#METABOLOMICS WORKBENCH bjj25_20200109_074752 DATATRACK_ID:1891 STUDY_ID:ST001332 ANALYSIS_ID:AN002221 PROJECT_ID:PR000910 VERSION 1 CREATED_ON March 24, 2020, 2:12 pm #PROJECT PR:PROJECT_TITLE LCMS lipid and acyl-carnitine analysis PR:PROJECT_SUMMARY Lipidomics and acyl-carnitine analysis. PR:INSTITUTE University of Cambridge PR:LABORATORY CMaLL PR:LAST_NAME Jenkins PR:FIRST_NAME Benjamin PR:ADDRESS Department of Biochemistry, University of Cambridge, c/o Level 4, Pathology PR:ADDRESS Building PR:EMAIL bjj25@medschl.cam.ac.uk PR:PHONE 07731103718 #STUDY ST:STUDY_TITLE LCMS lipid and acyl-carnitine analysis ST:STUDY_SUMMARY LCMS Lipidomics and acyl-carnitine analysis. ST:INSTITUTE University of Cambridge ST:LABORATORY CMaLL ST:LAST_NAME Jenkins ST:FIRST_NAME Benjamin ST:ADDRESS Department of Biochemistry, University of Cambridge, c/o Level 4, Pathology ST:ADDRESS Building ST:EMAIL bjj25@medschl.cam.ac.uk ST:PHONE 07731103718 #SUBJECT SU:SUBJECT_TYPE Invertebrate SU:SUBJECT_SPECIES Caenorhabditis elegans SU:TAXONOMY_ID 6239 #FACTORS #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 - 20190326_BJJ_Acyl_carnitines_Sample_1_HBS Treatment:HBS RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_010; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Acyl_carnitines_Sample_2_HB Treatment:HB RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_011; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Acyl_carnitines_Sample_3_HBS Treatment:HBS RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_012; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Acyl_carnitines_Sample_4_HB Treatment:HB RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_013; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Acyl_carnitines_Sample_5_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_014; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Acyl_carnitines_Sample_6_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_015; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Acyl_carnitines_Sample_7_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_016; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Acyl_carnitines_Sample_8_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_acyl_carnitines_017; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_1_HBS Treatment:HBS RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_20; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_2_HB Treatment:HB RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_21; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_3_HBS Treatment:HBS RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_22; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_4_HB Treatment:HB RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_23; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_5_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_24; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_6_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_25; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_7_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_26; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190326_BJJ_Lipids_Sample_8_BIG Treatment:BIG RAW_FILE_NAME=20190326_BJJ_Nick_Burton_LCMS_Lipids_02May2019_27; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_1_HB101 Treatment:HB101 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_009; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_2_HB101 Treatment:HB101 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_010; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_3_HB101 Treatment:HB101 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_011; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_4_PA14 Treatment:PA14 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_012; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_5_PA14 Treatment:PA14 