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MB Sample ID: SA011082
| Local Sample ID: | MaleRat3 |
| Subject ID: | SU000257 |
| Subject Type: | Animal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
| Genotype Strain: | Sprague-Dawley rat |
| Weight Or Weight Range: | 220-250g |
| Gender: | Male/Female |
| Animal Housing: | pathogen-free animal facility |
| Animal Light Cycle: | 12h light/dark cycle |
| Animal Feed: | free access to standard food |
| Animal Water: | free access to water |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000257 |
| Subject Type: | Animal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
| Genotype Strain: | Sprague-Dawley rat |
| Weight Or Weight Range: | 220-250g |
| Gender: | Male/Female |
| Animal Housing: | pathogen-free animal facility |
| Animal Light Cycle: | 12h light/dark cycle |
| Animal Feed: | free access to standard food |
| Animal Water: | free access to water |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| MaleRat3 | SA011082 | FL002700 | Male | Gender |
Collection:
| Collection ID: | CO000247 |
| Collection Summary: | Blood sample (3-5ml) was collected from the aorta of the rat under anesthesia and centrifuged to obtain serum |
| Collection Protocol Filename: | 1_COL_PROT.pdf |
| Sample Type: | Serum |
| Collection Method: | Blood sample (3-5ml) was collected from the aorta of the rat under anesthesia and centrifuged to obtain serum |
| Collection Location: | Lab |
| Collection Frequency: | Once |
| Collection Duration: | About 10~60 seconds |
| Volumeoramount Collected: | 3-5ml |
| Storage Conditions: | -80°C |
| Collection Vials: | 15ml Polypropylene tube |
| Storage Vials: | 1.5ml Eppendorf vial |
| Collection Tube Temp: | Room tempreture |
| Blood Serum Or Plasma: | Serum |
Treatment:
| Treatment ID: | TR000267 |
| Treatment Summary: | On the morning of sample collection day, each rat was deprived of food and put in metabolic cage for 24h urine collection. Then a blood sample (3-5ml) was collected from the aorta of the rat under anesthesia and centrifuged to obtain serum. All urine and serum samples were frozen at -80°C prior to analysis. |
| Treatment: | Intervention |
| Treatment Route: | Stay in metabolic cage for 24h urine collection |
| Animal Anesthesia: | Anesthesia using Pentobarbital Sodium |
| Animal Acclimation Duration: | ?3 days |
| Animal Fasting: | 24h |
| Animal Endp Euthanasia: | Died in anesthesia status |
| Animal Endp Tissue Coll List: | Blood |
| Animal Endp Tissue Proc Method: | Centrifuged to obtain serum |
| Animal Endp Clinical Signs: | Died |
Sample Preparation:
| Sampleprep ID: | SP000261 |
| Sampleprep Summary: | Amygdalin was dissolved in acetonitrile-water(1:1) to form 0.1mg/ml solution. Serum samples were unfrozen at room temperature. 500?l of serum sample was mixed with 20?l Amygdalin solution and 100?l 20% Trichloroacetic acid(TCA) solution. After centrifugation at about 14000rpm and -4?, the supernatant was injected into UPLC-TOF MS system. |
| Sampleprep Protocol Filename: | 2_PRE_PROT.pdf |
| Processing Method: | Precipitation |
| Processing Storage Conditions: | Precipitation at room tempreture; centrifugation at -4? |
| Extraction Method: | Precipitation by TCA |
| Extract Concentration Dilution: | 500?l of serum/620?l |
| Extract Storage: | 4? after centrifugation and during analysis |
| Sample Spiking: | 20?l Amygdalin solution/620?l |
Combined analysis:
| Analysis ID | AN000363 | AN000364 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Waters Acquity | Waters Acquity |
| Column | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Waters Synapt-HDMS | Waters Synapt-HDMS |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak Height | Peak Height |
Chromatography:
| Chromatography ID: | CH000265 |
| Chromatography Summary: | Untargeted UPLC-TOFMS |
| Methods Filename: | 3_CHR_PROT.pdf |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 24? |
| Flow Gradient: | Isocratic 2% B (01 min), linear gradient from 2% to 5% B (12 min), 5% to 12% B (25 min), 12% to 20% B (510 min), 20% to 30% B (1012 min), 30% to 50% B (1213 min), 50% to 100% B (1315 min), isocratic 100% B for 1min, then back to 2% B in 1 min and isocratic 2% B for 3min before next run |
| Flow Rate: | 0.5 ml/min |
| Injection Temperature: | 24? |
| Internal Standard: | Amygdalin |
| Sample Injection: | 8?L |
| Sampling Cone: | 40V |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Analytical Time: | 20min |
| Capillary Voltage: | Positive ion mode:3kV; Negative ion mode: 2.9kV |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000309 |
| Analysis ID: | AN000363 |
| Instrument Name: | Waters Synapt-HDMS |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3kV |
| Collision Energy: | 6V (trap)/4V (transfer) |
| Collision Gas: | Ar |
| Dry Gas Flow: | 900 L/h |
| Dry Gas Temp: | 450? |
| Ion Source Temperature: | 100? |
| Ionization: | ESI |
| Source Temperature: | 100? |
| Desolvation Gas Flow: | 900 L/h |
| Desolvation Temperature: | 450? |
| Acquisition Parameters File: | Inlet_Method_File_Name:_20100505 _Positive_MS_Mehod_File_Name:_20120221_pos_50-1500 |
| Processing Parameters File: | TR_0-18_50-1500_005_020_YES6_YES.mlm |
| MS ID: | MS000310 |
| Analysis ID: | AN000364 |
| Instrument Name: | Waters Synapt-HDMS |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Capillary Voltage: | 2.9kV |
| Collision Energy: | 6V (trap)/4V (transfer) |
| Collision Gas: | Ar |
| Dry Gas Flow: | 900 L/h |
| Dry Gas Temp: | 450? |
| Ion Source Temperature: | 100? |
| Ionization: | ESI |
| Source Temperature: | 100? |
| Desolvation Gas Flow: | 900 L/h |
| Desolvation Temperature: | 450? |
| Acquisition Parameters File: | Negative_MS_Mehod_File_Name:_20120221_neg_50-1500 |
| Processing Parameters File: | TR_0-18_50-1500_005_020_YES6_YES.mlm |
