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MB Sample ID: SA027895
| Local Sample ID: | P_6 |
| 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 |
Select appropriate tab below to view additional metadata details:
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 |
|---|---|---|---|---|
| P_6 | SA027895 | FL006917 | Pool serum from Sigma | Gene type |
| P_6 | SA027895 | FL006917 | - | 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 |