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_013; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_6_PA14 Treatment:PA14 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_014; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_7_P.LUMIN Treatment:P.Lumin RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_015; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_8_P.LUMIN Treatment:P.Lumin RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_016; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Acyl_carnitines_Sample_9_P.LUMIN Treatment:P.Lumin RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_acyl_carnitines_30May2019_017; Analysis=Acyl_carnitine SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_1_HB101 Treatment:HB101 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_046; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_2_HB101 Treatment:HB101 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_047; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_3_HB101 Treatment:HB101 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_048; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_4_PA14 Treatment:PA14 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_049; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_5_PA14 Treatment:PA14 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_050; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_6_PA14 Treatment:PA14 RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_051; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_7_P.lumin Treatment:P.Lumin RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_052; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_8_P.lumin Treatment:P.Lumin RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_053; Analysis=Lipids SUBJECT_SAMPLE_FACTORS - 20190510_BJJ_Lipids_Sample_9_P.lumin Treatment:P.Lumin RAW_FILE_NAME=20190510_BJJ_Nick_Burton_LCMS_Lipids_MSfilename_28May2019_054; Analysis=Lipids #COLLECTION CO:COLLECTION_SUMMARY LCMS lipid and acyl-carnitine analysis. CO:SAMPLE_TYPE C. elegans #TREATMENT TR:TREATMENT_SUMMARY Parental exposure to environmental stress can program adaptive changes in TR:TREATMENT_SUMMARY offspring in diverse organisms (1–4). The mechanisms by which parental TR:TREATMENT_SUMMARY exposure to environmental stresses can program predictive adaptive responses in TR:TREATMENT_SUMMARY offspring remain almost completely unknown. Here we report that the soil TR:TREATMENT_SUMMARY bacteria Pseudomonas vranovensis is a natural pathogen of the nematode TR:TREATMENT_SUMMARY Caenorhabditis elegans and that parental exposure of C. elegans to P. TR:TREATMENT_SUMMARY vranovensis promotes offspring resistance to infection. This adaptation can be TR:TREATMENT_SUMMARY transmitted transgenerationally such that infection of adults can enhance the TR:TREATMENT_SUMMARY immunity of their descendants four generations later. We find that parental TR:TREATMENT_SUMMARY infection by P. vranovensis results in increased expression of the cysteine TR:TREATMENT_SUMMARY synthases CYSL-1 and CYSL-2 and the regulator of hypoxia inducible factor RHY-1 TR:TREATMENT_SUMMARY in progeny, that the expression of these three genes in offspring is required TR:TREATMENT_SUMMARY for adaptation to P. vranovensis, and that the expression of these genes is TR:TREATMENT_SUMMARY regulated by the WD40 repeat protein WDR-23. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY C. elegans was prepared for LC-MS lipidomics and acyl-carnitine analysis as SP:SAMPLEPREP_SUMMARY previously described (3) with minor modifications. Briefly, ~40 µL of SP:SAMPLEPREP_SUMMARY concentrated embryos were re-suspended in 100 µL of water, then 0.4 mL of SP:SAMPLEPREP_SUMMARY chloroform was added to each sample followed by 0.2 mL of methanol containing SP:SAMPLEPREP_SUMMARY the stable isotope labelled acyl-carnitine internal standards SP:SAMPLEPREP_SUMMARY (Butyryl-L-carnitine-d7 at 5 µM and Hexadecanoyl-L-carnitine-d3 at 5 µM). The SP:SAMPLEPREP_SUMMARY samples were then homogenised by vortexing then transferred into a 2 mL SP:SAMPLEPREP_SUMMARY Eppendorf screw-cap tube. The original container was washed out with 0.5 mL of SP:SAMPLEPREP_SUMMARY chloroform: methanol (2: 1, respectively) and added to the appropriate 2 mL SP:SAMPLEPREP_SUMMARY Eppendorf screw-cap tube. This was followed by the addition of 150 µL of the SP:SAMPLEPREP_SUMMARY following stable isotope labelled internal standards (approximately 10 to 50 µM SP:SAMPLEPREP_SUMMARY in methanol): Ceramide_C16d31, LPC_(C14:0d42), PC_(C16:0d31 / C18:1), SP:SAMPLEPREP_SUMMARY PE_(C16:0d31 / C18:1), PG_(C16:0d31 / C18:1), PI_(C16:0d31 / C18:1), SP:SAMPLEPREP_SUMMARY PS_(C16:0d62), SM_(C16:0d31), TG_(45:0d29) and TG_(48:0d31). Then, 400 µL of SP:SAMPLEPREP_SUMMARY sterile water was added. The samples were vortexed for 1 min, and then SP:SAMPLEPREP_SUMMARY centrifuged at ~20,000 rpm for 5 minutes. For the intact lipid sample SP:SAMPLEPREP_SUMMARY preparation, 0.3 mL of the organic layer (the lower chloroform layer) was SP:SAMPLEPREP_SUMMARY collected into a 2 mL amber glass vial (Agilent Technologies, Santa Clara SP:SAMPLEPREP_SUMMARY California, USA) and dried down to dryness in an Eppendorf Concentrator Plus SP:SAMPLEPREP_SUMMARY system (Eppendorf, Stevenage, UK) run for 60 minutes at 45 °C. The dried lipid SP:SAMPLEPREP_SUMMARY samples were then reconstituted with 100 µL of 2:1:1 solution of propan-2-ol, SP:SAMPLEPREP_SUMMARY acetonitrile and water, respectively, and then vortexed thoroughly. The lipid SP:SAMPLEPREP_SUMMARY samples were then transferred into a 300 μL low-volume vial insert inside a 2 SP:SAMPLEPREP_SUMMARY mL amber glass auto-sample vial ready for liquid chromatography separation with SP:SAMPLEPREP_SUMMARY mass spectrometry detection (LC-MS) of intact lipid species. For the SP:SAMPLEPREP_SUMMARY acyl-carnitine sample preparation, 0.2 mL of the organic layer (the lower SP:SAMPLEPREP_SUMMARY chloroform layer) and 0.2 mL of the aqueous layer (the top water layer) were SP:SAMPLEPREP_SUMMARY mixed into a 2 mL amber glass vial and dried down to dryness. The dried SP:SAMPLEPREP_SUMMARY acyl-carnitine samples were then reconstituted with 100 µL of water and SP:SAMPLEPREP_SUMMARY acetonitrile (4: 1, respectively) and thoroughly vortexed. The acyl-carnitine SP:SAMPLEPREP_SUMMARY samples were then transferred into a 300 μL low-volume vial insert inside a 2 SP:SAMPLEPREP_SUMMARY mL amber glass auto-sample vial ready for liquid chromatography separation with SP:SAMPLEPREP_SUMMARY mass spectrometry detection (LC-MS) of the acyl-carnitine species. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Full chromatographic separation of intact lipids (4) was achieved using Shimadzu CH:CHROMATOGRAPHY_SUMMARY HPLC System (Shimadzu UK Limited, Milton Keynes, United Kingdom) with the CH:CHROMATOGRAPHY_SUMMARY injection of 10 µL onto a Waters Acquity UPLC® CSH C18 column; 1.7 µm, I.D. CH:CHROMATOGRAPHY_SUMMARY 2.1 mm X 50 mm, maintained at 55 °C. Mobile phase A was 6:4, acetonitrile and CH:CHROMATOGRAPHY_SUMMARY water with 10 mM ammonium formate. Mobile phase B was 9:1, propan-2-ol and CH:CHROMATOGRAPHY_SUMMARY acetonitrile with 10 mM ammonium formate. The flow was maintained at 500 µL per CH:CHROMATOGRAPHY_SUMMARY minute through the following gradient: 0.00 minutes_40% mobile phase B; 0.40 CH:CHROMATOGRAPHY_SUMMARY minutes_43% mobile phase B; 0.45 minutes_50% mobile phase B; 2.40 minutes_54% CH:CHROMATOGRAPHY_SUMMARY mobile phase B; 2.45 minutes_70% mobile phase B; 7.00 minutes_99% mobile phase CH:CHROMATOGRAPHY_SUMMARY B; 8.00 minutes_99% mobile phase B; 8.3 minutes_40% mobile phase B; 10 CH:CHROMATOGRAPHY_SUMMARY minutes_40% mobile phase B; 10.00 minutes_40% mobile phase B. The sample CH:CHROMATOGRAPHY_SUMMARY