#METABOLOMICS WORKBENCH javier_frontinan_20220425_002153 DATATRACK_ID:3217 STUDY_ID:ST002157 ANALYSIS_ID:AN003532 PROJECT_ID:PR001370 VERSION 1 CREATED_ON April 30, 2022, 7:29 am #PROJECT PR:PROJECT_TITLE Effect of long-term exposure to graphene on skin cell metabolism PR:PROJECT_TYPE MS HaCaT Cells PR:PROJECT_SUMMARY Graphene-derived materials are a family of nanomaterials with multiple potential PR:PROJECT_SUMMARY applications in different fields such as biomedicine. It is therefore essential PR:PROJECT_SUMMARY to understand their interaction with cellular barriers such as skin. In this PR:PROJECT_SUMMARY work we evaluated the metabolic changes in human skin cells (HaCaT) exposed to PR:PROJECT_SUMMARY different GRMs for 7 and 30 days. Objectives Endogenous metabolic profiles of PR:PROJECT_SUMMARY control and graphene-treated keratinocytes have been studied using ultra-high PR:PROJECT_SUMMARY performance liquid chromatography – mass spectrometry (UHPLC-MS). PR:PROJECT_SUMMARY Keratinocytes were treated with graphene oxide (GO) from two different suppliers PR:PROJECT_SUMMARY and with few layer graphene (FLG). Samples were collected one week and one month PR:PROJECT_SUMMARY after the start of the treatment. The general aim of the project was to evaluate PR:PROJECT_SUMMARY potential metabolic differences between: 1) Graphene-treated keratinocytes and PR:PROJECT_SUMMARY control keratinocytes at one week; 2) Graphene-treated keratinocytes and control PR:PROJECT_SUMMARY keratinocytes at one month; 3) Control keratinocytes at 1 month and 1 week; 4) PR:PROJECT_SUMMARY Graphene-treated keratinocytes at 1 month and 1 week. Experimental Procedures A PR:PROJECT_SUMMARY successful metabolic profiling experiment relies on the ability to determine PR:PROJECT_SUMMARY changes in an organism’s biofluid or tissue complement of metabolites. Mass PR:PROJECT_SUMMARY spectrometry coupled to ultra-high performance liquid chromatography (UHPLC-MS) PR:PROJECT_SUMMARY is well suited to such analyses due to its high sensitivity, large coverage over PR:PROJECT_SUMMARY different classes of metabolites, high throughput capacity, and wide dynamic PR:PROJECT_SUMMARY range. In this study, one UHPLC-MS based platform was used to analyse endogenous PR:PROJECT_SUMMARY analytes for inclusion in subsequent statistical analysis procedures used to PR:PROJECT_SUMMARY study metabolic differences between the groups of samples. PR:INSTITUTE University of Castilla-La Mancha PR:DEPARTMENT Medical Sciences PR:LABORATORY Group of Oxidative Stress and Neurodegeneration PR:LAST_NAME Frontiñán PR:FIRST_NAME Javier PR:ADDRESS F. Medicina Camino de Moledores s/n 13071 PR:EMAIL javier.frontinan@uclm.es PR:PHONE +34656967979 PR:FUNDING_SOURCE Graphene Flagship (EU) #STUDY ST:STUDY_TITLE Effect of long-term exposure to graphene on skin cell metabolism ST:STUDY_SUMMARY Graphene-derived materials are a family of nanomaterials with multiple potential ST:STUDY_SUMMARY applications in different fields such as biomedicine. It is therefore essential ST:STUDY_SUMMARY to understand their interaction with cellular barriers such as skin. In this ST:STUDY_SUMMARY work we evaluated the metabolic changes in human skin cells (HaCaT) exposed to ST:STUDY_SUMMARY different GRMs for 7 and 30 days. Objectives Endogenous metabolic profiles of ST:STUDY_SUMMARY control and graphene-treated keratinocytes have been studied using ultra-high ST:STUDY_SUMMARY performance liquid chromatography – mass spectrometry (UHPLC-MS). ST:STUDY_SUMMARY Keratinocytes were treated with graphene oxide (GO) from two different suppliers ST:STUDY_SUMMARY and with few layer graphene (FLG). Samples were collected one week and one month ST:STUDY_SUMMARY after the start of the treatment. The general aim of the project was to evaluate ST:STUDY_SUMMARY potential metabolic differences between: 1) Graphene-treated keratinocytes and ST:STUDY_SUMMARY control keratinocytes at one week; 2) Graphene-treated keratinocytes and control ST:STUDY_SUMMARY keratinocytes at one month; 3) Control keratinocytes at 1 month and 1 week; 4) ST:STUDY_SUMMARY Graphene-treated keratinocytes at 1 month and 1 week. Experimental Procedures A ST:STUDY_SUMMARY successful metabolic profiling experiment relies on the ability to determine ST:STUDY_SUMMARY changes in an organism’s biofluid or tissue complement of metabolites. Mass ST:STUDY_SUMMARY spectrometry coupled to ultra-high performance liquid chromatography (UHPLC-MS) ST:STUDY_SUMMARY is well suited to such analyses due to its high sensitivity, large coverage over ST:STUDY_SUMMARY different classes of metabolites, high throughput capacity, and wide dynamic ST:STUDY_SUMMARY range. In this study, one UHPLC-MS based platform was used to analyse endogenous ST:STUDY_SUMMARY analytes for inclusion in subsequent statistical analysis procedures used to ST:STUDY_SUMMARY study metabolic differences between the groups of samples. Results The oxidation ST:STUDY_SUMMARY degree and size of the GRMs is determinant in the effect on cell metabolism, as ST:STUDY_SUMMARY well as the exposure time. Thus, one of the materials used generated a change in ST:STUDY_SUMMARY the energy metabolism of the cells, significantly increasing the level of ST:STUDY_SUMMARY different Krebs cycle metabolites. ST:INSTITUTE University of Castilla-La Mancha ST:DEPARTMENT Medical Sciences ST:LABORATORY Group of Oxidative Stress and Neurodegeneration ST:LAST_NAME Frontinan ST:FIRST_NAME Javier ST:ADDRESS F. Medicina Camino de Moledores s/n 13071 ST:EMAIL javier.frontinan@uclm.es ST:PHONE +34656967979 ST:NUM_GROUPS 8 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Male SU:CELL_BIOSOURCE_OR_SUPPLIER CLS SU:CELL_STRAIN_DETAILS In vitro spontaneously transformed keratinocytes from histologically normal skin SU:CELL_PRIMARY_IMMORTALIZED HaCaT SU:CELL_PASSAGE_NUMBER Up to the 15th passage #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_001 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=REF_CCM_04; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_001.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_002 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=REF_CCM_04; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_002.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_003 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=OWL-MIX-074; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_003.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_004 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=OWL-MIX-074; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_004.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_005 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=AA_CCM; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_005.