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MB Sample ID: SA254479
| Local Sample ID: | KIND3 |
| Subject ID: | SU002622 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002622 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| KIND3 | SA254479 | FL032595 | LBPKI/KI | Genotype |
| KIND3 | SA254479 | FL032595 | ND | Treatment |
Collection:
| Collection ID: | CO002615 |
| Collection Summary: | Quick-frozen fresh tissue with liquid nitrogen |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR002634 |
| Treatment Summary: | WT and LBPKI/KI mice were fed with normal diet(ND) and high-fat diet(HFD) for 16 weeks respectively. WT mice were fasted for 24 hours or treated with NAC for 3h after a 24-hour fast. |
Sample Preparation:
| Sampleprep ID: | SP002628 |
| Sampleprep Summary: | Lipids were extracted according to MTBE method. Briefly, a 200-µL volume of water was added to 30 mg sample and vortexed for 5 s. Subsequently, 240 µL of precooling methanol was added and the mixture vortexed for 30 s. After that, 800 µL of MTBE was added and the mixture was ultrasound 20 min at 4℃ followed by sitting still for 30 min at room temperature. The solution was centrifuged at 14000g for 15min at 10℃ and the upper organic solvent layer was obtained and dried under nitrogen. |
Combined analysis:
| Analysis ID | AN004155 | AN004156 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | SHIMADZU UHPLC Nexera LC-30A | SHIMADZU UHPLC Nexera LC-30A |
| Column | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Relative intensity | Relative intensity |
Chromatography:
| Chromatography ID: | CH003075 |
| Instrument Name: | SHIMADZU UHPLC Nexera LC-30A |
| Column Name: | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0-2 minutes, B was maintained at 30%; 2–25 min, B changed linearly from 30% to 100%; 25–35 min, B maintained at 30%. |
| Flow Rate: | 300 μL/min |
| Solvent A: | 60% acetonitrile/40% water; 0.1% formic acid; 0.1Mm ammonium formate |
| Solvent B: | 10% acetonitrile/90% isopropanol; 0.1% formic acid; 0.1Mm ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003902 |
| Analysis ID: | AN004155 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | mass spectrometric analysis was performed using a Q Exactive mass spectrometer (Thermo Scientific). ESI source conditions were as follows: Heater Temp 300 °C, Sheath Gas Flow rate 45 arb, Aux Gas Flow Rate15 arb, Sweep Gas Flow Rate 1arb, Spray voltage 3.0KV, Capillary Temp 350 °C, S-Lens RF Level 50%, MS1scan ranges: 200-1800. The mass-charge ratio of lipid molecules and lipid fragments were obtained by collecting 10 fragment maps (MS 2scan, HCD) after each fullscan. The resolution of MS1 at M/Z 200 was 70,000 and that of MS2 at M/Z 200 was 17,500. LipidSearch was used for peak identification, peak extraction and lipid molecules identification (secondary identification). The main parameters were precursor tolerance: 5 ppm, Product Tolerance: 5 ppm, and Product Ion Threshold: 5%. The obtained data were subjected to quality control for subsequent data lipid difference analysis. |
| Ion Mode: | POSITIVE |
| MS ID: | MS003903 |
| Analysis ID: | AN004156 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | mass spectrometric analysis was performed using a Q Exactive mass spectrometer (Thermo Scientific). ESI source conditions were as follows: Heater Temp 300 °C, Sheath Gas Flow rate 45 arb, Aux Gas Flow Rate15 arb, Sweep Gas Flow Rate 1arb, Spray voltage 3.0KV, Capillary Temp 350 °C, S-Lens RF Level 50%, MS1scan ranges: 200-1800. The mass-charge ratio of lipid molecules and lipid fragments were obtained by collecting 10 fragment maps (MS 2scan, HCD) after each fullscan. The resolution of MS1 at M/Z 200 was 70,000 and that of MS2 at M/Z 200 was 17,500. LipidSearch was used for peak identification, peak extraction and lipid molecules identification (secondary identification). The main parameters were precursor tolerance: 5 ppm, Product Tolerance: 5 ppm, and Product Ion Threshold: 5%. The obtained data were subjected to quality control for subsequent data lipid difference analysis. |
| Ion Mode: | NEGATIVE |