injection needle was washed using 9:1, 2-propan-2-ol and acetonitrile with 0.1 % CH:CHROMATOGRAPHY_SUMMARY formic acid. The mass spectrometer used was the Thermo Scientific Exactive CH:CHROMATOGRAPHY_SUMMARY Orbitrap with a heated electrospray ionisation source (Thermo Fisher Scientific, CH:CHROMATOGRAPHY_SUMMARY Hemel Hempstead, UK). The mass spectrometer was calibrated immediately before CH:CHROMATOGRAPHY_SUMMARY sample analysis using positive and negative ionisation calibration solution CH:CHROMATOGRAPHY_SUMMARY (recommended by Thermo Scientific). Additionally, the heated electrospray CH:CHROMATOGRAPHY_SUMMARY ionisation source was optimised at 50:50 mobile phase A to mobile phase B for CH:CHROMATOGRAPHY_SUMMARY spray stability (capillary temperature; 380 °C, source heater temperature; 420 CH:CHROMATOGRAPHY_SUMMARY °C, sheath gas flow; 60 (arbitrary), auxiliary gas flow; 20 (arbitrary), sweep CH:CHROMATOGRAPHY_SUMMARY gas; 5 (arbitrary), source voltage; 3.5 kV. The mass spectrometer resolution was CH:CHROMATOGRAPHY_SUMMARY set to 25,000 with a full-scan range of m/z 100 to 1,800 Da, with continuous CH:CHROMATOGRAPHY_SUMMARY switching between positive and negative mode. Lipid quantification was achieved CH:CHROMATOGRAPHY_SUMMARY using the area under the curve (AUC) of the corresponding high resolution CH:CHROMATOGRAPHY_SUMMARY extracted ion chromatogram (with a window of ± 8 ppm) at the indicative CH:CHROMATOGRAPHY_SUMMARY retention time. The lipid analyte AUC relative to the associated internal CH:CHROMATOGRAPHY_SUMMARY standard AUC for that lipid class was used to semi-quantify and correct for any CH:CHROMATOGRAPHY_SUMMARY extraction/instrument variation. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Shimadzu 20AD CH:COLUMN_NAME Waters Acquity UPLC CSH C18(1.7 µm, 2.1 mm X 50 mm) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Exactive MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS The mass spectrometer used was the Thermo Scientific Exactive Orbitrap with a MS:MS_COMMENTS heated electrospray ionisation source (Thermo Fisher Scientific, Hemel MS:MS_COMMENTS Hempstead, UK). The mass spectrometer was calibrated immediately before sample MS:MS_COMMENTS analysis using positive and negative ionisation calibration solution MS:MS_COMMENTS (recommended by Thermo Scientific). Additionally, the heated electrospray MS:MS_COMMENTS ionisation source was optimised at 50:50 mobile phase A to mobile phase B for MS:MS_COMMENTS spray stability (capillary temperature; 380 °C, source heater temperature; 420 MS:MS_COMMENTS °C, sheath gas flow; 60 (arbitrary), auxiliary gas flow; 20 (arbitrary), sweep MS:MS_COMMENTS gas; 5 (arbitrary), source voltage; 3.5 kV. The mass spectrometer resolution was MS:MS_COMMENTS set to 25,000 with a full-scan range of m/z 100 to 1,800 Da, with continuous MS:MS_COMMENTS switching between positive and negative mode. Lipid quantification was achieved MS:MS_COMMENTS using the area under the curve (AUC) of the corresponding high resolution MS:MS_COMMENTS extracted ion chromatogram (with a window of ± 8 ppm) at the indicative MS:MS_COMMENTS retention time. The lipid analyte AUC relative to the associated internal MS:MS_COMMENTS standard AUC for that lipid class was used to semi-quantify and correct for any MS:MS_COMMENTS extraction/instrument variation. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Area ratio MS_METABOLITE_DATA_START Samples 20190326_BJJ_Acyl_carnitines_Sample_1_HBS 20190326_BJJ_Acyl_carnitines_Sample_2_HB 20190326_BJJ_Acyl_carnitines_Sample_3_HBS 20190326_BJJ_Acyl_carnitines_Sample_4_HB 20190326_BJJ_Acyl_carnitines_Sample_5_BIG 20190326_BJJ_Acyl_carnitines_Sample_6_BIG 20190326_BJJ_Acyl_carnitines_Sample_7_BIG 20190326_BJJ_Acyl_carnitines_Sample_8_BIG 20190326_BJJ_Lipids_Sample_1_HBS 20190326_BJJ_Lipids_Sample_2_HB 20190326_BJJ_Lipids_Sample_3_HBS 20190326_BJJ_Lipids_Sample_4_HB 20190326_BJJ_Lipids_Sample_5_BIG 20190326_BJJ_Lipids_Sample_6_BIG 20190326_BJJ_Lipids_Sample_7_BIG 20190326_BJJ_Lipids_Sample_8_BIG 20190510_BJJ_Acyl_carnitines_Sample_1_HB101 20190510_BJJ_Acyl_carnitines_Sample_2_HB101 20190510_BJJ_Acyl_carnitines_Sample_3_HB101 20190510_BJJ_Acyl_carnitines_Sample_4_PA14 20190510_BJJ_Acyl_carnitines_Sample_5_PA14 20190510_BJJ_Acyl_carnitines_Sample_6_PA14 20190510_BJJ_Acyl_carnitines_Sample_7_P.LUMIN 20190510_BJJ_Acyl_carnitines_Sample_8_P.LUMIN 20190510_BJJ_Acyl_carnitines_Sample_9_P.LUMIN 20190510_BJJ_Lipids_Sample_1_HB101 20190510_BJJ_Lipids_Sample_2_HB101 20190510_BJJ_Lipids_Sample_3_HB101 20190510_BJJ_Lipids_Sample_4_PA14 20190510_BJJ_Lipids_Sample_5_PA14 20190510_BJJ_Lipids_Sample_6_PA14 20190510_BJJ_Lipids_Sample_7_P.lumin 20190510_BJJ_Lipids_Sample_8_P.lumin 20190510_BJJ_Lipids_Sample_9_P.lumin Factors Treatment:HBS Treatment:HB Treatment:HBS Treatment:HB Treatment:BIG Treatment:BIG Treatment:BIG Treatment:BIG Treatment:HBS Treatment:HB Treatment:HBS Treatment:HB Treatment:BIG Treatment:BIG Treatment:BIG Treatment:BIG Treatment:HB101 Treatment:HB101 Treatment:HB101 Treatment:PA14 Treatment:PA14 Treatment:PA14 Treatment:P.Lumin Treatment:P.Lumin Treatment:P.Lumin Treatment:HB101 Treatment:HB101 Treatment:HB101 Treatment:PA14 Treatment:PA14 Treatment:PA14 Treatment:P.Lumin Treatment:P.Lumin Treatment:P.Lumin Carnitine_(C0) 0.05867 0.04902 0.0877 0.05589 0.03468 0.03369 0.03204 0.03238 0.368 0.047 0.162 0.04 0.06 0.096 0.067 0.064 0.038 Carnitine_(C04_dicarboxylic_acid) Carnitine_(C02) 0.03055 0.02733 0.0144 0.02042 0.03236 0.02483 0.02465 0.02523 0.001 0.024 0.004 0.044 0.034 0.022 0.036 0.036 0.042 Carnitine_(C04_OH) 0.001 Carnitine_(C04:1) Carnitine_(C06_OH) Carnitine_(C03) 0.03385 0.02841 0.01986 0.02205 0.05489 0.02262 0.03488 0.02556 0.002 0.009 0.004 0.016 0.022 0.008 0.011 0.013 0.016 Carnitine_(C03_OH) 0.00049 0.00039 0.00044 0.00035 0.001 Carnitine_(C06:1) Carnitine_(C03_dicarboxylic_acid) Carnitine_(C03_methyldicarboxylic_acid) Carnitine_(C05_dicarboxylic_acid) Carnitine_(C05_OH) 0.00088 0.00063 0.00072 0.00054 0.00035 0.00015 0.00023 0.00023 0.001 0.001 0.001 Carnitine_(C04) 0.00392 0.00354 0.00419 0.00309 0.00473 0.00327 0.0039 0.00331 0.001 0.001 0.001 0.001 0.002 0.001 0.002 0.002 0.002 Carnitine_(C03:1) Carnitine_(C05:1) 0.00293 0.00198 0.00288 0.00166 0.00206 0.00161 0.00143 0.00103 0.001 0.002 0.001 0.002 0.002 0.002 Carnitine_(C05) 0.00815 0.01132 0.00509 0.00661 0.01227 0.0102 0.01275 0.01389 0.002 0.001 0.002 0.001 0.001 0.001 0.001 0.002 Carnitine_(C06_dicarboxylic_acid) 0.00012 0.0003 0.00008 0.00009 0.00021 0.00022 0.0002 0.00021 Carnitine_(C06) 0.00367 0.0058 0.00194 0.00348 0.00507 0.00597 0.00544 0.00594 0.003 0.001 0.002 0.001 0.001 0.001 0.001 0.001 Carnitine_(C10:1_OH) Carnitine_(C08_dicarboxylic_acid) Carnitine_(C10:2) Carnitine_(C07) 0.007 0.00888 0.00431 0.00535 0.00381 0.00493 0.00356 0.005 0.002 0.001 0.001 0.001 0.001 0.001 0.001 0.001 Carnitine_(C08:1) 0.00177 0.00207 0.00144 0.00167 0.00125 0.00164 0.00124 0.00151 Carnitine_(C10_OH) 0.00027 0.00064 0.00012 0.00027 0.00032 0.00034 0.00034 0.00042 Carnitine_(C08) 0.00293 0.00375 0.00235 0.00238 0.00295 0.00325 0.00313 0.00299 0.001 0.001 Carnitine_(C10_dicarboxylic_acid) Carnitine_(C09) 0.00545 0.00672 0.00377 0.00441 0.00172 0.00229 0.00159 0.00185 0.001 0.001 0.017 0.017 0.009 Carnitine_(C10:1) 0.00152 0.00202 0.00124 0.00133 0.00109 0.00106 0.00097 0.00104 Carnitine_(C12:1_OH) 0.00103 0.00152 0.00073 0.00089 0.0008 0.0009 0.0008 0.0008 0.001 0.003 0.003 0.005 0.002 0.002 0.002 0.002 0.003 Carnitine_(C10) 0.00225 0.0023 0.00167 0.00175 0.00215 0.00209 0.00222 0.00174 Carnitine_(C12_OH) 0.00007 0.00022 0.00002 0.00009 0.00002 0.00001 Carnitine_(C14:1_OH) Carnitine_(C12:1) 0.00184 0.002 0.00155 0.00147 0.00544 0.0057 0.00706 0.00478 0.001 0.002 0.001 0.001 0.001 0.001 Carnitine_(C11) 0.00312 