#METABOLOMICS WORKBENCH swalmsle89_20170421_123359 DATATRACK_ID:891 STUDY_ID:ST000625 ANALYSIS_ID:AN000957 PROJECT_ID:PR000456 VERSION 1 CREATED_ON June 23, 2017, 10:34 am #PROJECT PR:PROJECT_TITLE A strategy for producing a searchable bronchoalveolar lavage fluid compound PR:PROJECT_TITLE database PR:PROJECT_TYPE MS PR:PROJECT_SUMMARY Comprehensive metabolomic databases for Lipid(+/-) and Aqueous(+) fractions of PR:PROJECT_SUMMARY BALF assembled from a mouse and human COPD experiment. PR:INSTITUTE University of Colorado, Denver PR:DEPARTMENT Pharmaceutical Sciences, Anschutz Medical Campus PR:LABORATORY Metabolomics and Mass Spectrometry PR:LAST_NAME Residorph PR:FIRST_NAME Nichole PR:ADDRESS 12850 East Montview Blvd, Denver, CO 80204 PR:EMAIL nichole.reisdorph@ucdenver.edu PR:PHONE - PR:FUNDING_SOURCE P20-HL113445 Metabolic Profiles of COPD phenotypes #STUDY ST:STUDY_TITLE Non targeted metabolomic analysis of mouse and human bronchoalveolar lavage ST:STUDY_TITLE fluid, Aqueous(+) experiment (part I) ST:STUDY_TYPE Comprehensive profile ST:STUDY_SUMMARY Mouse BALF was collected from C57BL/6 mice (n = 10). Five mice were exposed to ST:STUDY_SUMMARY ambient air for one day (air control) and five mice were exposed to CS for nine ST:STUDY_SUMMARY months (smoking). Human BALF was collected from subjects from the COPDGene ST:STUDY_SUMMARY cohort (n = 5). COPD diagnosis was based on the ratio of forced expiratory ST:STUDY_SUMMARY volume in 1 second to forced vital capacity (FEV1/FVC). Subjects were 45-70 ST:STUDY_SUMMARY years old, BMI 27-45, weight 76-125kg, and were categorized as follows: 1 male ST:STUDY_SUMMARY former smoker without COPD (FEV1/FVC = 0.82), 2 male current smokers without ST:STUDY_SUMMARY COPD (FEV1/FVC = 0.91 and 0.79), 1 female current smoker with moderate COPD ST:STUDY_SUMMARY (FEV1/FVC = 0.51), and 1 male current smoker with moderate COPD (FEV1/FVC = ST:STUDY_SUMMARY 0.64). ST:INSTITUTE University of Colorado Anschutz Medical Campus ST:DEPARTMENT Pharmaceutical Sciences ST:LABORATORY Metabolomics and Mass Spectrometry ST:LAST_NAME Residorph ST:FIRST_NAME Nichole ST:ADDRESS 12850 East Montview Blvd, Denver, CO 80204 ST:EMAIL nichole.reisdorph@ucdenver.edu ST:PHONE - ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 15 #SUBJECT SU:SUBJECT_TYPE Human;Animal SU:SUBJECT_SPECIES Homo sapiens;Mus musculus SU:TAXONOMY_ID 9606 | 10093 SU:GENOTYPE_STRAIN Animal: C57BL/6 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS 10031R HBAL_10031R Species:Human | SmokingStatus:Smoker | Phenotype:COPD Gender=F SUBJECT_SAMPLE_FACTORS 11833G HBAL_11833G Species:Human | SmokingStatus:Former smoker | Phenotype:Moderate Gender=M SUBJECT_SAMPLE_FACTORS 12558P HBAL_12558P Species:Human | SmokingStatus:Smoker | Phenotype:Healthy Gender=M SUBJECT_SAMPLE_FACTORS 14442G HBAL_14442G Species:Human | SmokingStatus:Smoker | Phenotype:Healthy Gender=M SUBJECT_SAMPLE_FACTORS 17800V HBAL_17800V Species:Human | SmokingStatus:Smoker | Phenotype:COPD Gender=M SUBJECT_SAMPLE_FACTORS MET106 MBAL_MET106 Species:Mouse | SmokingStatus:Smoker | Phenotype:Emphysema Gender=F SUBJECT_SAMPLE_FACTORS MET107 MBAL_MET107 Species:Mouse | SmokingStatus:Smoker | Phenotype:Emphysema Gender=F SUBJECT_SAMPLE_FACTORS MET108 MBAL_MET108 Species:Mouse | SmokingStatus:Smoker | Phenotype:Emphysema Gender=F SUBJECT_SAMPLE_FACTORS MET126 MBAL_MET126 