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MB Sample ID: SA169976
Local Sample ID: | S1_0104 |
Subject ID: | SU001906 |
Subject Type: | Synthetic sample |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001906 |
Subject Type: | Synthetic sample |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S1_0104 | SA169976 | FL020243 | Standard PCs | Treatments |
S1_0104 | SA169976 | FL020243 | no treatment | Treatment times |
S1_0104 | SA169976 | FL020243 | - | Air conditions |
Collection:
Collection ID: | CO001899 |
Collection Summary: | PC16:0/18:2, PC16:0/20:4, and PC16:0/22:6 were purchased from Avanti Polar Lipids (Albaster, AL). Each PC was oxidized by using several LPO systems, such as AAPH, autoxidation, AAPH+hemin and CuSO4+AsA. |
Sample Type: | Standard phosphadylcholines |
Treatment:
Treatment ID: | TR001919 |
Treatment Summary: | For in vitro experiments, a mixture containing 500 µM PC16:0/PUFAs (PC16:0/18:2, PC16:0/20:4, and PC16:0/22:6), 0.5% ethanol, and either 50 mM AAPH or 50 mM AAPH + 10 µM hemin was incubated in phosphate-buffered saline (PBS; pH 7.4) at 37 °C under normal or 18O2 air. For autoxidation, a solution of 500 nmol of PC16:0/18:2 in chloroform (1 mL) was immediately dried using nitrogen gas in a glass tube. The dried samples were incubated at 37 °C for 24 h and then resuspended in methanol (200 µL). For CuSO4 + AsA–induced peroxidation, a solution containing 500 µM PC16:0/18:2 and 500 µM CuSO4 + 1 mM AsA was incubated in PBS (pH 7.4) at 37 °C for 72 h. |
Sample Preparation:
Sampleprep ID: | SP001912 |
Sampleprep Summary: | For in vitro experiments, the reaction solutions were extracted using the modified Bligh and Dyer method. Briefly, a mixed solution of 1 mL methanol and 1 mL chloroform, containing 100 μM dibutylhydroxytoluene and ethylenediaminetetraacetic acid as antioxidants, and 100 nM PC15:0/18:1-d7 as an internal standard was added to the reaction solution. After 1 min vortex, the organic layer was collected in a glass tube. The extracted solution was dried under a stream of nitrogen gas, and the residue was dissolved in methanol (200 µL) and stored at −80 °C before injection into the HPLC column. |
Combined analysis:
Analysis ID | AN002968 | AN002969 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Nexera LC system (Shimadzu Co., Kyoto, Japan) | Nexera LC system (Shimadzu Co., Kyoto, Japan) |
Column | Inertsil ODS-P (150 x 2.1mm,3um) | Inertsil ODS-P (150 x 2.1mm,3um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
Ion Mode | POSITIVE | NEGATIVE |
Units | Normalized amount | Normalized amount |
Chromatography:
Chromatography ID: | CH002199 |
Instrument Name: | Nexera LC system (Shimadzu Co., Kyoto, Japan) |
Column Name: | Inertsil ODS-P (150 x 2.1mm,3um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002758 |
Analysis ID: | AN002968 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Individual oxPCs were identified by full-scan MS/data-dependent MS/MS or parallel reaction monitoring (PRM). The ionization conditions were as follows: ionization mode = positive, sheath gas flow rate = 40 arbitrary units, auxiliary gas flow rate = 10 arbitrary units, spray voltage = 2000 V, capillary temperature = 265 °C, S-lens level = 50, and heater temperature = 425 °C. The experimental conditions for full-scan MS were as follows: resolving power = 70000, automatic gain control target = 1 × 106, trap fill time = 100 ms, scan range = m/z 120–1500. The experimental conditions for MS/MS and PRM were as follows: resolving power = 17500, automatic gain control target = 1 × 106, trap fill time = 80 ms, isolation width = ±0.6 Da, fixed first mass = m/z 80, normalized collision energy = 20 or 35 eV, intensity threshold of precursor ions for MS/MS analysis = 3100, apex trigger = 2–4 s, and dynamic exclusion = 2 s. The intensity threshold of precursor ions for MS/MS analysis and the dynamic exclusion were set to 1 × 104 and 1 s, respectively. LC/HRMS/MS analysis was controlled using Xcalibur 4.2.47 software (Thermo Fisher Scientific). |
Ion Mode: | POSITIVE |
MS ID: | MS002759 |
Analysis ID: | AN002969 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Individual oxPCs were identified by full-scan MS/data-dependent MS/MS or parallel reaction monitoring (PRM). The ionization conditions were as follows: ionization mode = negative, sheath gas flow rate = 40 arbitrary units, auxiliary gas flow rate = 10 arbitrary units, spray voltage = 2000 V, capillary temperature = 265 °C, S-lens level = 50, and heater temperature = 425 °C. The experimental conditions for full-scan MS were as follows: resolving power = 70000, automatic gain control target = 1 × 106, trap fill time = 100 ms, scan range = m/z 120–1500. The experimental conditions for MS/MS and PRM were as follows: resolving power = 17500, automatic gain control target = 1 × 106, trap fill time = 80 ms, isolation width = ±0.6 Da, fixed first mass = m/z 80, normalized collision energy = 20 or 35 eV, intensity threshold of precursor ions for MS/MS analysis = 3100, apex trigger = 2–4 s, and dynamic exclusion = 2 s. The intensity threshold of precursor ions for MS/MS analysis and the dynamic exclusion were set to 1 × 104 and 1 s, respectively. LC/HRMS/MS analysis was controlled using Xcalibur 4.2.47 software (Thermo Fisher Scientific). |
Ion Mode: | NEGATIVE |