0.00188 0.00249 0.00265 0.00104 0.00233 0.00216 0.00063 0.001 0.001 0.001 Carnitine_(C14:2) Carnitine_(C18:2_OH) Carnitine_(C14_OH) 0.00022 0.0004 0.00011 0.00018 0.00004 0.00006 0.00006 0.00003 Carnitine_(C12) 0.0035 0.00283 0.00322 0.00293 0.00185 0.00172 0.00176 0.00146 0.001 Carnitine_(C16:1_OH) 0.08316 0.1195 0.07396 0.08075 0.08031 0.10019 0.09111 0.07739 0.001 0.001 Carnitine_(C14:1) 0.04742 0.06338 0.04217 0.04056 0.04661 0.04551 0.04644 0.03473 0.001 0.001 0.002 0.002 0.003 0.003 0.003 0.002 0.001 Carnitine_(C13) 0.00434 0.00268 0.00354 0.00319 0.00265 0.0031 0.00303 0.00231 0.001 0.001 0.001 0.001 0.001 Carnitine_(C16:2) 0.00278 0.00174 0.00219 0.00186 0.00301 0.00237 0.00336 0.00186 0.001 Carnitine_(C18:1_OH) 0.0022 0.00032 0.00032 0.00021 0.00053 0.00041 0.00045 0.00032 Carnitine_(C16_OH) 0.00009 0.00014 0.00007 0.00005 0.00001 0.00001 0.00001 Carnitine_(C18:3) 0.00637 0.0035 0.00566 0.00284 0.00263 0.00184 0.0022 0.00145 0.001 0.001 0.001 0.001 Carnitine_(C14) 0.01052 0.00862 0.01065 0.00703 0.00443 0.00416 0.00399 0.00301 0.002 0.001 0.002 0.001 0.001 0.001 0.001 Carnitine_(C22:5) Carnitine_(C18:2) 0.01423 0.0079 0.01513 0.00749 0.01005 0.00598 0.0085 0.0049 0.003 0.003 0.001 0.002 0.001 0.003 0.002 0.001 Carnitine_(C16:1) Carnitine_(C15) 0.01544 0.00816 0.01875 0.01017 0.0094 0.00806 0.00854 0.00603 0.005 0.001 0.005 0.001 0.003 0.002 0.004 0.003 0.001 Carnitine_(C18_OH) Carnitine_(C16) 0.00344 0.00338 0.00444 0.00335 0.00212 0.00207 0.00206 0.00173 Carnitine_(C18:1) 0.00507 0.00263 0.00641 0.00301 0.01969 0.01422 0.01945 0.01316 0.004 0.004 0.005 0.013 0.01 0.011 0.006 0.005 Carnitine_(C17) 0.00748 0.00493 0.01178 0.00497 0.00673 0.00555 0.00655 0.00432 0.007 0.001 0.005 0.001 0.005 0.003 0.006 0.004 0.002 Carnitine_(C18) 0.00211 0.00128 0.00407 0.00233 0.00142 0.00132 0.00145 0.00109 0.002 0.002 0.001 0.002 0.001 0.001 Carnitine_(C19) 0.00047 0.00025 0.00037 0.00035 0.00047 0.00045 0.00053 0.00046 Carnitine_(C20) 0.0006 0.00035 0.00065 0.00053 0.00038 0.00038 0.00034 0.00035 Cer_(d18:1_15:0) Cer_(d18:0_15:0) 0.001 0.001 0.001 0.001 Cer_(d18:0_16:1) Cer_(d18:1_17:0) 0.001 0.001 Cer_(d18:0_16:0) 0.001 Cer_(d18:1_18:1) 0.001 0.039 0.015 0.003 0.003 0.002 0.002 0.002 0.002 0.001 0.001 0.001 0.001 Cer_(d18:0_17:0) 0.001 0.001 0.001 0.001 0.001 0.001 0.001 Cer_(d18:0_18:1) 0.001 0.001 0.001 0.001 0.004 0.003 0.003 0.003 0.002 0.001 0.001 0.002 0.001 0.002 0.001 0.002 Cer_(d18:1_24:2) 0.001 0.001 0.001 0.001 0.001 0.001 Cer_(d18:0_18:0) 0.001 0.001 Cer_(d18:1_20:1) 0.144 0.091 0.116 0.126 0.256 0.178 0.238 0.178 0.136 0.047 0.128 0.064 0.122 0.09 0.148 0.084 0.094 Cer_(d18:1_22:1) 0.204 0.135 0.158 0.163 0.466 0.265 0.361 0.311 0.249 0.083 0.248 0.112 0.188 0.138 0.341 0.191 0.185 Cer_(d18:0_20:1) 0.022 0.019 0.018 0.019 0.058 0.032 0.04 0.037 0.024 0.012 0.023 0.022 0.028 0.018 0.031 0.021 0.02 Cer_(d18:0_20:0) 0.005 0.003 0.006 0.006 0.006 0.003 0.004 0.004 0.008 0.002 0.009 0.003 0.007 0.004 0.009 0.004 0.004 Cer_(d18:0_22:1) 0.009 0.008 0.007 0.006 0.013 0.01 0.011 0.009 0.01 0.002 0.009 0.003 0.006 0.003 0.016 0.009 0.007 Cer_(d18:1_24:1) 0.156 0.126 0.11 0.127 0.27 0.141 0.195 0.167 0.175 0.048 0.173 0.044 0.092 0.051 0.177 0.106 0.105 Cer_(d18:0_22:0) 0.006 0.004 0.006 0.006 0.01 0.005 0.007 0.006 0.012 0.003 0.013 0.003 0.007 0.005 0.02 0.01 0.008 Cer_(d18:0_24:1) 0.002 0.002 0.002 0.002 0.003 0.002 0.002 0.002 0.003 0.002 0.001 0.002 0.001 0.003 0.002 0.001 Cer_(d18:0_24:0) 0.003 0.002 0.003 0.002 0.005 0.003 0.003 0.003 0.006 0.001 0.005 0.001 0.003 0.002 0.005 0.003 0.002 Cer_(d18:1_26:1) 0.02 0.018 0.015 0.016 0.038 0.022 0.028 0.023 0.019 0.007 0.016 0.01 0.018 0.01 0.019 0.013 0.011 Cer_(d18:0_26:1) 0.001 0.001 0.001 0.001 0.002 0.001 0.001 0.001 0.001 0.001 0.001 0.002 0.002 0.001 0.002 0.002 0.002 Cer_(d18:0_26:0) 0.001 0.001 0.001 0.001 0.002 0.001 0.002 0.001 0.002 0.001 0.002 0.001 0.002 0.001 