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MB Sample ID: SA346777
| Local Sample ID: | DS2-039-015 |
| Subject ID: | SU003303 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Gender: | Not applicable |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003303 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Gender: | Not applicable |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| DS2-039-015 | SA346777 | FL040697 | Vehicle + high oleic acid diet | factor |
| DS2-039-015 | SA346777 | FL040697 | tumor | Sample source |
Collection:
| Collection ID: | CO003296 |
| Collection Summary: | Mice were randomized into groups with combinations of sunflower or coconut oil diet, and vehicle or CAY10566. B16F10 cells were implanted in the flank of C57BL/6 mice. Mice were treated with vehicle or CAY10566 at 30 mg/kg twice daily by oral gavage three days after tumor cell implantation (bilateral tumors in n=4 per group). Mice were euthanized and lungs were harvested 17 days after tumor cell injection/14 days after the start of treatment. |
| Sample Type: | Tumor cells |
Treatment:
| Treatment ID: | TR003312 |
| Treatment Summary: | All animal procedures were carried out in accordance with the IACUC-approved protocol of Cincinnati Children’s Hospital Medical Center. Animals were monitored daily by Veterinary Services. For subcutaneous tumor implantation, 1x10^6 B16F10 cells were mixed with matrigel 1:1 and implanted subcutaneously into NOD-SCID IL2Rgnull or C57BL/6 mice, respectively. CAY10566 was suspended in 0.5% Methyl Cellulose in water with 0.2% Tween 80 at 9 parts per 1 part DMSO stock. CAY10566 was administered at 30mg/kg via oral gavage, twice daily, with a drug holiday during the weekends. |
Sample Preparation:
| Sampleprep ID: | SP003310 |
| Sampleprep Summary: | For tumor analysis, tissue was weighed and placed into 1.5mL Eppendorf Safe-Lock tubes that were pre-filled with 1.0mm glass grinder beads (Biospec Products Inc). Cold methanol was then added to each tube so that all final tissue concentrations were 20 mg/mL. Samples were then homogenized via a “Next Advance” Bullet Blender for 5 minutes and subsequently vortexed for 30 minutes. Following vortex, insoluble material was pelleted by centrifugation (18000g, 10 min) and supernatants were isolated for analysis by UHPLC-MS. |
| Processing Storage Conditions: | 4℃ |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN005228 | AN005229 | AN005230 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish |
| Column | Phenomenex Kinetex C18 (30 x 2.1mm,1.7um) | Phenomenex Kinetex C18 (30 x 2.1mm,1.7um) | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| MS Type | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE | POSITIVE | NEGATIVE |
| Units | peak area | peak area | peak area |
Chromatography:
| Chromatography ID: | CH003955 |
| Chromatography Summary: | Negative C18 global lipidomics |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (30 x 2.1mm,1.7um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0-3 min 10-95% B at 0.3 mL/min, 3-4.2 min hold at 95% B at 0.3 mL/min, 4.2-4.3 min drop to 10% B at 0.45 mL/min, 4.3-4.9 min hold at 10%B while lowering flow rate to 0.4 mL/min, 4.9-5 min hold at 10%B while lowering flow rate to 0.3 mL/min. |
| Flow Rate: | 0.3-0.4 ml/min |
| Sample Injection: | 10 uL |
| Solvent A: | 75% water/25% acetonitrile; 5 mM ammonium acetate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 5 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003956 |
| Chromatography Summary: | Positive C18 global lipidomics |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (30 x 2.1mm,1.7um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0-3 min 30-100% B at 0.3 mL/min, 3-4.2 min hold at 100% B at 0.3 mL/min, 4.2-4.3 min 100-30% B at 0.4 mL/min, 4.3-4.9 min hold at 30%B and 0.4 mL/min, 4.9-5 min hold at 30%B while lowering flow rate to 0.3 mL/min |
| Flow Rate: | 0.3-0.4 ml/min |
| Sample Injection: | 10 uL |
| Solvent A: | 75% water/25% acetonitrile; 5 mM ammonium acetate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 5 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003957 |
| Chromatography Summary: | Negative C18 fatty acids |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0-1 min 25% B and 0.3 ml/min; 1-2 min 25-50% B and 0.3 mL/min; 2–8 min 50–90% B and 0.3 mL/min; 8–10 min 90–99% B and 0.3 mL/min; 10–14 min hold at 99% B and 0.3 mL/min; 14–14.1 min 99–25% B and 0.3 mL/min; 14.1–16.9 min hold at 25% B and 0.4 mL/min; 16.9–17 min hold at 25% B and resume flow of 0.3 mL/min |
| Flow Rate: | 0.3-0.4 ml/min |
| Solvent A: | 75% water/25% acetonitrile; 5 mM ammonium acetate |
| Solvent B: | 50% isopropanol/45% acetonitrile/5% water; 5 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004961 |
| Analysis ID: | AN005228 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004962 |
| Analysis ID: | AN005229 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004963 |
| Analysis ID: | AN005230 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control. |
| Ion Mode: | NEGATIVE |
