Body weight, height, waist/hip circumference and blood pressure were determined at the morning of surgery. Fat and lean body mass were estimated by bio-electrical impedance analyses (Bodystat 1500; Bodystat Ltd., Isle of Man, UK). Fasting venous blood samples were collected after an overnight fast for measurement of plasma glucose, serum insulin and HbA1C. Insulin sensitivity was assessed by the homeostatic model assessment index for insulin resistance (HOMA-IR), calculated from fasting glucose and insulin, according to the formula: fasting insulin (mU/l)*fasting glucose (mmol/l)/22.525 (link). Plasma glucose concentration was measured by the glucose oxidase method using an AU2700 analyser (Beckman Coulter, Brea, CA, USA). Serum insulin concentration was assessed by immunoassay (ADVIA Centaur Insulin IRI; Siemens Medical Solutions Diagnostics, Tarrytown, NY, USA). HbA1C was assessed by high performance liquid chromatography using a HA-8160 Hi-Auto A1C analyser (Menarini, Zaventem, Belgium).
Au2700 analyser
Manufactured by Beckman Coulter
Sourced in United States, United Kingdom
The AU2700 analyser is a automated clinical chemistry analyser designed for in-vitro diagnostic testing. It utilizes spectrophotometric and immunoturbidimetric methodologies to perform a variety of routine and specialty laboratory tests. The AU2700 is capable of processing a high volume of samples with high throughput and efficiency.
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7 protocols using au2700 analyser
1
Anthropometric and Metabolic Assessment
2
Cord Blood Vitamin D and Mineral Levels
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Following delivery, three milliliters of cord blood were collected from infants who met the study criteria to determine serum concentrations of 25(OH)D, calcium (Ca), magnesium (Mg), phosphorus (P), albumin, parathormone (PTH), and alkaline phosphatase (ALP). Blood samples were collected and sent to the laboratory in accordance with the cold chain protocol. Calcium, P, Mg, albumin, and ALP levels in serum samples were measured using enzymatic, kinetic, and end-point methods on a Beckman Coulter AU2700 analyser with original reagents (Brea, CA, USA). The electrochemiluminescence method was used on an analyzer to measure serum PTH levels (Beckman Coulter DXI Brea, CA, USA), as well as 25(OH)D (Roche Diagnostics Cobas e401 GMBH Mannheim, Germany). The global consensus defines vitamin D deficiency and insufficiency in children as <12 and 12–20 ng/mL, respectively, in the prevention and treatment of nutrition-related rickets [17 (link)]. Based on this consensus, three groups were formed based on cord 25(OH) D vitamin levels; Group 1: 25(OH)D < 12 ng/mL (Deficient); Group 2: 25(OH)D = 12–19 ng/mL (Insufficient); and Group 3: 25(OH)D = 20–100 ng/mL (Optimum).
3
Serum Biomarker Measurement in Sleep Apnea
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Blood samples were drawn from the patients between 8:00 AM and 9:00 AM, after they had undergone a baseline PSG. After 3 months of treatment, a further serum sample was obtained. Subjects were required to fast before blood samples were collected. The blood samples were centrifuged within 30 min at 4°C at 3000 g for 10 min. Serum samples were stored at −80°C until used in the assay. Serum IMA concentrations were measured with a colourimetric method in which reduced cobalt binding to albumin is measured, as described by Bar-Or et al. [14 (link)]. The results were expressed in absorbance units (ABSU). The assay has an intra-assay coefficient of variation (%CV) of <3.5% and an interassay %CV of <6.1%. The serum C-reactive protein (CRP) concentration was measured as highly sensitive CRP (CRP) in an immunoturbidimetric assay using the CRP reagent and a Beckman Coulter AU2700 analyser (Beckman Coulter, Inc., Fullerton, CA USA).