0.002 0.001 0.001 LPC_(12:0) LPC_(18:2) 0.399 0.176 0.52 0.273 0.325 0.215 0.201 0.189 0.657 0.11 0.636 0.092 0.298 0.16 0.339 0.144 0.131 LPC_(20:5) 0.673 0.189 0.864 0.439 0.438 0.286 0.345 0.279 0.725 0.109 0.718 0.11 0.309 0.181 0.776 0.431 0.296 LPE_(14:0) LPC_(14:0) 0.014 0.007 0.018 0.016 0.023 0.004 0.019 0.001 0.005 0.003 0.007 0.003 0.003 LPC_(16:1) 0.028 0.013 0.037 0.031 0.172 0.112 0.111 0.109 0.147 0.032 0.129 0.042 0.127 0.078 0.099 0.046 0.042 LPC_(17:1) 0.563 0.307 0.619 0.446 0.277 0.173 0.185 0.168 0.694 0.139 0.794 0.028 0.079 0.052 0.36 0.164 0.132 LPE_(15:0) 0.006 0.003 0.002 0.003 0.005 0.003 0.004 0.004 0.001 0.001 0.001 0.001 0.001 0.001 0.002 0.001 0.001 LPE_(18:2) 0.018 0.019 0.025 0.01 0.015 0.009 0.013 0.007 0.01 LPE_(16:1) 0.001 0.019 0.01 0.011 0.009 0.001 0.003 0.001 0.002 0.001 0.002 0.001 0.002 LPE_(17:1) 0.023 0.026 0.017 0.026 0.006 0.004 0.01 0.006 0.01 0.014 0.013 0.001 0.003 0.002 0.008 0.006 0.006 LPE_(18:3) 0.007 0.008 0.004 0.007 0.007 0.006 0.007 0.006 0.001 0.002 0.001 LPE_(20:4) 0.049 0.049 0.035 0.042 0.07 0.064 0.062 0.069 0.029 0.021 0.035 0.02 0.027 0.017 0.015 0.011 0.012 LPE_(20:5) 0.002 0.003 0.004 0.002 0.003 0.015 0.015 0.02 0.008 0.014 0.008 0.012 0.005 0.007 LPC_(15:0) 0.14 0.07 0.169 0.119 0.126 0.093 0.098 0.08 0.206 0.039 0.174 0.029 0.074 0.049 0.093 0.05 0.041 LPC_(16:0) 0.166 0.096 0.194 0.157 0.178 0.121 0.138 0.105 0.183 0.042 0.244 0.03 0.068 0.051 0.06 0.037 0.033 LPC_(18:3) 0.118 0.059 0.135 0.102 0.076 0.057 0.055 0.056 0.172 0.028 0.188 0.028 0.08 0.051 0.088 0.039 0.032 LPE_(16:0) 0.013 0.016 0.009 0.014 0.022 0.016 0.021 0.014 0.007 0.011 0.008 0.002 0.003 0.002 0.034 0.047 0.033 LPC_(13:0) LPC_(18:1) 0.725 0.39 0.81 0.541 2.543 1.594 1.62 1.457 1.91 0.357 2.216 0.772 2.163 1.415 1.457 0.694 0.613 LPC_(22:6) LPE_(18:1) 0.108 0.093 0.085 0.093 0.298 0.201 0.238 0.195 0.108 0.1 0.16 0.101 0.156 0.111 0.116 0.092 0.102 LPC_(22:5) 0.019 0.006 0.02 0.008 0.016 0.014 0.011 0.013 0.007 0.006 0.001 0.002 0.001 0.023 0.034 0.021 LPCO_(16:0) LPC_(17:0) 0.395 0.156 0.434 0.232 0.457 0.347 0.368 0.298 0.372 0.057 0.356 0.069 0.161 0.125 0.162 0.091 0.083 LPE_(22:6) 0.003 0.003 0.004 0.001 0.001 LPE_(17:0) 0.044 0.025 0.029 0.028 0.049 0.034 0.037 0.028 0.014 0.007 0.019 0.007 0.014 0.009 0.019 0.012 0.015 LPC_(20:3) 0.207 0.108 0.282 0.164 0.192 0.133 0.125 0.121 0.259 0.042 0.263 0.043 0.125 0.075 0.119 0.054 0.046 LPC_(20:4) 0.404 0.172 0.508 0.272 0.567 0.437 0.375 0.412 0.497 0.074 0.473 0.116 0.316 0.181 0.255 0.111 0.1 LPC_(22:4) 0.01 0.004 0.01 0.006 0.027 0.021 0.018 0.021 0.004 0.003 0.003 0.008 0.005 0.003 0.001 0.002 LPE_(12:0) LPE_(22:5) 0.004 0.004 0.005 0.005 0.003 0.002 0.004 LPC_(18:0) 0.237 0.176 0.226 0.234 0.239 0.185 0.195 0.174 0.17 0.04 0.185 0.036 0.085 0.057 0.093 0.043 0.046 LPE_(18:0) 0.048 0.029 0.036 0.036 0.034 0.023 0.025 0.022 0.021 0.008 0.023 0.006 0.012 0.006 0.019 0.01 0.012 LPEO_(16:0) LPE_(22:4) LPC_(19:0) 0.033 0.013 0.031 0.02 0.038 0.027 0.029 0.023 LPCO_(18:0) 0.026 0.007 0.012 0.01 0.043 0.024 0.033 0.02 0.028 0.006 0.027 0.006 0.015 0.01 0.018 0.008 0.008 LPE_(19:0) 0.01 0.006 0.008 0.008 0.005 0.005 0.003 0.003 0.011 0.002 0.012 0.005 0.011 0.008 0.009 0.005 0.005 LPE_(20:3) 0.055 0.019 0.035 0.034 0.019 0.014 0.013 0.017 0.006 0.003 0.007 0.002 0.004 0.002 0.005 0.003 0.003 LPEO_(18:0) 0.057 0.028 0.028 0.038 0.072 0.05 0.051 0.043 0.002 0.001 0.003 0.001 0.001 0.002 0.001 0.002 LPE_(20:0) 0.012 0.006 0.009 0.009 0.01 0.007 0.008 0.007 0.025 0.018 0.035 0.016 0.025 0.015 0.038 0.029 0.031 LPEO_(19:0) 0.002 0.001 0.001 0.001 0.005 0.004 0.005 0.003 LPE_(22:0) 0.007 0.007 0.007 0.005 0.006 0.004 0.004 0.005 0.009 0.003 0.01 0.003 0.005 0.004 0.008 0.004 0.005 LPE_(20:1) 0.002 0.003 0.001 0.003 0.002 0.002 LPE_(21:1) 0.006 0.003 0.006 0.005 0.029 0.02 0.015 0.015 0.005 0.002 0.005 0.001 0.002 0.001 0.006 0.004 0.005 