#METABOLOMICS WORKBENCH shancock_20250220_161950 DATATRACK_ID:5661 STUDY_ID:ST003763 ANALYSIS_ID:AN006178 PROJECT_ID:PR002346 VERSION 1 CREATED_ON February 21, 2025, 9:57 pm #PROJECT PR:PROJECT_TITLE Effect of SCD1 or FADS2 knockdown on fatty acid profile of PDAC cells. PR:PROJECT_SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive disease with few PR:PROJECT_SUMMARY treatment options and poor survivability. In this work we sought to characterise PR:PROJECT_SUMMARY changes in fatty acid profile following small interfering RNA knockdown of PR:PROJECT_SUMMARY stearoyl-CoA desaturase 1 (SCD1) or fatty acid desaturase 2 (FADS2). PR:INSTITUTE Victor Chang Cardiac Research Institute PR:LAST_NAME Hancock PR:FIRST_NAME Sarah PR:ADDRESS Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010, PR:ADDRESS Australia PR:EMAIL s.hancock@victorchang.edu.au PR:PHONE +61414537526 #STUDY ST:STUDY_TITLE Effect of SCD1 or FADS2 knockdown on fatty acid profile of PDAC cells. ST:STUDY_SUMMARY In this work we sought to characterise the impact of knockdown of stearoyl-CoA ST:STUDY_SUMMARY desaturase 1 (SCD1) and fatty acid desaturase 2 (FADS2) on Panc1 and MiaPaca2 ST:STUDY_SUMMARY PDAC cells by small interfering RNA (siRN). Panc1 cells exhibit significant Δ6 ST:STUDY_SUMMARY desaturase activity on saturated fatty acids producing n-10 monounsaturated ST:STUDY_SUMMARY fatty acids, whereas MiaPaCa2 exhibit little to no FADS2 activity. Knockdown of ST:STUDY_SUMMARY SCD1 (siSCD) in MiaPaCa2 cells produces a significant decrease in 16:1n-7 when ST:STUDY_SUMMARY compared with an siRNA negative control (siNEG). In Panc1 cells, siSCD results ST:STUDY_SUMMARY in a large increase in FADS2-derived 16:1n-10, whereas siFADS2 results in a ST:STUDY_SUMMARY large increase in SCD1-derived 16:1n-7. ST:INSTITUTE Victor Chang Cardiac Research Institute ST:LAST_NAME Hancock ST:FIRST_NAME Sarah ST:ADDRESS Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010, ST:ADDRESS Australia ST:EMAIL s.hancock@victorchang.edu.au ST:PHONE +61414537526 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 - Blank_Mia Sample source:Blank | group1:Blank | group2:Blank | batch:1 RAW_FILE_NAME(Raw file name)=Blank_Mia.mzML; protein=- SUBJECT_SAMPLE_FACTORS - Blank_p4 Sample source:Blank | group1:Blank | group2:Blank | batch:2 RAW_FILE_NAME(Raw file name)=Blank_p4.mzML; protein=- SUBJECT_SAMPLE_FACTORS - Blank_Panc1 Sample source:Blank | group1:Blank | group2:Blank | batch:1 RAW_FILE_NAME(Raw file name)=Blank_Panc1.mzML; protein=- SUBJECT_SAMPLE_FACTORS - FAMEs-mix_01 Sample source:FAMEs | group1:FAMEs | group2:FAMEs | batch:1 RAW_FILE_NAME(Raw file name)=FAMEs-mix_01.mzML; protein=- SUBJECT_SAMPLE_FACTORS - FAMEs-mix_02 Sample source:FAMEs | group1:FAMEs | group2:FAMEs | batch:1 RAW_FILE_NAME(Raw file name)=FAMEs-mix_02.mzML; protein=- SUBJECT_SAMPLE_FACTORS - iQC_01 Sample source:pooled QC | group1:iQC | group2:iQC | batch:1 RAW_FILE_NAME(Raw file name)=iQC_01.mzML; protein=- SUBJECT_SAMPLE_FACTORS - iQC_02 Sample source:pooled QC | group1:iQC | group2:iQC | batch:1 RAW_FILE_NAME(Raw file name)=iQC_02.mzML; protein=- SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siFADS2_p1 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siFADS2_p1.mzML; protein=137.2225383 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siFADS2_p2 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siFADS2_p2.mzML; protein=127.7411941 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siFADS2_p3 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siFADS2_p3.mzML; protein=186.4606517 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siNEG_p1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siNEG_p1.mzML; protein=194.8003146 