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MB Sample ID: SA267038
| Local Sample ID: | wt2-1n |
| Subject ID: | SU002798 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | Wild-type and ATP10A knockouts |
| Age Or Age Range: | 16-20 weeks |
| Weight Or Weight Range: | 23-28 grams |
| Animal Housing: | temperature and humidity-controlled facilities |
| Animal Light Cycle: | 12 h light/dark cycles |
| Animal Feed: | standard chow or 60% HFD (D12492, Research Diets) ad libitum, |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002798 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | Wild-type and ATP10A knockouts |
| Age Or Age Range: | 16-20 weeks |
| Weight Or Weight Range: | 23-28 grams |
| Animal Housing: | temperature and humidity-controlled facilities |
| Animal Light Cycle: | 12 h light/dark cycles |
| Animal Feed: | standard chow or 60% HFD (D12492, Research Diets) ad libitum, |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| wt2-1n | SA267038 | FL034956 | Wild-type | Genotype |
Collection:
| Collection ID: | CO002791 |
| Collection Summary: | Plasma was collected, via a retroorbital bleed or cardiac puncture, from 5-hr fasted mice and 5-hr fasted mice that had undergone an OGTT. |
| Sample Type: | Blood (plasma) |
| Collection Method: | retroorbital bleed or cardiac puncture |
| Storage Conditions: | -20℃ |
Treatment:
| Treatment ID: | TR002807 |
| Treatment Summary: | Mice were fasted for 5 hours (7AM-12PM). No additional treatments were administered. |
| Treatment: | WT vs KO |
| Treatment Compound: | n/a |
| Treatment Route: | n/a |
| Treatment Dose: | n/a |
| Treatment Dosevolume: | n/a |
| Treatment Doseduration: | n/a |
| Treatment Vehicle: | n/a |
| Animal Vet Treatments: | n/a |
| Animal Anesthesia: | n/a |
| Animal Acclimation Duration: | n/a |
| Animal Fasting: | 5 hour |
| Animal Endp Euthanasia: | n/a |
| Animal Endp Tissue Coll List: | Plasma was collected, via a retroorbital bleed or cardiac puncture |
| Animal Endp Clinical Signs: | n/a |
Sample Preparation:
| Sampleprep ID: | SP002804 |
| Sampleprep Summary: | 100 uL of plasma was collected from each mouse. An isotopically labeled lipid mixture (SPLASH LIPIDOMIX, Avanti) was added as an internal standard to each plasma sample. A liquid-liquid extraction was performed by adding 1 mL methyl tert-butyl ether (MTBE), vortexing, and subsequently centrifuging for 10 minutes at 10,000 rpm and 4˚C. The nonpolar, MTBE fraction (top liquid layer) containing the lipophilic components was removed and dried under vacuum centrifugation. Dried samples were stored at -80˚C until the day of MS analysis. For LC-IM-MS analysis, dried samples were resuspended in 100 µL IPA containing 40 µg/mL heptadecanoic acid and nonadecanoic acid, as well as 10 µg/mL glucosyl(β) sphingosine and N-heptadecanoyl-D-erythrosphingosine. |
| Processing Storage Conditions: | 4℃ |
| Extraction Method: | MTBE |
| Extract Enrichment: | vacuum centrifuge |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 100 uL IPA |
| Sample Derivatization: | n/a |
| Sample Spiking: | 14 heavy-labeled lipids prior to extraction; 4 odd-chain lipids during reconstitution |
| Subcellular Location: | n/a |
Combined analysis:
| Analysis ID | AN004369 | AN004370 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Agilent 6560 | Agilent 6560 |
| Column | Agilent ZORBAX RRHD Extend-C18 (50 x 2.1mm,1.8um) | Agilent ZORBAX RRHD Extend-C18 (50 x 2.1mm,1.8um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Agilent 6560 Ion Mobility | Agilent 6560 Ion Mobility |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Ion Abundances | Ion Abundances |
Chromatography:
| Chromatography ID: | CH003275 |
| Chromatography Summary: | HPLC-IM-MS/MS on an Agilent 6560 mass spectrometer using a ZORBAX Extend-C18 RPLC column (Phase A: 0.1% formic acid and 10 mM NH4CHOO in water, Phase B: 0.1% formic acid and 10 mM NH4CHOO in 60:36:4 isopropanol:acetonitrile:water). |
| Instrument Name: | Agilent 6560 |
| Column Name: | Agilent ZORBAX RRHD Extend-C18 (50 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | 70% B for 1 min, 70-86% B in 2.5 min, 86% B for 6.5 min, 86-100% B in 1 min, 100% B for 6 min, 100-70% B in 0.1 min, and 70% B for 1.9 min. |
| Flow Rate: | 300 ul/min |
| Solvent A: | Water (10mM Amm. Formate) 0.1% formic acid |
| Solvent B: | 60:36:4 IPA:ACN:H2O (10mM Amm. Formate) 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004116 |
| Analysis ID: | AN004369 |
| Instrument Name: | Agilent 6560 Ion Mobility |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Data alignment and biostatical analysis was performed using Progenesis QI (Waters). |
| Ion Mode: | POSITIVE |
| MS ID: | MS004117 |
| Analysis ID: | AN004370 |
| Instrument Name: | Agilent 6560 Ion Mobility |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Data alignment and biostatical analysis was performed using Progenesis QI (Waters). |
| Ion Mode: | NEGATIVE |
