol, by Roche (1 mentions)
Hdl c plus, by Roche (1 mentions)
Cobas c311 analyser, by Roche (1 mentions)
Cobas c501 analyser, by Roche (1 mentions)
Hdl c plus third generation, by Roche (1 mentions)
Integra 400 plus, by Roche (1 mentions)
11 protocols using cholesterol chod pap
1
Plasma Lipid Profile Analysis
2
Comprehensive Lipid and Carnitine Profiling
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Lipids from plasma were measured enzymatically on a Hitachi 917 system (Roche Diagnostics GmbH, Mannheim, Germany) using the cholesterol (CHOD-PAP) and TAG (GPO-PAP) kit from Roche Diagnostics, and the free cholesterol (Free Cholesterol FS), non-esterified fatty acid (NEFA FS,) and phospholipid kit (Phospholipids FS) from DiaSys (Diagnostic Systems GmbH, Holzheim, Germany). Lipoproteins were also separated from 2.5 μL of individual plasma samples by size exclusion chromatography (SEC), using a Superose 6 PC 3.2/300 column (GE Healthcare Bio-Sciences AB, Uppsala, Sweden). Lipoproteins were eluted as a fraction appearing in the exclusion volume of the sepharose column that contained chylomicrons (if present) together with VLDL, then LDL and last HDL. Total cholesterol was calculated after integration of the AUC in the individual chromatograms [52 (link),53 (link),54 (link)], generated by the enzymatic-colorimetric reaction Cholesterol CHOD-PAP (Roche Diagnostics).
L-carnitine, trimethyllysine, γ-butyrobetaine, acetylcarnitine, propionylcarnitine, valerylcarnitine, octanoylcarnitine, lauroylcarniitne, myristoylcarnitine, palmitoylcarnitine, betaine, choline and TMAO were analyzed in plasma by HPLC-MS/MS as described by Vernez et al. [55 (link)] with some modifications [56 (link)].
L-carnitine, trimethyllysine, γ-butyrobetaine, acetylcarnitine, propionylcarnitine, valerylcarnitine, octanoylcarnitine, lauroylcarniitne, myristoylcarnitine, palmitoylcarnitine, betaine, choline and TMAO were analyzed in plasma by HPLC-MS/MS as described by Vernez et al. [55 (link)] with some modifications [56 (link)].
3
Lipid and Glucose Profiling in Plasma
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Lipids from plasma were measured enzymatically on a Hitachi 917 system (Roche Diagnostics GmbH, Mannheim, Germany) using the cholesterol (Cholesterol CHOD-PAP, 11491458-216), and triacylglycerol (Triglycerides GPO-PAP, 11730711) kit from Roche Diagnostics, and the free cholesterol (Free Cholesterol FS, Ref 113609910930), non-esterified fatty acid (NEFA FS, Ref 157819910935) and phospholipid kit (Phospholipids FS, Ref 157419910930) from DiaSys (Diagnostic Systems GmbH, Holzheim, Germany). Plasma bile acid was measured enzymatically on a Roche Modular P chemistry analyzer (Roche Diagnostica), using the BA kit (Total Bile Acid Assy Kit, 05471605001) from Diazyme (Diazyme Laboratories, Gregg, CA, USA). The fatty acid composition was determined by GC/MS as previously described [47 (link)]. Glucose was measured on Hithachi 917 using the Glucose/HK kit (Roche Diagnostics, Ref 11876899-216). Fasting insulin was measured in two parallels of 10 μL plasma from each rat using a rat/mouse insulin 96 well plate assay ELISA kit (EZRMI-13K) from EMD Millipore (Billerica, MA, USA), according to the manufacturer’s instructions.
4
Lipid Profile Measurement Protocol
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The lipid profile, including total cholesterol (TC), triglycerides (TG), LDL-cholesterol (LDL-C) and HDL cholesterol (HDL-C), of all the subjects was obtained using a Roche/Hitachi automated system with commercial kits for CHOL (Cholesterol CHOD-PAP), LDL-C plus 2nd generation (LDL Cholesterol), HDL-C plus 3rd generation (HDL-Cholesterol) and TG (Triglyceride GPO-PAP) (all from Roche Diagnostics, Germany).
5
Liver Tissue Lipid Quantification
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Approximately 50 mg of frozen liver tissue was homogenized as described previously [7] (link), [17] (link). Both plasma and liver lipid levels were measured with enzymatic color tests (1489232, cholesterol CHOD-PAP, Roche, Basel,Switzerland; TR0100, TG GPO-trinder, Sigma Aldrich, Sigma Aldrich, St. Louis, MO, USA; 999-75406, NEFAC, ACS-ACOD, Wako Chemicals, Neuss, Germany) as was described before [7] (link), [17] (link).
6
Fasting Blood Biomarker Measurements
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Blood samples were taken after overnight fasting and immediately transported to the central laboratory of the Gynecologic Obstetrical University Hospital in Poznan for analysis. HbA1c level in whole blood was determined using the turbidimetric inhibition immunoassay (TINIA) (Tina-quant Hemoglobin A1c II test in a Cobas c311 analyser (Roche Diagnostics, Basel, Switzerland)).
The total serum cholesterol, HDL cholesterol and triglyceride (TG) levels were determined with Roche Diagnostics reagents (Cholesterol CHOD-PAP, HDL-C plus, and Triglycerides GPO-PAP, respectively) on a Cobas c501 analyser. The following formula was used to calculate the level of LDL cholesterol: LDL cholesterol = total cholesterol − HDL cholesterol − (TG/5).