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siNEG_p2 Sample source:MiaPaCa2 cells | group1:siNEG | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siNEG_p2.mzML; protein=167.7533723 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siNEG_p3 Sample source:MiaPaCa2 cells | group1:siNEG | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siNEG_p3.mzML; protein=220.8306083 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siSCD_p1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siSCD_p1.mzML; protein=90.33670901 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siSCD_p2 Sample source:MiaPaCa2 cells | group1:siSCD | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siSCD_p2.mzML; protein=100.3769553 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_CON_siSCD_p3 Sample source:MiaPaCa2 cells | group1:siSCD | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_CON_siSCD_p3.mzML; protein=125.186033 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siFADS2_p1 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siFADS2_p1.mzML; protein=142.8434602 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siFADS2_p2 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siFADS2_p2.mzML; protein=198.3417456 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siFADS2_p3 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siFADS2_p3.mzML; protein=94.43047716 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siNEG_p1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siNEG_p1.mzML; protein=156.6314238 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siNEG_p2 Sample source:MiaPaCa2 cells | group1:siNEG | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siNEG_p2.mzML; protein=242.7150091 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siNEG_p3 Sample source:MiaPaCa2 cells | group1:siNEG | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siNEG_p3.mzML; protein=103.3206361 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siSCD_p1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siSCD_p1.mzML; protein=116.9838096 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siSCD_p2 Sample source:MiaPaCa2 cells | group1:siSCD | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siSCD_p2.mzML; protein=160.4036837 SUBJECT_SAMPLE_FACTORS - MiaPaCa2_GEMR_siSCD_p3 Sample source:MiaPaCa2 cells | group1:siSCD | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=MiaPaCa2_GEMR_siSCD_p3.mzML; protein=81.62299067 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siFADS2_p1 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siFADS2_p1.mzML; protein=147.2135179 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siFADS2_p2 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siFADS2_p2.mzML; protein=159.3717429 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siFADS2_p3 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siFADS2_p3.mzML; protein=133.5628 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siFADS2_p4 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:2 RAW_FILE_NAME(Raw file name)=Panc1_CON_siFADS2_p4.mzML; protein=269.2377164 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siNEG_p1 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siNEG_p1.mzML; protein=194.0193044 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siNEG_p2 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siNEG_p2.mzML; protein=167.7244666 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siNEG_p3 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siNEG_p3.mzML; protein=134.929931 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siNEG_p4 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:2 RAW_FILE_NAME(Raw