#METABOLOMICS WORKBENCH Poonam27_20211124_230224 DATATRACK_ID:2946 STUDY_ID:ST002165 ANALYSIS_ID:AN003548 PROJECT_ID:PR001378 VERSION 1 CREATED_ON November 29, 2021, 2:16 am #PROJECT PR:PROJECT_TITLE Untargeted metabolomics following radiation exposure PR:PROJECT_TYPE Untargeted metabolic profiling PR:PROJECT_SUMMARY Comprehensive metabolomics profiling of mice urine after 5 Gy and 7.5 Gy total PR:PROJECT_SUMMARY body radiation exposure at 24 hours using LC-MS and NMR. PR:INSTITUTE Institute of Nuclear Medicine and Allied Sciences PR:DEPARTMENT Metabolomics facility PR:LABORATORY Metabolomics facility PR:LAST_NAME Rana PR:FIRST_NAME Poonam PR:ADDRESS DRDO, Lucknow Road, Timarpur, Delhi, Delhi, 110054, India PR:EMAIL poonam@inmas.drdo.in PR:PHONE +911123905319 PR:FUNDING_SOURCE INM 313, UGC-JRF #STUDY ST:STUDY_TITLE Untargeted metabolomics profiling of mice urine after 5 Gy and 7.5 Gy total body ST:STUDY_TITLE radiation exposure at 24 hours. ST:STUDY_TYPE Untargeted LCMS ST:STUDY_SUMMARY Untargeted metabolomics profiling of mice urine after 5 Gy and 7.5 Gy total body ST:STUDY_SUMMARY radiation exposure at 24 hours. Metabolic profiling of urine samples were ST:STUDY_SUMMARY performed by TripleTOF 5600+ (AB Sciex) in both ionization mode ST:INSTITUTE Institute of Nuclear Medicine and Allied Sciences, Defence Research and ST:INSTITUTE Development Organisation, INDIA ST:DEPARTMENT Institute of Nuclear Medicine and Allied Sciences ST:LABORATORY Metabolomics facility ST:LAST_NAME Rana ST:FIRST_NAME Poonam ST:ADDRESS DRDO, Lucknow Road, Timarpur ST:EMAIL poonamrana27@gmail.com ST:PHONE +911123905319 ST:NUM_GROUPS 3 ST:TOTAL_SUBJECTS 21 ST:NUM_MALES 21 #SUBJECT SU:SUBJECT_TYPE Other organism SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL6 SU:AGE_OR_AGE_RANGE 8-10 weeks SU:GENDER Male SU:SPECIES_GROUP 3 #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 - R_ QC5 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC6 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC7 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC8 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC9 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC10 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC11 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC12 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC13 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC14 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC15 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_ QC16 Treatment:QC SUBJECT_SAMPLE_FACTORS - R_Ctrl 1 Treatment:Control SUBJECT_SAMPLE_FACTORS - R_Ctrl 2 Treatment:Control SUBJECT_SAMPLE_FACTORS - R_Ctrl 3 Treatment:Control SUBJECT_SAMPLE_FACTORS - R_Ctrl 4 Treatment:Control SUBJECT_SAMPLE_FACTORS - R_Ctrl 5 Treatment:Control SUBJECT_SAMPLE_FACTORS - R_Ctrl 6 Treatment:Control SUBJECT_SAMPLE_FACTORS - R_Ctrl 7 Treatment:Control SUBJECT_SAMPLE_FACTORS - R_5 Gy 1 Treatment:5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_5 Gy 2 Treatment:5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_5 Gy 3 Treatment:5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_5 Gy 4 Treatment:5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_5 Gy 5 Treatment:5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_5 Gy 6 Treatment:5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_5 Gy 7 Treatment:5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_7.5 Gy 1 Treatment:7.5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_7.5 Gy 2 Treatment:7.5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_7.5 Gy 3 Treatment:7.5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_7.5 Gy 4 Treatment:7.5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_7.5 Gy 6 Treatment:7.5 Gy Irradiated SUBJECT_SAMPLE_FACTORS - R_7.5 Gy 7 Treatment:7.5 Gy Irradiated #COLLECTION CO:COLLECTION_SUMMARY Animals were acclimatized for 48 hrs in polypropylene cages. Urine samples were CO:COLLECTION_SUMMARY collected at 24 hrs post radiation exposure. CO:SAMPLE_TYPE Urine CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_TUBE_TEMP 4℃ #TREATMENT TR:TREATMENT_SUMMARY Random categorization was done into three groups: control, 5 Gy (sub-lethal) and TR:TREATMENT_SUMMARY 7.5 Gy (lethal) irradiated (seven animals in each group). Mice were kept for 12 TR:TREATMENT_SUMMARY hrs light and dark cycle with room temperature (19–23°C) and humidity TR:TREATMENT_SUMMARY (45–65%) conditions properly maintained. Mice were exposed to total-body gamma TR:TREATMENT_SUMMARY radiation (field size; 35x35 cm) of 5 Gy and 7.5 Gy through Tele 60Co unit γ- TR:TREATMENT_SUMMARY irradiation facility (Bhabhatron II, Panacea Medical, Bangalore, India) with TR:TREATMENT_SUMMARY dose rate of 1 Gy/min. Control mice were sham irradiated. TR:TREATMENT Radiation exposure TR:TREATMENT_DOSE 5 Gy and 7.5 Gy Gamma-60Cobalt irradiation #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Urine samples (50 uL) were added to 200ul of water containing internal standards SP:SAMPLEPREP_SUMMARY (caffeine, nitrobenzoic acid and acetaminophen) and were filtered through 3 kDa SP:SAMPLEPREP_SUMMARY filter. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACT_STORAGE On ice #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME ExionLC CH:COLUMN_NAME Kinetex EVO C18 column (Phenomenex, Torrance, CA, USA) having dimensions 100 mm CH:COLUMN_NAME x 2.1 mm, 1.7 um #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 5600 TripleTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Metabolites were identified by matching with, company provided library and MS:MS_COMMENTS Metlin public database (https://metlin.scripps.edu) MS:MS_RESULTS_FILE ST002165_AN003548_Results.txt UNITS:Intensity Has m/z:Yes Has RT:Yes RT units:Minutes #END