Return to study ST001258 main page
MB Sample ID: SA091413
Local Sample ID: | 7 2e6 Mf |
Subject ID: | SU001326 |
Subject Type: | Invertebrate |
Subject Species: | Brugia malayi |
Taxonomy ID: | 6279 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001326 |
Subject Type: | Invertebrate |
Subject Species: | Brugia malayi |
Taxonomy ID: | 6279 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
7 2e6 Mf | SA091413 | FL013172 | Mf | Group |
Collection:
Collection ID: | CO001320 |
Collection Summary: | All parasites were obtained from FR3 (Filariasis Research Reagent Resource Center; BEI Resources, Manassas, VA, USA) where they were isolated and separated by sex from infected gerbils (Meriones unguiculatus) or mosquitoes (Aedes aegypti). Worms were flash-frozen and shipped to the New York Blood Center for processing. Stages used for metabolomics analysis included L3 larvae from mosquitoes, adult male and female worms at 120dpi, and microfilaria. The number of worms per sample were 20 adult female worms, 40 adult males, 2X106 microfilariae, and 200 L3 larvae per biological replicate. Samples were washed in 1x PBS and run in triplicate. Adult male and female worms were picked individually from PBS and each biological was weighed. The microfilaria and L3 samples were spun down, the PBS pipetted off, and weighed directly into a metabolomics 2mL screw cap vial with total amounts ranging from 1.3 mg (adult males) to 15.8 mg (microfilaria). Metabolites were extracted and the data analyzed as described in the Supplementary Information. |
Sample Type: | Worms |
Treatment:
Treatment ID: | TR001341 |
Treatment Summary: | All parasites were obtained from FR3 (Filariasis Research Reagent Resource Center; BEI Resources, Manassas, VA, USA) where they were isolated and separated by sex from infected gerbils (Meriones unguiculatus) or mosquitoes (Aedes aegypti). Worms were flash-frozen and shipped to the New York Blood Center for processing. Stages used for metabolomics analysis included L3 larvae from mosquitoes, adult male and female worms at 120dpi, and microfilaria. The number of worms per sample were 20 adult female worms, 40 adult males, 2X106 microfilariae, and 200 L3 larvae per biological replicate. Samples were washed in 1x PBS and run in triplicate. Adult male and female worms were picked individually from PBS and each biological was weighed. The microfilaria and L3 samples were spun down, the PBS pipetted off, and weighed directly into a metabolomics 2mL screw cap vial with total amounts ranging from 1.3 mg (adult males) to 15.8 mg (microfilaria). Metabolites were extracted and the data analyzed as described in the Supplementary Information. |
Sample Preparation:
Sampleprep ID: | SP001334 |
Sampleprep Summary: | Metabolite extraction – The mass of the weighed worm samples was used to scale the metabolite extraction to a ratio of 16.5 mg / 1 mL extraction solvent. Freezing 80% acetonitrile was added directly to each vial containing the samples, along with zirconium disruption beads (0.5 mm, RPI) and homogenized for 3 min at 4°C in a BeadBlasterTM with a 30 sec on, 30 sec off pattern. The resulting lysate was centrifuged at 21,000 x g for 3 min, and 90% of the supernatant volume was transferred to a 1.5 mL microfuge tube for speed vacuum concentration, no heating. The dry extracts were resolublized in a volume of LCMS grade water 1/10th of that used for the homogenization step, sonicated in a water bath for 3 min, and transferred to a glass insert for analysis. |
Combined analysis:
Analysis ID | AN002087 | AN002088 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | HILIC | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
Column | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Waters Acquity BEH Phenyl (150 x 2.1mm,1.7um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
Ion Mode | UNSPECIFIED | UNSPECIFIED |
Units |
Chromatography:
Chromatography ID: | CH001524 |
Instrument Name: | Thermo Dionex Ultimate 3000 RS |
Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
Chromatography Type: | HILIC |
Chromatography ID: | CH001525 |
Instrument Name: | Thermo Dionex Ultimate 3000 RS |
Column Name: | Waters Acquity BEH Phenyl (150 x 2.1mm,1.7um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001938 |
Analysis ID: | AN002087 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Positive/Negative Polarity Switching |
Ion Mode: | UNSPECIFIED |
MS ID: | MS001939 |
Analysis ID: | AN002088 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Positive/Negative Polarity Switching |
Ion Mode: | UNSPECIFIED |