#METABOLOMICS WORKBENCH niemuthn_20210518_075653 DATATRACK_ID:2645 STUDY_ID:ST001807 ANALYSIS_ID:AN002930 VERSION 1 CREATED_ON 02-08-2024 #PROJECT PR:PROJECT_TITLE Energy starvation in Daphnia magna from exposure to a lithium cobalt oxide PR:PROJECT_TITLE nanomaterial PR:PROJECT_TYPE Untargeted MS PR:PROJECT_SUMMARY Our previous research demonstrated that energy metabolism is significantly PR:PROJECT_SUMMARY impacted in sediment-dwelling invertebrate Chironomus riparius upon exposure to PR:PROJECT_SUMMARY metal oxide nanomaterial lithium cobalt oxide (LCO), used as a cathode material PR:PROJECT_SUMMARY in lithium-ion batteries, an impact that is not replicated by ion controls. To PR:PROJECT_SUMMARY further explore metabolic impacts, we determined LCO’s effects on model PR:PROJECT_SUMMARY organism Daphnia magna using untargeted metabolomics. Our results show a PR:PROJECT_SUMMARY sublethal 1 mg/L 48 h LCO exposure causes significant impacts on D. magna PR:PROJECT_SUMMARY metabolites, while ion control exposure equivalent to released Li and Co had no PR:PROJECT_SUMMARY impact, showing the nano-specificity of LCO impacts. Specifically, metabolomic PR:PROJECT_SUMMARY analysis indicated alteration of metabolites related to amino acid metabolism PR:PROJECT_SUMMARY and starch, sucrose, and galactose metabolism as a result of LCO exposure. PR:INSTITUTE University of Wisconsin - Milwaukee PR:DEPARTMENT School of Freshwater Sciences PR:LABORATORY Rebecca Klaper PR:LAST_NAME Niemuth PR:FIRST_NAME Nicholas PR:ADDRESS 600 E Greenfield Ave, Milwaukee, WI, 53204, USA PR:EMAIL niemuthn@uwm.edu PR:PHONE 4143821763 PR:FUNDING_SOURCE National Science Foundation CHE-2001611 PR:DOI http://dx.doi.org/10.21228/M8711S PR:CONTRIBUTORS Nicholas J Niemuth, Becky J Curtis, Elizabeth D Laudadio, Jelena Sostare, Evan A PR:CONTRIBUTORS Bennett, Nicklaus J Neureuther, Mark R Viant, Robert J Hamers, Rebecca D Klaper #STUDY ST:STUDY_TITLE Untargeted metabolomics of Daphnia magna exposed to a lithium cobalt oxide ST:STUDY_TITLE nanomaterial ST:STUDY_TYPE Untargeted MS ST:STUDY_SUMMARY The goal of this project was to determine lithium cobalt oxide (LCO)’s effects ST:STUDY_SUMMARY on pathways in the model organism Daphnia magna through untargeted metabolomics. ST:INSTITUTE University of Wisconsin - Milwaukee ST:DEPARTMENT School of Freshwater Sciences ST:LABORATORY Rebecca Klaper ST:LAST_NAME Klaper ST:FIRST_NAME Rebecca ST:ADDRESS 600 E Greenfield Ave, Milwaukee, WI 53204 ST:EMAIL rklaper@uwm.edu ST:PHONE 4143821713 ST:SUBMIT_DATE 2021-05-18 #SUBJECT SU:SUBJECT_TYPE Invertebrate SU:SUBJECT_SPECIES Daphnia magna SU:TAXONOMY_ID 35525 SU:AGE_OR_AGE_RANGE neonates < 9 h old #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS ion3 Dm_polneg_Ion_1_14 characteristics:Ion Loading order=14; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_14_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_14_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_14_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_14_R4 SUBJECT_SAMPLE_FACTORS ion4 Dm_polneg_Ion_1_19 characteristics:Ion Loading order=19; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_19_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_19_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_19_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_19_R4 SUBJECT_SAMPLE_FACTORS ion5 Dm_polneg_Ion_1_24 characteristics:Ion Loading order=24; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_24_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_24_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_24_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_24_R4 SUBJECT_SAMPLE_FACTORS ion6 Dm_polneg_Ion_1_29 characteristics:Ion Loading order=29; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_29_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_29_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_29_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_29_R4 SUBJECT_SAMPLE_FACTORS ion7 Dm_polneg_Ion_1_34 characteristics:Ion Loading order=34; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_34_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_34_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_34_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_34_R4 SUBJECT_SAMPLE_FACTORS ion8 Dm_polneg_Ion_1_39 characteristics:Ion Loading order=39; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_39_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_39_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_39_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_39_R4 SUBJECT_SAMPLE_FACTORS ion1 Dm_polneg_Ion_1_4 characteristics:Ion Loading order=4; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_4_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_4_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_4_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_4_R4 SUBJECT_SAMPLE_FACTORS ion9 Dm_polneg_Ion_1_44 characteristics:Ion Loading order=44; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_44_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_44_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_44_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_44_R4 SUBJECT_SAMPLE_FACTORS ion10 Dm_polneg_Ion_1_49 characteristics:Ion Loading order=49; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_49_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_49_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_49_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_49_R4 