Carboxy-terminal telopeptides levels of collagen I (CTx) and insulin were measured by electrochemiluminescence (E170 Modular Analyser; Roche Diagnostics, Mannheim, Germany). Levels of total alkaline phosphatase (ALP), total calcium and phosphate, glucose, triglycerides, total cholesterol and high-density lipoprotein cholesterol (HDL) were assessed using a spectrophotometer (Olympus 5400, Olympus, Melville, NY, USA). Low-density lipoprotein cholesterol (LDL-cholesterol, mg/dL) was calculated as TC − (HDL + TG/5)19 (link). The HOMA-IR (homeostasis model assessment) insulin resistance index was calculated as fasting serum insulin in µIU/mL x (fasting serum glucose in mg/dL × 0.05551)/22.520 (link).
Modular e170 analyser
Manufactured by Roche
Sourced in Germany
The Modular E170 Analyser is a versatile laboratory instrument designed for automated immunoassay testing. It features a modular design and can perform a wide range of in-vitro diagnostic tests.
Automatically generated - may contain errors
Lab products found in correlation
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Ria kit, by Linco Research (1 mentions)
Ria kit, by Phoenix Pharmaceuticals (1 mentions)
Modular p analyser, by Roche (1 mentions)
Immulite 2000 immunoassay system, by Siemens (1 mentions)
9 protocols using modular e170 analyser
1
Bone Metabolism and Lipid Profile Assessment
2
Preoperative and Postoperative OGTT Measurements
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At the time of the preoperative and postoperative OGTT the following clinical information was ascertained: age, gender, past medical history, treatment, and duration of diabetes. Baseline clinical measurements consisted of weight, height, BMI, waist circumference, and systolic and diastolic blood pressure. Baseline biochemical measurements (total cholesterol, low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), and triglycerides) were performed within the local hospital accredited laboratory. Fasting and 2-hour glucose, fasting lipids (Roche Modular P800 Analyser), and fasting insulin and C-peptide (Roche E170 Modular Analyser) were also measured in the local hospital laboratory. All samples were collected on ice, centrifuged and separated within one hour of collection, and subsequently stored at −80°C until analysis.
3
Analytical Methodologies in Clinical Biochemistry
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All measurements were performed at the department of clinical chemistry and laboratory medicine (AKCL) of Leiden University Medical Center, which is accredited according to CCKL (National Coordination Committee for Quality Assurance for Health Care Laboratories in The Netherlands). Cortisol was measured using an ECLIA assay on a Modular E170 analyser from Roche (Roche Diagnostics, Almere, The Netherlands), ACTH and DHEAS on an Immulite 2000 Xpi analyser (Siemens Healthcare diagnostics, The Hague, The Netherlands) and HbA1c on a Primus Ultra 2 HPLC analyser (Trinity Biotech, Bray, Ireland), using boronate affinity separation. For each participant, all samples from one time series were measured within the same lot number and in the same batch. For this study, the precision and quality of all assayed analytes met or surpassed the level of desirable quality specifications[15 ]. For cortisol Randox controls (Cat. Nr. I/1160EC and 3/1165EC) were used and overall coefficients of variation (CV) for cortisol ranged between 2.4–5.1%, which was well below the desirable CV of 10.5%. For ACTH two levels of controls were used (C2000LACCM1 and C2000LACCM2) and the CV ranged between 3.8–7.7%, which was well below the desirable CV of 10%. In our laboratory the reference range for is ACTH is 3–75 ng/L, for cortisol 0.1–0.6 μmol/L, for HbA1c 20–42 mmol/mol Hb.
4
Menopausal Status and Symptom Assessment
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We made a diagnosis of perimenopausal or postmenopausal status according to the 2012 Stages of Reproductive Aging Workshop (STRAW) criteria [9 (link)].
Menopausal symptoms were assessed using the modified Kupperman Menopausal Index score (mKMI) [10 (link)].
For women with no menstruation, blood samples were collected for hormone tests. Women who still had an identifiable menstrual cycle were asked to undergo hormone tests on the second day of the cycle. The sample was collected from each subject the next morning between 8 and 11 am after overnight fast. Levels of E2, progesterone (P), testosterone (TE), LH and FSH were estimated by ELISA with Roche Modular E170 Analyser, Berlin, Germany. To analyze E2, there was a lower limit of detection of 18.35 pmol/L.
Menopausal symptoms were assessed using the modified Kupperman Menopausal Index score (mKMI) [10 (link)].
