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MB Sample ID: SA201231
| Local Sample ID: | 20210522-CH-3 |
| Subject ID: | SU002182 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002182 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 20210522-CH-3 | SA201231 | FL024477 | 3 times per week with 4 consecutive respectively intraperitoneal injection with 10% CCl4 (10 mL/kg of olive oil) | Treatment |
Collection:
| Collection ID: | CO002175 |
| Collection Summary: | Animals were killed at 24h post the last CCl4 treatment and the liver tissue samples were harvested for further assays. |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR002194 |
| Treatment Summary: | Prior to experimental, animals were acclimatized breeding to laboratory conditions for 1 week. The mice were randomly allocated into 6 groups (n=8 each), as follows: a control group (C); two acute hepatitis groups, low and high concentrations (AL/ AH); three chronic hepatitis groups, low, middle and high concentration (CL/ CM/ CH). Mice in chronic groups were treated 3 times per week with 4 consecutive respectively intraperitoneal (i.p) injection with 1% / 5% / 10% CCl4 (10 mL/kg, dissolved in olive oil). However, in the acute groups, mice were only intraperitoneally (i.p) injected with 1% / 5% CCl4 (10 mL/kg, dissolved in olive oil) at the 12th injection. During the previous 11 times they were treated the same as the healthy controls with an equal volume of olive oil. Animals were killed at 24h post the last CCl4 treatment and serum, stool and liver tissue samples were harvested for further assays. |
Sample Preparation:
| Sampleprep ID: | SP002188 |
| Sampleprep Summary: | 100-130 mg liver tissue samples were weighted, 1.2 mL of 80% ice-cold aqueous methanol, which contains 0.001 mg/mL internal standard, and approximately 1 g of glass beads (1.0 mm i.d.) was added into a 2 mL screw-cap plastic microvial. The Mini-BeadBeater-16 (Biospec Products, USA) was used for tissue disruption. Samples were homogenized 3 times for 2 min each, with cooling procedure in between, centrifuged at 20,000 g for 10 min at 4 °C, and the supernatants were mixed with 800 μL of ice-cold acetonitrile and centrifuged again for protein precipitation. The supernatant was filtered through a 0.22 µm organic phase membrane, blown dry in nitrogen, reconstituted in 100 µL of water and transferred to vials for metabolomics assay. |
Combined analysis:
| Analysis ID | AN003425 | AN003426 | AN003427 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | Reversed phase |
| Chromatography system | UltiMate 3000 (Thermo Fisher Scientific) | Agilent 1290 Infinity | Agilent 1290 Infinity |
| Column | Waters X-Bridge C18 (150 x 1.0mm,3.5um) | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Triple quadrupole | Triple quadrupole |
| MS instrument name | Thermo Q Exactive Plus Orbitrap | Agilent 6495 QQQ | Agilent 6495 QQQ |
| Ion Mode | NEGATIVE | POSITIVE | NEGATIVE |
| Units | pmol/10,000,000 cells | counts | counts |
Chromatography:
| Chromatography ID: | CH002534 |
| Chromatography Summary: | In this study, a newly developed precision-targeted metabolomics method with a UPLC-TQ/MS system (Agilent 1290 Infnity, Agilent Technologies, USA; Agilent 6495 QQQ, Agilent Technologies, USA) in a DMRM scan-mode was applied to analyze the metabolome of interest from trial samples (serum, liver tissues and stool). Briefly, the method was performed with an ACQUITY UPLC HSS T3 column (2.1 mm i.d. × 100 mm, 1.8 μm; Waters); mobile phase A and B were water and acetonitrile with 0.1% formic acid (v/v) respectively. The flow rate was at 0.3 mL/min and the column temperature was maintained at 40 ℃. The samples were placed in an auto-sampler maintained at 4 °C with a 5 μL injection volume. The optimized gradient-elution program, as follows: 0-2 min, 98% A; 2-10 min, 98-65% A; 10-12 min, 65-20% A; 12-14 min, 20-2% A; 14-30 min, 2% A. |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | 0-2 min, 98% A; 2-10 min, 98-65% A; 10-12 min, 65-20% A; 12-14 min, 20-2% A; 14-30 min, 2% A |
| Flow Rate: | 0.3 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: | MS003189 |
| Analysis ID: | AN003426 |
| Instrument Name: | Agilent 6495 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Agilent MassHunter Workstation Data Acquisition Agilent MassHunter |
| Ion Mode: | POSITIVE |
| MS ID: | MS003190 |
| Analysis ID: | AN003427 |
| Instrument Name: | Agilent 6495 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Agilent MassHunter Workstation Data Acquisition Agilent MassHunter |
| Ion Mode: | NEGATIVE |