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_006 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=AA_CCM; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_006.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_007 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Equilibration; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_007.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_008 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Equilibration; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_008.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_009 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=BLK; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_009.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_010 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Equilibration; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_010.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_011 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Pool 01; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_011.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_012 Exposure time:Week | Nanomaterial:GO | Replicate:N1 COD.OWL=Owl-1909-011; COD.EXT=1w GO N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_012.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_013 Exposure time:Month | Nanomaterial:FLG | Replicate:N4 COD.OWL=Owl-1909-074; COD.EXT=1m FLG N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_013.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_014 Exposure time:Week | Nanomaterial:FLG | Replicate:N1 COD.OWL=Owl-1909-031; COD.EXT=1w FLG N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_014.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_015 Exposure time:Week | Nanomaterial:GO | Replicate:N3 COD.OWL=Owl-1909-013; COD.EXT=1w GO N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_015.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_016 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=SeraM-04 01; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_016.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_017 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Pool 02; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_017.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_018 Exposure time:Month | Nanomaterial:GO- | Replicate:N1 COD.OWL=Owl-1909-056; COD.EXT=1m GO- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_018.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_019 Exposure time:Month | Nanomaterial:GO | Replicate:N5 COD.OWL=Owl-1909-055; COD.EXT=1m GO N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_019.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_020 Exposure time:Week | Nanomaterial:C | Replicate:N1 COD.OWL=Owl-1909-001; COD.EXT=1w C N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_020.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_021 Exposure time:Week | Nanomaterial:GO g- | Replicate:N1 COD.OWL=Owl-1909-026; COD.EXT=1w GOg- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_021.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_022 Exposure time:Month | Nanomaterial:GO g | Replicate:N4 COD.OWL=Owl-1909-064; COD.EXT=1m GOg N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_022.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_023 Exposure time:Week | Nanomaterial:GO g | Replicate:N1 COD.OWL=Owl-1909-021; COD.EXT=1w GOg N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_023.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_024 Exposure time:Month | Nanomaterial:FLG | Replicate:N1 COD.OWL=Owl-1909-071; COD.EXT=1m FLG N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_024.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_025 Exposure time:Week | Nanomaterial:C | Replicate:N4 COD.OWL=Owl-1909-004; COD.EXT=1w C N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_025.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_026 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=SeraM-04 02; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_026.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_027 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Pool 03; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_027.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_028 Exposure time:Month | Nanomaterial:GO g | Replicate:N2 COD.OWL=Owl-1909-062; COD.EXT=1m GOg N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_028.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_029 Exposure time:Month | Nanomaterial:GO | Replicate:N2 COD.OWL=Owl-1909-052; COD.EXT=1m GO N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_029.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_030 Exposure time:Month | Nanomaterial:C | Replicate:N4 COD.OWL=Owl-1909-044; COD.EXT=1m C N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_030.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_031 Exposure time:Month | Nanomaterial:GO g- | Replicate:N1 COD.OWL=Owl-1909-066; COD.EXT=1m GOg- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_031.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_032 Exposure time:Week | Nanomaterial:FLG- | Replicate:N1 COD.OWL=Owl-1909-036; COD.EXT=1w FLG- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_032.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_033 Exposure time:Week | Nanomaterial:GO g | Replicate:N5 COD.OWL=Owl-1909-025; COD.EXT=1w GOg N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_033.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_034 Exposure time:Week | Nanomaterial:GO g | Replicate:N4 COD.OWL=Owl-1909-024; COD.EXT=1w GOg N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_034.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_035 Exposure time:Month | Nanomaterial:GO g | Replicate:N5 COD.OWL=Owl-1909-065; COD.EXT=1m GOg N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_035.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_036 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=SeraM-04 03; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_036.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_037 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Pool 04; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_037.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_038 Exposure time:Month | Nanomaterial:FLG | Replicate:N5 COD.OWL=Owl-1909-075; COD.EXT=1m FLG N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_038.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_039 Exposure time:Week | Nanomaterial:C | Replicate:N5 COD.OWL=Owl-1909-005; COD.EXT=1w C N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_039.