SUBJECT_SAMPLE_FACTORS ion2 Dm_polneg_Ion_1_9 characteristics:Ion Loading order=9; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_Ion_1_9_R1; RAW_FILE_NAME=Dm_polneg_Ion_1_9_R2; RAW_FILE_NAME=Dm_polneg_Ion_1_9_R3; RAW_FILE_NAME=Dm_polneg_Ion_1_9_R4 SUBJECT_SAMPLE_FACTORS ion3 Dm_polpos_Ion_1_14 characteristics:Ion Loading order=14; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_14_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_14_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_14_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_14_R4 SUBJECT_SAMPLE_FACTORS ion4 Dm_polpos_Ion_1_19 characteristics:Ion Loading order=19; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_19_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_19_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_19_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_19_R4 SUBJECT_SAMPLE_FACTORS ion5 Dm_polpos_Ion_1_24 characteristics:Ion Loading order=24; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_24_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_24_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_24_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_24_R4 SUBJECT_SAMPLE_FACTORS ion6 Dm_polpos_Ion_1_29 characteristics:Ion Loading order=29; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_29_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_29_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_29_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_29_R4 SUBJECT_SAMPLE_FACTORS ion7 Dm_polpos_Ion_1_34 characteristics:Ion Loading order=34; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_34_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_34_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_34_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_34_R4 SUBJECT_SAMPLE_FACTORS ion8 Dm_polpos_Ion_1_39 characteristics:Ion Loading order=39; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_39_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_39_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_39_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_39_R4 SUBJECT_SAMPLE_FACTORS ion1 Dm_polpos_Ion_1_4 characteristics:Ion Loading order=4; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_4_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_4_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_4_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_4_R4 SUBJECT_SAMPLE_FACTORS ion9 Dm_polpos_Ion_1_44 characteristics:Ion Loading order=44; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_44_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_44_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_44_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_44_R4 SUBJECT_SAMPLE_FACTORS ion10 Dm_polpos_Ion_1_49 characteristics:Ion Loading order=49; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_49_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_49_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_49_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_49_R4 SUBJECT_SAMPLE_FACTORS ion2 Dm_polpos_Ion_1_9 characteristics:Ion Loading order=9; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_Ion_1_9_R1; RAW_FILE_NAME=Dm_polpos_Ion_1_9_R2; RAW_FILE_NAME=Dm_polpos_Ion_1_9_R3; RAW_FILE_NAME=Dm_polpos_Ion_1_9_R4 SUBJECT_SAMPLE_FACTORS low3 Dm_polneg_LCO_01_12 characteristics:LCO 0.1 mg/L Loading order=12; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_12_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_12_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_12_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_12_R4 SUBJECT_SAMPLE_FACTORS low4 Dm_polneg_LCO_01_17 characteristics:LCO 0.1 mg/L Loading order=17; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_17_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_17_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_17_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_17_R4 SUBJECT_SAMPLE_FACTORS low1 Dm_polneg_LCO_01_2 characteristics:LCO 0.1 mg/L Loading order=2; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_2_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_2_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_2_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_2_R4 SUBJECT_SAMPLE_FACTORS low5 Dm_polneg_LCO_01_22 characteristics:LCO 0.1 mg/L Loading order=22; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_22_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_22_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_22_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_22_R4 SUBJECT_SAMPLE_FACTORS low6 Dm_polneg_LCO_01_27 characteristics:LCO 0.1 mg/L Loading order=27; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_27_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_27_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_27_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_27_R4 SUBJECT_SAMPLE_FACTORS low7 Dm_polneg_LCO_01_32 characteristics:LCO 0.1 mg/L Loading order=32; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_32_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_32_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_32_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_32_R4 SUBJECT_SAMPLE_FACTORS low8 Dm_polneg_LCO_01_37 characteristics:LCO 0.1 mg/L Loading order=37; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_37_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_37_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_37_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_37_R4 