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MB Sample ID: SA080914
Local Sample ID: | 0.1_NC_48h_2 |
Subject ID: | SU001235 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Not applicable |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001235 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Not applicable |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
0.1_NC_48h_2 | SA080914 | FL012365 | 48 | hours |
0.1_NC_48h_2 | SA080914 | FL012365 | - | Doxorubicin dosage(μM) |
Collection:
Collection ID: | CO001229 |
Collection Summary: | Cells were washed with cold PBS twice and then permeabilized with cold 80% methanol followed by a 20-minute incubation at -80℃. The cell lysates were centrifuges at 14000g for 5 mins and the supernatant were collected and evaporated to dryness. |
Sample Type: | Cultured cells |
Treatment:
Treatment ID: | TR001250 |
Treatment Summary: | MCF-7 cells were exposed to 10 μM doxorubicin for 4 hr followed by subsequent withdrawal of the drug. Meanwhile, MCF-7 cells continuously exposed to a low dosage of doxorubicin at 0.1 μM for 96 hr. Doxorubicin-treated cells and control cells were collected at different time. |
Sample Preparation:
Sampleprep ID: | SP001243 |
Sampleprep Summary: | MCF-7 cells were harvested and permeabilized with cold (80% (v/v) methanol: water solution followed by a 20-min incubation at -80 ℃. A stock solution of 4-Chloro-DL-phenylalanine was pre-spiked into the methanol aqueous buffer to a final concentration of 1.5 μg/mL, which subsequently served as an internal standard (IS). The cell lysates were centrifuged at 14,000g for 5 min at 4 ℃, and the resultant supernatant were collected and evaporated to dryness. The samples were reconstituted in 100 μL of water (LC-MS grade) before analysis on an HPLC−MS/MS, and 20 μL was used for analysis. For each group of samples subjected to metabolomics analysis, five biological replicates were prepared. |
Combined analysis:
Analysis ID | AN001935 |
---|---|
Analysis type | MS |
Chromatography type | HILIC |
Chromatography system | Shimadzu LC-30A |
Column | Waters XBridge Amide (100 x 4.6mm,3.5um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | ABI Sciex 5600 TripleTOF |
Ion Mode | NEGATIVE |
Units | normalized peak area |
Chromatography:
Chromatography ID: | CH001404 |
Chromatography Summary: | The mobile phase consisted of solvent A (5 mM ammonium acetate, pH=9, and 5% acetonitrile) and solvent B (acetonitrile). The gradient was set as follows: 0–3 min, 85% B; 3–6 min, 85–30% B; 6–15 min, 30–2% B; 15–18 min, 2% B; 18–19 min, 2–85% B; 19–26 min, 85% B. The flow rate was set at 0.4 mL/min. |
Instrument Name: | Shimadzu LC-30A |
Column Name: | Waters XBridge Amide (100 x 4.6mm,3.5um) |
Flow Gradient: | 0-3 min, 85% B; 3-6 min, 85-30% B; 6-15 min, 30-2% B; 15-18 min, 2% B; 18-19 min, 2-85% B; 19-26 min, 85% B. |
Flow Rate: | 0.4 mL/min |
Solvent A: | 5% acetonitrile/95% water; 5 mM ammonium acetate, pH 9 |
Solvent B: | 100% acetonitrile |
Chromatography Type: | HILIC |
MS:
MS ID: | MS001791 |
Analysis ID: | AN001935 |
Instrument Name: | ABI Sciex 5600 TripleTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The MS data was acquired in the negative ion mode using a data-dependent acquisition approach. The electrospray ionization (ESI) source conditions were used as follows: TOF MS scan, m/z 50 – 1000 Da; Product ion scan, m/z 50 – 900 Da; Ion Source Gas 1 (Gas 1), 50 psi; Ion Source Gas 2 (Gas 2), 30 psi; Curtain gas, 30 psi; Source temperature, 500 ℃; Ion Spray Voltage Floating, -4500 V; Declustering potential (DP), -100V; Collision energy (CE), -35 V; CE spread, 10 V. MS data acquisition was controlled by Analyst TF 1.6.1 (AB SCIEX, Framingham, MA, USA). The accurate mass was calibrated by Calibration Delivery System. |
Ion Mode: | NEGATIVE |