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_040 Exposure time:Week | Nanomaterial:GO | Replicate:N4 COD.OWL=Owl-1909-014; COD.EXT=1w GO N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_040.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_041 Exposure time:Month | Nanomaterial:GO | Replicate:N1 COD.OWL=Owl-1909-051; COD.EXT=1m GO N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_041.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_042 Exposure time:Month | Nanomaterial:GO g | Replicate:N1 COD.OWL=Owl-1909-061; COD.EXT=1m GOg N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_042.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_043 Exposure time:Month | Nanomaterial:C- | Replicate:N1 COD.OWL=Owl-1909-046; COD.EXT=1m C- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_043.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_044 Exposure time:Week | Nanomaterial:FLG | Replicate:N3 COD.OWL=Owl-1909-033; COD.EXT=1w FLG N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_044.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_045 Exposure time:Week | Nanomaterial:GO- | Replicate:N1 COD.OWL=Owl-1909-016; COD.EXT=1w GO- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_045.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_046 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=SeraM-04 04; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_046.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_047 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Pool 05; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_047.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_048 Exposure time:Week | Nanomaterial:C- | Replicate:N1 COD.OWL=Owl-1909-006; COD.EXT=1w C- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_048.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_049 Exposure time:Month | Nanomaterial:C | Replicate:N2 COD.OWL=Owl-1909-042; COD.EXT=1m C N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_049.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_050 Exposure time:Month | Nanomaterial:C | Replicate:N3 COD.OWL=Owl-1909-043; COD.EXT=1m C N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_050.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_051 Exposure time:Week | Nanomaterial:FLG | Replicate:N5 COD.OWL=Owl-1909-035; COD.EXT=1w FLG N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_051.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_052 Exposure time:Month | Nanomaterial:FLG | Replicate:N3 COD.OWL=Owl-1909-073; COD.EXT=1m FLG N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_052.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_053 Exposure time:Month | Nanomaterial:FLG | Replicate:N2 COD.OWL=Owl-1909-072; COD.EXT=1m FLG N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_053.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_054 Exposure time:Week | Nanomaterial:FLG | Replicate:N2 COD.OWL=Owl-1909-032; COD.EXT=1w FLG N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_054.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_055 Exposure time:Month | Nanomaterial:GO | Replicate:N4 COD.OWL=Owl-1909-054; COD.EXT=1m GO N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_055.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_056 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=SeraM-04 05; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_056.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_057 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Pool 06; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_057.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_058 Exposure time:Week | Nanomaterial:FLG | Replicate:N4 COD.OWL=Owl-1909-034; COD.EXT=1w FLG N4; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_058.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_059 Exposure time:Month | Nanomaterial:C | Replicate:N1 COD.OWL=Owl-1909-041; COD.EXT=1m C N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_059.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_060 Exposure time:Month | Nanomaterial:C | Replicate:N5 COD.OWL=Owl-1909-045; COD.EXT=1m C N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_060.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_061 Exposure time:Week | Nanomaterial:C | Replicate:N3 COD.OWL=Owl-1909-003; COD.EXT=1w C N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_061.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_062 Exposure time:Month | Nanomaterial:FLG- | Replicate:N1 COD.OWL=Owl-1909-076; COD.EXT=1m FLG- N1; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_062.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_063 Exposure time:Week | Nanomaterial:GO | Replicate:N5 COD.OWL=Owl-1909-015; COD.EXT=1w GO N5; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_063.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_064 Exposure time:Month | Nanomaterial:GO g | Replicate:N3 COD.OWL=Owl-1909-063; COD.EXT=1m GOg N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_064.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_065 Exposure time:Week | Nanomaterial:GO g | Replicate:N2 COD.OWL=Owl-1909-022; COD.EXT=1w GOg N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_065.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_066 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=SeraM-04 06; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_066.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_067 Exposure time:- | Nanomaterial:- | Replicate:- COD.OWL=Pool 07; COD.EXT=-; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_067.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_068 Exposure time:Week | Nanomaterial:GO g | Replicate:N3 COD.OWL=Owl-1909-023; COD.EXT=1w GOg N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_068.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_069 Exposure time:Month | Nanomaterial:GO | Replicate:N3 COD.OWL=Owl-1909-053; COD.EXT=1m GO N3; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_069.