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MB Sample ID: SA028151
| Local Sample ID: | S_60exp3 |
| 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 |
|---|---|---|---|---|
| S_60exp3 | SA028151 | FL006976 | Sensitive | Artemisinin resistant |
| S_60exp3 | SA028151 | FL006976 | WT | 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 |
