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MB Sample ID: SA267018
Local Sample ID: | ko5-1p |
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, |
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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 |
---|---|---|---|---|
ko5-1p | SA267018 | FL034955 | ATP10A-knockout | 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 |