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MB Sample ID: SA151947
| Local Sample ID: | F6 |
| Subject ID: | SU001735 |
| Subject Type: | Yeast |
| Subject Species: | Saccharomyces cerevisiae |
| Taxonomy ID: | 4932 |
| Cell Strain Details: | BY4741 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001735 |
| Subject Type: | Yeast |
| Subject Species: | Saccharomyces cerevisiae |
| Taxonomy ID: | 4932 |
| Cell Strain Details: | BY4741 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| F6 | SA151947 | FL017875 | HPLC fraction 6 | Treatment |
Collection:
| Collection ID: | CO001728 |
| Collection Summary: | Yeast cultures were grown in SC-sulfate media and were harvested at OD600 0.4-0.8 by vacuum filtration then washed with 100mM ammonium acetate - TEA (pH 8.0). Metabolites were extracted by the addition of pre-chilled 10mL 90% methanol, 10mM ammonium acetate - TEA (pH 8.0). Cell debris was removed by centrifugation at 4000g for 10-15 minutes. Extracts were subsequently passed through 3 kDa MWCO filters. Filtrate was diluted 2-fold with water and lyophilized. Lyophilized metabolite extracts were resuspended with water and pH was adjusted to 10 with TEA. The extract was then passed through a mixed-mode anion exchange solid phase extraction cartridge and fractionated by reverse-phase HPLC on a C18 column. One column volume of water plus 0.1% formic acid and eight column volumes of methanol were passed through the column. Fractions that eluted during the aqueous phase were then lyophilized. |
| Sample Type: | Yeast cells |
Treatment:
| Treatment ID: | TR001748 |
| Treatment Summary: | Active fractions were run on a normal-phase HPLC on an amide column and fractions were collected. Fractions 4, 5, and 6 were subjected to metabolomics analyses. |
Sample Preparation:
| Sampleprep ID: | SP001741 |
| Sampleprep Summary: | Fractions of interest were lyophilized then reconstituted in water plus 0.1% formic acid such that samples reached a final concentration of 20 mg/mL. |
Combined analysis:
| Analysis ID | AN002706 | AN002707 | AN002708 | AN002709 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | Normal phase | Normal phase |
| Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
| Column | Thermo Hypersil Gold aQ (150 x 2.1mm,1.9um) | Thermo Hypersil Gold aQ (150 x 2.1mm,1.9um) | PolyLC Polyhydroxyethyl A (200 x 2.1mm,5um) | PolyLC Polyhydroxyethyl A (200 x 2.1mm,5um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
| Units | Peak Area | Peak Area | Peak Area | Peak Area |
Chromatography:
| Chromatography ID: | CH001997 |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Thermo Hypersil Gold aQ (150 x 2.1mm,1.9um) |
| Column Temperature: | 30 |
| Flow Gradient: | 0-2min 0% B, 2-13min 0-60% B, 13-15min 60-90% B, 15-16min 90-0% B, 16-20min 0% B |
| Flow Rate: | 0.2 mL/min |
| Solvent A: | 100% water; 0.1% formic acid; 1mM ammonium acetate |
| Solvent B: | 20% water/80% acetonitrile; 0.1% formic acid; 1 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH001998 |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | PolyLC Polyhydroxyethyl A (200 x 2.1mm,5um) |
| Column Temperature: | 30 |
| Flow Gradient: | 0-2min 80% B, 2-13min 80-40% B, 13-15min 40-10% B, 15-16min 10-80% B, 16-20min 80% B |
| Flow Rate: | 0.2 mL/min |
| Solvent A: | 100% water; 0.1% formic acid; 1mM ammonium acetate |
| Solvent B: | 20% water/80% acetonitrile; 0.1% formic acid; 1 mM ammonium acetate |
| Chromatography Type: | Normal phase |
MS:
| MS ID: | MS002503 |
| Analysis ID: | AN002706 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS1 data was acquired at a resolution of 140,000 over a scan range of 65-850 m/z. Data was analyzed with Thermo Compound Discoverer software. |
| Ion Mode: | POSITIVE |
| MS ID: | MS002504 |
| Analysis ID: | AN002707 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS1 data was acquired at a resolution of 140,000 over a scan range of 65-850 m/z. Data was analyzed with Thermo Compound Discoverer software. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS002505 |
| Analysis ID: | AN002708 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS1 data was acquired at a resolution of 140,000 over a scan range of 65-850 m/z. Data was analyzed with Thermo Compound Discoverer software. |
| Ion Mode: | POSITIVE |
| MS ID: | MS002506 |
| Analysis ID: | AN002709 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS1 data was acquired at a resolution of 140,000 over a scan range of 65-850 m/z. Data was analyzed with Thermo Compound Discoverer software. |
| Ion Mode: | NEGATIVE |
