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MB Sample ID: SA087927
| Local Sample ID: | Blank-30012018 |
| Subject ID: | SU001300 |
| Subject Type: | Cultured cells |
| Subject Species: | Plasmodium falciparum |
| Taxonomy ID: | 5833 |
| Genotype Strain: | 3D7 |
| Cell Strain Details: | MR4-3D7 |
| Subject Comments: | All were MACS seperated Trophozoite infected red blood cells |
| Cell Counts: | 1x10^8 per technical replicate |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001300 |
| Subject Type: | Cultured cells |
| Subject Species: | Plasmodium falciparum |
| Taxonomy ID: | 5833 |
| Genotype Strain: | 3D7 |
| Cell Strain Details: | MR4-3D7 |
| Subject Comments: | All were MACS seperated Trophozoite infected red blood cells |
| Cell Counts: | 1x10^8 per technical replicate |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Blank-30012018 | SA087927 | FL012939 | No treatment | Drug treatment |
Collection:
| Collection ID: | CO001294 |
| Collection Summary: | Aspiration of 4 mL media, transfer of sample (1 mL) to 1.5 mL tubes |
| Sample Type: | Cultured cells |
| Collection Method: | Aspiration of 4 mL media, transfer of sample (1 mL) to 1.5 mL tubes |
| Collection Location: | Biological safety hood |
| Collection Frequency: | once after 2.5 hours treatment |
| Collection Duration: | Collection and extraction occurs in under 20 minutes per plate |
| Volumeoramount Collected: | 1 mL containing 1x10^8 infected cells |
| Storage Conditions: | On ice |
| Collection Vials: | 1.5 mL eppendorf |
| Storage Vials: | 1.5 mL eppendorf |
| Tissue Cell Quantity Taken: | 1 mL containing 1x10^8 infected cells |
Treatment:
| Treatment ID: | TR001315 |
| Treatment Summary: | Infected Red Blood cells were exposed to Antimalarial Compounds, associated with sample names. No treatment (as in the case of blanks) denotes these samples did not undergo the treatment incubation (blanks are made at the point of extraction in this case), whereas treated with no drug relates that these samples (no drug or ND) underwent the same treatment incubation as other treated samples, but in this case as a negative control with no drug administered. |
| Treatment: | Abiotic |
| Treatment Compound: | Atovaquone, Dihydroartemisinin (DHA), Ferroquin (FQ), GNF-Pf-5660, KAI407, Lumefantrine (LMF), Methylene Blue, MMV00017 (Naphthoquine), MMV000442, MMV00787, MMV007181, MMV019555, MMV020746, MMV021735, MMV022224, MMV022478, MMV027496, MMV030666, MMV085071, MMV665794, MMV665939, MMV667491, MMV668311, MMV675939, Mefloquine (MQ), Piperaquine (PPQ), WLL |
| Treatment Route: | Addition to media |
| Treatment Dose: | 10xIC50 |
| Treatment Dosevolume: | Always less than 10 uL per 5 mL sample |
| Treatment Doseduration: | 2.5 hours |
| Treatment Vehicle: | Stocks kept in either DMSO or Water |
| Cell Storage: | Temperature/Gas Mixture controlled incubator |
| Cell Growth Container: | 6-well plate |
| Cell Growth Config: | 5 mL per well, each with 1x10^8 cells |
| Cell Media: | RPMI 1640 w/ Albumax, Gentamycin, Hypoxanthine, HEPES, Sodium Bicarbonate |
| Cell Harvesting: | PBS Wash followed by 90% methanol extraction |
Sample Preparation:
| Sampleprep ID: | SP001308 |
| Sampleprep Summary: | Samples washed in PBS, spun down, aspirated, extracted in 90% methanol with labelled aspartate, then dried, resuspended in water labelled with chlorpropamide |
| Sampleprep Protocol Filename: | Metabolite_Extraction_for_LCMS_2017_Llinas.pdf |
| Processing Method: | Wash, Lysis, Drying, resuspension |
| Processing Storage Conditions: | On ice |
| Extraction Method: | 90% Methanol Lysis |
| Extract Cleanup: | Centrifugation, pellet removal |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 100 uL water with chlorpropamide |
| Sample Spiking: | Labelled Aspartate, Chlorpropamide |
Combined analysis:
| Analysis ID | AN002050 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Waters XSelect HSS T3 XP (100 x 2.1mm,2.5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Exactive Plus Orbitrap |
| Ion Mode | NEGATIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH001489 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters XSelect HSS T3 XP (100 x 2.1mm,2.5um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001902 |
| Analysis ID: | AN002050 |
| Instrument Name: | Thermo Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Blank subtraction, RSD cutoffs both by RSD per sample and pooled QCs to eliminate metabolites affected by time-dependency |
| Ion Mode: | NEGATIVE |
