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MB Sample ID: SA071205
| Local Sample ID: | C-1FSK7-U-00 |
| Subject ID: | SU001090 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001090 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| C-1FSK7-U-00 | SA071205 | FL011188 | CHEAR 2016-1407 Study Sample | Analysis type |
Collection:
| Collection ID: | CO001084 |
| Collection Summary: | Each study participant’s spot urine samples have been collected, at baseline, with potential for post- intervention urine collection for the remainder of the study period. Samples will be frozen, stored and shipped at -80°C to the CHEAR Laboratory Network Hub (PI Dr. Robert Wright) with a Material Transfers Agreement in place for the biomarkers specified on the lab feasibility hub including phthalates, phenols, environmental tobacco smoke biomarkers (e.g. cotinine), and trace metals as well as applicable metabolomic studies. |
| Sample Type: | Urine |
Treatment:
| Treatment ID: | TR001104 |
| Treatment Summary: | This study will use a cross-sectional design and nested case control comparison of inner-city children with poorly controlled asthma versus those with well-controlled asthma. Participants will be eligible for the study if they attend one of the inner-city schools sampled in SICAS-2 and have a physician diagnosis of asthma. SICAS2 is a factorial randomized double-blinded, placebo-controlled classroom air cleaner intervention [High Efficiency Particulate Air (HEPA) filter/purifying units]/ integrated pest management (IPM) intervention. Baseline Clinical measures: Prior to randomization (a) at a clinic visit we will assess clinical characteristics (asthma symptoms, health care utilization, home characteristics); pulmonary function, FeNO, and allergen sensitivities, and (b) we will assess and sample the school/classroom environment and product use. Parental/child report of personal care product use and dietary history will be ascertained from each study participant. Clinical Follow-up: Enrolled students with asthma, randomized to treatment group by school (IPM) and classroom (HEPA) will be followed during the academic school year. Follow-up phone health outcomes surveys (e.g., symptoms, health care use; time-activity) will be conducted every 2 months after baseline, yielding evenly-spaced follow-up measures during the school year and 1 after school ends. Home environmental measures will be collected twice by dust sampling. Follow-up school/classroom visits will be conducted twice in opposite seasons to assess/collect week-long environmental classroom allergen/mold/NO2/pollution/endotoxin levels in the dust and air. Classroom/School/Home Exposure Data: The CHEAR resources will support critical analysis of urinary biomarkers at pre-intervention (baseline) during the academic school year prior to randomization and intervention. Urine specimens have been collected from 103 children with asthma at baseline over the first 2 years of the intervention study. These specimens can be linked to classroom/school allergen, mold and air sampling and school environmental questionnaires as well as enrolled students and longitudinal health outcome measures including pulmonary function testing and FeNO measurements. |
Sample Preparation:
| Sampleprep ID: | SP001097 |
| Sampleprep Summary: | Urine samples were thawed on ice, vortexed, and specific gravity (SpG) was measured. Samples were diluted with LC-MS grade water to the lowest measured SpG with ultrapure water (HILIC-positive only). Aliquots of 20 μL of the diluted urine were prepared for analysis with the LC-HRMS. A third 20 μL aliquot from each sample was combined for use as a pooled quality control sample (‘LQC’). When aliquoting was complete, the LQC sample was re-aliquoted into 20μL samples. All aliquots were returned to -80°C until analysis. Extraction was performed immediately prior to LC-HRMS analysis. All sample aliquots were thawed on ice, combined with 180uL of acetonitrile containing internal standards. Samples were then centrifuged and 80μL of supernatant transferred to an LC vial for analysis. Following the same protocol matrix blank (replacing the urine with H2O) and multiple LQCs were extracted. |