SUBJECT_SAMPLE_FACTORS low9 Dm_polneg_LCO_01_42 characteristics:LCO 0.1 mg/L Loading order=42; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_42_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_42_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_42_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_42_R4 SUBJECT_SAMPLE_FACTORS low10 Dm_polneg_LCO_01_47 characteristics:LCO 0.1 mg/L Loading order=47; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_47_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_47_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_47_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_47_R4 SUBJECT_SAMPLE_FACTORS low2 Dm_polneg_LCO_01_7 characteristics:LCO 0.1 mg/L Loading order=7; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_01_7_R1; RAW_FILE_NAME=Dm_polneg_LCO_01_7_R2; RAW_FILE_NAME=Dm_polneg_LCO_01_7_R3; RAW_FILE_NAME=Dm_polneg_LCO_01_7_R4 SUBJECT_SAMPLE_FACTORS low3 Dm_polpos_LCO_01_12 characteristics:LCO 0.1 mg/L Loading order=12; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_12_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_12_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_12_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_12_R4 SUBJECT_SAMPLE_FACTORS low4 Dm_polpos_LCO_01_17 characteristics:LCO 0.1 mg/L Loading order=17; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_17_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_17_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_17_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_17_R4 SUBJECT_SAMPLE_FACTORS low1 Dm_polpos_LCO_01_2 characteristics:LCO 0.1 mg/L Loading order=2; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_2_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_2_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_2_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_2_R4 SUBJECT_SAMPLE_FACTORS low5 Dm_polpos_LCO_01_22 characteristics:LCO 0.1 mg/L Loading order=22; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_22_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_22_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_22_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_22_R4 SUBJECT_SAMPLE_FACTORS low6 Dm_polpos_LCO_01_27 characteristics:LCO 0.1 mg/L Loading order=27; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_27_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_27_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_27_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_27_R4 SUBJECT_SAMPLE_FACTORS low7 Dm_polpos_LCO_01_32 characteristics:LCO 0.1 mg/L Loading order=32; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_32_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_32_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_32_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_32_R4 SUBJECT_SAMPLE_FACTORS low8 Dm_polpos_LCO_01_37 characteristics:LCO 0.1 mg/L Loading order=37; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_37_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_37_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_37_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_37_R4 SUBJECT_SAMPLE_FACTORS low9 Dm_polpos_LCO_01_42 characteristics:LCO 0.1 mg/L Loading order=42; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_42_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_42_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_42_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_42_R4 SUBJECT_SAMPLE_FACTORS low10 Dm_polpos_LCO_01_47 characteristics:LCO 0.1 mg/L Loading order=47; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_47_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_47_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_47_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_47_R4 SUBJECT_SAMPLE_FACTORS low2 Dm_polpos_LCO_01_7 characteristics:LCO 0.1 mg/L Loading order=7; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_01_7_R1; RAW_FILE_NAME=Dm_polpos_LCO_01_7_R2; RAW_FILE_NAME=Dm_polpos_LCO_01_7_R3; RAW_FILE_NAME=Dm_polpos_LCO_01_7_R4 SUBJECT_SAMPLE_FACTORS high3 Dm_polneg_LCO_1_13 characteristics:LCO 1 mg/L Loading order=13; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_13_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_13_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_13_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_13_R4 SUBJECT_SAMPLE_FACTORS high4 Dm_polneg_LCO_1_18 characteristics:LCO 1 mg/L Loading order=18; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_18_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_18_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_18_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_18_R4 SUBJECT_SAMPLE_FACTORS high5 Dm_polneg_LCO_1_23 characteristics:LCO 1 mg/L Loading order=23; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_23_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_23_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_23_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_23_R4 SUBJECT_SAMPLE_FACTORS high6 Dm_polneg_LCO_1_28 characteristics:LCO 1 mg/L Loading order=28; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_28_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_28_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_28_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_28_R4 SUBJECT_SAMPLE_FACTORS high1 Dm_polneg_LCO_1_3 characteristics:LCO 1 mg/L Loading