file name)=Panc1_CON_siNEG_p4.mzML; protein=194.1686899 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siSCD_p1 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siSCD_p1.mzML; protein=145.9809695 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siSCD_p2 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siSCD_p2.mzML; protein=135.2898928 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siSCD_p3 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_CON_siSCD_p3.mzML; protein=113.8309455 SUBJECT_SAMPLE_FACTORS - Panc1_CON_siSCD_p4 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:2 RAW_FILE_NAME(Raw file name)=Panc1_CON_siSCD_p4.mzML; protein=201.4310037 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siFADS2_p1 Sample source:Panc1 cells | group1:siFADS2 | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siFADS2_p1.mzML; protein=156.9619387 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siFADS2_p2 Sample source:Panc1 cells | group1:siFADS2 | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siFADS2_p2.mzML; protein=115.608538 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siFADS2_p4 Sample source:Panc1 cells | group1:siFADS2 | group2:GEMR | batch:2 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siFADS2_p4.mzML; protein=235.2658543 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siNEG_p1 Sample source:Panc1 cells | group1:siNEG | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siNEG_p1.mzML; protein=155.7219314 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siNEG_p2 Sample source:Panc1 cells | group1:siNEG | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siNEG_p2.mzML; protein=117.3880465 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siNEG_p4 Sample source:Panc1 cells | group1:siNEG | group2:GEMR | batch:2 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siNEG_p4.mzML; protein=213.408657 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siSCD_p1 Sample source:Panc1 cells | group1:siSCD | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siSCD_p1.mzML; protein=124.3823145 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siSCD_p2 Sample source:Panc1 cells | group1:siSCD | group2:GEMR | batch:1 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siSCD_p2.mzML; protein=93.13830126 SUBJECT_SAMPLE_FACTORS - Panc1_GEMR_siSCD_p4 Sample source:Panc1 cells | group1:siSCD | group2:GEMR | batch:2 RAW_FILE_NAME(Raw file name)=Panc1_GEMR_siSCD_p4.mzML; protein=170.7392053 #COLLECTION CO:COLLECTION_SUMMARY Samples were obtained from established immortalised adherent human pancreatic CO:COLLECTION_SUMMARY ductal adenocarcinoma cells (Panc1 & MiaPaCa2). Cells were cultured in high CO:COLLECTION_SUMMARY glucose DMEM containing 4.5 g/L glucose and 4 mM glutamine and were supplemented CO:COLLECTION_SUMMARY with 10% FCS. Cells were cultured under 5% CO2 at 37ºC and routinely passaged CO:COLLECTION_SUMMARY every 2-3 days (when ~80-90% confluent). Cell lines were regularly screened for CO:COLLECTION_SUMMARY mycoplasma infection. CO:SAMPLE_TYPE Pancreas #TREATMENT TR:TREATMENT_SUMMARY Gemcitabine (GEMC) resistant (GEMR) cells were produced by serial treatment with TR:TREATMENT_SUMMARY escalating doses of GEMC for approximately 12 weeks. The final concentration of TR:TREATMENT_SUMMARY GEMC for Panc1 cells was 150 nM, resulting in a half-maximal inhibitory TR:TREATMENT_SUMMARY concentration of 720.5 nM (approximately 2.5 times increase over control (CON) TR:TREATMENT_SUMMARY cells, 273.3 nM). The final concentration of GEMC for MiaPaCa2 cells was 64 nM, TR:TREATMENT_SUMMARY resulting in a half-maximal inhibitory concentration of 92.2 nM (approximately TR:TREATMENT_SUMMARY 3x increase over control (CON) cells, 30.6 nM). Knockdown experiments were TR:TREATMENT_SUMMARY performed in adherent cells grown in 6 well plates for 72 h. Cells were seeded TR:TREATMENT_SUMMARY at 1-3105 cells/well and allowed to adhere overnight. Cells were transfected TR:TREATMENT_SUMMARY with Silencer Select siRNA targeting ACACA (s883), FASN (s5030), SCD TR:TREATMENT_SUMMARY (s12505,s12504) FADS2 (s18025,s18023) or negative control (Silencer Select TR:TREATMENT_SUMMARY Negative Control No. 1) using Lipofectamine RNAiMAX transfection reagent as per TR:TREATMENT_SUMMARY the manufacturer's instructions (ThermoFisher Scientific). Sample groups include TR:TREATMENT_SUMMARY source - blank, fatty acid methyl ester (FAMEs) reference standard, pooled TR:TREATMENT_SUMMARY internal quality controls (iQC), MiaPaCa2 & Panc1 cells group1 - siRNA knockdown TR:TREATMENT_SUMMARY groups, negative control (siNEG), stearoyl-CoA desaturase 1 (siSCD) and fatty TR:TREATMENT_SUMMARY acid desaturase 2 (siFADS2). group2 - control (CON) or gemcitabine-resistant TR:TREATMENT_SUMMARY (GEMR) cells batch - two different batches were process to ensure a minimum n>2 #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Lipids were extracted from cells grown to 80-90% confluency in 6 well plates SP:SAMPLEPREP_SUMMARY using a modified methyl tert-butyl ether (MTBE) method. Adherent cells were SP:SAMPLEPREP_SUMMARY washed with ice-cold PBS and scraped into methanol containing 0.01% butylated SP:SAMPLEPREP_SUMMARY hydroxytoluene and 100 pmol each of internal standards (phosphatidylcholine SP:SAMPLEPREP_SUMMARY 19:0/19:0, phosphatidylethanolamine 17:0/17:0, phosphatidylserine 17:0/17:0, SP:SAMPLEPREP_SUMMARY lysophosphatidylcholine 17:0, lysophosphatidylethanolamine 14:0, SP:SAMPLEPREP_SUMMARY phosphatidylglycerol 17:0/17:0, phosphatidic acid 17:0/17:0, ceramide SP:SAMPLEPREP_SUMMARY d18:1/17:0, dihydrosphingomyelin 12:0, cholesteryl ester 22:1, diacylglycerol SP:SAMPLEPREP_SUMMARY 17:0/17:0, D5-triacylglycerol 16:0/16:0/16:0 and cardiolipin SP:SAMPLEPREP_SUMMARY 14:0/14:0/14:0/14:0, and 20 nmol of methyl nonadecanoate). An empty well was SP:SAMPLEPREP_SUMMARY also scraped for use in background subtraction. To this, MTBE was added at a SP:SAMPLEPREP_SUMMARY ratio of 3:1 v/v, and extracts were rotated overnight at 4ºC. Following SP:SAMPLEPREP_SUMMARY overnight rotation, 1 part of ice-cold 150 mM ammonium acetate was added, SP:SAMPLEPREP_SUMMARY samples were vortexed vigorously and centrifuged at 2000 x g for 5 min. The SP:SAMPLEPREP_SUMMARY upper organic phase containing lipids was transferred into a 1.5 mL autosampler SP:SAMPLEPREP_SUMMARY vial and dried under N2 at 37ºC Dried lipids were reconstituted in SP:SAMPLEPREP_SUMMARY chloroform:methanol:water (60:30:4.5 v/v/v), transferred to a sleeved vial, and SP:SAMPLEPREP_SUMMARY stored at -20ºC until analysis. The aqueous phase containing the protein pellet SP:SAMPLEPREP_SUMMARY was dried under N2 and digested with 1M NaOH overnight at 4ºC. Digested protein SP:SAMPLEPREP_SUMMARY was diluted 1:2 v/v with water and protein concentration was determined using a SP:SAMPLEPREP_SUMMARY Pierce BCA assay kit (ThermoFisher Scientific) Following MTBE extraction of SP:SAMPLEPREP_SUMMARY lipids, an aliquot was taken and hydrolysed in 0.6 M KOH in 75% v/v methanol at SP:SAMPLEPREP_SUMMARY 60ºC for 30 min. Samples were then cooled to room temperature and neutralised SP:SAMPLEPREP_SUMMARY with 25% v/v acetic acid. Water and n-hexane (3:2 v/v) were added separately and SP:SAMPLEPREP_SUMMARY samples were vortexed vigorously before being centrifuged at 2000 x g for 5 min. SP:SAMPLEPREP_SUMMARY The upper phase containing hydrolysed fatty acids was removed, and the aqueous SP:SAMPLEPREP_SUMMARY