The total serum cholesterol, HDL cholesterol and triglyceride (TG) levels were determined with Roche Diagnostics reagents (Cholesterol CHOD-PAP, HDL-C plus, and Triglycerides GPO-PAP, respectively) on a Cobas c501 analyser. The following formula was used to calculate the level of LDL cholesterol: LDL cholesterol = total cholesterol − HDL cholesterol − (TG/5).
7
Serum Lipid Profiles in Children
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Fasting blood samples were taken from the children’s antecubital fossa. All blood samples were analysed for serum TC, high-density lipoprotein (HDL) cholesterol and triacylglycerol (TAG). TC and TAG were analysed using an enzymatic colorimetric test (Cholesterol CHOD-PAP, Roche Diagnostics, Mannheim, Germany). HDL cholesterol was measured using the same method after precipitation and centrifugation. LDL cholesterol was calculated from the serum TC, TAG and HDL concentrations expressed in mmol/L using the Friedewald formula [16 (link)], which is considered valid if TAG concentrations do not exceed 4.52 mmol/L [17 (link)]. Lipid levels were classified according to guidelines for cardiovascular health and risk reduction in children and adolescents by the US National Heart, Lung, and Blood Institute [18 ]. Cut-off points for acceptable and high levels, respectively, were <4.40 mmol/L and ≥5.18 mmol/L for total cholesterol, <2.85 mmol/L and ≥3.37 mmol/L for LDL cholesterol, and <0.85 mmol/L and ≥1.13 mmol/L for TAG [18 ].
8
Plasma Lipid and Cytokine Profiling
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Plasma cholesterol, triglycerides, and free fatty acid (FFA) levels were determined by commercially available kits, according to the manufacturer's instructions (cholesterol: Cholesterol CHOD-PAP, Roche, Woerden, Netherlands; triglycerides: Hitachi, Roche, Woerden, Netherlands; FFA: Diagnostic Systems, Holzheim, Germany).
Plasma TNF-α, mouse KC (mKC; CXCL1), IFN-γ, and IL-6 levels were measured using a Meso Scale Discovery (Gaithersburg, USA) 10-Plex MULTI-SPOT Mouse Cytokine Assay for plasma, according to the manufacturer's instructions. Plasma concentrations of IL-1β, IL-5, and IL-12p70 were under the detection limit of the assay. Plasma serum amyloid A (SAA) was measured by ELISA (Tridelta, Maynooth, Ireland) according to the manufacturer's instructions.
Plasma TNF-α, mouse KC (mKC; CXCL1), IFN-γ, and IL-6 levels were measured using a Meso Scale Discovery (Gaithersburg, USA) 10-Plex MULTI-SPOT Mouse Cytokine Assay for plasma, according to the manufacturer's instructions. Plasma concentrations of IL-1β, IL-5, and IL-12p70 were under the detection limit of the assay. Plasma serum amyloid A (SAA) was measured by ELISA (Tridelta, Maynooth, Ireland) according to the manufacturer's instructions.
9
Lipid Extraction and Analysis Protocol
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Liver and heart lipids were extracted from frozen samples according to Bligh and Dyer [29 (link)], evaporated under nitrogen, and redissolved in isopropanol before analysis. Lipids from liver, heart and plasma were measured enzymatically on a Hitachi 917 system (Roche Diagnostics GmbH, Mannheim, Germany) using the cholesterol (Cholesterol CHOD-PAP, 11,491,458–216), and triacylglycerol (Triglycerides GPO-PAP, 11,730,711) kit from Roche Diagnostics, and the free cholesterol (Free Cholesterol FS, Ref 113,609,910,930), non-esterified fatty acid (NEFA FS, Ref 157,819,910,935) and phospholipid kit (Phospholipids FS, Ref 157,419,910,930) from DiaSys (Diagnostic Systems GmbH, Holzheim, Germany). L-carnitine, trimethyl-lysine, γ-butyrobetaine, palmitoylcarnitine and acetylcarnitine were analysed in plasma samples, some pooled from 2 to 3 animals (n = 3–6), by HPLC-MS/MS as described by Vernez et al. [30 (link)] with some modifications [31 (link)].
10
Biochemical Analyses of Metabolic Markers
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Measurements of glucose, HbA1c, total cholesterol (TC), triglycerides (TG), high density lipoprotein (HDL) and LDL were done using the standard procedures and available commercial kits in a fully automated system (COBAS integra 400 plus).
Cholesterol CHOD-PAP, triglycerides GPO-PAP, LDL-C plus second generation, HDL-C plus third generation, reagents (Roche Diagnostics, Indianapolis, IN) were used on the Chemistry Analyzer to determine levels of TC, TG, LDL-cholesterol and HDL-cholesterol, respectively. All assays were done following the recommended procedures for instrument operation, calibration, quality control, and assay guidelines. The instrument was calibrated using calibrator for automated systems (Roche Diagnostics) for glucose, TCand TAG, and calibrator for automated systems lipids (Roche Diagnostics) for LDL-cholesterol and HDL-cholesterol.
Results were expressed for all parameters in mg/dl. Except for HbA1c, where it was expressed as percentage of glycosylated hemoglobin [16 (link)].
Cholesterol CHOD-PAP, triglycerides GPO-PAP, LDL-C plus second generation, HDL-C plus third generation, reagents (Roche Diagnostics, Indianapolis, IN) were used on the Chemistry Analyzer to determine levels of TC, TG, LDL-cholesterol and HDL-cholesterol, respectively. All assays were done following the recommended procedures for instrument operation, calibration, quality control, and assay guidelines. The instrument was calibrated using calibrator for automated systems (Roche Diagnostics) for glucose, TCand TAG, and calibrator for automated systems lipids (Roche Diagnostics) for LDL-cholesterol and HDL-cholesterol.
Results were expressed for all parameters in mg/dl. Except for HbA1c, where it was expressed as percentage of glycosylated hemoglobin [16 (link)].
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