Fasting blood samples of all patients were taken from the antecubital vein after an overnight fasting period of at least 8 h. Biochemical parameters were studied from plasma samples. Plasma glucose levels were measured using the enzymatic reference method with hexokinase (Beckman Coulter AU5800), and plasma HbA1c levels were measured by high-performance liquid chromatography (HPLC) and mass spectroscopy method (Premier HB9210). Low-density lipoprotein (LDL) was measured using enzymatic colorimetric assay (Beckman Coulter AU5800), and plasma creatinine was assessed using the kinetic Jaffé method (Beckman Coulter AU5800). Urine protein level was measured by the protein error of indicator method (IQ 200/iChem velocity).
Au5800
Manufactured by Beckman Coulter
Sourced in United States, Japan, Germany, United Kingdom, China, Italy, Canada
The AU5800 is a chemistry analyzer designed for high-throughput clinical laboratory testing. It features advanced optics and automation to provide reliable and efficient sample processing. The core function of the AU5800 is to perform a variety of clinical chemistry tests, including immunoassays, on patient samples.
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Cobas 6000, by Roche (9 mentions)
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Au680, by Beckman Coulter (7 mentions)
Cobas 8000, by Roche (6 mentions)
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659 protocols using au5800
1
Fasting Blood Biomarker Profiling
2
Analytical Performance of Immunoassay Tests
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Precision studies were performed in qualitative (one-point calibration against cutoff calibrator) and semi-quantitative (five-point calibration) modes on the Beckman Coulter AU480, AU680, AU5800, and DxC 700 AU clinical analyzers. Cross-reactivity: Cross-reactivity studies were performed in qualitative and semi-quantitative modes on the Beckman Coulter AU480, AU680, AU5800, and DxC 700 AU clinical analyzers. Proof-of-principle cross-reactivity testing was performed and crossreactivity was similar across all four analyzer platforms. Method Comparison: Method comparison studies were performed in qualitative and semi-quantitative modes on the Beckman Coulter AU480, AU680, AU5800, and DxC 700 AU clinical analyzers. Clinical samples containing hydrocodone and hydromorphone were confirmed by LC/MS.
3
Comprehensive Metabolic Profiling in Adults
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BMI was calculated by the standard formula [BMI = weight (kg) / height (m 2 )]. The systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured after 15 min of rest using a mercury sphygmomanometer [17] . Following overnight fasting, blood samples were obtained from all participants through venipuncture. Fasting plasma glucose (FPG), triglycerides (TG), total cholesterol (TC), low density lipoproteincholesterol (LDL-C), high density lipoprotein-cholesterol (HDL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and serum creatinine (Scr) were measured using an automatic biochemistry analyzer (AU5800; Beckman Coulter, America). The estimated glomerular filtration rate (GFR) was calculated as the endogenous creatinine clearance using the Cockcroft equation for males: GFR = [140 -age (years) * body weight (kg)] / [0.818 * Scr (µmol/L)]. The GFR was multiplied by 0.85 for females. HbA1c (Glycated Hemoglobin, Type A1c) was determined using high-performance liquid chromatography (HA-8180; ARKRAY, Japan). The urinary albumin concentration was measured by immunophelometry (AU5800; Beckman Coulter, America). The urinary albumin-to-creatinine ratio (ACR) was determined as ACR = urine albumin (mg) / urine creatinine (g). All measurements were repeated twice.
4
Metabolic Biomarker Profiling in Fasting Participants
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Blood samples will be obtained from participants’ antecubital vein in a supine position during the morning in a fasting state. Blood parameters will include insulin, glucose, triglycerides, total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine transaminase (ALT), and γ-glutamyl transferase (γ-GT). Glucose levels will be measured by spectrophotometric techniques (AU5800, Beckman Coulter, Brea, California, USA). Insulin will be assessed by chemiluminescence immunoassay with paramagnetic particles (UniCel DxI 800, Beckman Coulter, Brea, California, USA). Triglycerides, total cholesterol, and HDL-C will be automatically evaluated by spectrophotometric techniques (AU5800, Beckman Coulter, Brea, California, USA). LDL-C will be considered as . ALT and γ-GT will be calculated by absorption spectrophotometric techniques (Beckman Coulter, Brea, California, USA). Insulin glucose ratio (insulin/glucose), LDL-C/HDL-C ratio (LDL-C/HDL-C), and triglycerides/HDL-C ratio (triglycerides/HDL-C) will also be determined.
5
Biomarkers for Acute Coronary Syndrome
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Plasma fasting blood glucose (FBG), triglyceride (TG), high‐density lipoprotein cholesterol (HDL‐C), and LDL cholesterol (LDL‐C) concentrations were measured by standard laboratory procedures in an automatic biochemistry analyzer (AU5800; Beckman Coulter) using detection kits (Beckman Coulter). In addition, plasma CK‐MB, myoglobin (MYO), and high‐sensitive cardiac troponin I (hs‐cTnI) were measured by a UniCel DxI 800 analyzer (Beckman Coulter) with commercial detection kits (Beckman Coulter).
For this study, we used archived plasma samples of healthy controls and patients with ACS that had been frozen at −80°C and never previously thawed. The determination of sd‐LDL‐C concentrations was performed by a fully automated homogeneous method (Denka Seiken Co., Ltd.) in an automated biochemistry analyzer (AU5800; Beckman Coulter) as previously described.20 The ratio of sd‐LDL‐C/LDL‐C was calculated as measured sd‐LDL‐C (mmol/L)/ LDL‐C (mmol/L).18 All the laboratory measurements of patients with ACS and healthy subjects were under no treatment after admission.
