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MB Sample ID: SA257702
| Local Sample ID: | MCF001027_wt3_ep40 |
| Subject ID: | SU002662 |
| Subject Type: | Fish |
| Subject Species: | Danio rerio |
| Taxonomy ID: | 7955 |
| Genotype Strain: | WT/PMM2 mutant sa10150 |
| Age Or Age Range: | 6 dpf |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002662 |
| Subject Type: | Fish |
| Subject Species: | Danio rerio |
| Taxonomy ID: | 7955 |
| Genotype Strain: | WT/PMM2 mutant sa10150 |
| Age Or Age Range: | 6 dpf |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| MCF001027_wt3_ep40 | SA257702 | FL033089 | 40µM Epalrestat | Sample treatment |
Collection:
| Collection ID: | CO002655 |
| Collection Summary: | 10 embryos per sample were homogenized by sonication in extraction buffer (80% MeOH, IS). The metabolites were then precipitated at -80 °C overnight, after which the samples were centrifuged at 15,000 rpm, 20 min, 4 °C. Next, the supernatant was transferred to a fresh Eppendorf tube. The samples were centrifuged again at 15,000 rpm, 20 min, 4 °C to remove any leftover impurities and transferred to a fresh MS vial. Finally, the samples were analyzed by LC/MS |
| Sample Type: | Larvae |
| Storage Conditions: | -80? |
Treatment:
| Treatment ID: | TR002674 |
| Treatment Summary: | Zebrafish were treated with vehicle or 10 or 40 microM epalrestat |
Sample Preparation:
| Sampleprep ID: | SP002668 |
| Sampleprep Summary: | 10 embryos per sample were homogenized by sonication in extraction buffer (80% MeOH, IS). The metabolites were then precipitated at -80 °C overnight, after which the samples were centrifuged at 15,000 rpm, 20 min, 4 °C. Next, the supernatant was transferred to a fresh Eppendorf tube. The samples were centrifuged again at 15,000 rpm, 20 min, 4 °C to remove any leftover impurities and transferred to a fresh MS vial. Finally, the samples were analyzed by LC/MS |
| Processing Storage Conditions: | -80? |
| Extraction Method: | 80 % MeOH with Internal standards (e.g. d27 myristic acid) |
Combined analysis:
| Analysis ID | AN004221 | AN004222 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | HILIC |
| Chromatography system | Waters Acquity | Agilent Infinity |
| Column | C-18: Acquity UPLC -HSS T3 1. 8 µm; 2.1 x 150 mm, Waters | Poroshell 120 HILIC-Z PEEK Column 2.1x 150mm, 5 um |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Focus |
| Ion Mode | NEGATIVE | NEGATIVE |
| Units | AUC | AUC |
Chromatography:
| Chromatography ID: | CH003130 |
| Chromatography Summary: | Reverse phase, IP, C18 column |
| Instrument Name: | Waters Acquity |
| Column Name: | C-18: Acquity UPLC -HSS T3 1. 8 µm; 2.1 x 150 mm, Waters |
| Column Temperature: | 40 |
| Flow Gradient: | The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B until 2 min post injection. A linear gradient to 37% B was carried out until 7 min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the gradient returned to 5% B. The chromatography was stopped at 40 min. |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 100% water; 10mM tributylamine; 15mM acetic acid |
| Solvent B: | 100% methanol |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003131 |
| Chromatography Summary: | HILIC |
| Instrument Name: | Agilent Infinity |
| Column Name: | Poroshell 120 HILIC-Z PEEK Column 2.1x 150mm, 5 um |
| Column Temperature: | 25 |
| Flow Gradient: | A linear gradient was carried out starting with 90% solvent A and 10% solvent B. From 2 to 12 min the gradient changed to 60% B. The gradient was kept on 60% B for 3 minutes and followed by a decrease to 10% B. The chromatography was stopped at 25 min |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 100% acetonitrile |
| Solvent B: | 100% water; 10 mM sodium acetate, pH 9.3 |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003968 |
| Analysis ID: | AN004221 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | El-Maven polly, ThemoFisher Xcalibur, Metabolites were annotated using in-house metabolite library- elution time and m/z were considered |
| Ion Mode: | NEGATIVE |
| MS ID: | MS003969 |
| Analysis ID: | AN004222 |
| Instrument Name: | Thermo Q Exactive Focus |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | El-Maven polly, ThemoFisher Xcalibur Metabolites were annotated using in-house metabolite library- elution time and m/z were considered |
| Ion Mode: | NEGATIVE |