| Processing Storage Conditions: | On ice |
Combined analysis:
| Analysis ID | AN001716 | AN001717 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | Reversed phase |
| Chromatography system | Agilent 1290 Infinity II | Agilent 1290 Infinity II |
| Column | SeQuant ZIC-pHILIC (100 x 2.1mm,3.5um) | Agilent Zorbax Eclipse Plus C18 (50 x 2.1mm, 1.8 um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Agilent 6550 QTOF | Agilent 6550 QTOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak Intensity | Peak Intensity |
Chromatography:
| Chromatography ID: | CH001212 |
| Chromatography Summary: | Sample extracts were analyzed using an ultra-high performance liquid chromatography (UHPLC) 1290 Infinity II system (including 0.3 µm inline filter, Agilent Technologies, Santa Clara, USA) with 1260 Infinity II isocratic pump (including 1:100 splitter) coupled to a 6550 iFunnel quadrupole-time time of flight (Q-TOF) mass spectrometer with a dual AJS electrospray ionization source (Agilent Technologies, Santa Clara, USA). Samples were maintained at 5C in the autosampler module. For polar metabolites separation, 2 uL of sample was injected onto a HILIC SeQuant® ZIC®-HILIC column (100 mm × 2.1 mm, 100 Å, 3.5 µm particle size, Merck, Darmstadt, Germany) with a guard fitting (14 mm × 1 mm, 5.0 µm particle size, Merck, Darmstadt, Germany) maintained at 25C. Separation occurred using Mobile phase A consisted of water with 0.1% formic acid and Mobile phase B consisted of Acetonitrile with 0.1% formic acid at a flow rate of 0.3 ml/min as described in Table 1. Data was acquired with a mass range of 40-1200 m/z. Solvent gradients were as follows: 95% solvent B, hold for 1.5 min; linear decrease to 40% solvent B at 12 minutes; hold for 2 min, linear decrease to 25% solvent B at 14.2 min, hold for 2.8 min; increase to 95% solvent B at 18 min, hold for 7 min. |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | SeQuant ZIC-pHILIC (100 x 2.1mm,3.5um) |
| Column Temperature: | 25C |
| Flow Rate: | 0.3 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Analytical Time: | 25 min |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH001213 |
| Chromatography Summary: | Sample extracts were analyzed using an ultra-high performance liquid chromatography (UHPLC) 1290 Infinity II system (including 0.3 µm inline filter, Agilent Technologies, Santa Clara, USA) with 1260 Infinity II isocratic pump (including 1:100 splitter) coupled to a 6550 iFunnel quadrupole-time time of flight (Q-TOF) mass spectrometer with a dual AJS electrospray ionization source (Agilent Technologies, Santa Clara, USA). Samples were maintained at 5C in the autosampler module. For nonpolar metabolites separation, 2 uL of sample sandwiched between 10 uL of water was injected onto a Zorbax Eclipse Plus C18, RRHD column (50 mm × 2.1 mm, 1.8 µm particle size, Agilent Technologies, Santa Clara, USA) coupled to a guard column (5 mm × 2 mm, 1.8 µm Agilent Technologies, Santa Clara, USA) maintained at 50C. Separation occurred using Mobile phase A consisted of water with 0.1% formic acid and Mobile phase B consisted of 2-propanol:ACN (90:10, v/v) with 0.1% formic acid at a flow rate of 0.4 ml/min as described in Table 2. Data was acquired with a mass range of 50-1200 m/z. Solvent gradients were as follows: 5% solvent B; linear increase to 98% solvent B at 13.5 min, hold for 1.5 min; decrease to 5% solvent B at 15.5 min, hold for 3.5 min. |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | Agilent Zorbax Eclipse Plus C18 (50 x 2.1mm, 1.8 um) |
| Column Temperature: | 50C |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 90% isopropanol/10% acetonitrile; 0.1% formic acid |
| Analytical Time: | 19 min |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001588 |
| Analysis ID: | AN001716 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 250C |
| Capillary Voltage: | 3000 |
| MS ID: | MS001589 |
| Analysis ID: | AN001717 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 250C |
| Capillary Voltage: | -3000 |
