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MB Sample ID: SA209449
Local Sample ID: | 9 |
Subject ID: | SU002266 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Combined analysis:
Analysis ID | AN003570 | AN003571 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Flow induction analysis | Flow induction analysis |
Chromatography system | Shimazdu LC-30AD | Shimazdu LC-30AD |
Column | none | none |
MS Type | ESI | ESI |
MS instrument type | Triple quadrupole | Triple quadrupole |
MS instrument name | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap |
Ion Mode | UNSPECIFIED | UNSPECIFIED |
Units | nmol/g | nmol/g |
MS:
MS ID: | MS003327 |
Analysis ID: | AN003570 |
Instrument Name: | ABI Sciex 5500 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | The Lipidyzer (SCIEX), a QTRAP system with SelexION ion mobility, was used for targeted profiling as described previously (Contrepois et al., 2018). In brief, flow injection analysis was performed with a LC-30AD (Shimazdu) operating at 8 μL/min (50 μL injection volume) using a running solution that consisted of 10 mM ammonium acetate in dichloromethane:MeOH (50:50). DMS separates lipids based on the principle that each lipid class has a different head group dipole moment and thus mobility in the DMS aperture (Schneider et al., 2010). The lipid molecular species were identified and quantified using multiple reaction monitoring (MRM) and positive/negative switching. Two acquisition methods were employed covering 10 lipid classes across positive and negative mode. Method 1 had SelexION voltages turned on while Method 2 had SelexION voltages turned off. Method 1 employed an isocratic flow of 8 μL/min for 7.9 min, followed by a 2 minute wash at 30 μL/min. Method 2 employed an isocratic flow of 8 μL/min for 6 minutes, followed by 2 minute wash at 30 μL/min. Each lipid was acquired throughout 20 cycles. Lipid classes targeted in positive mode: SM, DAG, CE, CER, and TAG. Lipid classes targeted in negative mode: LPE, LPC, PC, PE, and FFA. Lipids were quantified using the LWM software, which compares endogenous lipids to the known concentrations of structurally most similar spiked-in lipid standards and reports all detected lipids in nmol/g. Data analysis. Data were downloaded from the Lipidyzer LWM and merged and processed in R. In brief, Excel files (LWM output) were read with the “loadWorkbook” package. From all samples, lipid concentrations determined in a blank control (sample processed in parallel without the addition of cells) were subtracted to correct for background signals. The data set was further filtered accepting only lipid species that detected in at least 25% of all samples. Missing values were imputed by drawing from a random distribution. |
Ion Mode: | UNSPECIFIED |
MS ID: | MS003328 |
Analysis ID: | AN003571 |
Instrument Name: | ABI Sciex 5500 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | The Lipidyzer (SCIEX), a QTRAP system with SelexION ion mobility, was used for targeted profiling as described previously (Contrepois et al., 2018). In brief, flow injection analysis was performed with a LC-30AD (Shimazdu) operating at 8 μL/min (50 μL injection volume) using a running solution that consisted of 10 mM ammonium acetate in dichloromethane:MeOH (50:50). DMS separates lipids based on the principle that each lipid class has a different head group dipole moment and thus mobility in the DMS aperture (Schneider et al., 2010). The lipid molecular species were identified and quantified using multiple reaction monitoring (MRM) and positive/negative switching. Two acquisition methods were employed covering 10 lipid classes across positive and negative mode. Method 1 had SelexION voltages turned on while Method 2 had SelexION voltages turned off. Method 1 employed an isocratic flow of 8 μL/min for 7.9 min, followed by a 2 minute wash at 30 μL/min. Method 2 employed an isocratic flow of 8 μL/min for 6 minutes, followed by 2 minute wash at 30 μL/min. Each lipid was acquired throughout 20 cycles. Lipid classes targeted in positive mode: SM, DAG, CE, CER, and TAG. Lipid classes targeted in negative mode: LPE, LPC, PC, PE, and FFA. Lipids were quantified using the LWM software, which compares endogenous lipids to the known concentrations of structurally most similar spiked-in lipid standards and reports all detected lipids in nmol/g. Data analysis. Data were downloaded from the Lipidyzer LWM and merged and processed in R. In brief, Excel files (LWM output) were read with the “loadWorkbook” package. From all samples, lipid concentrations determined in a blank control (sample processed in parallel without the addition of cells) were subtracted to correct for background signals. The data set was further filtered accepting only lipid species that detected in at least 25% of all samples. Missing values were imputed by drawing from a random distribution. |
Ion Mode: | UNSPECIFIED |