order=3; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_3_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_3_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_3_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_3_R4 SUBJECT_SAMPLE_FACTORS high7 Dm_polneg_LCO_1_33 characteristics:LCO 1 mg/L Loading order=33; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_33_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_33_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_33_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_33_R4 SUBJECT_SAMPLE_FACTORS high8 Dm_polneg_LCO_1_38 characteristics:LCO 1 mg/L Loading order=38; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_38_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_38_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_38_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_38_R4 SUBJECT_SAMPLE_FACTORS high9 Dm_polneg_LCO_1_43 characteristics:LCO 1 mg/L Loading order=43; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_43_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_43_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_43_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_43_R4 SUBJECT_SAMPLE_FACTORS high10 Dm_polneg_LCO_1_48 characteristics:LCO 1 mg/L Loading order=48; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_48_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_48_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_48_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_48_R4 SUBJECT_SAMPLE_FACTORS high2 Dm_polneg_LCO_1_8 characteristics:LCO 1 mg/L Loading order=8; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_LCO_1_8_R1; RAW_FILE_NAME=Dm_polneg_LCO_1_8_R2; RAW_FILE_NAME=Dm_polneg_LCO_1_8_R3; RAW_FILE_NAME=Dm_polneg_LCO_1_8_R4 SUBJECT_SAMPLE_FACTORS high3 Dm_polpos_LCO_1_13 characteristics:LCO 1 mg/L Loading order=13; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_13_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_13_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_13_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_13_R4 SUBJECT_SAMPLE_FACTORS high4 Dm_polpos_LCO_1_18 characteristics:LCO 1 mg/L Loading order=18; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_18_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_18_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_18_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_18_R4 SUBJECT_SAMPLE_FACTORS high5 Dm_polpos_LCO_1_28 characteristics:LCO 1 mg/L Loading order=28; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_28_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_28_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_28_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_28_R4 SUBJECT_SAMPLE_FACTORS high1 Dm_polpos_LCO_1_3 characteristics:LCO 1 mg/L Loading order=3; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_3_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_3_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_3_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_3_R4 SUBJECT_SAMPLE_FACTORS high6 Dm_polpos_LCO_1_33 characteristics:LCO 1 mg/L Loading order=33; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_33_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_33_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_33_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_33_R4 SUBJECT_SAMPLE_FACTORS high7 Dm_polpos_LCO_1_38 characteristics:LCO 1 mg/L Loading order=38; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_38_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_38_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_38_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_38_R4 SUBJECT_SAMPLE_FACTORS high8 Dm_polpos_LCO_1_43 characteristics:LCO 1 mg/L Loading order=43; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_43_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_43_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_43_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_43_R4 SUBJECT_SAMPLE_FACTORS high9 Dm_polpos_LCO_1_48 characteristics:LCO 1 mg/L Loading order=48; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_48_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_48_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_48_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_48_R4 SUBJECT_SAMPLE_FACTORS high2 Dm_polpos_LCO_1_8 characteristics:LCO 1 mg/L Loading order=8; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_LCO_1_8_R1; RAW_FILE_NAME=Dm_polpos_LCO_1_8_R2; RAW_FILE_NAME=Dm_polpos_LCO_1_8_R3; RAW_FILE_NAME=Dm_polpos_LCO_1_8_R4 SUBJECT_SAMPLE_FACTORS qc1 Dm_polneg_QC_1 characteristics:Quality control Loading order=1; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_1_R1; RAW_FILE_NAME=Dm_polneg_QC_1_R2; RAW_FILE_NAME=Dm_polneg_QC_1_R3; RAW_FILE_NAME=Dm_polneg_QC_1_R4 SUBJECT_SAMPLE_FACTORS qc10 Dm_polneg_QC_10 characteristics:Quality control Loading order=10; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_10_R1; RAW_FILE_NAME=Dm_polneg_QC_10_R2; RAW_FILE_NAME=Dm_polneg_QC_10_R3; RAW_FILE_NAME=Dm_polneg_QC_10_R4 SUBJECT_SAMPLE_FACTORS qc11 Dm_polneg_QC_11 characteristics:Quality control Loading order=11; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_11_R1; RAW_FILE_NAME=Dm_polneg_QC_11_R2; RAW_FILE_NAME=Dm_polneg_QC_11_R3; RAW_FILE_NAME=Dm_polneg_QC_11_R4 SUBJECT_SAMPLE_FACTORS qc12 Dm_polneg_QC_12 characteristics:Quality control Loading