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_070 Exposure time:Week | Nanomaterial:C | Replicate:N2 COD.OWL=Owl-1909-002; COD.EXT=1w C N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_070.raw SUBJECT_SAMPLE_FACTORS - 190905_19_0009_UCLM_CCM_071 Exposure time:Week | Nanomaterial:GO | Replicate:N2 COD.OWL=Owl-1909-012; COD.EXT=1w GO N2; RAW_FILE_NAME=190905_19_0009_UCLM_CCM_071.raw #COLLECTION CO:COLLECTION_SUMMARY HaCaT cells treated with GO1, GO2 and FLG CO:SAMPLE_TYPE Keratinocytes CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Cells were treated with 5 ug/mL of the different GRMs for 7 days and 30 days (1 TR:TREATMENT_SUMMARY treatment/week) #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Keratinocytes were defrosted on ice and proteins were precipitated from the SP:SAMPLEPREP_SUMMARY lysed cell samples by adding the extraction solvent spiked with metabolites not SP:SAMPLEPREP_SUMMARY detected in unspiked cell extracts. These metabolites, considered as internal SP:SAMPLEPREP_SUMMARY standards, were tryptophan-d5, Anthranilic acid-(ring-13C6), Phenylthiohydantoin SP:SAMPLEPREP_SUMMARY (PTH)-valine, Glycocholic-2,2,4,4-d4 acid (Sigma Aldrich). Then, cell extracts SP:SAMPLEPREP_SUMMARY were incubated at -20 ˚C for 1 hour and after that, samples were vortexed and SP:SAMPLEPREP_SUMMARY centrifuged at 18,000 x g for 10 minutes at 4 ºC. Supernatants were collected SP:SAMPLEPREP_SUMMARY and kept on ice. A second extraction was performed from the remaining pellets, SP:SAMPLEPREP_SUMMARY following the same steps described above. Supernatants obtained from the second SP:SAMPLEPREP_SUMMARY extraction were collected and put together with the supernatants of the first SP:SAMPLEPREP_SUMMARY extraction. Finally, these supernatants were dried under vacuum, reconstituted SP:SAMPLEPREP_SUMMARY in water, resuspended with agitation for 10 minutes, centrifuged at 18,000 x g SP:SAMPLEPREP_SUMMARY for 5 minutes at 4ºC, and transferred to vials for UHPLC-MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Chromatography was performed on a 1.0-mm internal diameter x 150 mm Acquity HSS CH:CHROMATOGRAPHY_SUMMARY T3 1.7 µm column (Waters Corp., Milford, MA) using an ACQUITY UPLC system CH:CHROMATOGRAPHY_SUMMARY (Waters Corp.). The column was maintained at 40 ºC. Samples (2µL) were CH:CHROMATOGRAPHY_SUMMARY injected onto the column at a flow rate of 100 µL/min, for a total run time of CH:CHROMATOGRAPHY_SUMMARY 25 min. The following linear elution gradient was used: 100% solvent A (10mM CH:CHROMATOGRAPHY_SUMMARY tributylamine + 15mM acetic acid + 0.2% methanol in water), to which solvent B CH:CHROMATOGRAPHY_SUMMARY (methanol) was added incrementally to reach a concentration of 4% B after 1.5 CH:CHROMATOGRAPHY_SUMMARY min, increasing to 20% B at 3 min, 25% B at 8 min, 50% B at 10 min, 55% at 15 CH:CHROMATOGRAPHY_SUMMARY min, and 100% B over the next 5 min, and returning to the initial composition CH:CHROMATOGRAPHY_SUMMARY over the final 5 min. CH:CHROMATOGRAPHY_TYPE Ion pairing CH:INSTRUMENT_NAME Waters Acquity H-Class CH:COLUMN_NAME Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Waters LCT premier MS:INSTRUMENT_TYPE Time of flight MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS The eluent was introduced into the mass spectrometer (LCT Premier (Waters Corp.) MS:MS_COMMENTS by electrospray ionization, with capillary and cone voltages set in the negative MS:MS_COMMENTS ion mode to 2800 and 100 V, respectively. The nebulization gas was set to 600 MS:MS_COMMENTS L/hour and 300ºC. The cone gas was fixed at 50 L/hour, and the source MS:MS_COMMENTS temperature was maintained at 120ºC. Centroid data were acquired over the mass MS:MS_COMMENTS range of 50-1000 Da, using an accumulation time of 0.2 seconds per spectrum. All MS:MS_COMMENTS spectra were mass corrected in real time by reference to leucine enkephalin, MS:MS_COMMENTS infused at 10 µL/minute through an independent reference electrospray, sampled MS:MS_COMMENTS every 10 seconds. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Log2 (fold-change) MS_METABOLITE_DATA_START Samples 190905_19_0009_UCLM_CCM_020 190905_19_0009_UCLM_CCM_070 190905_19_0009_UCLM_CCM_061 190905_19_0009_UCLM_CCM_025 190905_19_0009_UCLM_CCM_039 190905_19_0009_UCLM_CCM_012 190905_19_0009_UCLM_CCM_071 190905_19_0009_UCLM_CCM_015 190905_19_0009_UCLM_CCM_040 190905_19_0009_UCLM_CCM_063 190905_19_0009_UCLM_CCM_023 190905_19_0009_UCLM_CCM_065 190905_19_0009_UCLM_CCM_068 190905_19_0009_UCLM_CCM_034 190905_19_0009_UCLM_CCM_033 190905_19_0009_UCLM_CCM_014 190905_19_0009_UCLM_CCM_054 190905_19_0009_UCLM_CCM_044 190905_19_0009_UCLM_CCM_058 190905_19_0009_UCLM_CCM_051 190905_19_0009_UCLM_CCM_059 190905_19_0009_UCLM_CCM_049 190905_19_0009_UCLM_CCM_050 190905_19_0009_UCLM_CCM_030 190905_19_0009_UCLM_CCM_060 190905_19_0009_UCLM_CCM_041 190905_19_0009_UCLM_CCM_029 190905_19_0009_UCLM_CCM_069 190905_19_0009_UCLM_CCM_055 190905_19_0009_UCLM_CCM_019 190905_19_0009_UCLM_CCM_042 190905_19_0009_UCLM_CCM_028 190905_19_0009_UCLM_CCM_064 190905_19_0009_UCLM_CCM_022 190905_19_0009_UCLM_CCM_035 190905_19_0009_UCLM_CCM_024 190905_19_0009_UCLM_CCM_053 190905_19_0009_UCLM_CCM_052 190905_19_0009_UCLM_CCM_013 190905_19_0009_UCLM_CCM_038 Factors Exposure time:Week | Nanomaterial:C | Replicate:N1 Exposure time:Week | Nanomaterial:C | Replicate:N2 Exposure time:Week | Nanomaterial:C | Replicate:N3 Exposure time:Week | Nanomaterial:C | Replicate:N4 Exposure time:Week | Nanomaterial:C | Replicate:N5 Exposure time:Week | Nanomaterial:GO | Replicate:N1 Exposure time:Week | Nanomaterial:GO | Replicate:N2 Exposure time:Week | Nanomaterial:GO | Replicate:N3 Exposure time:Week | Nanomaterial:GO | Replicate:N4 Exposure time:Week | Nanomaterial:GO | Replicate:N5 Exposure time:Week | Nanomaterial:GO g | Replicate:N1 Exposure time:Week | Nanomaterial:GO g | Replicate:N2 Exposure time:Week | Nanomaterial:GO g | Replicate:N3 Exposure time:Week | Nanomaterial:GO g | Replicate:N4 Exposure time:Week | Nanomaterial:GO g | Replicate:N5 Exposure time:Week | Nanomaterial:FLG | Replicate:N1 Exposure time:Week | Nanomaterial:FLG | Replicate:N2 Exposure time:Week | Nanomaterial:FLG | Replicate:N3 Exposure time:Week | Nanomaterial:FLG | Replicate:N4 Exposure time:Week | Nanomaterial:FLG | Replicate:N5 Exposure time:Month | Nanomaterial:C | Replicate:N1 Exposure time:Month | Nanomaterial:C | Replicate:N2 Exposure time:Month | Nanomaterial:C | Replicate:N3 Exposure time:Month | Nanomaterial:C | Replicate:N4 Exposure time:Month | Nanomaterial:C | Replicate:N5 Exposure time:Month | Nanomaterial:GO | Replicate:N1 Exposure time:Month | Nanomaterial:GO | Replicate:N2 Exposure time:Month | Nanomaterial:GO | Replicate:N3 Exposure time:Month | Nanomaterial:GO | Replicate:N4 Exposure time:Month | Nanomaterial:GO | Replicate:N5 Exposure time:Month | Nanomaterial:GO g | Replicate:N1 Exposure time:Month | Nanomaterial:GO g | Replicate:N2 Exposure time:Month | Nanomaterial:GO g | Replicate:N3 Exposure time:Month | Nanomaterial:GO g | Replicate:N4 Exposure time:Month | Nanomaterial:GO g | Replicate:N5 Exposure time:Month | Nanomaterial:FLG | Replicate:N1 Exposure time:Month | Nanomaterial:FLG | Replicate:N2 Exposure time:Month | Nanomaterial:FLG | Replicate:N3 Exposure time:Month | Nanomaterial:FLG | Replicate:N4 Exposure time:Month | Nanomaterial:FLG | Replicate:N5 L-Leucine 082111098 0614364186 1446385234 