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MB Sample ID: SA178308

Local Sample ID:160
Subject ID:SU002004
Subject Type:Invertebrate
Subject Species:Drosophila melanogaster
Taxonomy ID:7227
Gender:Male and female

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Combined analysis:

Analysis ID AN003130 AN003131 AN003132
Analysis type MS MS MS
Chromatography type HILIC HILIC HILIC
Chromatography system Agilent 1260 Agilent 1260 Agilent 1260
Column Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um)
MS Type ESI ESI ESI
MS instrument type Other QTOF QTOF
MS instrument name ABI Sciex 5500 QTrap Agilent 6520 QTOF Agilent 6520 QTOF
Ion Mode UNSPECIFIED POSITIVE NEGATIVE
Units Peak counts Peak counts

Chromatography:

Chromatography ID:CH002312
Chromatography Summary:Targeted LC/MS-MS 1st column. LC-MS/MS experiments were performed on an Agilent 1260 LC (Agilent Technologies, Santa Clara, CA)-AB Sciex QTrap 5500 mass spectrometer (AB Sciex, Toronto, ON, Canada) system at the University of Washington Northwest Metabolomics Research Center (UWNMRC). Each sample was injected twice, 10 µL for analysis using negative ionization mode and 2 µL for analysis using positive ionization mode. Both chromatographic separations were performed in hydrophilic interaction chromatography (HILIC) mode on two Waters XBridge BEH Amide columns (150 x 2.1 mm, 2.5 µm particle size, Waters Corporation, Milford, MA) connected in parallel. The flow rate was 0.300 mL/min, auto-sampler temperature was kept at 4 °C, and the column compartment was set at 40 °C. The mobile phase was composed of Solvents A (5 mM ammonium acetate in 90%H2O/ 10% acetonitrile + 0.2% acetic acid) and B (5 mM ammonium acetate in 90%acetonitrile/ 10% H2O + 0.2% acetic acid). After the initial 2 min isocratic elution of 90% B, the percentage of Solvent B decreased to 50% at t=5 min. The composition of Solvent B maintained at 50% for 4 min (t=9 min), and then the percentage of B gradually went back to 90%, to prepare for the next injection. Targeted data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The LC-MS system was controlled by Analyst 1.5 software (AB Sciex). The extracted MRM peaks were integrated using MultiQuant 2.1 software (AB Sciex). Samples were spiked with 13C internal standards, and two types of LC-MS/MS quality control (QC) samples were run at 11 evenly-spaced intervals throughout the run to track potential drift in the assay. One QC sample was a pool of 10 fly samples, and the other was a sample of human serum. The CV for these QCs was 8.2% and 7.7% respectively
Instrument Name:Agilent 1260
Column Name:Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um)
Column Temperature:40
Flow Rate:0.300 mL/min
Internal Standard:13C
Solvent A:90% water/10% acetonitrile; 0.2% acetic acid; 5 mM ammonium acetate
Solvent B:90% acetonitrile/ 10% water; 0.2% acetic acid; 5 mM ammonium acetate
Chromatography Type:HILIC
  
Chromatography ID:CH002313
Chromatography Summary:Targeted LC/MS-MS 1st column. LC-MS/MS experiments were performed on an Agilent 1260 LC (Agilent Technologies, Santa Clara, CA)-AB Sciex QTrap 5500 mass spectrometer (AB Sciex, Toronto, ON, Canada) system at the University of Washington Northwest Metabolomics Research Center (UWNMRC). Each sample was injected twice, 10 µL for analysis using negative ionization mode and 2 µL for analysis using positive ionization mode. Both chromatographic separations were performed in hydrophilic interaction chromatography (HILIC) mode on two Waters XBridge BEH Amide columns (150 x 2.1 mm, 2.5 µm particle size, Waters Corporation, Milford, MA) connected in parallel. The flow rate was 0.300 mL/min, auto-sampler temperature was kept at 4 °C, and the column compartment was set at 40 °C. The mobile phase was composed of Solvents A (5 mM ammonium acetate in 90%H2O/ 10% acetonitrile + 0.2% acetic acid) and B (5 mM ammonium acetate in 90%acetonitrile/ 10% H2O + 0.2% acetic acid). After the initial 2 min isocratic elution of 90% B, the percentage of Solvent B decreased to 50% at t=5 min. The composition of Solvent B maintained at 50% for 4 min (t=9 min), and then the percentage of B gradually went back to 90%, to prepare for the next injection. Targeted data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The LC-MS system was controlled by Analyst 1.5 software (AB Sciex). The extracted MRM peaks were integrated using MultiQuant 2.1 software (AB Sciex). Samples were spiked with 13C internal standards, and two types of LC-MS/MS quality control (QC) samples were run at 11 evenly-spaced intervals throughout the run to track potential drift in the assay. One QC sample was a pool of 10 fly samples, and the other was a sample of human serum. The CV for these QCs was 8.2% and 7.7% respectively
Instrument Name:Agilent 1260
Column Name:Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um)
Column Temperature:40
Flow Rate:0.300 mL/min
Internal Standard:13C
Solvent A:90% water/10% acetonitrile; 0.2% acetic acid; 5 mM ammonium acetate
Solvent B:90% acetonitrile/ 10% water; 0.2% acetic acid; 5 mM ammonium acetate
Chromatography Type:HILIC
  
Chromatography ID:CH002314
Chromatography Summary:Targeted LC/MS-MS 1st column. LC-MS/MS experiments were performed on an Agilent 1260 LC (Agilent Technologies, Santa Clara, CA)-AB Sciex QTrap 5500 mass spectrometer (AB Sciex, Toronto, ON, Canada) system at the University of Washington Northwest Metabolomics Research Center (UWNMRC). Each sample was injected twice, 10 µL for analysis using negative ionization mode and 2 µL for analysis using positive ionization mode. Both chromatographic separations were performed in hydrophilic interaction chromatography (HILIC) mode on two Waters XBridge BEH Amide columns (150 x 2.1 mm, 2.5 µm particle size, Waters Corporation, Milford, MA) connected in parallel. The flow rate was 0.300 mL/min, auto-sampler temperature was kept at 4 °C, and the column compartment was set at 40 °C. The mobile phase was composed of Solvents A (5 mM ammonium acetate in 90%H2O/ 10% acetonitrile + 0.2% acetic acid) and B (5 mM ammonium acetate in 90%acetonitrile/ 10% H2O + 0.2% acetic acid). After the initial 2 min isocratic elution of 90% B, the percentage of Solvent B decreased to 50% at t=5 min. The composition of Solvent B maintained at 50% for 4 min (t=9 min), and then the percentage of B gradually went back to 90%, to prepare for the next injection. Targeted data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The LC-MS system was controlled by Analyst 1.5 software (AB Sciex). The extracted MRM peaks were integrated using MultiQuant 2.1 software (AB Sciex). Samples were spiked with 13C internal standards, and two types of LC-MS/MS quality control (QC) samples were run at 11 evenly-spaced intervals throughout the run to track potential drift in the assay. One QC sample was a pool of 10 fly samples, and the other was a sample of human serum. The CV for these QCs was 8.2% and 7.7% respectively
Instrument Name:Agilent 1260
Column Name:Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um)
Column Temperature:40
Flow Rate:0.300 mL/min
Internal Standard:13C
Solvent A:90% water/10% acetonitrile; 0.2% acetic acid; 5 mM ammonium acetate
Solvent B:90% acetonitrile/ 10% water; 0.2% acetic acid; 5 mM ammonium acetate
Chromatography Type:HILIC
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