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MB Sample ID: SA298952

Local Sample ID:71_T3
Subject ID:SU002893
Subject Type:Mammal
Subject Species:Sus scrofa
Taxonomy ID:9823
Age Or Age Range:~4 months
Weight Or Weight Range:35-45 kg
Gender:Male
Animal Animal Supplier:TOPIGS TN70, Tojapigs, the Netherlands
Animal Light Cycle:12 hours
Animal Water:ad libitum
Species Group:Mammals

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Combined analysis:

Analysis ID AN004533
Chromatography ID CH003406
MS ID MS004280
Analysis type MS
Chromatography type Reversed phase
Chromatography system Thermo Dionex Ultimate 3000
Column Waters ACQUITY UPLC BEH C18 (150 x 2.1mm,1.7um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Focus
Ion Mode NEGATIVE
Units Peak area

Chromatography:

Chromatography ID:CH003406
Chromatography Summary:10 µl of each sample was loaded into a Dionex UltiMate 3000 LC System (Thermo Scientific Bremen, Germany) equipped with a Waters ACQUITY UPLC BEH C18 (150 x 2.1mm,1.7um) column coupled to a Q Exactive Orbitrap mass spectrometer (Thermo Scientific) operating in negative ion mode. A step gradient was carried out using solvent A (10 mM TBA and 15 mM acetic acid) and solvent B (100% methanol). The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B until 2 min post injection. A linear gradient to 37% B was carried out until 7 min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the gradient returned to 5% B. The chromatography was stopped at 40 min. The flow was kept constant at 0.25 mL/min and the column was placed at 40°C throughout the analysis. The MS operated in full scan mode (m/z range: [70.0000-1050.0000]) using a spray voltage of 4.80 kV, capillary temperature of 300°C, sheath gas at 40.0, auxiliary gas at 10.0. The AGC target was set at 3.0E+006 using a resolution of 140000, with a maximum IT fill time of 512 ms. Data collection was performed using the Xcalibur software (Thermo Scientific). The data analyses were performed by integrating the peak areas (El-Maven - Polly - Elucidata).
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:Waters ACQUITY UPLC BEH C18 (150 x 2.1mm,1.7um)
Column Temperature:40°C
Flow Gradient:The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B until 2 min post injection. A linear gradient to 37% B was carried out until 7 min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the gradient returned to 5% B. The chromatography was stopped at 40 min.
Flow Rate:0.25 ml/min
Solvent A:100% water; 10 mM TBA; 15 mM acetic acid
Solvent B:100% methanol
Chromatography Type:Reversed phase
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