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MB Sample ID: SA093083

Local Sample ID:MMV688179_1
Subject ID:SU001346
Subject Type:Cultured cells
Subject Species:Trypanosoma brucei brucei
Taxonomy ID:5702
Genotype Strain:427

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Subject ID:SU001346
Subject Type:Cultured cells
Subject Species:Trypanosoma brucei brucei
Taxonomy ID:5702
Genotype Strain:427


Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
MMV688179_1SA093083FL013324MMV688179Drug treatment


Collection ID:CO001340
Collection Summary:Trypanosoma brucei brucei (T.b.b) bloodstream forms (strain 427) were maintained in vitro in 5-10 ml cultures at 37 °C and 5% CO2 in Creeks minimal media supplemented with 10% HMI-9. The cultures were passaged every 2-3 days and cells were grown to a maximum density of 2x106cells/ml.
Sample Type:Cultured cells


Treatment ID:TR001361
Treatment Summary:For drug-induced metabolic perturbation experiments, cells were sub-cultured at 10e6cells/ml in 20 ml volume with drugs added at 0.5 µM concentration and incubated for a further 5 hours until cell density reached ~2x10e6cells/ml. Cells were then used for metabolite quenching and extraction.

Sample Preparation:

Sampleprep ID:SP001354
Sampleprep Summary:Metabolism was rapidly quenched by rapidly cooling cultures to 4°C in a dry ice and ethanol bath. Cultures were then centrifuged for 10 minutes at 1250g at 4°C. The supernatant was discarded and cells were washed with 1 ml of cold PBS by centrifugation for 1 minute at 2100g at 4°C. The supernatant was removed and cells were extracted with 100 µl extraction solvent containing chloroform: methanol: water (1:3:1 v/v) followed by vortexing at 4 °C for 1 hour. The resulting suspension was centrifuged for 10 minutes at 2100g at 4°C and the supernatant was transferred to glass vials and stored at -80 °C until analysis by liquid chromatography and high-resolution mass spectrometry.

Combined analysis:

Analysis ID AN002115
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Dionex Ultimate 3000 RS
Column SeQuant ZIC-pHILIC (150 x 4.6mm,5um)
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Units Peak height


Chromatography ID:CH001548
Chromatography Summary:Metabolite extracts were analysed using hydrophilic interaction (HILIC) liquid chromatography (LC) and high resolution mass spectrometry on an Orbitrap system.
Chromatography Comments:Dionex RSLC3000 UHPLC (Thermo). Column additional info: Merck Sequant ZIC-pHILIC 5 µm polymer metal-free(150 x 4.6 mm)
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:SeQuant ZIC-pHILIC (150 x 4.6mm,5um)
Column Temperature:25°C
Flow Gradient:linear gradient -time, %B as follows: 0min- 80%, 15min- 50%, 18min- 5%, 21min- 5%, 24min- 80%, 32min- 80%.
Flow Rate:300 μL/min
Internal Standard:internal standards (CHAPS, CAPS, PIPES and TRIS; all at 1 µM)
Sample Injection:10 μL
Solvent A:100% water; 20 mM ammonium carbonate
Solvent B:100% acetonitrile
Chromatography Type:HILIC


MS ID:MS001970
Analysis ID:AN002115
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Comments:The raw LC-MS data was processed using IDEOM.
Capillary Temperature:300°C
Capillary Voltage:+50 V
Spray Voltage:4kV
Resolution Setting:35000
Scan Range Moverz:85- 1275 m/z