0946129261 0769052461 0993390967 0929253355 1995960482 1084588225 1218493312 0873790217 08644051 1840345892 1028031702 1254007268 0850680623 1045381649 1155284935 0895702002 0857649294 0701389996 0818321511 079023769 0782607899 1075983463 1095778744 1043061865 1511902787 1447082492 0729706285 1019135897 1119829651 1461501974 1423954291 1027516362 1445444835 063085022 0910337698 1111787439 0386381245 L-Asparagine 1432776353 0783619934 1591048853 0783005084 1013159221 1846232172 1075082241 3207784517 1107186744 129344355 0897545177 1024084015 2554949202 1293053198 1536527701 1124484431 097474866 1134320522 1086016146 1111004373 1130294354 0988015255 0765511305 0845618659 2455916099 0703408065 138165536 1637888663 1186130707 089309092 109887936 109731979 1565926842 1691575472 1300755591 1272540696 0801789635 0858022837 0989108624 063222618 L-Aspartic acid 0394285389 0183256644 075159538 0487008012 0442953724 0545938861 0556373139 0696420934 0861015719 0631061965 0540073496 0393296104 0739110914 0713262151 042686766 0724681279 0682085237 0517687254 0616306354 0401322488 0204405834 0359298236 113328319 1082251667 1671660038 0504185524 0675205427 2542572902 23181921 1220704585 0776290951 0871565794 263890983 1816527078 284226466 0985719047 0257480668 2324584002 2306155083 0588908774 L-Glutamine 1161209854 0860093175 0470994076 1225086597 0721396275 1422491216 1328854951 0605247385 0787939473 0937912977 0630894966 1246665895 0403808539 0636615536 0351262918 0903176947 1421890769 0335412019 0847863515 1025292695 0946581891 081097993 0926229295 0882688693 165987103 1091269684 0867644982 100995308 1497643443 1946966987 0941373426 0858594618 1157720079 1125934618 1684047161 107807843 075824549 1109206332 1650912507 1291716286 L-Lysine 1008450214 0695614771 0796069326 103483887 1103414096 0724819594 0789002413 2214212029 0772248262 0844175104 0857812401 0912177796 1341863387 12283594 0965125198 1020339633 1341845426 1268204301 0514093548 0792908922 0534939971 1340459286 0989748766 1006300084 1511850707 1081455912 1095802532 1423582028 2084704389 0578973244 1120961562 0907804039 1188667915 1373479247 1152683291 1603231935 1063993904 0990106056 0838889646 0707422065 L-Methionine 0887109243 0760864093 102019977 1019278193 0907113175 1287094767 0989227709 1287330474 1093767303 1420320714 1049905614 1108713468 1743933459 1198990566 1450499396 10046515 1105180829 0876438825 0788845723 0900718757 0702399434 1100310848 0901275926 0954560052 2777309006 1218403196 1006634113 2330460132 1693104388 0713735507 1036508449 1047567878 16119489 1318187127 1157488716 1142439711 0588993339 0997367005 1126212623 0459916387 L-Phenylalanine 0844860651 0563332729 1107392303 0934202239 0801273173 109741552 0726481185 1494852075 1159399582 1296636088 0979816507 0930743313 1639117155 1094401046 1296745547 082954874 0824030238 0897604898 084785738 0786003077 0744446651 0685938955 0679946229 0812914283 1111089001 109856943 0942315905 1610639297 138883728 0786403507 1032432161 1050234462 1604333307 1489866069 1065829521 1373514921 0627083717 0764539768 1110938134 0411983865 L-Tyrosine 0909246166 0687557431 0934944093 1017602277 1029122244 1165823555 0924308567 1430737253 1362985226 1167435582 0976533983 1007086871 1420416606 1402273956 1311992975 0900277903 1169561554 093238639 0818090413 0985271936 0701005489 0904237448 0916058775 091623331 1294043052 1498275447 1024248638 1654707831 202943437 0763629874 1385890095 1031918269 1688651713 1818318962 141751454 1158061492 0767863528 1049769251 0989489337 0459908378 L-Tryptophan 0796055865 0641689074 1267755556 0761657603 0774263106 1149229495 0922926562 2011475809 1047987707 1376086353 0916795876 0934228473 1580549123 0954863874 1283411322 0906787194 0984473911 0971138232 0885195386 0864363943 070163464 0741224583 077687388 0665737237 095940313 1035036319 0986345128 1833080112 1555339329 0750449383 0928480676 0992476305 1543818603 1585479878 0921167101 1170701595 0622324769 0902138854 1084285502 0340882738 Glutathione 0768922415 0559520392 1254861372 073031231 0611669811 1037554632 1085456374 1355824066 1078140448 070056026 0872907554 0947300666 1565641882 1107244869 095693212 0790203831 0596584142 0869078896 0687106133 0478303637 0158124962 0445140536 0690835432 0818351037 1928188285 081469461 093694808 1335931878 1554576647 1338691253 1158114144 1140832466 2068404121 1770733067 1517055127 0505183835 0235380942 0909155355 1362169225 0783474604 Oxidized glutathione 0095789946 0638691587 0378358919 0503461429 0447393508 1114192871 0873219447 0364682943 1272066617 0889103452 2780691896 0856464964 0662954564 07588588 0532301976 0761060029 041927642 0544303572 0809528567 1048942533 0653653609 0208590854 0477434946 1041286696 1429124791 0739881702 0980756221 1460701364 0814837632 0743838809 148306767 1303611189 1972159301 0976559876 0616692607 044369386 081676634 0678616254 0786600574 0325871886 S-Adenosylhomocysteine 0917901335 0940501707 1408715342 0858766228 1075497587 1289218877 1185978016 1700031563 1004755574 0377432359 0826481556 0553310104 0238284585 1388936489 0447273328 1114119086 1293101136 1202701316 0927298962 0910878224 027784364 0958272545 1268145538 1237599302 1482734466 1474596579 1158709082 1671510931 1089417237 1701982001 1499128057 171670973 2578414163 1493978623 1055171334 0546890886 160036382 0915393771 0912822703 Acetylglycine 0793824067 0545959192 1249844431 0777545093 0736995731 1143449299 0853368782 23154353 1102432948 1293666483 0882078598 0869632286 1606104016 1032457198 1267012025 0893519069 0854748192 09768688 0865201058 084579436 0602358144 0667376553 0689704784 0604914397 0931357264 1005119206 1023224555 1713119562 1722789485 076941365 0797432356 1028421144 1509272921 1967287546 088863868 1213408145 0580775421 0794205175 1056878537 0310957641 Pyroglutamic acid 1274874525 1218732396 0768273432 107784367 0724389807 1299871905 1831601196 101401159 0960305135 1195560578 1196070312 1833254403 0869264735 102041841 0989189278 1170416848 1988271974 0687657788 1026959378 0783411358 0822977453 0797733536 0676109511 0855433204 0913723299 1107095307 1083395377 1218329271 1213148055 087354983 1087378713 1112369104 1156871274 1180843362 0896553846 1354737243 0744591866 0788955452 1004597415 0707513449 3-Hydroxyglutaric acid 0847659628 0940888555 0622118174 0897676308 0922559365 1557203663 0829703709 108504803 1239679997 1388279723 1134188147 0694926512 093751478 0671047099 0829687035 1181284821 0639010827 1095449643 1235496421 119027936 1568972823 1096486572 0626254668 0808411297 0600951119 1286472275 1196800794 0894832849 0928136307 0405706172 0762671166 0861909116 0570680074 0647357369 0314199742 1986952938 1218697542 0482082729 0716027634 0152539421 Fructose 1,6-bisphosphate 015956543 