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MB Sample ID: SA028147
Local Sample ID: | R2_60exp1 |
Subject ID: | SU000568 |
Subject Type: | Cells |
Subject Species: | Plasmodium falciparum |
Taxonomy ID: | 5833 |
Genotype Strain: | Cam3.IIR539T (Cambodian isolate, artemisinin resistant) R1 | Cam3.IIC580Y (Cambodian isolate, artemisinin resistant) R2 | Cam3.IIrev (Cambodian isolate, artemisinin sensitive) S | uninfected red blood cells |
Cell Strain Details: | Artemisinin resistant strain (R1) | Artemisinin resistant strain (R2) | Artemisinin sensitive strain (S) | uninfected red blood cells |
Cell Primary Immortalized: | Primary | Primary | Primary | Primary |
Cell Counts: | 5 x 10e7 per sample (purified infected cells) | 5 x 10e7 per sample (purified infected cells) | 5 x 10e7 per sample (purified infected cells) | 5 x 10e7 per sample |
Species Group: | Microorganism |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000568 |
Subject Type: | Cells |
Subject Species: | Plasmodium falciparum |
Taxonomy ID: | 5833 |
Genotype Strain: | Cam3.IIR539T (Cambodian isolate, artemisinin resistant) R1 | Cam3.IIC580Y (Cambodian isolate, artemisinin resistant) R2 | Cam3.IIrev (Cambodian isolate, artemisinin sensitive) S | uninfected red blood cells |
Cell Strain Details: | Artemisinin resistant strain (R1) | Artemisinin resistant strain (R2) | Artemisinin sensitive strain (S) | uninfected red blood cells |
Cell Primary Immortalized: | Primary | Primary | Primary | Primary |
Cell Counts: | 5 x 10e7 per sample (purified infected cells) | 5 x 10e7 per sample (purified infected cells) | 5 x 10e7 per sample (purified infected cells) | 5 x 10e7 per sample |
Species Group: | Microorganism |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
R2_60exp1 | SA028147 | FL006974 | Resistant | Artemisinin resistant |
R2_60exp1 | SA028147 | FL006974 | C580Y | PfKelch13 mutation |
Collection:
Collection ID: | CO000562 |
Collection Summary: | None |
Sample Type: | Cell |
Collection Method: | Plasmodium falciparum parasites (two clonal artemisnin resistant |
Treatment:
Treatment ID: | TR000582 |
Treatment Summary: | None |
Sample Preparation:
Sampleprep ID: | SP000575 |
Sampleprep Summary: | Metabolism was quenched by washing with ice cold PBS (1 mL). Cells were then pelleted by centrifugation (6000 xg for 3 min) and metabolites were extracted from 5 x 107 uninfected red blood cells and 5 x 107 parasitised red blood cells reticulocytes by addition of 140 µL methanol containing internal standards (CHAPS, CAPS, PIPES and TRIS; 1 µM) and left for 1 hour at 4 °C with automatic vortex. After mixing, cellular debris was removed by centrifugation (16,000 rpm for 10 mins) and the supernatant was kept at -20 °C prior to analysis. |
Processing Method: | Lysis with automatic vortex at 4°C |
Processing Storage Conditions: | On ice or 4°C |
Extraction Method: | Methanol |
Extract Storage: | -20°C |
Sample Spiking: | Internal standards (CHAPS, CAPS, PIPES and TRIS; all at 1 µM) mixed in extraction solvent |
Cell Type: | Artemisinin resistant and sensitive parasitised red blood cells and uninfected red blood cells |
Combined analysis:
Analysis ID | AN000832 | AN000833 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | HILIC | HILIC |
Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
Column | ZIC-pHILIC (Merck Sequant) | ZIC-pHILIC (Merck Sequant) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
Ion Mode | POSITIVE | NEGATIVE |
Units | peak height | Peak height |
Chromatography:
Chromatography ID: | CH000595 |
Chromatography Summary: | Untargeted HILIC method |
Instrument Name: | Thermo Dionex Ultimate 3000 RS |
Column Name: | ZIC-pHILIC (Merck Sequant) |
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 |
Injection Temperature: | 4 °C |
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:
MS ID: | MS000733 |
Analysis ID: | AN000832 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
Ion Mode: | POSITIVE |
Capillary Temperature: | 300°C |
Capillary Voltage: | +50 V |
Fragmentation Method: | None |
Spray Voltage: | 4kV |
MS ID: | MS000734 |
Analysis ID: | AN000833 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
Capillary Temperature: | 300°C |
Capillary Voltage: | +50 V |
Fragmentation Method: | None |
Spray Voltage: | 3.5kV |