order=12; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_12_R1; RAW_FILE_NAME=Dm_polneg_QC_12_R2; RAW_FILE_NAME=Dm_polneg_QC_12_R3; RAW_FILE_NAME=Dm_polneg_QC_12_R4 SUBJECT_SAMPLE_FACTORS qc13 Dm_polneg_QC_13 characteristics:Quality control Loading order=13; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_13_R1; RAW_FILE_NAME=Dm_polneg_QC_13_R2; RAW_FILE_NAME=Dm_polneg_QC_13_R3; RAW_FILE_NAME=Dm_polneg_QC_13_R4 SUBJECT_SAMPLE_FACTORS qc14 Dm_polneg_QC_14 characteristics:Quality control Loading order=14; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_14_R1; RAW_FILE_NAME=Dm_polneg_QC_14_R2; RAW_FILE_NAME=Dm_polneg_QC_14_R3; RAW_FILE_NAME=Dm_polneg_QC_14_R4 SUBJECT_SAMPLE_FACTORS qc2 Dm_polneg_QC_2 characteristics:Quality control Loading order=2; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_2_R1; RAW_FILE_NAME=Dm_polneg_QC_2_R2; RAW_FILE_NAME=Dm_polneg_QC_2_R3; RAW_FILE_NAME=Dm_polneg_QC_2_R4 SUBJECT_SAMPLE_FACTORS qc3 Dm_polneg_QC_3 characteristics:Quality control Loading order=3; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_3_R1; RAW_FILE_NAME=Dm_polneg_QC_3_R2; RAW_FILE_NAME=Dm_polneg_QC_3_R3; RAW_FILE_NAME=Dm_polneg_QC_3_R4 SUBJECT_SAMPLE_FACTORS qc4 Dm_polneg_QC_4 characteristics:Quality control Loading order=4; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_4_R1; RAW_FILE_NAME=Dm_polneg_QC_4_R2; RAW_FILE_NAME=Dm_polneg_QC_4_R3; RAW_FILE_NAME=Dm_polneg_QC_4_R4 SUBJECT_SAMPLE_FACTORS qc5 Dm_polneg_QC_5 characteristics:Quality control Loading order=5; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_5_R1; RAW_FILE_NAME=Dm_polneg_QC_5_R2; RAW_FILE_NAME=Dm_polneg_QC_5_R3; RAW_FILE_NAME=Dm_polneg_QC_5_R4 SUBJECT_SAMPLE_FACTORS qc6 Dm_polneg_QC_6 characteristics:Quality control Loading order=6; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_6_R1; RAW_FILE_NAME=Dm_polneg_QC_6_R2; RAW_FILE_NAME=Dm_polneg_QC_6_R3; RAW_FILE_NAME=Dm_polneg_QC_6_R4 SUBJECT_SAMPLE_FACTORS qc7 Dm_polneg_QC_7 characteristics:Quality control Loading order=7; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_7_R1; RAW_FILE_NAME=Dm_polneg_QC_7_R2; RAW_FILE_NAME=Dm_polneg_QC_7_R3; RAW_FILE_NAME=Dm_polneg_QC_7_R4 SUBJECT_SAMPLE_FACTORS qc8 Dm_polneg_QC_8 characteristics:Quality control Loading order=8; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_8_R1; RAW_FILE_NAME=Dm_polneg_QC_8_R2; RAW_FILE_NAME=Dm_polneg_QC_8_R3; RAW_FILE_NAME=Dm_polneg_QC_8_R4 SUBJECT_SAMPLE_FACTORS qc9 Dm_polneg_QC_9 characteristics:Quality control Loading order=9; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_QC_9_R1; RAW_FILE_NAME=Dm_polneg_QC_9_R2; RAW_FILE_NAME=Dm_polneg_QC_9_R3; RAW_FILE_NAME=Dm_polneg_QC_9_R4 SUBJECT_SAMPLE_FACTORS qc1 Dm_polpos_QC_1 characteristics:Quality control Loading order=1; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_1_R1; RAW_FILE_NAME=Dm_polpos_QC_1_R2; RAW_FILE_NAME=Dm_polpos_QC_1_R3; RAW_FILE_NAME=Dm_polpos_QC_1_R4 SUBJECT_SAMPLE_FACTORS qc10 Dm_polpos_QC_10 characteristics:Quality control Loading order=10; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_10_R1; RAW_FILE_NAME=Dm_polpos_QC_10_R2; RAW_FILE_NAME=Dm_polpos_QC_10_R3; RAW_FILE_NAME=Dm_polpos_QC_10_R4 SUBJECT_SAMPLE_FACTORS qc11 Dm_polpos_QC_11 characteristics:Quality control Loading order=11; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_11_R1; RAW_FILE_NAME=Dm_polpos_QC_11_R2; RAW_FILE_NAME=Dm_polpos_QC_11_R3; RAW_FILE_NAME=Dm_polpos_QC_11_R4 SUBJECT_SAMPLE_FACTORS qc12 Dm_polpos_QC_12 characteristics:Quality control Loading order=12; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_12_R1; RAW_FILE_NAME=Dm_polpos_QC_12_R2; RAW_FILE_NAME=Dm_polpos_QC_12_R3; RAW_FILE_NAME=Dm_polpos_QC_12_R4 SUBJECT_SAMPLE_FACTORS qc13 Dm_polpos_QC_13 characteristics:Quality control Loading order=13; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_13_R1; RAW_FILE_NAME=Dm_polpos_QC_13_R2; RAW_FILE_NAME=Dm_polpos_QC_13_R3; RAW_FILE_NAME=Dm_polpos_QC_13_R4 SUBJECT_SAMPLE_FACTORS qc14 Dm_polpos_QC_14 characteristics:Quality control Loading order=14; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_14_R1; RAW_FILE_NAME=Dm_polpos_QC_14_R2; RAW_FILE_NAME=Dm_polpos_QC_14_R3; RAW_FILE_NAME=Dm_polpos_QC_14_R4 SUBJECT_SAMPLE_FACTORS qc2 Dm_polpos_QC_2 characteristics:Quality control Loading order=2; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_2_R1; RAW_FILE_NAME=Dm_polpos_QC_2_R2; RAW_FILE_NAME=Dm_polpos_QC_2_R3; RAW_FILE_NAME=Dm_polpos_QC_2_R4 SUBJECT_SAMPLE_FACTORS qc3 Dm_polpos_QC_3 characteristics:Quality control Loading order=3; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_3_R1; RAW_FILE_NAME=Dm_polpos_QC_3_R2; RAW_FILE_NAME=Dm_polpos_QC_3_R3; RAW_FILE_NAME=Dm_polpos_QC_3_R4 SUBJECT_SAMPLE_FACTORS qc4 Dm_polpos_QC_4 characteristics:Quality control Loading order=4; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_4_R1; RAW_FILE_NAME=Dm_polpos_QC_4_R2; RAW_FILE_NAME=Dm_polpos_QC_4_R3; RAW_FILE_NAME=Dm_polpos_QC_4_R4 SUBJECT_SAMPLE_FACTORS qc5 Dm_polpos_QC_5 characteristics:Quality control Loading order=5; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_5_R1; RAW_FILE_NAME=Dm_polpos_QC_5_R2; RAW_FILE_NAME=Dm_polpos_QC_5_R3; RAW_FILE_NAME=Dm_polpos_QC_5_R4 SUBJECT_SAMPLE_FACTORS qc6 Dm_polpos_QC_6 characteristics:Quality control Loading order=6; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_6_R1; RAW_FILE_NAME=Dm_polpos_QC_6_R2; RAW_FILE_NAME=Dm_polpos_QC_6_R3; RAW_FILE_NAME=Dm_polpos_QC_6_R4 SUBJECT_SAMPLE_FACTORS qc7 Dm_polpos_QC_7 characteristics:Quality control Loading order=7; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_7_R1; RAW_FILE_NAME=Dm_polpos_QC_7_R2; RAW_FILE_NAME=Dm_polpos_QC_7_R3; RAW_FILE_NAME=Dm_polpos_QC_7_R4 