0605260207 0216781548 1613266436 059691649 0073867277 3049302918 0436471385 5379967255 0494399634 1840885686 0174188567 0180036914 0855575556 1161822758 2085213279 3393904637 1123313315 0269361186 1874422966 1003508989 192418355 0658844327 0617604158 183411559 4788129955 0442327428 1080136118 0525799894 2010903465 0123422525 027588276 0317865857 4851226459 3032394765 0548941721 0196994166 2-Phosphoglyceric acid 0899171346 0360361822 0356714846 0609826113 0168838813 0740886707 0640755634 0648274873 1654354374 0629707589 1453367602 1098756514 1527344572 1148301484 2533720191 0442267316 0506791156 015319501 0193040744 0090376566 0259284212 0478974358 0224434028 0289018957 2075362327 0404421355 0910671748 3205831882 195695922 0375704895 0173733637 0669959795 2474042056 2602041404 1334050656 4143057318 0231960132 0329968702 0905666392 009210401 ADP 0685278549 0351444403 0382941457 0478216084 0430125873 0889619138 0527861639 0578984722 0985420517 029915841 0991383914 0327958158 1224512117 0392916133 0841967609 0661216267 0198146549 0528710639 0312266998 0542089073 0909066153 0467178813 0642860054 1057505691 1950477887 0775095389 0943288045 2267117387 183591736 0821086902 048778038 0632410148 1823824377 1160843491 1475964042 0908400113 0922746055 0793839226 1014183421 0282877642 Adenosine triphosphate 0702309107 0432961677 0271024932 1204266616 0613315858 0930021922 0531931993 0403578336 1755687977 0623853128 0548013785 0637746424 0395331117 1668913759 0963640751 0660678426 0285874565 0357328524 1120005876 0788305987 1170584535 046401931 0771016894 1176466752 1299395525 1134413557 0888212258 1658541331 1759826182 0901040608 1456374274 1296400451 1887847349 2019914551 1429016819 1004220272 0912482236 1079939756 1153627329 0382469032 Adenosine monophosphate 0687728588 0309059497 0338627021 0195126042 0258722347 0849238372 0335296605 0770053041 050018859 0194919405 1989685474 0137075924 2142913164 0105489117 0570135258 0745047511 0113121521 0536944243 0167154421 0362074624 0912183166 0380215104 0461186971 0957789595 1619760261 0430978096 0948005962 2845673572 1705001541 0831538537 0140418798 025475319 1425168216 0785386005 1357055999 1069461903 0860405176 0530602101 1089531661 024897112 Alpha-D-Glucose 1079178126 0995198266 1150235702 1232057001 1209528819 1081651669 1157699252 1770493987 137722545 1224509492 1101259031 1071548204 1236539632 1179063391 1147287015 1078580885 1148677353 1273409007 0951539265 1020374463 0972800098 1209831701 0927475308 0950748793 1778516049 1553833364 1423200797 126882153 113087562 054860313 108941831 1180731853 0966164431 0867541481 0784464922 1431931821 1062581684 0858287645 0762948312 0695624168 Glucose 6-phosphate 0417906673 0313556669 0644018655 0996999928 0467544244 1264547661 0397665263 0768862051 1410965437 0477756082 1575001911 0605167125 2396587045 0684669318 0865368166 0789076205 036233439 0663649879 0525063916 0790511029 1203561863 0303726906 0501721497 0988035554 1275968264 1021258326 0908699633 0847770806 0837998281 0935242113 0727125794 0921959394 147635415 0742933511 0952513834 1098990826 0901704121 0477280196 0939583509 L-Ribulose 0805935481 0622646187 0496659982 0311307896 0292147185 0860716059 0898268556 0760017237 0743523043 04138793 1283279788 0767192322 2077695706 0396179133 095514789 0717682674 0306127048 0854668247 0337045806 0419943695 0792434962 0234400246 0152790122 0502674746 1530581554 0513655897 0643384838 1285973435 1062959258 1037073506 0449609496 05618877 1467809707 087096549 0651662046 0880585391 0612282974 0278500305 0931652784 0603791735 D-Ribose 5-phosphate 0839444963 0775492502 0511607706 0363948915 0346950324 0809040434 0944732273 0833170897 0808665009 0455626094 1376582505 0917399235 2252917264 0479783822 0086711282 0771377482 0297336203 0888077464 0340734418 0435564912 0976361387 0287723953 0174956095 0581303976 1300098339 0585902355 0678129226 0905211569 0889796372 0922733973 0527627661 0614237504 145979937 0862254366 0605231564 0906061638 0607140095 0231614954 0938543756 0688907271 NADP 0798137794 053263321 0460832554 0558681357 0644033432 1087486615 0751029453 100483133 1138619696 0734693494 0828781846 0572498899 1106717648 0577090895 0837428917 0867537146 0417414017 0993025021 0440322188 0843926005 095383964 0512567946 0469522741 0862594856 1215826314 0840260757 1060238168 2095213064 187825937 0600461864 056728189 0783367145 1490824735 1117884399 1035592794 088507488 0893371747 0607971505 0817495671 0303526695 NADPH 0668363466 054995505 0453055345 0408896043 0485530113 1174472383 0732136306 0922894836 0849279058 0654773012 0818127671 0491914971 1045228357 0398463027 0738991452 08999905 0319243153 0866759315 0510210659 0817429021 0954206622 0403722753 0311268565 080275027 1026911598 0700498498 088149689 1647108269 149639848 0687800454 0488744182 0850141659 1496616477 1165984481 0855655481 0831399345 0812295805 0445943009 0892918946 0245699454 Succinic acid 0790823953 0813086839 0357870602 0462461423 0602387837 1107737728 1074297744 1902397063 0591101939 0936756572 1482719442 1161179445 0949062185 0505133178 0742652375 1042143529 1248648964 10186945 0702832859 0639964014 0723281546 0505502644 0752473376 0731748836 0958364001 0739405765 1410429354 1167004076 1652025808 1192633379 0702126726 0946081966 1474270966 1791795083 1425879244 054374073 040405909 0837500739 0987133336 0326491141 Malic acid 055080164 05425972 0298502698 0628452559 0336556152 1026615979 0992950272 0769520872 1115150665 0673604296 1113720182 088907617 0926814969 0983414706 0326403224 084743334 0645789371 0670881638 0771605035 0466877405 0401171182 0302125839 0583611523 0774518654 1343948563 0523940955 0897355607 1192074853 1459029861 12729362 1080399828 1364020217 1719635947 221694239 1294676111 0655094598 0342745465 0736728869 1035807067 0431089305 Oxoglutaric acid 0837573996 0956735536 0264112376 0935082749 0915721478 1135182943 1029257534 0878293563 1184701733 1492005258 1169035105 1211713314 0606899608 141364004 1595967942 0971040686 0602357134 0701763746 096327441 1121774405 1345251069 0870049446 0469030041 0590211802 0762418446 1205523946 0863082133 0650003278 071317167 0846339401 1160413403 1137291089 1052648852 1454148235 0428870707 1341079106 1338288001 0397493029 0612966929 0621826878 Fumaric acid 0770029508 0615858437 0276030912 0888670843 042090955 0914567321 1153897901 1043337926 1323356397 0642808901 1062804096 0988654455 1013414136 1400861473 0367275305 0953713315 0983775095 0831264262 0650007419 0419392704 032625454 0452236526 0832514352 0709282131 2217812564 0672707153 0995407793 1490811902 1910652454 1772342515 1247228846 1644503272 2736464562 3051486905 1847320602 0877765237 0363858619 0924903936 1224011534 0622973168 FAD 0903510275 0825497986 1237887484 