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MB Sample ID: SA184290
| Local Sample ID: | O_1 |
| Subject ID: | SU002035 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 53 ± 15 |
| Weight Or Weight Range: | 116 ± 25 |
| Gender: | Male and female |
| Human Inclusion Criteria: | (i) body mass index (BMI)>35kg/m2 plus GFR 30–60ml/min and proteinuria >1 g/24h or GFR>60ml/min and proteinuria >2.5 g/24h despite receiving maximally tolerated doses of renin-angiotensin-aldosterone system (RAAS) blocker and (ii) BMI>40kg/m2 with a GFR>30ml/min and proteinuria >0.5 g/24h despite receiving maximally tolerated doses of RAAS blocker. For the renal function criterion, eGFR was used. The follow-up time was 24 months. |
| Human Exclusion Criteria: | 1) Patients who had participated or were participating in another clinical trial or had taken an experimental drug in the last 28 days. 2) Patients with renal transplantation and/or chronic replacement therapy (hemodialysis and/or peritoneal dialysis). 3) Subjects with poorly controlled blood pressure (SBP > 170 mmHg or DBP > 110 mmHg). 4) Patients with a history of cardiovascular events in the past six months. 5) Patients treated with immunosuppressants. 6) Subjects with a history of renovascular disease, autoimmune diseases, cancer, drug use, or obstructive uropathy. 6) Patients who did not sign the informed consent. 7) Patients who were pregnant or lactating. eGFR was used to establish the renal function criteria. Patients were monitored for 24 months. |
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Subject:
| Subject ID: | SU002035 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 53 ± 15 |
| Weight Or Weight Range: | 116 ± 25 |
| Gender: | Male and female |
| Human Inclusion Criteria: | (i) body mass index (BMI)>35kg/m2 plus GFR 30–60ml/min and proteinuria >1 g/24h or GFR>60ml/min and proteinuria >2.5 g/24h despite receiving maximally tolerated doses of renin-angiotensin-aldosterone system (RAAS) blocker and (ii) BMI>40kg/m2 with a GFR>30ml/min and proteinuria >0.5 g/24h despite receiving maximally tolerated doses of RAAS blocker. For the renal function criterion, eGFR was used. The follow-up time was 24 months. |
| Human Exclusion Criteria: | 1) Patients who had participated or were participating in another clinical trial or had taken an experimental drug in the last 28 days. 2) Patients with renal transplantation and/or chronic replacement therapy (hemodialysis and/or peritoneal dialysis). 3) Subjects with poorly controlled blood pressure (SBP > 170 mmHg or DBP > 110 mmHg). 4) Patients with a history of cardiovascular events in the past six months. 5) Patients treated with immunosuppressants. 6) Subjects with a history of renovascular disease, autoimmune diseases, cancer, drug use, or obstructive uropathy. 6) Patients who did not sign the informed consent. 7) Patients who were pregnant or lactating. eGFR was used to establish the renal function criteria. Patients were monitored for 24 months. |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| O_1 | SA184290 | FL022670 | O | Genotype |
Collection:
| Collection ID: | CO002028 |
| Collection Summary: | First morning void and 24-hour urine samples, were collected, aliquoted and stored at -80°C until extraction. 24-h urine samples were prepared collecting all the urine after the first morning void until the first urine in the next day. |
| Sample Type: | Urine |
| Collection Frequency: | First void and 24h urines |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002047 |
| Treatment Summary: | CKD patients with severe obesity and non-CKD patients with severe obesity underwent bariatric surgery. |
Sample Preparation:
| Sampleprep ID: | SP002041 |