For women with no menstruation, blood samples were collected for hormone tests. Women who still had an identifiable menstrual cycle were asked to undergo hormone tests on the second day of the cycle. The sample was collected from each subject the next morning between 8 and 11 am after overnight fast. Levels of E2, progesterone (P), testosterone (TE), LH and FSH were estimated by ELISA with Roche Modular E170 Analyser, Berlin, Germany. To analyze E2, there was a lower limit of detection of 18.35 pmol/L.
5
Thyroid function and environmental exposures
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Blood samples were analysed at the RD&E Blood Sciences department for serum thyroid stimulating hormone (TSH), free thyroxine (FT4) and thyroid peroxidase antibodies (TPO-Ab) using the electrochemiluminescent immunoassay, run on the Modular E170 Analyser (Roche, Burgess Hill, UK). Intra-assay coefficients of variations were < 5.3% for both TSH and FT4. The manufacturer’s population reference ranges were: TSH 0.35–4.5 mIU/l and FT4 11–24 pmol/l. Serum TPO-Ab titre above 34 IU/l was considered positive.
Urine samples were analysed for iodine, perchlorate, and thiocyanate concentration using ion chromatography-mass spectrometry at the Iodine Research Laboratory (Boston University, Boston, Massachusetts) [15 (link)]. The inter-assay coefficient of variation ranges from 3.1–8.2% for the three analytes.
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6
Plasma Biomarker Analysis in Subjects
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Each sample was collected into a cooled plastic tube containing 100 μl of 5% EDTA. The plasma was obtained after centrifugation for 5 min at 2000 g at 4 °C. Plasma was separated and frozen within half an hour of being drawn from the subject, and was stored at -20 °C until analysed.
C-peptide was measured in serum using ECLIA (electrochemiluminiscence immunoassay, Modular E 170 analyser, Roche). The measuring range of the kit (defined by the lower detection limit and the maximum of the master curve) was 0.003-13.3 nmol/l or 0.01-40.0 ng/ml for plasma. Intra-and inter-assay coefficients of variation were 1.5% and 2.3%, respectively.
Blood glucose was measured using the enzymatic reference method with hexokinase (Cobas Integra 400 plus analyser, Roche). The measuring range of the kit was 0.12-40 mmol/l (2.16-720 mg/dl). Intra-set and inter-set reproducibility were 1.7% and 2.6%, respectively.
Cortisol was determined by the conventional RIA method.
C-peptide was measured in serum using ECLIA (electrochemiluminiscence immunoassay, Modular E 170 analyser, Roche). The measuring range of the kit (defined by the lower detection limit and the maximum of the master curve) was 0.003-13.3 nmol/l or 0.01-40.0 ng/ml for plasma. Intra-and inter-assay coefficients of variation were 1.5% and 2.3%, respectively.
Blood glucose was measured using the enzymatic reference method with hexokinase (Cobas Integra 400 plus analyser, Roche). The measuring range of the kit was 0.12-40 mmol/l (2.16-720 mg/dl). Intra-set and inter-set reproducibility were 1.7% and 2.6%, respectively.
Cortisol was determined by the conventional RIA method.
7
Plasma Biomarker Measurement Protocol
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Each sample was collected into a cooled plastic tube containing 100 μl of 5%EDTA. The plasma was obtained after centrifugation for 5 min at 2000 rpm at 4 °C. Plasma was separated and frozen within half an hour of being drawn from the subject, and was stored at -20 °C until analyzed.
C-peptide was measured in serum using ECLIA (electrochemiluminiscence immunoassay, Modular E 170 analyser, Roche). The measuring range of the kit (defined by the lower detection limit and the maximum of the master curve) was 0.003-13.3 nmol/l or 0.01-40.0 ng/ml for plasma. Intra-and inter-assay coefficients of variation were 1.5 % and 2.3 %, respectively.
Blood glucose was measured using the enzymatic reference method with hexokinase (Cobas Integra 400 plus analyser, Roche). The measuring range of the kit was 0.12-40 mmol/l (2.16-720 mg/dl). Intraset and inter-set reproducibility were 1.7 % and 2.6 %, respectively.
Total plasma ghrelin was determined by a commercially available RIA kit (Linco Research, Inc., St. Charles, Missouri, USA, detection range: 93-6000 pg/ml), and orexin A was also measured by a RIA kit (Phoenix Pharmaceuticals, Inc., detection range 10-1280 pg/ml).