1028437218 1153879109 1065599249 0603555612 1722601532 0682583491 0933673097 0688958914 056103301 0874328077 098715111 0634416794 1297179671 0766092915 1129861167 0633865143 1101499416 1134949445 1335727975 1492765594 1503788336 1468427013 1383145668 1142452942 0656364223 1388607369 0730123422 086021325 1045651366 1232835918 1426072678 0995948511 1274300126 112216359 1246626493 1016766049 054998829 Guanosine diphosphate 081979939 0524286803 0581805427 0547339549 0646361272 1091624664 0620895522 1011791889 0895222454 0597563714 0831604141 045381016 1129767619 0466288273 0871933942 0872745245 0371175896 0745528241 0380720315 0697450741 0739532327 0558121025 066643008 1074578856 16123162 09081142 1097957978 2300317298 1900948242 0733245006 0562966002 0815120308 1748899248 1434839177 1453260165 0987126987 0876710667 0798589861 1009398 0312062622 Guanosine triphosphate 08704166 0659416215 0407586984 1487895828 1005372292 1019347806 0696476618 0654136463 2033056607 1193091214 0506979627 1027811938 0372908925 2112250564 1101219991 0848073748 0553332663 0567747391 1375163195 1045581174 0870249621 057616758 0856772002 1262820316 1101260439 1462453711 1069362659 1864933905 2080237883 0819603316 2052584561 1863384397 2465727355 2997373338 1444614936 115107969 0966392266 1250979111 1138891734 0440789285 cis-Aconitic acid 0430703275 0532574866 0489005834 0973683498 0594235577 1436478075 0757595424 0929035561 1606823838 0778678349 0847334277 0704966831 0822777938 0968674087 1077207824 0908274214 1089205119 0570168163 0693124743 0749741695 0599185185 0608763441 0702026287 0924604945 1082100007 1139126156 1552338519 0135624466 1519991241 0682663989 0834801961 1718390831 2264267101 1747522871 1151739405 118917325 0604562784 0900188222 1056483688 0137061364 NADH 059239692 0791151326 0270960723 0479369634 1483663875 4173815259 0768772381 0386525307 14319058 0897961698 2434638406 0526751526 0217849699 0257926968 086063375 4205995998 499088389 3771930187 0848832659 3391675595 1186501122 0775970469 1388949289 8653844105 0442153851 1086644175 1578880188 1562065735 3235256911 0571035376 1266457682 0498248099 0831322185 0330632559 0271023543 0543995166 1222880637 2056825258 0569477922 0101589292 NAD 0858461915 0678875062 1112914818 1023148618 0937557943 0856244708 077535848 1214771462 1192281316 1043151433 0809235552 0695922447 0876234527 1010928632 0874399465 086557443 0707290803 0911502516 1046364864 0752902006 0842366752 0973971322 0933964918 1053860545 1065627407 1175424196 1237907903 1253896648 1481099246 0820693798 1015365774 1133259676 1482454918 1518102481 1037650939 1260614251 0858382886 0954903386 1131525817 0403627969 D-Lactic acid 1164397553 0947419333 1383581393 0941941762 0920584323 138936788 1079731354 1441029335 0828337018 1024054528 1100583511 1171109128 1405896553 1009087876 0735783349 0990077404 1118036156 1101302436 1070526042 0863365284 0919394795 1042646724 0797139246 0849524456 1088765742 1224640978 1013105069 0834810398 1102546591 1036424699 0975753007 0967624057 140618885 1159021768 1102742215 1062661241 083747303 0752737955 0717505575 0650152048 5-Methyltetrahydrofolic acid 0895185273 0833968621 2636416674 0660389222 0774193762 0988691804 0960659798 2954735943 0945279252 0986230181 1471824415 0835846618 3135639317 0510785904 1350464449 0560579276 0822574665 1332470634 0539862755 0502343546 0588355731 0901289534 0981518512 0852832022 1067328979 1052864437 1659609071 2104877802 5721066185 050503784 0420121852 0378169606 2372068137 2496463074 2190708327 0513818045 0336425482 0580578832 0424897748 Hydroxyphenyllactic acid 1163980818 085453551 0782424904 1114633054 1217777043 157007625 0905751683 2180002807 1239414282 1293531056 1255978103 0938302621 1782146103 123256532 1224681203 1062969841 0906256734 1278251605 0879093224 1131037404 0930788715 1103629064 063316392 0940931868 0935970169 1474338366 0905108026 1150562764 203784766 074362892 075507487 0610200259 1191445578 1577974413 0559942022 1655096453 1277212902 042303366 0542148062 0320712366 Trehalose 0490139598 0342030397 099005943 0922189852 0725700288 0382544334 0509256718 1856123395 0601969088 0678534358 0540201189 3359807947 2980451031 1068225696 1155536433 0495178904 1051625926 1235453261 0529092459 080194253 0573552716 0841809548 1411347315 1444954612 1997815452 0974536734 0983870197 157353454 1989950337 0973013391 0977922102 1177734503 2153918547 2165819436 175346806 1127415368 0757298537 1441029947 1150088558 112729152 Sorbitol 1368840944 1231939752 1154072669 0474602038 1044336488 5070291488 0806571215 076505088 0500719649 0556847661 1282451991 1742246515 0948453867 0620850388 0540861822 2375742022 1509195693 1067957451 0691415583 0580791811 1076778604 097475105 049828669 0464707342 073258108 0667908957 1803180644 1052925759 0929412219 0882831611 0787579236 1995319191 1350925896 0555064631 0554056752 0805986115 0889830192 0439965699 0380481102 0322535525 Coenzyme A 0834476643 0665443488 0862867466 0734779985 0642903699 0943273722 1232253388 1091519213 0938773229 0755684525 0631936404 1122898446 2055008296 0702488843 0928504879 0847335315 0706689516 0737696222 038523055 0594132618 0261212443 0586917826 0614830083 0999850969 2425585707 0684313171 0944028489 2661291102 2117323178 0914113779 0595659758 1065625155 3191986483 1736508093 1263284311 1040892603 0453476162 0730325652 1361443532 0423232223 Cytidine 1265297077 0874365184 1073206337 1129004552 108256442 1017679464 0576682541 1037463215 07094382 0715973749 0766416776 0465408926 0636000234 0705324949 0341533255 1156174694 0809323513 0947836225 0896183995 1030510462 1013954565 157904236 2016617733 2044313233 1199585769 110597096 0966225485 1404231199 1930960147 1051835962 049046791 0695301659 131448592 1308734099 1589239068 1104307121 1229441197 1776923328 150248777 063896445 Guanosine 1653540179 0841201711 200932812 2336896761 2363237589 2170232592 083523239 4989017821 1281924331 1620193912 0155694982 076778944 1268538534 2259557821 1463233704 2670618172 1590595476 2088944821 1074884337 1638888384 0710718851 2564506031 2093022309 1826309937 1119816975 2416612809 2485635394 1290859781 2206456981 0390237589 1726889619 2027732817 1403131131 2692921396 0871317372 1276213194 0911765299 1798632115 1235843981 0264857966 Deoxyguanosine 5293209839 120090623 0749334101 1781761225 3780904551 1167274268 1111946347 1932676883 0468303138 0453438352 0034388235 042242281 0079032923 5147036369 1953755717 6802879512 1573238794 3158359059 1244368914 0909143473 420956669 1040700098 3273524915 1324598093 1565654947 3354709215 2256837297 0670866624 2649399127 7366004244 1318820759 1027780612 0915277735 1086267967 1084929823 8350338233 4876262146 301093396 Cytidine monophosphate 065736762 0326993587 0602071197 