SUBJECT_SAMPLE_FACTORS qc8 Dm_polpos_QC_8 characteristics:Quality control Loading order=8; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_8_R1; RAW_FILE_NAME=Dm_polpos_QC_8_R2; RAW_FILE_NAME=Dm_polpos_QC_8_R3; RAW_FILE_NAME=Dm_polpos_QC_8_R4 SUBJECT_SAMPLE_FACTORS qc9 Dm_polpos_QC_9 characteristics:Quality control Loading order=9; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_QC_9_R1; RAW_FILE_NAME=Dm_polpos_QC_9_R2; RAW_FILE_NAME=Dm_polpos_QC_9_R3; RAW_FILE_NAME=Dm_polpos_QC_9_R4 SUBJECT_SAMPLE_FACTORS control2 Dm_polneg_C_11 characteristics:untreated Loading order=11; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_11_R1; RAW_FILE_NAME=Dm_polneg_C_11_R2; RAW_FILE_NAME=Dm_polneg_C_11_R3; RAW_FILE_NAME=Dm_polneg_C_11_R4 SUBJECT_SAMPLE_FACTORS control3 Dm_polneg_C_16 characteristics:untreated Loading order=16; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_16_R1; RAW_FILE_NAME=Dm_polneg_C_16_R2; RAW_FILE_NAME=Dm_polneg_C_16_R3; RAW_FILE_NAME=Dm_polneg_C_16_R4 SUBJECT_SAMPLE_FACTORS control4 Dm_polneg_C_21 characteristics:untreated Loading order=21; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_21_R1; RAW_FILE_NAME=Dm_polneg_C_21_R2; RAW_FILE_NAME=Dm_polneg_C_21_R3; RAW_FILE_NAME=Dm_polneg_C_21_R4 SUBJECT_SAMPLE_FACTORS control5 Dm_polneg_C_26 characteristics:untreated Loading order=26; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_26_R1; RAW_FILE_NAME=Dm_polneg_C_26_R2; RAW_FILE_NAME=Dm_polneg_C_26_R3; RAW_FILE_NAME=Dm_polneg_C_26_R4 SUBJECT_SAMPLE_FACTORS control6 Dm_polneg_C_31 characteristics:untreated Loading order=31; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_31_R1; RAW_FILE_NAME=Dm_polneg_C_31_R2; RAW_FILE_NAME=Dm_polneg_C_31_R3; RAW_FILE_NAME=Dm_polneg_C_31_R4 SUBJECT_SAMPLE_FACTORS control7 Dm_polneg_C_36 characteristics:untreated Loading order=36; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_36_R1; RAW_FILE_NAME=Dm_polneg_C_36_R2; RAW_FILE_NAME=Dm_polneg_C_36_R3; RAW_FILE_NAME=Dm_polneg_C_36_R4 SUBJECT_SAMPLE_FACTORS control8 Dm_polneg_C_41 characteristics:untreated Loading order=41; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_41_R1; RAW_FILE_NAME=Dm_polneg_C_41_R2; RAW_FILE_NAME=Dm_polneg_C_41_R3; RAW_FILE_NAME=Dm_polneg_C_41_R4 SUBJECT_SAMPLE_FACTORS control1 Dm_polneg_C_6 characteristics:untreated Loading order=6; ion mode=negative; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polneg_C_6_R1; RAW_FILE_NAME=Dm_polneg_C_6_R2; RAW_FILE_NAME=Dm_polneg_C_6_R3; RAW_FILE_NAME=Dm_polneg_C_6_R4 SUBJECT_SAMPLE_FACTORS control2 Dm_polpos_C_11 characteristics:untreated Loading order=11; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_11_R1; RAW_FILE_NAME=Dm_polpos_C_11_R2; RAW_FILE_NAME=Dm_polpos_C_11_R3; RAW_FILE_NAME=Dm_polpos_C_11_R4 SUBJECT_SAMPLE_FACTORS control3 Dm_polpos_C_16 characteristics:untreated Loading order=16; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_16_R1; RAW_FILE_NAME=Dm_polpos_C_16_R2; RAW_FILE_NAME=Dm_polpos_C_16_R3; RAW_FILE_NAME=Dm_polpos_C_16_R4 SUBJECT_SAMPLE_FACTORS control4 Dm_polpos_C_21 characteristics:untreated Loading order=21; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_21_R1; RAW_FILE_NAME=Dm_polpos_C_21_R2; RAW_FILE_NAME=Dm_polpos_C_21_R3; RAW_FILE_NAME=Dm_polpos_C_21_R4 SUBJECT_SAMPLE_FACTORS control5 Dm_polpos_C_26 characteristics:untreated Loading order=26; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_26_R1; RAW_FILE_NAME=Dm_polpos_C_26_R2; RAW_FILE_NAME=Dm_polpos_C_26_R3; RAW_FILE_NAME=Dm_polpos_C_26_R4 SUBJECT_SAMPLE_FACTORS control6 Dm_polpos_C_31 characteristics:untreated Loading order=31; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_31_R1; RAW_FILE_NAME=Dm_polpos_C_31_R2; RAW_FILE_NAME=Dm_polpos_C_31_R3; RAW_FILE_NAME=Dm_polpos_C_31_R4 SUBJECT_SAMPLE_FACTORS control7 Dm_polpos_C_36 characteristics:untreated Loading order=36; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_36_R1; RAW_FILE_NAME=Dm_polpos_C_36_R2; RAW_FILE_NAME=Dm_polpos_C_36_R3; RAW_FILE_NAME=Dm_polpos_C_36_R4 SUBJECT_SAMPLE_FACTORS control8 Dm_polpos_C_41 characteristics:untreated Loading order=41; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_41_R1; RAW_FILE_NAME=Dm_polpos_C_41_R2; RAW_FILE_NAME=Dm_polpos_C_41_R3; RAW_FILE_NAME=Dm_polpos_C_41_R4 SUBJECT_SAMPLE_FACTORS control9 Dm_polpos_C_46 characteristics:untreated Loading order=46; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_46_R1; RAW_FILE_NAME=Dm_polpos_C_46_R2; RAW_FILE_NAME=Dm_polpos_C_46_R3; RAW_FILE_NAME=Dm_polpos_C_46_R4 SUBJECT_SAMPLE_FACTORS control1 Dm_polpos_C_6 characteristics:untreated Loading order=6; ion mode=positive; organism=Daphnia magna; characteristics: strain=WT; characteristics: genotype=wild type; characteristics: tissue=whole daphnids; characteristics: age=< 9 h old; characteristics: exposure=48 h; RAW_FILE_NAME=Dm_polpos_C_6_R1; RAW_FILE_NAME=Dm_polpos_C_6_R2; RAW_FILE_NAME=Dm_polpos_C_6_R3; RAW_FILE_NAME=Dm_polpos_C_6_R4 #COLLECTION CO:COLLECTION_SUMMARY Daphnid cultures D. magna were cultured at a density of 20 animals per liter CO:COLLECTION_SUMMARY MHRW, 16 h:8 h light:dark photoperiod, and a temperature of 20 °C. For CO:COLLECTION_SUMMARY metabolomics, cultures were fed daily on suspensions of unicellular green alga, CO:COLLECTION_SUMMARY Chlorella vulgaris (7.84 × 107 cells/mL) at 2 mL/L. Algae was supplemented CO:COLLECTION_SUMMARY daily by 50 µL/L of dried bakers yeast (1mg/mL stock, Sigma–Aldrich). CO:COLLECTION_SUMMARY Cultures were maintained