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MB Sample ID: SA027813
Local Sample ID: | S_3 |
Subject ID: | SU000552 |
Subject Type: | Serum |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BLKsJ-db/db (disease model) and C57BLKsJ wild type (wild type) |
Age Or Age Range: | 14 weeks; drug administration since the 5th week |
Weight Or Weight Range: | 37-58 g for db/db mice; 18-25 g for wild type mice |
Gender: | Male |
Species Group: | Mammal |
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Subject:
Subject ID: | SU000552 |
Subject Type: | Serum |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BLKsJ-db/db (disease model) and C57BLKsJ wild type (wild type) |
Age Or Age Range: | 14 weeks; drug administration since the 5th week |
Weight Or Weight Range: | 37-58 g for db/db mice; 18-25 g for wild type mice |
Gender: | Male |
Species Group: | Mammal |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S_3 | SA027813 | FL006907 | C57BLKsJ-db/db | Gene type |
S_3 | SA027813 | FL006907 | Group 2, DG extract low dose | Phenotypic grouping and treaments |
Collection:
Collection ID: | CO000546 |
Collection Summary: | Mice were fasting over night and then sacrified to collect blood. |
Sample Type: | Serum |
Collection Location: | Wuhan Institute of Drug and Device Control, China |
Storage Conditions: | Frozen -80oC |
Collection Vials: | Eppendorf tube 1 mL |
Storage Vials: | Eppendorf tube 0.5 mL |
Collection Tube Temp: | Room temperature |
Blood Serum Or Plasma: | Serum |
Treatment:
Treatment ID: | TR000566 |
Treatment Summary: | C57BLKsJ-db/db mice were classified randomly into 8 groups. Seven groups were with 9~10 mice for each and one group with 4 mice. Mice were treated with DG extract (5 g/Kg), DG extract (1 g/Kg), KK01 (50 mg/Kg), KK01 (20 mg/Kg), metformin (5 mg/Kg); rotaglitazone (5 mg/Kg); and saline. Group treated with KK01(20 mg/Kg) and the group with 4 mice treated by saline were used intraperitoneal injection. Other groups were in intragastric administration. Also, a group with ten C57BLKsJ mice fedding with saline was used as healthy control. The drug administration were started with the 5th week of the age and lasted for 9 weeks. |
Treatment Route: | Intragastric administration for all except for group of KK01 low-dose (20 mg/Kg), and 4 db/db mice treated with saline which were treated by intraperitoneal injection. |
Treatment Dose: | 5 g/Kg for DG high dose; 1 g/Kg for DG low dose; 50 mg/Kg for KK01 high dose; 20 mg/Kg (i.p.) for KK01 low dose; 5 mg/Kg for metformin, 5 mg /Kg for rosiglitazone |
Treatment Dosevolume: | 0.5 mL |
Treatment Doseduration: | 9 weeks |
Treatment Vehicle: | saline |
Animal Fasting: | Over night |
Animal Endp Tissue Coll List: | liver, kidney, brain, white fat, brown fat, pancreas |
Sample Preparation:
Sampleprep ID: | SP000559 |
Sampleprep Summary: | In lipidomics analysis, samples were first thawed on ice, and then mixed by vortex for 4 min at 4,000 rpm, followed by centrifugation for 4 min at 4, 000 rcf to spin down particulate. Pooled mouse serum from Sigma Aldrich (S7273) were used as QC samples. A 30 µL aliquot of each serum sample (including QC sample and real sample) was transferred to a 2.0 mL Eppendorf Lo-Bind tube and extracted by 600 µL of 2:1 dichloromethane: methanol (DCM) via vortex-mixing for 2 min at 4,000 rpm. Then, 120 µL of H2O was added and repeated the vortex-mixing for 1 min at 4,000 rpm. After placing the sample at room temperature for 10 min, the tube was centrifuged at 16,000 rcf for 10 minutes at 10 °C. A 370 µL aliquot of the lower lipid-rich DCM layer was transferred to a new labeled 2.0 mL Eppendorf Lo-Bind tubes and then lyophilized to complete dryness overnight. For immediate analysis, 300 µL of acetonitrile:isopropanol:H2O (65:30:5, v/v/v) was added to reconstitute the dried tissue extract, and the samples were thoroughly mixed on a multi-tube vortexer for 10 min at 5000 rpm and centrifuged at 16,000 rcf for 4 min. The supernatants were transferred to labeled autosampler vials for data acquisition by LC-MS. |
Extraction Method: | cold 2:1 dichloromethane: methanol vortex extraction |
Extract Storage: | -80 oC |
Sample Resuspension: | acetonitrile:isopropanol:H2O (65:30:5, v/v/v) |
Sample Spiking: | 1,2-diheptadecanoyl-sn-glycero-3-phosphocholine |
Combined analysis:
Analysis ID | AN000808 | AN000809 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Waters Acquity | Waters Acquity |
Column | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Orbitrap | Thermo Orbitrap |
Ion Mode | POSITIVE | NEGATIVE |
Units | m/z | m/z |
Chromatography:
Chromatography ID: | CH001295 |
Instrument Name: | Waters Acquity |
Column Name: | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
Column Pressure: | 6000-10000 |
Column Temperature: | 50C |
Flow Rate: | 0.25 mL/min |
Injection Temperature: | 4C |
Internal Standard: | 1,2-diheptadecanoyl-sn-glycero-3-phosphocholine |
Solvent A: | 60% acetonitrile/40% water; 0.1% formic acid; 10 mM ammonium formate |
Solvent B: | 90% isopropanol/10% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
Analytical Time: | 30 min |
Weak Wash Solvent Name: | 60:40 water/acetonitrile in 10 mM ammonium formate with 0.1% formic acid |
Weak Wash Volume: | 1200 uL |
Strong Wash Solvent Name: | 90:10 isopropanol/acetonitrile in 10 mM ammonium formate with 0.1% formic acid |
Strong Wash Volume: | 600 uL |
Sample Loop Size: | 10 uL |
Sample Syringe Size: | 100 uL |
Randomization Order: | Yes |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS000715 |
Analysis ID: | AN000808 |
Instrument Name: | Thermo Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
Ion Mode: | POSITIVE |
Capillary Temperature: | 250 oC |
Collision Energy: | 35 |
Fragmentation Method: | CID |
Ion Source Temperature: | 400 oC |
Ion Spray Voltage: | 4.0 kV |
Ionization: | ES+ |
Mass Accuracy: | 10 ppm |
Desolvation Gas Flow: | 50 arbitary unit |
Desolvation Temperature: | 400 oC |
Resolution Setting: | 30000 |
Scan Range Moverz: | 120-2000 m/s |
MS ID: | MS000716 |
Analysis ID: | AN000809 |
Instrument Name: | Thermo Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
Capillary Temperature: | 250 oC |
Collision Energy: | 35 |
Fragmentation Method: | CID |
Ion Source Temperature: | 400 oC |
Ion Spray Voltage: | 4.0 kV |
Ionization: | ES- |
Mass Accuracy: | 10 ppm |
Desolvation Gas Flow: | 50 arbitary unit |
Desolvation Temperature: | 400 oC |
Resolution Setting: | 30000 |
Scan Range Moverz: | 120-2000 m/s |