0359832958 0387540828 0738032218 0370825063 0722707698 0545823679 0288401209 1083380832 0183293836 1758087584 0112952934 0561357623 0683182793 0129922054 0563404302 0155674268 0615590971 2014876325 0733665524 0567369993 1100060581 2065532302 0478841517 0705442735 154632411 1662295041 109092385 0190654526 0416710164 1635668572 0962085522 1020400662 1544385249 1157575459 0498338728 1256269915 04396013 Cytidine triphosphate 1075068956 0577254578 032867198 2062308077 1057033238 0942884003 0539040405 0665757192 1680227404 0701883753 0613750406 0860638924 0427373786 2275368249 101450322 0896244151 0709179015 0438753821 1732291331 0827377785 1049668136 1141004229 1205949052 161356478 1709995563 1066878524 0767518079 1440895283 1827703664 1625885877 1922535725 1438114719 2082474104 3823465095 1453966316 1355506498 1439678671 1339476704 1443721249 0785706754 Uridine 5'-monophosphate 0609259898 0307756539 0598896053 0265028732 0288670633 0837360652 0407188487 0794765506 0538896432 0355923687 121614471 0154352696 1831829711 0096998178 0477308996 0636556474 0134256244 0594946455 0186439434 0397337949 0846993374 0347966764 0434069664 092362767 2537666942 0437696551 0938287455 1947000687 1602925556 1235891594 0176024076 0412585943 1572743208 0824700793 1308663485 094273185 0543560954 0492113674 1185057193 0516768762 Inosine 1410810623 0888789927 1271843826 1772791383 191073112 1371772467 0927950212 2226220686 1584665045 1233962819 0234981368 0915649676 1049353492 1997714942 147894427 1507982697 1169767798 1628175165 1075896554 1196542746 0811331241 143792757 1408833616 1550884472 1119292909 1855895034 1943859143 1302176076 1744131661 0425036498 1849685899 1747883214 127877281 2084774162 1035651758 1215968597 0834975687 1367744745 1064972749 0357117164 Xanthosine 2732899626 109668889 0992081863 1339827385 2520357589 3867208548 11295833 3217399814 0811922896 1903482695 0045193384 0571198735 0676266217 1213475372 145777796 3807164913 1769078049 1897418903 0745477606 1900685534 0582833247 1328431721 2056200628 2848258902 041291125 2338339905 2571501741 1340872634 1864194331 0319177693 0869103091 134791101 0773781254 183762998 0388379962 104745162 0953422185 1838826321 1461770494 0082744408 Uridine diphosphate glucose 052541291 0497993589 0850244136 0734747814 0727579976 0840264201 0680404454 100250147 1002981533 0878095605 0765734236 0826266699 1103474688 1208270634 1038836122 0753362281 0591531235 0874574562 0704061822 080963951 0611336339 0603748936 107033665 0091193774 2823755499 1013328085 0944591689 1566706065 2123057698 153590119 1609352576 1211890491 2567423909 2442101797 1881627803 0667824643 0478392815 1516221413 1157162771 0771032882 ADP-glucose 0663254864 1096350928 1613351699 0938608489 0779975367 1105913623 129128651 1134696537 1055135069 1758854518 0939591276 1146174949 1513765328 1060036474 0984730945 1010082618 0708476118 0790425516 1859274466 0869255414 1499107452 0779893071 0725943158 0229928647 1575334402 1069317031 0824250635 1877758604 1169124993 0765331709 1071233659 1352054657 2452643686 1488207738 0941812386 1066559805 0801770162 0798730537 1135805072 0305487617 Uracil 1453884858 1174446257 1865520262 0221547452 1391383171 1613361952 1141049111 2207756357 0292367353 140905393 0984006392 0466315251 1908759069 0137109478 0759755829 1624813767 1592599962 1976310627 043075446 1676800141 0549958971 0624757029 0447565644 0444640877 1190562803 0907812731 0846620363 0628030525 0887309456 0532856886 0522926667 0543679232 0437249739 0409994259 0625460816 1090905166 0590509586 0514013662 0426683723 0297686188 dGTP 0708398485 0457031981 027408546 1218071687 0627916422 0925675464 0537654969 0399994969 1790757234 0635747294 0552023437 064623967 0408697749 1710145019 0961646269 0669828222 0281459565 0367557795 1179817826 0788987119 116539519 0465707535 0774327092 1189981346 13124734 1170468481 0899771545 1733652604 1746991778 0904565836 1496150159 1325916805 1972343159 2071414774 1446281277 1001644485 0899393502 1069589673 1171490416 0397038898 Folic acid 0778908036 1006245993 0979317876 0774421618 1265971297 2021616416 1247687701 11934463 1743209922 0676828881 2373233768 0281412 1481402744 1630815016 0706580448 3229367955 0651881364 0921232163 0860094192 0963689264 0580471632 0436445967 0848319218 1957816221 1558302004 1012658103 1050768416 0497163048 132756736 1577759959 1146654735 1337348244 0723290627 1670863897 0748553624 0702573506 0606942718 049427664 Pantothenic acid 0947243198 0895161535 016236349 076084801 0675736098 1311135954 1276818169 0401303551 1020687136 108198718 1360115034 1552934268 0826047881 1401049291 1557458807 0916946898 1914185129 0211816405 0886855985 079267672 0906209383 0762464192 0475862993 0546820855 1012502795 1339601085 0957893211 1043536383 0970084686 0800620347 1650268807 1250545338 1293511317 1343501606 0836965989 1928017495 0880010155 0624326693 0667991254 0491715755 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name HMDB KEGG L-Leucine HMDB00687 C00123 L-Asparagine HMDB00168 C00152 L-Aspartic acid HMDB00191 C00049 L-Glutamine HMDB00641 C00064 L-Lysine HMDB00182 C00047 L-Methionine HMDB00696 C00073 L-Phenylalanine HMDB00159 C00079 L-Tyrosine HMDB00158 C00082 L-Tryptophan HMDB00929 C00078 Glutathione HMDB00125 C00051 Oxidized glutathione HMDB03337 C00127 S-Adenosylhomocysteine HMDB00939 C00021 Acetylglycine HMDB00532 Pyroglutamic acid HMDB00267 C01879 3-Hydroxyglutaric acid HMDB00428 Fructose 1,6-bisphosphate HMDB01058 C00354 2-Phosphoglyceric acid HMDB00362 ADP HMDB01341 C00008 Adenosine triphosphate HMDB00538 C00002 Adenosine monophosphate HMDB00045 C00020 Alpha-D-Glucose HMDB03345 C00267 Glucose 6-phosphate HMDB01401 C00092 L-Ribulose HMDB03371 C00310 D-Ribose 5-phosphate HMDB01548 C00117 NADP HMDB00217 C00006 NADPH HMDB00221 C00005 Succinic acid HMDB00254 C00042 Malic acid HMDB00744 C00711 Oxoglutaric acid HMDB00208 C00026 Fumaric acid HMDB00134 C00122 FAD HMDB01248 C00016 Guanosine diphosphate HMDB01201 C00035 Guanosine triphosphate HMDB01273 C00044 cis-Aconitic acid HMDB00072 C00417 NADH HMDB01487 C00004 NAD HMDB00902 C00003 D-Lactic acid HMDB01311 C00256 5-Methyltetrahydrofolic acid HMDB01396 C00440 Hydroxyphenyllactic acid HMDB00755 C03672 Trehalose HMDB00975 C01083 Sorbitol HMDB00247 C00794 Coenzyme A HMDB01423 C00010 Cytidine HMDB00089 C00475 Guanosine HMDB00133 C00387 Deoxyguanosine HMDB00085 C00330 Cytidine monophosphate HMDB00095 C00055 Cytidine triphosphate HMDB00082 C00063 Uridine 5'-monophosphate HMDB00288 C00105 Inosine HMDB00195 C00294 Xanthosine HMDB00299 C01762 Uridine diphosphate glucose HMDB00286 C00029 ADP-glucose HMDB06557 C00498 Uracil HMDB00300 C00106 dGTP HMDB01440 C00286 Folic acid HMDB00121 C00504 Pantothenic acid HMDB00210 C00864 METABOLITES_END #END