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MB Sample ID: SA021491
Local Sample ID: | 10jan12_58-r001.d |
Subject ID: | SU000443 |
Subject Type: | Animal |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000443 |
Subject Type: | Animal |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
10jan12_58-r001.d | SA021491 | FL005293 | T1D good glycemic control | treatment |
Collection:
Collection ID: | CO000437 |
Collection Summary: | At 5:00 AM after an overnight fast, baseline blood samples were collected from study participants. Plasma samples were stored at 80°C until analysis. |
Sample Type: | Blood. Plasma was isolated for MS analysis. |
Treatment:
Treatment ID: | TR000457 |
Treatment Summary: | Participants were admitted to the Clinical Research Unit at St Mary’s Hospital (Rochester, Minnesota) the evening before the study and spent overnight in the Clinical Research Unit. The participants were given a standard meal on the evening of the admission after which they fasted overnight. Participants with T1D were treated with insulin as per their usual individual programs. At 5:00 AM after an overnight fast, baseline blood samples were collected from study participants. Plasma samples were stored at 80°C until analysis. |
Sample Preparation:
Sampleprep ID: | SP000450 |
Sampleprep Summary: | Plasma quality-control samples used in the study were prepared from pooled plasma spiked with a selection of metabolites to mimic elevated levels of metabolites during I− (insulin withdrawn) condition. Plasma was spiked with a standard mixture (3:1 ratio of plasma to spiking solution) containing 100 μg/mL niacin, hypoxanthine, leucine, isoleucine, phenylalanine, tryptophan, citric acid, glucose, hippuric acid, and taurocholic acid dissolved in 1:1 acetonitrile/water. All plasma samples (200 μL) were thawed on ice at 4°C followed by deproteinization with methanol (1:4 ratio of plasma to methanol) and vortexed for 10 s, followed by incubation at −20°C for 2 h. The samples were then centrifuged at 15,871g for 30 min at 4°C. The supernatants were lyophilized (Savant, Holbrook, NY) and stored at −20°C prior to analysis. The samples were reconstituted in 50% H2O/acetonitrile and passed through a Microcon YM3 filter (Millipore Corporation). The supernatants were transferred to analytical vials, stored in the autosampler at 4°C, and analyzed within 48 h of reconstitution in buffer. |
Combined analysis:
Analysis ID | AN000667 | AN000668 | AN000669 | AN000670 |
---|---|---|---|---|
Analysis type | MS | MS | MS | MS |
Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
Chromatography system | Agilent 6220 | Agilent 6220 | Agilent 6220 | Agilent 6220 |
Column | none | none | none | none |
MS Type | ESI | ESI | ESI | ESI |
MS instrument type | TOF | TOF | TOF | TOF |
MS instrument name | Agilent 6220 TOF | Agilent 6220 TOF | Agilent 6220 TOF | Agilent 6220 TOF |
Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
Units | intensity | intensity | intensity | intensity |
Chromatography:
Chromatography ID: | CH000482 |
Chromatography Summary: | The liquid chromatography platform consisted of an Acquity UPLC system (Waters, Milford, MA). Plasma metabolite separation was achieved using both hydrophilic interaction chromatography (ethylene-bridged hybrid 2.1 × 150 mm, 1.7 μm; Waters) and reversed-phase liquid chromatography C18 (high-strength silica 2.1 × 150 mm, 1.8 μm; Waters). For each column, the run time was 20 min at a flow rate of 400 μL/min. Reverse-phase chromatography was performed using 99% solvent A (5 mmol/L NH4 acetate, 0.1% formic acid, and 1% acetonitrile) to 100% solvent B (95% acetonitrile with 0.1% formic acid). The gradient was 0 min, 0% B; 1 min, 0% B; 3 min, 5% B; 13.0 min, 100% B; 16 min, 100% B; 16.5 min, 0% B; and 20 min, 0% B. Other LC parameters were injection volume 5 μL and column temperature 50°C. Each sample was injected in triplicate with blank injections between each sample. Quality controls and standards were run at the beginning and end of the sequence. |