For this study, we used archived plasma samples of healthy controls and patients with ACS that had been frozen at −80°C and never previously thawed. The determination of sd‐LDL‐C concentrations was performed by a fully automated homogeneous method (Denka Seiken Co., Ltd.) in an automated biochemistry analyzer (AU5800; Beckman Coulter) as previously described.
6
Standardized Biomarker Measurements for Epidemiological Studies
7
Urine and Blood Biomarker Analysis
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Urine was collected by participants using containers designed by the researchers and was stored at +4 °C in the refrigerator before measurement. Blood samples were collected by trained nurses. After being centrifuged, all the blood samples were stored at −20 °C in the refrigerator before measurement. Urine and plasma osmolality were measured with the method of the freezing point using an osmotic pressure molar concentration meter (SMC 30C, Tianhe, Tianjin, China).
The electrolyte concentrations of sodium, potassium, chloride, calcium, magnesium, and phosphate in the urine and blood were assessed using an automatic biochemical analyzer (AU5800, Beckman, Brea, CA, USA) with the ion-selective electrode potentiometer method. Urine specific gravity (USG) was tested with the uric dry-chemistry method by a trained technician using an automatic urinary sediment analyzer (H-800, Dirui, Jilin, China).
Blood glucose (BG) was measured with an automatic biochemical analyzer (AU5800, Beckman, Brea, CA, USA), with the ion-selective electrode potentiometer method.
The electrolyte concentrations of sodium, potassium, chloride, calcium, magnesium, and phosphate in the urine and blood were assessed using an automatic biochemical analyzer (AU5800, Beckman, Brea, CA, USA) with the ion-selective electrode potentiometer method. Urine specific gravity (USG) was tested with the uric dry-chemistry method by a trained technician using an automatic urinary sediment analyzer (H-800, Dirui, Jilin, China).
Blood glucose (BG) was measured with an automatic biochemical analyzer (AU5800, Beckman, Brea, CA, USA), with the ion-selective electrode potentiometer method.
8
Kidney Oxidative Stress Measurements
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A total of 90 min after the left kidney was exposed by laparotomy, 5 ml of blood was drawn from the inferior vena cava and collected into heparinized tubes. Serum urea and creatinine levels were measured by a spectrophotometric method using a commercial kit (Beckman Coulter® AU 5800, USA). Blood urea nitrogen (BUN) levels were calculated from urea values. The serum levels of tumor necrosis factor-alpha (TNF-α) and cystatin C (Cys C) were determined via enzyme-linked immunosorbent assay using commercial kits (BioVendor, Brno, Czech Republic and Invitrogen, Camarillo, CA, USA). All assays were studied twice, and the mean value was recorded.
A half of the left kidney was washed with 0.9% NaCl and stored at −80°C until homogenization. Kidney tissues were homogenized with 1:10 phosphate tamponade at two steps using a homogenizer (Janke and Kunkel Ultra-Turrax® T25, Germany) and a sonicator (UW-2070, Bandelin Electronic GmbH and Co, Germany). The total antioxidant status (TAS) and total oxidant status (TOS) of the tissues were analyzed using status assay kits (Reel Assay Diagnostics, Turkey) in accordance with the modified Erel method.[7 (link),8 (link)] Protein levels of the homogenates were measured using spectrophotometry (Beckman Coulter® AU 5800, USA). TAS and TOS results were expressed as mmol Trolox Eq/gr and μmol H2O2 Eq/gr, respectively.
A half of the left kidney was washed with 0.9% NaCl and stored at −80°C until homogenization. Kidney tissues were homogenized with 1:10 phosphate tamponade at two steps using a homogenizer (Janke and Kunkel Ultra-Turrax® T25, Germany) and a sonicator (UW-2070, Bandelin Electronic GmbH and Co, Germany). The total antioxidant status (TAS) and total oxidant status (TOS) of the tissues were analyzed using status assay kits (Reel Assay Diagnostics, Turkey) in accordance with the modified Erel method.[7 (link),8 (link)] Protein levels of the homogenates were measured using spectrophotometry (Beckman Coulter® AU 5800, USA). TAS and TOS results were expressed as mmol Trolox Eq/gr and μmol H2O2 Eq/gr, respectively.
9
Biomarker Assessment in Longitudinal Study
10
Serum Biomarkers in Clinical Assessments
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After BP measurements, participants remained in the seated position and blood samples were drawn via venipuncture from a vein in the antecubital fossa using a disposable cannula (S-Monovette, Sarstedt, Nürmbrecht, Germany). After collection, blood samples were immediately processed and transported to the diagnostic laboratories of the University Hospital Erlangen for further analyses. Blood samples were used to determine serum concentrations of fasting glucose, triglycerides, total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and liver biochemistry (alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma glutamyl transpeptidase (GGT)), which were measured photometrically (Clinical Chemistry Analyzer AU700 or AU5800, Beckman Coulter, Brea, CA, USA), and high-sensitivity C-reactive protein (hsCRP) and glycated hemoglobin A1c (HbA1c) using turbidimetric immunoassays (Clinical Chemistry Analyzer AU700 or AU5800, Beckman Coulter, Brea, CA, USA, and COBAS Integra 400, Roche Diagnostics, Mannheim, Germany, respectively).
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