Return to study ST001715 main page
MB Sample ID: SA160850
Local Sample ID: | Naphthalene24_c18_01_hfqe.mzXML |
Subject ID: | SU001792 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Genotype Strain: | Pooled Liver S9 fractions |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001792 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Genotype Strain: | Pooled Liver S9 fractions |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Naphthalene24_c18_01_hfqe.mzXML | SA160850 | FL018917 | Naphthalene24 | Precursor |
Naphthalene24_c18_01_hfqe.mzXML | SA160850 | FL018917 | 24 | Time |
Collection:
Collection ID: | CO001785 |
Collection Summary: | At the specified time point, S9 Reactions were extracted with 2x volume acetonitrile, and spun down at 14,000 rpm for 10 minutes and supernatants were transferred to autosampler vials. Samples were kept at -20°C until instrumental analysis. |
Sample Type: | Liver |
Storage Conditions: | Described in summary |
Treatment:
Treatment ID: | TR001805 |
Treatment Summary: | Xenobiotic precursors were incubated with pooled human liver S9 fractions with an NADPH regenerating system and other required cofactors and collected at specified time points as described in the associated manuscript. |
Sample Preparation:
Sampleprep ID: | SP001798 |
Sampleprep Summary: | Each test compound stock solution was diluted to 0.3 mM with water prior to incubation with S9 enzymes. The NADPH regenerating system was reconstituted with addition of 3.5 ml of water to make a nal volume of 5 ml. Cofactors were combined to form a 4X cofactor stock as follows, prior to addition into the reaction mixture: 10 mM UDPGA, 2 mM GSH, 2 mg/ml PAPS, 0.1 mM acetyl-CoA, and NADPH regenerating system (1 mM NADP, 5 mM glucose-6-phosphate, 1 unit glucose-6-phosphate dehydrogenase). Reactions were carried out at 30 °C on 96-well plates (Fig. 1). S9 fraction was diluted 10-fold in water immediately before mixing with 0.2 M Tris-Cl, pH 7.5/2 mM MgCl2 and 0.3 mM of the xenobiotic solution in a 1:1:1 ratio (15 µL each). and incubated at 30 °C for 5 min. To start the reaction, 15 µL of 4X cofactor stock was added, and incubation was carried out at 30 °C for the indicated times. To terminate the reaction, we added a three-fold volume of acetonitrile, covered the plate with paralm, vortexed, and froze at − 20 °C to precipitate insoluble materials such as protein. After thawing and centrifugation of the incubation plate, the supernatants were transferred into polypropylene autosampler vials, which were stored at − 20 °C until instrumental analysis |
Combined analysis:
Analysis ID | AN002792 | AN002793 | AN002794 | AN002795 |
---|---|---|---|---|
Analysis type | MS | MS | MS | MS |
Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
Column | Waters XBridge BEH Amide (50mm x 2.1mm,2.5um) | Waters XBridge BEH Amide (50mm x 2.1mm,2.5um) | Higgins Analytical Targa C18 (50 x 2.1mm,3um) | Higgins Analytical Targa C18 (50 x 2.1mm,3um) |
MS Type | ESI | ESI | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Fusion Tribrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap | Thermo Fusion Tribrid Orbitrap |
Ion Mode | POSITIVE | POSITIVE | NEGATIVE | NEGATIVE |
Units | relative intensity | relative intensity | relative intensity | relative intensity |
Chromatography:
Chromatography ID: | CH002065 |
Chromatography Summary: | HILIC/ESI+ |
Chromatography Comments: | HFQE |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Waters XBridge BEH Amide (50mm x 2.1mm,2.5um) |
Chromatography Type: | HILIC |
Chromatography ID: | CH002066 |
Chromatography Summary: | C18/ESI- |
Chromatography Comments: | HFQE |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Higgins Analytical Targa C18 (50 x 2.1mm,3um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002588 |
Analysis ID: | AN002792 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | PRM MSMS, ddMSMS and MS1 data collection, mzmine2 |
Ion Mode: | POSITIVE |
MS ID: | MS002589 |
Analysis ID: | AN002793 |
Instrument Name: | Thermo Fusion Tribrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | ddMSMS and MS1 data collection. mzmine2 |
Ion Mode: | POSITIVE |
MS ID: | MS002590 |
Analysis ID: | AN002794 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | PRM MSMS, ddMSMS and MS1 data collection. mzmine2 |
Ion Mode: | NEGATIVE |
MS ID: | MS002591 |
Analysis ID: | AN002795 |
Instrument Name: | Thermo Fusion Tribrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | ddMSMS and MS1 data collection. mzmine2 |
Ion Mode: | NEGATIVE |