Summary of Study ST001276

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000861. The data can be accessed directly via it's Project DOI: 10.21228/M8D392 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001276
Study TitleDevelopment and Characterisation of a Novel Class of Aroyl Guanidine Containing Anti-Trypanosomal Compounds
Study SummaryThe mode of action of a novel class of aroyl guanidine containing anti-Trypanosomal compounds was investigated using an unbiased metabolomics approach. Trypanosoma brucei parasites were incubated for five hours with the test compounds at 1 µM concentration. Analysis of cell pellets allowed reproducible detection of diverse metabolites from a range of metabolic pathways.
Institute
Monash University
Last NameCreek
First NameDarren
AddressMonash Institute of Pharmaceutical Sciences, 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Emaildarren.creek@monash.edu
Phone+61 (0) 3 9903 9249
Submit Date2019-11-16
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2020-01-13
Release Version1
Darren Creek Darren Creek
https://dx.doi.org/10.21228/M8D392
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000861
Project DOI:doi: 10.21228/M8D392
Project Title:Development and Characterisation of a Novel Class of Aroyl Guanidine Containing Anti-Trypanosomal Compounds
Project Summary:The mode of action of a novel class of aroyl guanidine containing anti-Trypanosomal compounds was investigated using an unbiased metabolomics approach. Trypanosoma brucei parasites were incubated for five hours with the test compounds at 1 µM concentration. Analysis of cell pellets allowed reproducible detection of diverse metabolites from a range of metabolic pathways.
Institute:Monash University
Last Name:Creek
First Name:Darren
Address:Monash Institute of Pharmaceutical Sciences, 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Email:darren.creek@monash.edu
Phone:+61 (0) 3 9903 9249

Subject:

Subject ID:SU001348
Subject Type:Cultured cells
Subject Species:Trypanosoma brucei brucei
Taxonomy ID:5702
Genotype Strain:427
Species Group:Unicellular parasites

Factors:

Subject type: Cultured cells; Subject species: Trypanosoma brucei brucei (Factor headings shown in green)

mb_sample_id local_sample_id Drug treatment
SA093199P_DMSO_1DMSO
SA093200P_DMSO_2DMSO
SA093201P_DMSO_3DMSO
SA093202P_DMSO_4DMSO
SA093203P_112_2MIPS112
SA093204P_112_3MIPS112
SA093205P_112_4MIPS112
SA093206P_112_1MIPS112
SA093207P_116_3MIPS116
SA093208P_116_1MIPS116
SA093209P_116_2MIPS116
SA093210P_116_4MIPS116
SA093211P_12828_3MIPS12828
SA093212P_12828_2MIPS12828
SA093213P_12828_1MIPS12828
SA093214P_12828_4MIPS12828
SA093215P_14554_1MIPS14554
SA093216P_14554_4MIPS14554
SA093217P_14554_2MIPS14554
SA093218P_14554_3MIPS14554
SA093219P_15_3MIPS15
SA093220P_15_1MIPS15
SA093221P_15_2MIPS15
SA093222P_15_4MIPS15
SA093223P_8664_4MIPS8664
SA093224P_8664_1MIPS8664
SA093225P_8664_2MIPS8664
SA093226P_8664_3MIPS8664
SA093227P_9560_4MIPS9560
SA093228P_9560_1MIPS9560
SA093229P_9560_2MIPS9560
SA093230P_9560_3MIPS9560
SA093231P_9880_2MIPS9880
SA093232P_9880_1MIPS9880
SA093233P_9880_3MIPS9880
SA093234P_9880_4MIPS9880
Showing results 1 to 36 of 36

Collection:

Collection ID:CO001342
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:

Treatment ID:TR001363
Treatment Summary:For drug-induced metabolic perturbation experiments, cells were sub-cultured at 10e6cells/ml in 20 ml volume with drugs added at 1 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:SP001356
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.

Chromatography:

Chromatography ID:CH001550
Chromatography Summary:Metabolite extracts were analysed using hydrophilic interaction (HILIC) liquid chromatography (LC) and high resolution mass spectrometry on an Orbitrap system.
Chromatography Comments: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

Analysis:

Analysis ID:AN002117
Analysis Type:MS
Chromatography ID:CH001550
Num Factors:9
Num Metabolites:585
Rt Units:Minutes
Units:Peak height
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