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MB Sample ID: SA216768

Local Sample ID:FIT01-204-neg
Subject ID:SU002337
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Age Or Age Range:6-17 years
Weight Or Weight Range:Non-exacerbation-prone IQR: 30-68 kg; Exacerbation-prone IQR: 42-63 kg
Height Or Height Range:Non-exacerbation-prone IQR: 131-165 cm; Exacerbation-prone IQR: 140-163 cm
Gender:Male and female
Human Race:18 white, 189 Black, 7 more than one race, 1 Asian
Human Ethnicity:2 Hispanic, 113 Not Hispanic
Human Medications:92 on high-dose inhaled corticosteroids; 94 on long-acting beta adrenergic receptor agonists; 111 on Montelukast
Human Smoking Status:24 exposed to tobacco smoke
Human Inclusion Criteria:1) current or historical evidence of >=12% reversibility in forced expiratory volume in one second (FEV1) relative to baseline after bronchodilator administration; 2) airway hyperresponsiveness to methacholine, with a provocative concentration of methacholine causing a 20% drop in FEV1
Human Exclusion Criteria:1) premature birth before 35 weeks of gestation; 2) other chronic airway disorders that could mimic asthma.

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Subject:

Subject ID:SU002337
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Age Or Age Range:6-17 years
Weight Or Weight Range:Non-exacerbation-prone IQR: 30-68 kg; Exacerbation-prone IQR: 42-63 kg
Height Or Height Range:Non-exacerbation-prone IQR: 131-165 cm; Exacerbation-prone IQR: 140-163 cm
Gender:Male and female
Human Race:18 white, 189 Black, 7 more than one race, 1 Asian
Human Ethnicity:2 Hispanic, 113 Not Hispanic
Human Medications:92 on high-dose inhaled corticosteroids; 94 on long-acting beta adrenergic receptor agonists; 111 on Montelukast
Human Smoking Status:24 exposed to tobacco smoke
Human Inclusion Criteria:1) current or historical evidence of >=12% reversibility in forced expiratory volume in one second (FEV1) relative to baseline after bronchodilator administration; 2) airway hyperresponsiveness to methacholine, with a provocative concentration of methacholine causing a 20% drop in FEV1
Human Exclusion Criteria:1) premature birth before 35 weeks of gestation; 2) other chronic airway disorders that could mimic asthma.

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
FIT01-204-negSA216768FL026296YesThreeorMoreBursts
FIT01-204-negSA216768FL026296YesICShighdose
FIT01-204-negSA216768FL026296Genetic R01Study
FIT01-204-negSA216768FL02629618.77AgeatEnrollment
FIT01-204-negSA216768FL02629618.77AsthmaDurationYears
FIT01-204-negSA216768FL026296MSex
FIT01-204-negSA216768FL026296Not HispanicEthnicity
FIT01-204-negSA216768FL026296BlackRace
FIT01-204-negSA216768FL026296171.05Height
FIT01-204-negSA216768FL02629656.1Weight
FIT01-204-negSA216768FL02629619.17BMI
FIT01-204-negSA216768FL026296YesLABA
FIT01-204-negSA216768FL026296YesMontelukast

Collection:

Collection ID:CO002330
Collection Summary:Patients were not fasted prior to blood collection, but did withhold controller medication for 24 hours prior to blood collection. Blood was collected via venipuncture into EDTA-coated tubes. Plasma was isolated via centrifugation and stored at -80 *C.
Sample Type:Blood (plasma)
Storage Conditions:-80℃
Collection Vials:EDTA-coated tubes
Storage Vials:Eppendorf tubes
Collection Tube Temp:Room temperature
Additives:EDTA

Treatment:

Treatment ID:TR002349
Treatment Summary:Plasma samples were not treated with anything prior to metabolite extraction.

Sample Preparation:

Sampleprep ID:SP002343
Sampleprep Summary:50 uL of plasma was mixed with 200 uL of a 50:50 mixture of methanol and acetonitirile, vortexed, incubated on ice for 30 min, then centrifuged at 20,000 g for 10 min at 4 *C. 200 uL of the supernatant was collected for analysis.
Extraction Method:50:50 methanol:acetonitrile for a 1:2:2 samples:methanol:acetonitrile final
Extract Cleanup:centrifuged at 20,000 g for 10 min at 4 *C
Extract Storage:-80℃

Combined analysis:

Analysis ID AN003677 AN003678
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Vanquish Horizon Binary ultrahigh performance liquid chromatography system Vanquish Horizon Binary ultrahigh performance liquid chromatography system
Column SeQuant ZIC-HILIC SeQuant ZIC-HILIC
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 POSITIVE NEGATIVE
Units peak area peak area

Chromatography:

Chromatography ID:CH002727
Chromatography Summary:Equilibration time was 4 min at 90% B, with 3.5 min at 0.35 mL/min, then 0.5 min at 0.25 mL/min. After sample injection, a gradient ran from 90% to 20% B over 17.5 min at 0.25 mL/min, then flow was increased to 0.35 mL/min for 1 min and held at 20% B at 0.35 mL/min for 3.5 min.
Instrument Name:Vanquish Horizon Binary ultrahigh performance liquid chromatography system
Column Name:SeQuant ZIC-HILIC
Column Pressure:15-180 bar
Column Temperature:40
Flow Rate:0.25-0.35 mL/min
Sample Injection:2 uL
Solvent A:100% water; 0.1% formic acid
Solvent B:100% acetonitrile; 0.1% formic acid
Analytical Time:1.85-14.5 min
Chromatography Type:HILIC

MS:

MS ID:MS003428
Analysis ID:AN003677
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Data were processed using Compound Discoverer 3.2 (Thermo). Minimum peak intensity was 5E5 for positive mode. Retention times were aligned across runs, quality control (QC) pooled sample areas were normalized per metabolite using a cubic spline method, requiring <50% relative standard deviation after correction for compound retention. Missing values were gap-filled based on real peak detection and metabolites with max sample area <5-fold that of the blank were censored.
Ion Mode:POSITIVE
Capillary Temperature:275
Ion Spray Voltage:+3.5 kV
Automatic Gain Control:1E6
  
MS ID:MS003429
Analysis ID:AN003678
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Data were processed using Compound Discoverer 3.2 (Thermo). Minimum peak intensity was 2E5 for negative mode. Retention times were aligned across runs, quality control (QC) pooled sample areas were normalized per metabolite using a cubic spline method, requiring <50% relative standard deviation after correction for compound retention. Missing values were gap-filled based on real peak detection and metabolites with max sample area <5-fold that of the blank were censored.
Ion Mode:NEGATIVE
Capillary Temperature:275
Ion Spray Voltage:-2.75 kV
Automatic Gain Control:1E6
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