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MB Sample ID: SA051140
Local Sample ID: | S19 |
Subject ID: | SU000915 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Age Or Age Range: | 13 - 66 |
Gender: | Female and Male |
Human Ethnicity: | Brazillian |
Human Inclusion Criteria: | Retrospective serum samples from leprosy patients diagnosed or not with type 1 reaction that were not receiving multidrug therapy or corticosteroid therapy at the time of blood collection. |
Human Exclusion Criteria: | Retrospective serum samples from leprosy patients that were under multidrug therapy and/or corticosteroid therapy at the time of blood collection were excluded from the study. |
Subject Comments: | The patients exhibited borderline tuberculoid, borderline borderline and borderline lepromatous forms of leprosy and was diagnosed or not with type 1 reaction. |
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Subject:
Subject ID: | SU000915 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Age Or Age Range: | 13 - 66 |
Gender: | Female and Male |
Human Ethnicity: | Brazillian |
Human Inclusion Criteria: | Retrospective serum samples from leprosy patients diagnosed or not with type 1 reaction that were not receiving multidrug therapy or corticosteroid therapy at the time of blood collection. |
Human Exclusion Criteria: | Retrospective serum samples from leprosy patients that were under multidrug therapy and/or corticosteroid therapy at the time of blood collection were excluded from the study. |
Subject Comments: | The patients exhibited borderline tuberculoid, borderline borderline and borderline lepromatous forms of leprosy and was diagnosed or not with type 1 reaction. |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S19 | SA051140 | FL009850 | No | Type 1 reaction (T1R) |
Collection:
Collection ID: | CO000909 |
Collection Summary: | The whole blood (10 mL) was collected in a Vacutainer tube from Becton Dickinson (BD). After collection of the whole blood, it was allowed the blood to clot by leaving it undisturbed at room temperature for 30 minutes. The clot was removed by centrifuging at 1,000–2,000 x g for 10 minutes in a refrigerated centrifuge (4 C). The resulting supernatant is designated serum. The serum samples were aliquoted in 100 microliters and stored in a serum bank at -80C. These samples were collected in the Leprosy Outpatient Unit (Oswaldo Cruz Foundation, Rio de Janeiro, Brazil) and then they were sent in dry ice to Colorado State University, where it was processed and analyzed for metabolomics. At Colorado State University the samples were stored at -80C until processing. |
Sample Type: | Serum |
Collection Location: | Leprosy Outpatient Unit (Oswaldo Cruz Foundation, Rio de Janeiro, Brazil) |
Collection Duration: | 30 minutes |
Storage Conditions: | -80C |
Additives: | no |
Blood Serum Or Plasma: | Serum |
Treatment:
Treatment ID: | TR000929 |
Treatment Summary: | No treatment of the patients was performed |
Sample Preparation:
Sampleprep ID: | SP000922 |
Sampleprep Summary: | Serum samples (7 serum samples from leprosy patients with T1R) were pooled prior the extraction. These pooled serum samples were incubated with 3 volumes of cold methanol (100%) for 1 hour at -20C. After this incubation, the samples were centrifuged for 30 minutes at 18,000 xg at 4C to clear the supernatant. The supernatant was then transferred and dried under vacuum. Samples were re-suspended in 80 μl of 50% methanol. Prior to the injections on the LC-MS instrument, the samples were centrifuged for 30 minutes at 18,000 xg at 4oC. |
Sampleprep Protocol ID: | SP000100 |
Sampleprep Protocol Filename: | 060613_Methanol_Serum_Extraction |
Sampleprep Protocol Comments: | Use always nitrile gloves. Do not use gloves with powder. Use pipette tips with aerosol barrier. |
Processing Method: | Homogenization and solvent removal with Speed Vac |
Processing Storage Conditions: | on ice |
Extraction Method: | Cold Methanol extraction (3:1 methanol/water) |
Extract Enrichment: | no enrichment was performed |
Extract Cleanup: | Centrifugation |
Extract Storage: | -80C |
Sample Resuspension: | 50% methanol |
Sample Derivatization: | no derivatization process was performed |
Sample Spiking: | Samples were spiked with the following commercial standards: arachidonic acid, docosahexaenoic acid, eicosapentaenoic acid and leukotriene B4 |
Combined analysis:
Analysis ID | AN001437 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 |
Column | Waters Xbridge BEH C18 XP (100 x 2.1mm,2.5um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6510 QTOF |
Ion Mode | NEGATIVE |
Units | arbitrary units |
Chromatography:
Chromatography ID: | CH001008 |
Chromatography Summary: | This analysis was performed only on negative mode. The solvent gradient used for all chromatography was 2% solvent B for 3 min followed by a 15 min linear gradient of 2% to 98% solvent B, which was held for 0.3 min. Then returned to starting conditions over 1.2 min. |
Instrument Name: | Agilent 1290 |
Column Name: | Waters Xbridge BEH C18 XP (100 x 2.1mm,2.5um) |
Column Temperature: | 50C |
Flow Gradient: | 0.25 mL/min |
Flow Rate: | 0.25 mL/min |
Injection Temperature: | 4C |
Sample Injection: | 20uL |
Solvent A: | 89% water/5% acetonitrile/5% isopropanol; 5 mM ammonium acetate |
Solvent B: | 50% acetonitrile/50% isopropanol; 5 mM ammonium acetate |
Capillary Voltage: | 2.8 kV |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001327 |
Analysis ID: | AN001437 |
Instrument Name: | Agilent 6510 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
Capillary Voltage: | 2.8kV |
Collision Energy: | 20eV |
Collision Gas: | Nitrogen |
Dry Gas Flow: | 10 L/min |
Dry Gas Temp: | 310C |
Fragment Voltage: | 120V |
Nebulizer: | 45 psig |
Scanning Range: | 2 spectra per second |
Skimmer Voltage: | 50V |