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MB Sample ID: SA299718

Local Sample ID:20210331-C8-2-24h
Subject ID:SU002901
Subject Type:Mammal
Subject Species:Rattus norvegicus
Gender:Female
Species Group:Mammals

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Subject:

Subject ID:SU002901
Subject Type:Mammal
Subject Species:Rattus norvegicus
Gender:Female
Species Group:Mammals

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
20210331-C8-2-24hSA299718FL036384ControlTreatment

Collection:

Collection ID:CO002894
Collection Summary:Whole blood was collected about 5 hours, 1 day, 2 days, 3 days, and 5 days before and after irradiation, and tissue samples were taken on the fifth day。
Sample Type:Blood (whole)

Treatment:

Treatment ID:TR002910
Treatment Summary:After anesthesia with 10% chloral hydrate solution (0.25 ml/100 g) by intraperitoneal injection, the small animal radiation therapy simulator (X-RAD SmART) was used to irradiate the whole lung tissue using a 40×40 mm lead collimator. Whole chest X-ray irradiation of 10 Gy, 20 Gy and 35 Gy in each dose group was given by anterior-anterior-field cross-irradiation, and the dose rate was about 2.7 Gy/min. The control group was also anesthetized, but not irradiated

Sample Preparation:

Sampleprep ID:SP002907
Sampleprep Summary:Plasma:Add 200 μL ice acetonitrile containing internal standard (internal standard and its concentration are shown in Table 1-3) to 50 μL plasma, swirl and mix well for 2 min, centrifuge at 13,000 rpm and 4 ℃ for 15 min, and divide the supernatant into two parts in parallel (100 μL each for determination and analysis of positive and negative ion modes of LC-MS). Freeze-dried and stored in a -80 ℃ refrigerator. Before metabolomics analysis, the samples were redissolved with 50 μL 25% acetonitrile/water (v/v) solution。 Tissue:Weigh about 20 mg lung tissue sample powder into 2 mL EP tube, add 1.5 mL 80% methanol/water (v/v) solution containing internal standard (internal standard and its concentration are shown in Table 2-1), swirl and mix well for 2 min, then add 3 zirconium beads into the sample and grind at 45 hz for 2 times, 5 min each time. The supernatant was divided into two parts in parallel (600 μL each for determination of positive and negative ion modes of LC-MS) at an interval of 15 s, and then centrifuged at 13,000 rpm for 15 min at 4 ℃. The supernatant was freeze-dried and stored in a refrigerator at -80 ℃. Before metabolomics analysis, samples were redissolved with 100 μL 80% methanol/water (v/v) solution.

Combined analysis:

Analysis ID AN004545 AN004546
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo TSQ Vantage HPLC-MS/MS Thermo TSQ Vantage HPLC-MS/MS
Column Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um)
MS Type ESI ESI
MS instrument type Triple quadrupole Triple quadrupole
MS instrument name Thermo TSQ Vantage Thermo TSQ Vantage
Ion Mode NEGATIVE POSITIVE
Units peak area peak area

Chromatography:

Chromatography ID:CH003414
Chromatography Summary:negtive ion mode
Instrument Name:Thermo TSQ Vantage HPLC-MS/MS
Column Name:Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um)
Column Temperature:55
Flow Gradient:0~1 min, 2% B; 1-20 min, 2~100% B; 20~24min, 100% B; 24~24.5min, 100~2% B; 24.5~34 min, 2% B
Flow Rate:0.35 mL/min
Solvent A:100% water; 0.1% formic acid
Solvent B:100% acetonitrile; 0.1% formic acid
Chromatography Type:Reversed phase
  
Chromatography ID:CH003415
Chromatography Summary:positive ion mode
Instrument Name:Thermo TSQ Vantage HPLC-MS/MS
Column Name:Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um)
Column Temperature:50
Flow Gradient:0~0.5min, 5% B; 0.5 ~24min, 5~100% B; 24-28 min, 40-100% B; 28~28.5min, 100~5% B; 28.5~48 min, 5% B
Flow Rate:0.35 mL/min
Solvent A:water(0.1% formic acid )
Solvent B:acetonitrile(0.1% formic acid )
Chromatography Type:Reversed phase

MS:

MS ID:MS004292
Analysis ID:AN004545
Instrument Name:Thermo TSQ Vantage
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:The temperature of ion source and ion transport tube is 350 ℃, the electrospray voltage in positive and negative ion mode is -2,500 V respectively, the sheath pressure is 35 Arb, and the auxiliary pressure is 10 Arb. Data acquisition and pre-processing and instrument interface control are performed in Xcalibur (Thermo, USA) software.
Ion Mode:NEGATIVE
  
MS ID:MS004293
Analysis ID:AN004546
Instrument Name:Thermo TSQ Vantage
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:The temperature of ion source and ion transport tube is 350 ℃, the electrospray voltage in positive and negative ion mode is 3,000 V respectively, the sheath pressure is 35 Arb, and the auxiliary pressure is 10 Arb. Data acquisition and pre-processing and instrument interface control are performed in Xcalibur (Thermo, USA) software.
Ion Mode:POSITIVE
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