using third or fourth brood neonates less than 24 h CO:COLLECTION_SUMMARY old. Ten replicates were prepared for each treatment: MHRW control, 1 mg/L LCO, CO:COLLECTION_SUMMARY and ion control (66 μg/L Li as LiCl and 15 μg/L Co as CoCl2; equivalent to CO:COLLECTION_SUMMARY ions released by 1 mg/L LCO over 48 h).1,2 Twenty neonates per replicate (< 9 CO:COLLECTION_SUMMARY h old) were transferred to 100 ml beakers 48 hours prior to exposure and fed CO:COLLECTION_SUMMARY proportionate amounts of food for 48 hours. At the end of the 48-hour feeding CO:COLLECTION_SUMMARY period, daphnids were transferred to 200 ml control or exposure beakers. CO:COLLECTION_SUMMARY Exposures were carried out for 48 h without food per standard OECD guidelines CO:COLLECTION_SUMMARY for D. magna. At the end of the exposure daphnids were collected and transferred CO:COLLECTION_SUMMARY (20 pooled animals per replicate) into labeled Precellys tubes using a fine CO:COLLECTION_SUMMARY sable brush and flash frozen in liquid nitrogen.At the end of the exposure CO:COLLECTION_SUMMARY daphnids were collected and transferred (20 pooled animals per replicate) into CO:COLLECTION_SUMMARY labeled Precellys tubes using a fine sable brush and flash frozen in liquid CO:COLLECTION_SUMMARY nitrogen. Samples were stored at -80 °C and shipped to the University of CO:COLLECTION_SUMMARY Birmingham, UK on dry ice. CO:SAMPLE_TYPE Whole animals CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS Precellys tubes #TREATMENT TR:TREATMENT_SUMMARY Ten replicates were prepared for each treatment: MHRW control, 1 mg/L LCO, and TR:TREATMENT_SUMMARY ion control (66 μg/L Li as LiCl and 15 μg/L Co as CoCl2; equivalent to ions TR:TREATMENT_SUMMARY released by 1 mg/L LCO over 48 h).1,2 Twenty neonates per replicate (< 9 h TR:TREATMENT_SUMMARY old) were transferred to 100 ml beakers 48 hours prior to exposure and fed TR:TREATMENT_SUMMARY proportionate amounts of food for 48 hours. At the end of the 48-hour feeding TR:TREATMENT_SUMMARY period, daphnids were transferred to 200 ml control or exposure beakers. TR:TREATMENT_SUMMARY Exposures were carried out for 48 h without food per standard OECD guidelines TR:TREATMENT_SUMMARY for D. magna. TR:TREATMENT_COMPOUND Lithium boalt oxide (LCO) nanosheets TR:TREATMENT_DOSEVOLUME 0.1 and 1 mg/L TR:TREATMENT_DOSEDURATION 48 h #SAMPLEPREP SP:SAMPLEPREP_SUMMARY all solvents were chilled to 4 ºC. A mixture of 320 µL of HPLC grade MeOH and SP:SAMPLEPREP_SUMMARY 128 µL of HPLC grade H2O were added to each sample tube and kept on ice. Tubes SP:SAMPLEPREP_SUMMARY were then placed in a Precellys 24 homogeniser for 2 × 10s bursts at 6400 rpm. SP:SAMPLEPREP_SUMMARY The homogenised mixture was then transferred into a clean 1.8 mL glass vial SP:SAMPLEPREP_SUMMARY (Fisher TUL 520 006 J) using a Pasteur pipette. 320 µL (32 µL/mg) CHCl3 (HPLC SP:SAMPLEPREP_SUMMARY grade) and 160 µL (16 µL/mg) dH2O (HPLC grade) were then added to each vial. SP:SAMPLEPREP_SUMMARY These vials were vortexed on full power for 15 s each to thoroughly mix polar SP:SAMPLEPREP_SUMMARY and non-polar solvents. Vials were then left on ice for on ice for 10 min to SP:SAMPLEPREP_SUMMARY allow initial phase separation. Vials were then centrifuged at 4000 rpm at 4 ºC SP:SAMPLEPREP_SUMMARY for 10 min to ensure complete phase separation. Centrifuged vials were allowed SP:SAMPLEPREP_SUMMARY to come to room temperature by setting them on the lab bench for 5 min. Samples SP:SAMPLEPREP_SUMMARY were then visibly biphasic, with protein debris separating the upper (polar) and SP:SAMPLEPREP_SUMMARY lower (non-polar) layers. A 500 uL Hamilton syringe was then used to remove the SP:SAMPLEPREP_SUMMARY polar phase (~ 2 × 150 µL aliquots) into 2 clean 1.5 mL Eppendorf tubes (one SP:SAMPLEPREP_SUMMARY for positive, one for negative ion analysis). Polar samples were then dried SP:SAMPLEPREP_SUMMARY using a Speed Vac concentrator and stored at -80 °C until analysis. Sample SP:SAMPLEPREP_SUMMARY Preparation and Direct Infusion Mass Spectrometry Metabolomics For positive ion SP:SAMPLEPREP_SUMMARY analysis, 30 µL of 4 ºC 80:20 methanol:water plus 0.25% formic acid was added SP:SAMPLEPREP_SUMMARY to each of the frozen, dried extracts, and each sample vortexed for 30s. For SP:SAMPLEPREP_SUMMARY negative ion analysis 30 µL of 4 ºC 80:20 methanol:water plus 20 mM ammonium SP:SAMPLEPREP_SUMMARY acetate was added to each of the frozen, dried extracts, and each sample SP:SAMPLEPREP_SUMMARY vortexed for 30 s. Samples were then centrifuged at 4000 g at 4 ºC for 10 mins. SP:SAMPLEPREP_SUMMARY For both positive and negative ion analyses, samples were randomized and 5 µL SP:SAMPLEPREP_SUMMARY of sample supernatant was pipetted into a pre-washed 96-well sample plate in SP:SAMPLEPREP_SUMMARY quadruplicate. Three quality control (QC) samples (a mixture with equal volume SP:SAMPLEPREP_SUMMARY from all samples) and a blank were also included on each plate. Loaded plates SP:SAMPLEPREP_SUMMARY were covered with a foil seal using heat sealer and loaded into a TriVersa SP:SAMPLEPREP_SUMMARY Nano-Mate® nanoelectro-spray ion source (Advion) with the cooler set at 10 ºC. SP:SAMPLEPREP_SUMMARY Non-targeted analysis was carried out on polar fractions by direct infusion mass SP:SAMPLEPREP_SUMMARY spectrometry (DIMS) using an LTQ Orbitrap Elite (Thermo Fisher Scientific). 