Abx pentra 400 analyzer

All patients were scheduled to follow-up clinical and laboratory parameters at their respective district hospitals at 3-month intervals. All blood and urine samples were analyzed with ABX Pentra 400 analyzer (HORIBA ABX S.A.S., France) located at these hospitals. All biochemistry analyses were validated according to the standard protocol of Department of Medical Sciences, Ministry of Public Health, Thailand. Serum creatinine was measured by the enzymatic method. It was standardized with standard reference material (SRM 967) by commutability study every 6 months [17 (link)]. Two out of twelve subgroups of both patient groups were randomly selected and assigned to collect 24-h urine urea nitrogen and sodium.

A protocol of laboratory investigation is summarized in Table 4. Prior to each hospital visit, all subjects will be instructed to have an 8-hour overnight fast. Plasma glucose, lipid profile, HbA1C, albumin, electrolytes, and urine analysis will be measured in the next morning. After centrifugation, plasma and urine samples will be centrifuged and transported at 4°C and analyzed within 24 hours at the laboratory Department of the District 2 Hospital. All blood chemistries will be measured with ABX Pentra 400 analyzer (HORIBA ABX S.A.S., France). Urine protein and creatinine will be measured with the pyrogallol red colorimetric method with the same analyzer. All biochemistry analyses will be validated according to the standard protocol of Department of Medical Sciences, Ministry of Public Health, Thailand. Serum creatinine will be measured by the enzymatic method. Serum creatinine measurement will be standardized with standard reference material (SRM 967) by commutability study at every 6 months [14 (link)].
Two out of twelve subgroups of both groups will be randomly selected and assigned to collect 24-hour urine. An aliquot of 24-hour urine of each sample will be sent for urine chemistry analysis protein, creatinine, urea, and sodium at Laboratory Department of Division of Nephrology, Chulalongkorn University, Bangkok, Thailand.

The ABX Pentra 400 analyzer (Horiba Ltd, Kyoto, Japan) is a compact bench top clinical biochemistry analyzer [20] (link). The configuration of the open channel was based on the Roche Total Mycophenolic Acid® assay package insert. Briefly, the analyzer added 3 µL of sample to 180 µL of R1 (cycle 1; a cycle every 12 s). After mixing and incubation for 5 min, 19 µL of R2 was added and mixed (cycle 26). At this point, bichromatic measurements (340/405 nm) were performed. The slope of the reaction rate is determined between cycles 49 and 62. The measuring temperature (37 °C) was controlled by an air bath. The assay was calibrated with 6-point calibration at 0, 1, 3, 5, 10 and 15 mg/L (Roche Total MPA Calibrators; Roche Diagnostics) using a logit/log4 calculation mode. All samples above 15 mg/L were manually diluted (1:3) with the diluent (equivalent to the 0 mg/L calibrator) from the Roche Total MPA Calibrators.
The assay was bidirectionally interfaced to the Laboratory Information System (LIS) using barcoded tubes. The reagents were stored in the integrated refrigerated reagent compartment (2–10 °C).

Blood samples were collected using ethylenediaminetetraacetic acid (EDTA) tubes after overnight fasting. Samples were centrifuged (2000 rpm, 4 °C) for 20 min to separate plasma and were stored at −80 °C. Blood samples were shipped to the Interclinical Biochemical Laboratory at Sechenov University for biochemical analyses, including the measurements of circulating glucose, creatinine, uric acid, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), plasma triglycerides (TGs), and thyroid hormones. Elevated glucose concentrations were categorized as >100 mg/dL [26 (link)]. Total cholesterol, LDL-C, VLDL-C, and TG were abnormal if >200 mg/dL, >100 mg/dL, >30 mg/dL, and >150 mg/dL, respectively [27 (link)]. Abnormal HDL-C was defined as <40 mg/dL in men and <50 mg/dL in women [27 (link)]. Abnormal serum creatinine was defined as >110 μmol/L for men and <45 or >90 μmol/L for women [28 ]. Abnormal uric acid was defined as >6.99 mg/dL and >5.6 mg/L in men and women, respectively [27 (link)]. Glucose, serum lipids, creatinine and uric acid were determined using an ABX Pentra 400 analyzer (Horiba ABX SAS, Montpellier, France). Extra plasma aliquots were shipped to the Laboratory of Pharmacokinetics and Metabolomics Analysis, Institute of Translational Medicine and Biotechnology at Sechenov University for metabolomics profiling.

Blood samples were collected in 9 ml serum collection tubes coated with clot activator (Vacuette®, Med-Kjemi AS, Norway). All samples were directly transported to the laboratory within a maximum of 120 min following sample collection without any prior chilling. Samples were analysed to determine the following parameters: iron (Fe), inorganic phosphate (P), copper (Cu), zinc (Zn), calcium (Ca), magnesium (Mg) and ceruloplasmin (Cp) levels. Levels of Fe and P were assessed by a colorimetric method (ABX Pentra 400 Analyzer, Horiba). Cu, Zn, Ca, Mg and Cp were determined by Atomic absorption spectrometry (AAnalyst 300 Perkin Elmer). Due to limited financial resources, blood sampling was restricted to Farms 2 and 3.
Tissue samples were stored at −20 °C until analysis. Inductively coupled plasma mass spectrometry (ICP-MS) was performed by SYNLAB.vet GmbH (Berlin, Germany) to determine Zn concentrations in liver and kidney samples.

The blood samples were collected 30 h after surgery, under anesthesia, by cardiac puncture [13 (link)]. The serum samples obtained after centrifugation were immediately frozen at − 20 °C before being analyzed at the phenotyping department of the Toulouse Anexplo platform.
The biochemical determinations (ALP, ALT, AST, serum creatinine, serum lactates, and BUN) were carried out with the ABX Pentra 400 analyzer (Horiba ABX, Montpellier, France). Assessment of serum cytokine concentrations of IL-6 and IL-10 was performed using the Luminex technique (LXAMSM-02 2-plex Mouse Luminex Magnetic Assay, IL6, IL10, Bio-Techne, Abingdon, United Kingdom).