Kits from Immunotech (France) were used to measure LH and FSH (IRMA kit). Cortisol was measured using RIA kit from Immunotech (France). Sex hormones bounding globulin (SHBG) were measured by the IRMA kit Immunotech (France).
C-peptide was measured in serum using ECLIA (electrochemiluminiscence immunoassay, Modular E 170 analyser, Roche). The measuring range of the kit (defined by the lower detection limit and the maximum of the master curve) was 0.003-13.3 nmol/l or 0.01-40.0 ng/ml for plasma. Intra-and inter-assay coefficients of variation were 1.5 % and 2.3 %, respectively.
Blood glucose was measured using the enzymatic reference method with hexokinase (Cobas Integra 400 plus analyser, Roche). The measuring range of the kit was 0.12-40 mmol/l (2.16-720 mg/dl). Intraset and inter-set reproducibility were 1.7 % and 2.6 %, respectively.
Total plasma ghrelin was determined by a commercially available RIA kit (Linco Research, Inc., St. Charles, Missouri, USA, detection range: 93-6000 pg/ml), and orexin A was also measured by a RIA kit (Phoenix Pharmaceuticals, Inc., detection range 10-1280 pg/ml).
Kits from Immunotech (France) were used to measure LH and FSH (IRMA kit). Cortisol was measured using RIA kit from Immunotech (France). Sex hormones bounding globulin (SHBG) were measured by the IRMA kit Immunotech (France).
8
Biomarker Quantification from Blood Samples
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Each sample was collected into a plastic tube containing 100 µl of 5% EDTA. Plasma was obtained after centrifugation for 5 min at 2000 rpm at 4 °C, then separated and frozen within half an hour of being drawn from the subject, and stored at -20 °C until analysed.
C-peptide was measured in serum using ECLIA (electrochemiluminiscence immunoassay, Modular E 170 analyser, Roche). The measuring range of the kit (defined by the lower detection limit and the maximum of the master curve) was 0.003-13.3 nmol/l or 0.01-40.0 ng/ml for plasma. Intra-and inter-assay coefficients of variation were 1.5% and 2.3%, respectively.
Blood glucose was measured using the enzymatic reference method with hexokinase (Cobas Integra 400 plus analyser, Roche). The measuring range of the kit was 0.12-40 mmol/l (2.16-720 mg/dl). Intra-set and inter-set reproducibility were 1.7% and 2.6%, respectively. Cortisol was measured using an RIA kit (Immunotech, France). Melatonin was measured using an RIA kit (Labor Diagnostika Nord GmbH and Co. KG, Germany). Sensitivity for melatonin was 2 pg/ml, intra-assay and inter-assay coefficients of variation were 9.8-12.1% and 9.6-12.3%, respectively.
C-peptide was measured in serum using ECLIA (electrochemiluminiscence immunoassay, Modular E 170 analyser, Roche). The measuring range of the kit (defined by the lower detection limit and the maximum of the master curve) was 0.003-13.3 nmol/l or 0.01-40.0 ng/ml for plasma. Intra-and inter-assay coefficients of variation were 1.5% and 2.3%, respectively.
Blood glucose was measured using the enzymatic reference method with hexokinase (Cobas Integra 400 plus analyser, Roche). The measuring range of the kit was 0.12-40 mmol/l (2.16-720 mg/dl). Intra-set and inter-set reproducibility were 1.7% and 2.6%, respectively. Cortisol was measured using an RIA kit (Immunotech, France). Melatonin was measured using an RIA kit (Labor Diagnostika Nord GmbH and Co. KG, Germany). Sensitivity for melatonin was 2 pg/ml, intra-assay and inter-assay coefficients of variation were 9.8-12.1% and 9.6-12.3%, respectively.
9
Maternal Biomarker Assay Protocol
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All maternal serum samples were assayed for hCG, progesterone, PAPP-A and hsCRP using commercial assays. Maternal serum progesterone and hCG assays were performed on a Modular E170 Analyser (Roche Diagnostics, Vilvoorde, Belgium) with an electrochemiluminescence competitive method. hsCRP assay was performed using an immunoturbidimetric method on a Modular P Analyser (Roche Diagnostics) with a quantitation limit of 0.5 mg/l. Maternal serum PAPP-A assay was performed on a IMMULITE 2000 immunoassay system (Siemens, Brussels, Belgium) with an enzyme-labelled chemiluminescent immunometric method.
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