21 SP:SAMPLEPREP_SUMMARY overlapping selected ion monitoring (SIM) windows were collected covering m/z SP:SAMPLEPREP_SUMMARY values from 50 to 620. SP:PROCESSING_STORAGE_CONDITIONS 4℃ SP:EXTRACT_STORAGE 4℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Non-targeted analysis was carried out on polar fractions by direct infusion mass CH:CHROMATOGRAPHY_SUMMARY spectrometry (DIMS) using an LTQ Orbitrap Elite (Thermo Fisher Scientific). 21 CH:CHROMATOGRAPHY_SUMMARY overlapping selected ion monitoring (SIM) windows were collected covering m/z CH:CHROMATOGRAPHY_SUMMARY values from 50 to 620. CH:INSTRUMENT_NAME LTQ Orbitrap Elite (Thermo Fisher Scientific) CH:COLUMN_NAME none CH:CHROMATOGRAPHY_TYPE None (Direct infusion) #ANALYSIS AN:LABORATORY_NAME Environmental Metabolomics Research Laboratory AN:ANALYSIS_TYPE MS AN:OPERATOR_NAME Elena Sostare #MS MS:INSTRUMENT_NAME Thermo Orbitrap Elite Hybrid Ion Trap-Orbitrap MS:INSTRUMENT_TYPE LTQ-FT MS:MS_TYPE ESI MS:MS_COMMENTS For both positive and negative ion analyses, samples were randomized and 5 µL MS:MS_COMMENTS of sample supernatant was pipetted into a pre-washed 96-well sample plate in MS:MS_COMMENTS quadruplicate. Three quality control (QC) samples (a mixture with equal volume MS:MS_COMMENTS from all samples) and a blank were also included on each plate. Loaded plates MS:MS_COMMENTS were covered with a foil seal using heat sealer and loaded into a TriVersa MS:MS_COMMENTS Nano-Mate® nanoelectro-spray ion source (Advion) with the cooler set at 10 ºC. MS:MS_COMMENTS Non-targeted analysis was carried out on polar fractions by direct infusion mass MS:MS_COMMENTS spectrometry (DIMS) using an LTQ Orbitrap Elite (Thermo Fisher Scientific). 21 MS:MS_COMMENTS overlapping selected ion monitoring (SIM) windows were collected covering m/z MS:MS_COMMENTS values from 50 to 620. Data are available in the NIH National Metabolomics Data MS:MS_COMMENTS Repository (NMDR). The Galaxy pipeline at the University of Birmingham was used MS:MS_COMMENTS to process raw data collected. SIM windows were assembled into single spectra MS:MS_COMMENTS for each sample (SIM-Stitching). Filtering A signal to noise ratio (SNR) of 10 MS:MS_COMMENTS was selected to filter out background noise from the data. A replicate filter MS:MS_COMMENTS was applied to retain only peaks found in at least 3 out of 4 technical MS:MS_COMMENTS replicates, and samples aligned across biological samples. A blank filter was MS:MS_COMMENTS applied to only retain peaks if they are a specified % larger than blank values. MS:MS_COMMENTS Finally a sample filter was applied to keep only those peaks found in greater MS:MS_COMMENTS than 80% of biological samples. Missing-value imputation, normalization, and MS:MS_COMMENTS quality assessment Probabilistic quotient normalization (PQN) was applied to MS:MS_COMMENTS normalize the DIMS metabolomics data to account for differences in dilution MS:MS_COMMENTS between samples. A K-nearest neighbor (KNN) algorithm was then applied to impute MS:MS_COMMENTS missing values. A generalized-log transformation was then applied to stabilize MS:MS_COMMENTS the technical variance of the DIMS measurements. To assess data quality, the MS:MS_COMMENTS median relative standard deviation (RSD) was measured across technical MS:MS_COMMENTS replicates and an RSD cutoff value of 10 was specified. Data analysis Univariate MS:MS_COMMENTS ANOVAs were carried out on metabolite data with a false discovery rate (FDR) MS:MS_COMMENTS correction to account for the large number of possible comparisons. Peaks were MS:MS_COMMENTS annotated using the Functional Analysis tool for MS peaks on the MetaboAnalyst MS:MS_COMMENTS 5.0 online web server.4 Peak list files were uploaded containing m/z values and MS:MS_COMMENTS FDR corrected p-values obtained by the processing above, and analyzed in the MS:MS_COMMENTS respective (positive or negative) ion mode with a 5.0 ppm mass tolerance. For MS:MS_COMMENTS enrichment analysis, the Mummichog algorithm was applied with a p-value cutoff MS:MS_COMMENTS of p < 0.1 and analyzed against the KEGG database for Homo sapiens and MS:MS_COMMENTS Drosophila melanogaster. References (1) Bozich, J.; Hang, M.; Hamers, R.; MS:MS_COMMENTS Klaper, R. Core Chemistry Influences the Toxicity of Multicomponent Metal Oxide MS:MS_COMMENTS Nanomaterials, Lithium Nickel Manganese Cobalt Oxide, and Lithium Cobalt Oxide MS:MS_COMMENTS to Daphnia Magna. Environ. Toxicol. Chem. 2017, 36 (9), 2493–2502. MS:MS_COMMENTS https://doi.org/10.1002/etc.3791. (2) Niemuth, N. J. N. J.; Curtis, B. J. B. J.; MS:MS_COMMENTS Hang, M. N. M. N.; Gallagher, M. J. M. J.; Fairbrother, D. H. H.; Hamers, R. J. MS:MS_COMMENTS R. J.; Klaper, R. D. R. D. Next-Generation Complex Metal Oxide Nanomaterials MS:MS_COMMENTS Negatively Impact Growth and Development in the Benthic Invertebrate Chironomus MS:MS_COMMENTS Riparius upon Settling. Environ. Sci. Technol. 2019, 53 (7), 3860–3870. MS:MS_COMMENTS https://doi.org/10.1021/acs.est.8b06804. (3) Davidson, R. L.; Weber, R. J. M.; MS:MS_COMMENTS Liu, H.; Sharma-Oates, A.; Viant, M. R. Galaxy-M: A Galaxy Workflow for MS:MS_COMMENTS Processing and Analyzing Direct Infusion and Liquid Chromatography Mass MS:MS_COMMENTS Spectrometry-Based Metabolomics Data. Gigascience 2016, 5 (1), 10. MS:MS_COMMENTS https://doi.org/10.1186/s13742-016-0115-8. (4) Xia, J.; Psychogios, N.; Young, MS:MS_COMMENTS N.; Wishart, D. S. MetaboAnalyst: A Web Server for Metabolomic Data Analysis and MS:MS_COMMENTS Interpretation. Nucleic Acids Res. 2009, 37 (Web Server), W652–W660. MS:MS_COMMENTS https://doi.org/10.1093/nar/gkp356. MS:ION_MODE NEGATIVE MS:MS_RESULTS_FILE ST001807_AN002930_Results.txt UNITS:arbitrary units Has m/z:Yes #END