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MB Sample ID: SA151188
| Local Sample ID: | 2P_002 |
| Subject ID: | SU001717 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001717 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 2P_002 | SA151188 | FL017690 | Control | Treatment |
Collection:
| Collection ID: | CO001710 |
| Collection Summary: | Samples of both plasma and liver were collected from healthy patients and patients with chronic liver diseases. |
| Sample Type: | Blood (plasma) |
Treatment:
| Treatment ID: | TR001730 |
| Treatment Summary: | Healthy patients (control) versus CLD and HCC for plasma. |
Sample Preparation:
| Sampleprep ID: | SP001723 |
| Sampleprep Summary: | Extraction of plasma lipids is based on the “Maytash'' method [1] which was subsequently modified. Extraction is carried out using a bi-phasic solvent system of cold methanol, methyl tert-butyl ether (MTBE), and water. In more detail, cold methanol (225 µL) containing a mixture of odd chain and deuterated lipid internal standards [lysoPE(17:1), lysoPC(17:0), PC(12:0/13:0), PE(17:0/17:0), PG(17:0/17:0), sphingosine (d17:1), d7-cholesterol, SM(17:0), C17 ceramide, d3-palmitic acid, MG(17:0/0:0/0:0), DG(18:1/2:0/0:0), DG(12:0/12:0/0:0), and d5-TG(17:0/17:1/17:0)] is added to a 20 µL sample aliquot, which is placed into a 1.5 mL Eppendorf tube, and the tube is vortexed for 10 s. Then, 750 µL of cold MTBE containing CE(22:1) (internal standard) are added, followed by vortexing for 10 s. and shaking for 6 min. at 4ºC. Phase separation is induced by adding 188 µL of mass spec-grade water. After vortexing for 20 s. The sample is centrifuged at 14,000 rpm for 2 min. The upper organic phase is collected in two 300 µL aliquots. One is stored at -20ºC as a backup and the other is evaporated to dryness in a SpeedVac. Dried extracts are resuspended using a mixture of methanol/toluene (9:1, v/v) (60 µL) containing an internal standard [12-[[(cyclohexylamino)carbonyl]amino]-dodecanoic acid (CUDA)] used as a quality control. |
Combined analysis:
| Analysis ID | AN002685 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent |
| Column | Waters ACQUITY UPLC CSH C18 |
| MS Type | EI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6530 QTOF |
| Ion Mode | POSITIVE |
| Units | counts per second |
Chromatography:
| Chromatography ID: | CH001979 |
| Chromatography Summary: | The LC/QTOFMS analyses are performed using an Agilent 1290 Infinity LC system (G4220A binary pump, G4226A autosampler, and G1316C Column Thermostat) coupled to either an Agilent 6530 (positive ion mode) or an Agilent 6550 mass spectrometer equipped with an ion funnel (iFunnel) (negative ion mode). Lipids are separated on an Acquity UPLC CSH C18 column (100 x 2.1 mm; 1.7 µm) maintained at 65°C at a flow-rate of 0.6 mL/min. Solvent pre-heating (Agilent G1316) was used. The mobile phases consist of 60:40 acetonitrile:water with 10 mM ammonium formate and 0.1% formic acid (A) and 90:10 propan-2-ol:acetonitrile with 10 mM ammonium formate and 0.1% formic acid. The gradient is as follows: 0 min 85% (A); 0–2 min 70% (A); 2–2.5 min 52% (A); 2.5–11 min 18% (A); 11–11.5 min 1% (A); 11.5–12 min 1% (A); 12–12.1 min 85% (A); 12.1–15 min 85% (A). A sample volume of 3 µL is used for the injection. Sample temperature is maintained at 4°C in the autosampler. |
| Instrument Name: | Agilent |
| Column Name: | Waters ACQUITY UPLC CSH C18 |
| Column Temperature: | 65 |
| Flow Gradient: | 0 min 85% (A); 0-2 min 70% (A); 2-2.5 min 52% (A); 2.5-11 min 18% (A); 11-11.5 min 1% (A); 11.5-12 min 1% (A); 12-12.1 min 85% (A); 12.1-15 min 85% (A). |
| Flow Rate: | 0.6 mL/min |
| Solvent A: | 60% acetonitrile/40% water; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002484 |
| Analysis ID: | AN002685 |
| Instrument Name: | Agilent 6530 QTOF |
| Instrument Type: | QTOF |
| MS Type: | EI |
| MS Comments: | The quadrupole/time-of-flight (QTOF) mass spectrometers are operated with electrospray ionization (ESI) performing full scan in the mass range m/z 65–1700 in positive (Agilent 6530, equipped with a JetStreamSource) and negative (Agilent 6550, equipped with a dual JetStream Source) modes producing both unique and complementary spectra. Instrument parameters are as follows (positive mode) Gas Temp 325°C, Gas Flow 8 l/min, Nebulizer 35 psig, Sheath Gas 350°C, Sheath Gas Flow 11, Capillary Voltage 3500 V, Nozzle Voltage 1000V, Fragmentor 120V, Skimmer 65V. Data (both profile and centroid) are collected at a rate of 2 scans per second. In negative ion mode, Gas Temp 200°C, Gas Flow 14 l/min, Fragmentor 175V, with the other parameters identical to positive ion mode. For the 6530 QTOF, a reference solution generating ions of 121.050 and 922.007 m/z in positive mode and 119.036 and 966.0007 m/z in negative mode, and these are used for continuous mass correction. For the 6550, the reference solution is introduced via a dual spray ESI, with the same ions and continuous mass correction. Samples are injected (1.7 μl in positive mode and 5 μl in negative ion mode) with a needle wash for 20 seconds (wash solvent is isopropanol). The valve is switched back and forth during the run for washing; this has been shown to be essential for reducing carryover of less polar lipids. |
| Ion Mode: | POSITIVE |