4
Anthropometric and Metabolic Measurements
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Body weight, height, waist/hip circumference and blood pressure were determined at the morning of surgery. Fat and lean body mass were estimated by bio-electrical impedance analyses (Bodystat 1500; Bodystat Ltd., Isle of Man, U.K.). Fasting venous blood samples were collected after an overnight fast for measurement of plasma glucose, serum insulin and glycated hemoglobin. Plasma glucose was measured by the glucose oxidase method using an AU2700 analyser (Beckman Coulter, Brea, CA, USA). Serum insulin was assessed by an immunoassay (ADVIA Centaur Insulin IRI; Siemens Medical Solutions Diagnostics, Tarrytown, NY, USA). Glycated hemoglobin was assessed by high performance liquid chromatography using a HA-8160 Hi-Auto A1C analyser (Menarini, Zaventem, Belgium). Blood cell counts were done with ADVIA 2120 Hematology System (Siemens) and are expressed as cells/L.
5
Biochemical Analysis of Blood Samples
6
Comparison of MELD-Na Calculations
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Each patient´s medical records were reviewed to gather clinical and demographic data, including sodium results from arterial blood gas analysis and from the blood chemistry. The arterial blood gas analysis was determined using the ABL835 FLEX (Radiometer Medical ApS, Denmark) between 2015 and 2018, and with the ABL90 FLEX (Radiometer Medical ApS, Denmark) during 2019; both analysers use direct ISE technique. The blood chemistry analysis was performed with the AU2700 analyser (Beckman Coulter, Brea, USA) via indirect ISE. We calculated the MELD-Na based on the results provided by the ABL835 FLEX/ABL90 FLEX [MELD-Na(D)] and by the AU2700 analyser [MELD-Na(I)], and we then calculated the difference [MELD-Na(D) − MELD-Na(I)] between them.
7
Comprehensive Metabolic Assessment Protocol
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Anthropometric measurements Weight, height and waist and hip circumferences were measured. Participants then rested for 5 min before BP was determined from an average of three measures.
Biochemical measurements Blood samples were collected and analysed using the Olympus AU2700 analyser (Beckman Coulter, High Wycombe, UK) with standard proprietary reagents as follows: glucose with hexokinase, total cholesterol and HDL-cholesterol with cholesterol esterase/ oxidase and triacylglycerol with glycerol kinase. LDLcholesterol was calculated according to the Friedewald formula. Insulin was measured using radioimmunoassay (Invitrogen, Paisley, UK). HOMA-IR was calculated using fasting glucose and insulin concentrations [24] . NEFA was measured using a NEFA assay kit (Randox Daytona, County Antrim, UK). Fasting NEFA and insulin concentrations were used to estimate adipose tissue insulin resistance (adipo-IR) [25] .
OGTT After fasting blood samples were collected, a 75 g glucose solution was consumed within 5 min and post-ingestion blood samples were drawn at 30, 60, 90 and 120 min. Glucose, insulin and NEFA responses were calculated as AUC. Matsuda index was calculated to estimate whole-body insulin sensitivity; indices of hepatic insulin resistance and skeletal muscle insulin sensitivity were determined as previously described [26, 27] .
Biochemical measurements Blood samples were collected and analysed using the Olympus AU2700 analyser (Beckman Coulter, High Wycombe, UK) with standard proprietary reagents as follows: glucose with hexokinase, total cholesterol and HDL-cholesterol with cholesterol esterase/ oxidase and triacylglycerol with glycerol kinase. LDLcholesterol was calculated according to the Friedewald formula. Insulin was measured using radioimmunoassay (Invitrogen, Paisley, UK). HOMA-IR was calculated using fasting glucose and insulin concentrations [24] . NEFA was measured using a NEFA assay kit (Randox Daytona, County Antrim, UK). Fasting NEFA and insulin concentrations were used to estimate adipose tissue insulin resistance (adipo-IR) [25] .
OGTT After fasting blood samples were collected, a 75 g glucose solution was consumed within 5 min and post-ingestion blood samples were drawn at 30, 60, 90 and 120 min. Glucose, insulin and NEFA responses were calculated as AUC. Matsuda index was calculated to estimate whole-body insulin sensitivity; indices of hepatic insulin resistance and skeletal muscle insulin sensitivity were determined as previously described [26, 27] .
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