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention_index quantified_m/z Carnitine_(C0) 0.75 162.11247 Carnitine_(C04_dicarboxylic_acid) 0.81 262.12851 Carnitine_(C02) 0.84 204.12303 Carnitine_(C04_OH) 0.85 248.14925 Carnitine_(C04:1) 0.85 230.13869 Carnitine_(C06_OH) 0.87 276.18054 Carnitine_(C03) 0.88 218.13869 Carnitine_(C03_OH) 0.88 234.1336 Carnitine_(C06:1) 0.88 258.16999 Carnitine_(C03_dicarboxylic_acid) 0.9 248.11286 Carnitine_(C03_methyldicarboxylic_acid) 0.9 262.15851 Carnitine_(C05_dicarboxylic_acid) 0.9 276.14416 Carnitine_(C05_OH) 0.9 262.16489 Carnitine_(C04) 0.91 232.15434 Carnitine_(C03:1) 1.05 216.12303 Carnitine_(C05:1) 1.07 244.15434 Carnitine_(C05) 2.63 246.16999 Carnitine_(C06_dicarboxylic_acid) 2.66 290.15981 Carnitine_(C06) 2.69 260.18564 Carnitine_(C10:1_OH) 2.69 330.2275 Carnitine_(C08_dicarboxylic_acid) 2.7 317.18384 Carnitine_(C10:2) 2.74 298.23767 Carnitine_(C07) 2.75 274.20129 Carnitine_(C08:1) 2.77 286.20129 Carnitine_(C10_OH) 2.8 332.24315 Carnitine_(C08) 2.83 288.21694 Carnitine_(C10_dicarboxylic_acid) 2.87 346.22241 Carnitine_(C09) 2.91 302.23259 Carnitine_(C10:1) 2.91 314.23259 Carnitine_(C12:1_OH) 2.93 358.2588 Carnitine_(C10) 3 316.24824 Carnitine_(C12_OH) 3 360.27445 Carnitine_(C14:1_OH) 3.05 386.2901 Carnitine_(C12:1) 3.09 342.26389 Carnitine_(C11) 3.11 330.26389 Carnitine_(C14:2) 3.2 354.30027 Carnitine_(C18:2_OH) 3.2 440.33705 Carnitine_(C14_OH) 3.21 388.30575 Carnitine_(C12) 3.26 344.27954 Carnitine_(C16:1_OH) 3.45 414.3214 Carnitine_(C14:1) 3.46 370.29519 Carnitine_(C13) 3.5 358.29519 Carnitine_(C16:2) 3.5 396.31084 Carnitine_(C18:1_OH) 3.5 442.3527 Carnitine_(C16_OH) 3.6 416.33705 Carnitine_(C18:3) 3.6 422.32649 Carnitine_(C14) 3.63 372.31084 Carnitine_(C22:5) 3.7 474.35779 Carnitine_(C18:2) 3.8 424.34214 Carnitine_(C16:1) 3.85 389.32649 Carnitine_(C15) 3.88 386.32649 Carnitine_(C18_OH) 4.05 444.36835 Carnitine_(C16) 4.2 400.34214 Carnitine_(C18:1) 4.2 426.35779 Carnitine_(C17) 4.4 414.35779 Carnitine_(C18) 4.8 428.37344 Carnitine_(C19) 5 442.38909 Carnitine_(C20) 5.3 456.40474 Cer_(d18:1_15:0) 3.55 524.5037 Cer_(d18:0_15:0) 3.64 526.5194 Cer_(d18:0_16:1) 3.95 538.5194 Cer_(d18:1_17:0) 4 552.535 Cer_(d18:0_16:0) 4.05 540.535 Cer_(d18:1_18:1) 4.14 564.535 Cer_(d18:0_17:0) 4.3 554.5507 Cer_(d18:0_18:1) 4.3 566.5507 Cer_(d18:1_24:2) 4.31 645.606 Cer_(d18:0_18:0) 4.38 568.5663 Cer_(d18:1_20:1) 4.47 592.5663 Cer_(d18:1_22:1) 4.54 620.5976 Cer_(d18:0_20:1) 4.61 594.582 Cer_(d18:0_20:0) 4.66 596.5976 Cer_(d18:0_22:1) 4.8 622.6133 Cer_(d18:1_24:1) 4.8 648.6289 Cer_(d18:0_22:0) 4.9 624.6289 Cer_(d18:0_24:1) 5.03 650.6446 Cer_(d18:0_24:0) 5.13 652.6602 Cer_(d18:1_26:1) 5.23 676.6602 Cer_(d18:0_26:1) 5.24 678.6759 Cer_(d18:0_26:0) 5.32 680.6915 LPC_(12:0) 0.39 440.2772 LPC_(18:2) 0.41 520.3398 LPC_(20:5) 0.41 542.3241 LPE_(14:0) 0.41 426.2615 LPC_(14:0) 0.43 468.3085 LPC_(16:1) 0.43 494.3241 LPC_(17:1) 0.43 508.3398 LPE_(15:0) 0.43 440.2772 LPE_(18:2) 0.43 478.2928 LPE_(16:1) 0.45 452.2772 LPE_(17:1) 0.45 466.2928 LPE_(18:3) 0.45 476.2772 LPE_(20:4) 0.45 502.2928 LPE_(20:5) 0.45 500.2772 LPC_(15:0) 0.46 482.3241 LPC_(16:0) 0.49 496.3398 LPC_(18:3) 0.49 518.3241 LPE_(16:0) 0.49 454.2928 LPC_(13:0) 0.4 454.2928 LPC_(18:1) 0.5 522.3554 LPC_(22:6) 0.5 568.3398 LPE_(18:1) 0.52 480.3085 LPC_(22:5) 0.53 570.3554 LPCO_(16:0) 0.54 482.3605 LPC_(17:0) 0.54 510.3554 LPE_(22:6) 0.55 526.2928 LPE_(17:0) 0.56 468.3085 LPC_(20:3) 0.58 546.3554 LPC_(20:4) 0.59 544.3398 LPC_(22:4) 0.59 572.3711 LPE_(12:0) 0.39 398.2302 LPE_(22:5) 0.6 528.3085 LPC_(18:0) 0.61 524.3711 LPE_(18:0) 0.64 482.3241 LPEO_(16:0) 0.65 440.3435 LPE_(22:4) 0.69 530.3241 LPC_(19:0) 0.71 538.3867 LPCO_(18:0) 0.73 510.3918 LPE_(19:0) 0.73 496.3398 LPE_(20:3) 0.74 504.3085 LPEO_(18:0) 0.77 468.3448 LPE_(20:0) 0.91 510.3554 LPEO_(19:0) 0.93 482.3605 LPE_(22:0) 1.41 538.3867 LPE_(20:1) 0.67 508.3398 LPE_(21:1) 0.89 521.3481 METABOLITES_END #END