Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA208979

Local Sample ID:	51
Subject ID:	SU002261
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Age Or Age Range:	8 - 10 weeks
Gender:	Male

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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
51	SA208979	FL025493	CTL	Factor
51	SA208979	FL025493	D7	Factor
51	SA208979	FL025493	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).

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