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MB Sample ID: SA208989
Local Sample ID: | 139 |
Subject ID: | SU002261 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Age Or Age Range: | 8 - 10 weeks |
Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002261 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Age Or Age Range: | 8 - 10 weeks |
Gender: | Male |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
139 | SA208989 | FL025495 | HDR | Factor |
139 | SA208989 | FL025495 | D1 | Factor |
139 | SA208989 | FL025495 | post | Factor |
Collection:
Collection ID: | CO002254 |
Collection Summary: | Urine was collected after irradiation |
Sample Type: | Urine |
Collection Method: | Spot Urine |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR002273 |
Treatment Summary: | Male 8 – 10 week old C57BL/6 mice were obtained from Charles River Laboratories (Frederick, MD, USA) and were irradiated using the FLASH irradiator at the Radiological Research Accelerator Facility [16] (Figure S1). This novel irradiator is based on a Clinac 2100C (Varian Medical Systems, Corona, CA, USA) where the pulse delivery is controlled using in house software. All irradiations were performed using 9 MeV electrons with no scatterer or flattening filter. For these experiments, mice were placed in a 72 mm x 41mm x41mm acrylic box in which air holes had been drilled (The Container Store, Coppell, TX, USA). For 0.7 Gy/min irradiations, mice (n=6) were individually placed at 120 cm above the clinac head and irradiation delivered at 3.25 pulses per second. In this configuration, 3 Gy was delivered in 580 pulses. For 7 Gy/sec mice (n=6) were individually placed 20cm above the clinac head (Figure S1) and dose delivered at 180 pulses/sec after allowing 20 sec where the acceleration and electron source were both on but operated asynchronously so that no beam is delivered. In this configuration, 3 Gy was delivered in 78 pulses. Dosimetry was performed prior to irradiation using a NIST-traceable Advanced Marcus Ion Chamber (AMIC) and Unidos E electrometer (PTW, Freiburg, Germany). Verification of dosimetry was performed using OBT3 radiochromic film (Ashland Specialty Chemicals, Wayne, NJ, USA). |
Sample Preparation:
Sampleprep ID: | SP002267 |
Sampleprep Summary: | urine (20 μl) was deproteinized (80 μl 50% cold acetonitrile) with internal standards (2 μM debrisoquine [M+H]+ = 176.1188; 30 μM 4-nitrobenzoic acid [M-H]- = 166.0141), vortexed, incubated on ice (10 min), and centrifuged for 10 min (max speed, 4 °C). A 1 μl aliquot of each sample was combined for a quality control (QC) sample. |
Processing Storage Conditions: | -80℃ |
Combined analysis:
Analysis ID | AN003563 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity |
Column | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Waters Synapt G2 S QTOF |
Ion Mode | POSITIVE |
Units | peak area |
Chromatography:
Chromatography ID: | CH002634 |
Chromatography Summary: | Mobile phases consisted of the following: solvent A (water/0.1% formic acid [FA]) and solvent B (acetonitrile/0.1% FA). The gradient for urine was (solvent A and B) 4.0 min 5% B, 4.0 min 20% B, 5.1 min 95% B, and 1.9 min 5% B at a flow rate of 0.5 ml/min, column temp 40 °C. |
Instrument Name: | Waters Acquity |
Column Name: | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) |
Column Temperature: | 40 |
Flow Gradient: | 4.0 min 5% B, 4.0 min 20% B, 5.1 min 95% B, and 1.9 min 5% B |
Flow Rate: | 0.5 ml/min |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003320 |
Analysis ID: | AN003563 |
Instrument Name: | Waters Synapt G2 S QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | Samples were injected (2 μl) into a Waters Acquity Ultra Performance Liquid Chromatography (UPLC) with a BEH C18 1.7 μm, 2.1 x 50 mm column and coupled to a Xevo® G2-S quadrupole time-of-flight (QTOF) MS (Waters, Milford, MA, USA). Positive and negative electrospray ionization (ESI) data-independent modes were used for data acquisition with leucine enkephalin ([M+H]+ = 556.2771, [M-H]- = 554.2615) as Lock-Spray®. Operating conditions for ESI were: capillary voltage 2.75 kV, cone voltage 30 V, desolvation temperature 500°C, desolvation gas flow 1000 L/Hr. |
Ion Mode: | POSITIVE |