Return to study ST001094 main page
MB Sample ID: SA074538
Local Sample ID: | sample.8 |
Subject ID: | SU001138 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001138 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
sample.8 | SA074538 | FL011447 | 2PAHs | Treatment |
sample.8 | SA074538 | FL011447 | 30 min | Time |
Collection:
Collection ID: | CO001132 |
Collection Summary: | For the global metabolomics with a lipid focus, immediately following treatment, cell monolayers were washed 3 times with cold PBS, and cells were scraped and fixed in 500 μL 70% MeOH. Samples were stored immediately at -80°C until pretreatment for solid phase extraction (SPE). |
Collection Protocol Filename: | AKB_Untargeted-protocol.docx |
Collection Protocol Comments: | For AKB study 1 protocol, the collection, treatment, and sample prep are all in the same file. |
Sample Type: | Epithelial cells |
Treatment:
Treatment ID: | TR001152 |
Treatment Summary: | For the global metabolomics with a focus on the lipids: Cells (passage <20) were maintained in CMRL 1066 media containing 10% FBS and 1% glutamate in a humidified atmosphere at 37˚ C, 5% CO2, and 95% air (Osgood et al. 2013). Once cells were grown to confluence, cells were serum deprived for 24 hr prior to treatment with the binary 1:1 LMW PAH mixture of 1-MeA and Flthn at a final dose of 40 μM. This mixture has been used previously in our lab (PMC5808746) to represent two LMW PAH’s common in secondhand smoke and environmental exposures. |
Treatment Protocol Filename: | AKB_Untargeted-protocol.docx |
Treatment Protocol Comments: | For AKB study 1 protocol, the collection, treatment, and sample prep are all in the same file. |
Sample Preparation:
Sampleprep ID: | SP001145 |
Sampleprep Summary: | The hydrophobic fraction was extracted from the samples using an organic liquid-liquid extraction technique with methyl tert-butyl ether (MTBE) and water (Cruickshank-Quinn et al. 2014; Yang et al. 2013). The upper hydrophobic layer was transferred to a new tube. The hydrophobic layer was then dried under nitrogen, reconstituted in 200 µL of methanol, and stored at -80 °C until LCMS analysis. |
Sampleprep Protocol Filename: | AKB_Targeted-protocol.docx |
Sampleprep Protocol Comments: | For AKB study 1 protocol, the collection, treatment, and sample prep are all in the same file. |
Combined analysis:
Analysis ID | AN001780 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 |
Column | Agilent Zorbax Rapid Resolution HD SB-C18 |
MS Type | ESI |
MS instrument type | TOF |
MS instrument name | Agilent 6210 TOF |
Ion Mode | POSITIVE |
Units | abundance/counts |
Chromatography:
Chromatography ID: | CH001258 |
Instrument Name: | Agilent 1290 |
Column Name: | Agilent Zorbax Rapid Resolution HD SB-C18 |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001644 |
Analysis ID: | AN001780 |
Instrument Name: | Agilent 6210 TOF |
Instrument Type: | TOF |
MS Type: | ESI |
Ion Mode: | POSITIVE |