Instrument Name: | Agilent 6220 |
Column Name: | none |
Column Temperature: | 50 |
Flow Gradient: | 0 min, 0% B; 1 min, 0% B; 3 min, 5% B; 13.0 min, 100% B; 16 min, 100% B; 16.5 min, 0% B; and 20 min, 0% B. |
Flow Rate: | 400 µL/min |
Solvent A: | 1% acetonitrile/99% water; 0.1% formic acid; 5 mM ammonium acetate |
Solvent B: | 95% acetonitrile/5% water; 0.1% formic acid |
Chromatography Type: | HILIC |
Chromatography ID: | CH000483 |
Chromatography Summary: | The liquid chromatography platform consisted of an Acquity UPLC system (Waters, Milford, MA). Plasma metabolite separation was achieved using both hydrophilic interaction chromatography (ethylene-bridged hybrid 2.1 × 150 mm, 1.7 μm; Waters) and reversed-phase liquid chromatography C18 (high-strength silica 2.1 × 150 mm, 1.8 μm; Waters). For each column, the run time was 20 min at a flow rate of 400 μL/min. Reverse-phase chromatography was performed using 99% solvent A (5 mmol/L NH4 acetate, 0.1% formic acid, and 1% acetonitrile) to 100% solvent B (95% acetonitrile with 0.1% formic acid). The gradient was 0 min, 0% B; 1 min, 0% B; 3 min, 5% B; 13.0 min, 100% B; 16 min, 100% B; 16.5 min, 0% B; and 20 min, 0% B. Other LC parameters were injection volume 5 μL and column temperature 50°C. Each sample was injected in triplicate with blank injections between each sample. Quality controls and standards were run at the beginning and end of the sequence. |
Instrument Name: | Agilent 6220 |
Column Name: | none |
Column Temperature: | 50 |
Flow Gradient: | 0 min, 0% B; 1 min, 0% B; 3 min, 5% B; 13.0 min, 100% B; 16 min, 100% B; 16.5 min, 0% B; and 20 min, 0% B. |
Flow Rate: | 400 µL/min |
Solvent A: | 99% water/1% acetonitrile; 0.1% formic acid; 5 mM ammonium acetate |
Solvent B: | 95% acetonitrile/5% water; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS000593 |
Analysis ID: | AN000667 |
Instrument Name: | Agilent 6220 TOF |
Instrument Type: | TOF |
MS Type: | ESI |
Ion Mode: | POSITIVE |
Capillary Temperature: | 300°C |
Capillary Voltage: | 3.5 kV |
Fragment Voltage: | 150 V |
Mass Accuracy: | <5 parts per million and ~20000 respectively |
Nebulizer: | nebulizer gas 325°C |
Octpole Voltage: | 250 V |
Scan Range Moverz: | 50-1200 |
Scanning Cycle: | 0.5 s |
Skimmer Voltage: | 58 V |
MS ID: | MS000594 |
Analysis ID: | AN000668 |
Instrument Name: | Agilent 6220 TOF |
Instrument Type: | TOF |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
Capillary Temperature: | 300°C |
Capillary Voltage: | 3.5 kV |
Fragment Voltage: | 150 V |
Mass Accuracy: | <5 parts per million and ~20000 respectively |
Nebulizer: | nebulizer gas 325°C |
Octpole Voltage: | 250 V |
Scan Range Moverz: | 50-1200 |
Scanning Cycle: | 0.5 s |
Skimmer Voltage: | 58 V |
MS ID: | MS000595 |
Analysis ID: | AN000669 |
Instrument Name: | Agilent 6220 TOF |
Instrument Type: | TOF |
MS Type: | ESI |
Ion Mode: | POSITIVE |
Capillary Temperature: | 300°C |
Capillary Voltage: | 3.5 kV |
Fragment Voltage: | 150 V |
Mass Accuracy: | <5 parts per million and ~20000 respectively |
Nebulizer: | nebulizer gas 325°C |
Octpole Voltage: | 250 V |
Scan Range Moverz: | 50-1200 |
Scanning Cycle: | 0.5 s |
Skimmer Voltage: | 58 V |
MS ID: | MS000596 |
Analysis ID: | AN000670 |
Instrument Name: | Agilent 6220 TOF |
Instrument Type: | TOF |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
Capillary Temperature: | 300°C |
Capillary Voltage: | 3.5 kV |
Fragment Voltage: | 150 V |
Mass Accuracy: | <5 parts per million and ~20000 respectively |
Nebulizer: | nebulizer gas 325°C |
Octpole Voltage: | 250 V |
Scan Range Moverz: | 50-1200 |
Scanning Cycle: | 0.5 s |
Skimmer Voltage: | 58 V |