#METABOLOMICS WORKBENCH niemuthn_20210518_075653 DATATRACK_ID:2645 STUDY_ID:ST001807 ANALYSIS_ID:AN002929
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                      	POSITIVE
MS:MS_RESULTS_FILE               	ST001807_AN002929_Results.txt	UNITS:arbitrary units	Has m/z:Yes
#END