Species:Mouse | SmokingStatus:Smoker | Phenotype:Emphysema Gender=F SUBJECT_SAMPLE_FACTORS MET128 MBAL_MET128 Species:Mouse | SmokingStatus:Smoker | Phenotype:Emphysema Gender=F SUBJECT_SAMPLE_FACTORS MET133 MBAL_MET133 Species:Mouse | SmokingStatus:Non-Smoker | Phenotype:Healthy Gender=F SUBJECT_SAMPLE_FACTORS MET141 MBAL_MET141 Species:Mouse | SmokingStatus:Non-Smoker | Phenotype:Healthy Gender=F SUBJECT_SAMPLE_FACTORS MET142 MBAL_MET142 Species:Mouse | SmokingStatus:Non-Smoker | Phenotype:Healthy Gender=F SUBJECT_SAMPLE_FACTORS MET143 MBAL_MET143 Species:Mouse | SmokingStatus:Non-Smoker | Phenotype:Healthy Gender=F SUBJECT_SAMPLE_FACTORS MET144 MBAL_MET144 Species:Mouse | SmokingStatus:Non-Smoker | Phenotype:Healthy Gender=F #COLLECTION CO:COLLECTION_SUMMARY Mouse BALF collection was performed using a total of 1.0 mL PBS divided into CO:COLLECTION_SUMMARY three washes and stored at -80°C until analysis. Human BALF was collected in CO:COLLECTION_SUMMARY the right middle lobe and lingual by instilling two aliquots of 40 mL and one CO:COLLECTION_SUMMARY aliquot of 50 mL of sterile saline per lobe (i.e., 130 mL per lobe, total CO:COLLECTION_SUMMARY volume = 260 mL per subject), which is withdrawn by gentle manual suction CO:COLLECTION_SUMMARY and immediately placed on ice. Aliquots for metabolomics analysis were frozen at CO:COLLECTION_SUMMARY -80 °C and stored until sample preparation and MS analysis. CO:SAMPLE_TYPE BALF CO:COLLECTION_METHOD Broncheoalveolar lavage #TREATMENT TR:TREATMENT_SUMMARY None #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 200uL of BALF was prepared using methanol assisted protein precipitation SP:SAMPLEPREP_SUMMARY followed by methyl-tert butyl ether (MTBE) liquid-liquid extraction. The aqueous SP:SAMPLEPREP_SUMMARY fraction was dried down in a SpeedVac at 45°C, then reconstituted in 50 µL of SP:SAMPLEPREP_SUMMARY 95:5 water:acetonitrile. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Analysis of the aqueous fraction was by reversed-phase chromatography using an CH:CHROMATOGRAPHY_SUMMARY Agilent 1200 series pump with an Agilent Zorbax Narrow Bore RRHT SB-AQ (1.8 CH:CHROMATOGRAPHY_SUMMARY micron, 2.1 x 100 mm, 80Å) analytical column and an Agilent Zorbax SB-AQ (5 CH:CHROMATOGRAPHY_SUMMARY micron, 2.1 x 12.5 mm) guard column. Sample injection volumes were 10 µL. A CH:CHROMATOGRAPHY_SUMMARY flow rate of 0.3ml/min was used with the following mobile phases: mobile phase A CH:CHROMATOGRAPHY_SUMMARY was water with 0.1% formic acid, and mobile phase B was 90:10 acetonitrile:water CH:CHROMATOGRAPHY_SUMMARY with 0.1% formic acid. Gradient elution was as follows: 0-3 minutes 2% B, 3-5 CH:CHROMATOGRAPHY_SUMMARY minutes 2-40% B, 5-20 minutes 40-100% B, 20-30 minutes 100% B, 30-30.01 100-2% CH:CHROMATOGRAPHY_SUMMARY B, 30.01-40 minutes 2% B. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1200 CH:COLUMN_NAME Agilent Zorbax Narrow Bore RRHT RRHD SB-AQ (100 x 2.1mm, 1.8um) CH:COLUMN_NAME analytical column and an Agilent Zorbax SB-AQ (5 micron, 2.1 x 12.5 mm) guard CH:COLUMN_NAME column CH:FLOW_RATE 0.3ml/min #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6520 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:CAPILLARY_VOLTAGE 4000V MS:DESOLVATION_GAS_FLOW 10L/min MS:DESOLVATION_TEMPERATURE 300C MS:MS_RESULTS_FILE ST000625_AN000957_Results.txt UNITS:peak intensity #END