#METABOLOMICS WORKBENCH hwelhaven_20220421_073140 DATATRACK_ID:3212 STUDY_ID:ST002146 ANALYSIS_ID:AN003515 PROJECT_ID:PR001360 VERSION 1 CREATED_ON April 21, 2022, 9:29 am #PROJECT PR:PROJECT_TITLE Welhaven_JBMR+_data PR:PROJECT_SUMMARY This experiment investigated metabolomic differences between male and female PR:PROJECT_SUMMARY musculoskeletal tissues. PR:INSTITUTE Montana State University PR:LAST_NAME Welhaven PR:FIRST_NAME Hope PR:ADDRESS Culbertson Hall, 100 PR:EMAIL hwelhaven@gmail.com PR:PHONE 4066961526 PR:FUNDING_SOURCE National Science Foundation (CMMI 1554708, CMMI 2120239) and the National PR:FUNDING_SOURCE Institutes of Health (NIAMS R01AR073964, NIGMS P20GM103474, NIH R03AG068680 #STUDY ST:STUDY_TITLE Metabolomic differences detected in C57BL/6 mice ST:STUDY_SUMMARY Mice were euthanized at 20 weeks of age, and various tissues were obtained for ST:STUDY_SUMMARY metabolomic analyses including whole bone, bone marrow, and isolated cortical ST:STUDY_SUMMARY bone. Metabolites were extracted and underwent LC-MS analysis using an Agilent ST:STUDY_SUMMARY LC 1290 coupled to an Agilent 6538 QTOF ST:INSTITUTE Montana State University ST:LAST_NAME Welhaven ST:FIRST_NAME Hope ST:ADDRESS Culbertson Hall, 100 ST:EMAIL hwelhaven@gmail.com ST:PHONE 4066961526 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 20 ST:NUM_MALES 10 ST:NUM_FEMALES 10 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57Bl/6 SU:AGE_OR_AGE_RANGE 20 weeks (at euthanasia) SU:GENDER Male and female SU:ANIMAL_ANIMAL_SUPPLIER Jackson Labratories SU:ANIMAL_LIGHT_CYCLE Standard light dark cycle SU:ANIMAL_FEED PicoLab Rodent Diet 20, 20% protein #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - H36 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H36 SUBJECT_SAMPLE_FACTORS - H35 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H35 SUBJECT_SAMPLE_FACTORS - H34 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H34 SUBJECT_SAMPLE_FACTORS - H33 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H33 SUBJECT_SAMPLE_FACTORS - H32 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H32 SUBJECT_SAMPLE_FACTORS - H31 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H31 SUBJECT_SAMPLE_FACTORS - H27 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H27 SUBJECT_SAMPLE_FACTORS - H30 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H30 SUBJECT_SAMPLE_FACTORS - H29 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H29 SUBJECT_SAMPLE_FACTORS - H28 Sex:F | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H28 SUBJECT_SAMPLE_FACTORS - H10 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H10 SUBJECT_SAMPLE_FACTORS - H8 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H8 SUBJECT_SAMPLE_FACTORS - H5 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H5 SUBJECT_SAMPLE_FACTORS - H9 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H9 SUBJECT_SAMPLE_FACTORS - H13 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H13 SUBJECT_SAMPLE_FACTORS - H4 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H4 SUBJECT_SAMPLE_FACTORS - H6 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H6 SUBJECT_SAMPLE_FACTORS - H12 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H12 SUBJECT_SAMPLE_FACTORS - H11 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H11 SUBJECT_SAMPLE_FACTORS - H7 Sex:M | Tissue:isolated_bone RAW_FILE_NAME=Welhaven_H7 #COLLECTION CO:COLLECTION_SUMMARY Samples were collected following mouse euthanasia. The humerus was obtained and CO:COLLECTION_SUMMARY bone marrow was flushed. CO:SAMPLE_TYPE Bone #TREATMENT TR:TREATMENT_SUMMARY Mice were housed at Montana State University after being obtained from Jackson TR:TREATMENT_SUMMARY labs. During this time, mice were kept on a light/dark cycle and fed a standard TR:TREATMENT_SUMMARY chow diet ad libitum. Upon 20 weeks of age, mice were euthanized. No additional TR:TREATMENT_SUMMARY factors such as injury, diet, or exercise occurred. Mice were simply used to TR:TREATMENT_SUMMARY analyze sex differences as well as differences in metabolism between TR:TREATMENT_SUMMARY musculoskeletal tissues. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Once euthanized, tissues were dissected and harvested. Samples underwent a SP:SAMPLEPREP_SUMMARY methanol:acetone extraction and underwent rounds of centrifugation. Once SP:SAMPLEPREP_SUMMARY metabolites were isolated, samples were dried via vacuum concentration and SP:SAMPLEPREP_SUMMARY resuspended with acetonitrile:water for LC-MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Samples were analyzed using an Agilent 1290 UPLC coupled to an Agilent 6538 CH:CHROMATOGRAPHY_SUMMARY QTOF. All samples were run in normal phase using a Cogent Diamond Hydride HILIC CH:CHROMATOGRAPHY_SUMMARY column (150 x 2.1 mm). Solvents: 0.1% formic acid in water (A), 0.1% formic acid CH:CHROMATOGRAPHY_SUMMARY in acetonitrile (B). Elution gradient: 100 to 25% solvent B over 17 minutes, 0 CH:CHROMATOGRAPHY_SUMMARY to 70% solvent A over 20 minutes, at 21 minutes, B increases from 25% to 100% CH:CHROMATOGRAPHY_SUMMARY till 25 minutes while A decreases from 70% to 0% till 25 minutes. The last 4 CH:CHROMATOGRAPHY_SUMMARY minutes act as a wash. Total run time = 25 minute method (per sample) CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 6538 CH:COLUMN_NAME Cogent Diamond Hydride HILIC 150 × 2.1 mm column #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6538 QTOF MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS - MS:MS_RESULTS_FILE ST002146_AN003515_Results.txt UNITS:Peak Intensiy Has m/z:Yes Has RT:No RT units:No RT data #END