| Sampleprep Summary: | 100 µL of each urine sample were mixed in a 100 µL IS (internal standard) mixture with 0.4 mM of methionine sulfone and 2 mM of paracetamol containing 5% of acetonitrile, then, samples were vortexed. Some CKD patients showed an elevated proteinuria and in order to avoid capillary collapse, a Merck Milipore filters (Ultrafiltration Device with Ultracel YM-T membrane, product 4104) were used. Samples mixed with IS mixture were added to filters and centrifuged (2000 g, 60 min, 4 °C). Then, 90 µl of each sample was transferred to a glass vial. Samples were stored at -20°C until analysis. Stability and reproducibility of the system were checked with quality control (QC) samples. Four types of QC samples were prepared combining 60 µl of each sample after filtration: 1. QC from OD and OD BS samples (QC CKD). 2. QC from O and O BS samples (QC Control). 3. QC from OD, OD BS and O and OD BS samples (QC CC). 4. QC from Healthy 1H and Healthy 24H samples (QC Healthy). QC samples were vortex-mixed, transferred to a vial and stored at -20°C until analysis. |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | On ice |
Combined analysis:
| Analysis ID | AN003187 |
|---|---|
| Analysis type | MS |
| Chromatography type | None (Direct infusion) |
| Chromatography system | none |
| Column | none |
| MS Type | EI |
| MS instrument type | CE-TOF-MS |
| MS instrument name | Agilent 6224 TOF |
| Ion Mode | POSITIVE |
| Units | Area |
Chromatography:
| Chromatography ID: | CH002355 |
| Chromatography Summary: | Analysis were performed in a capillary electrophoresis (CE) system coupled to a TOF-MS analyser (TOF: time-of-flight) purchased from Agilent Technologies (CE: 7100, MS: 6224). In coupling process, steath liquid was supplied by an ISO pump (Agilent 1200) to increase the volatility and compensate the volume for MS through an electrospray source. A fused silica capillary was used to perform metabolite separation (100 cm total length x 50 μm i.d. x 360 μm o.d., provided by Agilent Technologies). Capillary was conditioned with water, BGE and NaOH. Before analysis, capillary was rinsed for 5 min at 950 mbar with BGE and was applied a voltage of 30kV for 10 s to displace BGE ions. After conditioning, sample was loaded applying 50 mbar for 50 s, then, BGE was applied at 100 mbar for 10 s. In the capillary was applied a pressure of 25 mbar and a voltage of 30 kV to achieve metabolite separation. A positively charged spray is formed by a flow of 0.6 mL min−1 (1:100 split) when separated compounds leave the capillary from the auxiliary liquid system with a nebulization pressure of 10 psig with nitrogen and a capillary voltage of 3500 V. A hot nitrogen flow of 10 mL min−1 at 200 °C was applied to dry the spray. |
| Instrument Name: | none |
| Column Name: | none |
| Chromatography Type: | None (Direct infusion) |
MS:
| MS ID: | MS002965 |
| Analysis ID: | AN003187 |
| Instrument Name: | Agilent 6224 TOF |
| Instrument Type: | CE-TOF-MS |
| MS Type: | EI |
| MS Comments: | Ions in gaseous state were directed to the TOF using two different voltages for the fragmentor (125 V and 200 V) and voltages of 65 V and 750 V for the skimmer and the octopole, respectively. Mass range was set from 70 to 1050 Da at a scanning rate of 1.02 scans per second. Data was collected in ESI positive ion mode. MassHunter Workstation version B.06.01 (Agilent Technologies) was used to monitor CE-MS analysis. Voltage of 125 V in the fragmentor was applied to ordinary samples and QC control samples. Data acquired at 125 V, equivalent to MS1, provided information about adducts, isotopes, multimers and fragments ions with low intensity. Voltage of 200 V in the fragmentor was applied to four QC (covering the four types of QC prepared). Data acquired at 200 V is similar to data obtained in 10 eV MS/MS, although has been previously reported that some fragment ions could differ. An equilibration QC was prepared to adapt and prepare the equipment for the conditions of the samples to be analysed. Equilibration QC was prepared as a pool of the four types of QC used to cover completely the biological diversity of the samples. Equilibration QC was analysed before samples. QC samples were disposed throughout the instrumental analysis. |
| Ion Mode: | POSITIVE |