phase was washed with a second volume of n-hexane and combined. The combined SP:SAMPLEPREP_SUMMARY organic phase was then dried under N2 at 37°C. Dried hydrolysed fatty acids SP:SAMPLEPREP_SUMMARY were derivatised with an AMP+ MaxSpec kit (Cayman Chemical, Ann Arbor, MI, USA) SP:SAMPLEPREP_SUMMARY as per manufacturer instructions. Following this, AMP-derivatised fatty acids SP:SAMPLEPREP_SUMMARY were re-extracted using MTBE:water (1:1 v/v), and the upper MTBE phase was dried SP:SAMPLEPREP_SUMMARY under N2 before being resuspended in methanol. Samples were stored at -20°C SP:SAMPLEPREP_SUMMARY until analysis. A 37-component fatty acid methyl ester standard (Merck Life SP:SAMPLEPREP_SUMMARY Sciences) containing an additional 3.184 nmol of methyl nonadecanoate was SP:SAMPLEPREP_SUMMARY concurrently subjected to the same derivatisation procedure and used as an SP:SAMPLEPREP_SUMMARY external quality control for fatty acid identification by LC-MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity II CH:COLUMN_NAME Thermo Accucore C30 (150 x 2.1mm,2.6um) CH:SOLVENT_A Water (0.1% formic acid) CH:SOLVENT_B Acetonitrile (0.1% formic acid) CH:FLOW_GRADIENT 0-3.25 min increase from 50% B to 56% B, 3.25-6.5 min increase to 58% B, 6.5-7.5 CH:FLOW_GRADIENT min increase to 80% B, 7.5-9.5 min increase to 100% B, hold at 100% B for 1.5 CH:FLOW_GRADIENT mins, re-equilibrated at 50% B for 4.5 mins. CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 30 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6560 Ion Mobility MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS AMP-derivatised fatty acids were detected as positive ions in QTOF-only mode on MS:MS_COMMENTS an Agilent 6560 IM-MS QTOF mass spectrometer (Agilent Technologies). Source MS:MS_COMMENTS conditions were as follows: capillary voltage of 3.5 kV, nozzle voltage of 1 kV MS:MS_COMMENTS and fragmentor at 360 V. Nitrogen was used as the sheath and collision gas, with MS:MS_COMMENTS drying gas set at 5 L/min and sheath gas at 12 L/min (both gases heated to MS:MS_COMMENTS 300°C) and the nebuliser pressure set at 20 psi. Ions were acquired in Auto MS:MS_COMMENTS MS/MS mode at an MS1 mass range of m/z 100-1000 and MS2 mass range of m/z MS:MS_COMMENTS 50-600. Spectra were acquired in MS1 mode at 5 spectra/s and 200 ms/spectrum, MS:MS_COMMENTS and in MS2 mode at 10 spectra/s and 100 ms/spectrum. Precursor ions were MS:MS_COMMENTS isolated using a window of 1.3 Th, acquisition time of 50 ms/spectra, and MS:MS_COMMENTS collision energy of 45 eV. Calibration mix A was infused throughout and masses MS:MS_COMMENTS were automatically detected and corrected using a 100 ppm window and minimum MS:MS_COMMENTS height of 1x10^3 counts to ensure 5 ppm mass accuracy, with m/z 121.050873 and MS:MS_COMMENTS 922.009798 used as reference ions. AMP-derivatised fatty acids as [M+H]+ were MS:MS_COMMENTS aligned using Mass Profinder v10.0 (Agilent Technologies) with an RT tolerance MS:MS_COMMENTS of 0.5 min and MS1 tolerance of 10 ppm. Lipids were identified using a custom MS:MS_COMMENTS personal compound database and library (PCDL) generated from the RT of external MS:MS_COMMENTS quality control samples (attached) and MS/MS spectra were manually inspected to MS:MS_COMMENTS ensure correct MUFA isomer identification. Using this method, FA 18:1 isomers MS:MS_COMMENTS were only partially separated and so these data were not analysed further. Data MS:MS_COMMENTS were exported, blank subtracted and quantified from internal standards before MS:MS_COMMENTS being normalised to total protein. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/mg protein MS_METABOLITE_DATA_START Samples MiaPaCa2_CON_siFADS2_p1 MiaPaCa2_CON_siFADS2_p2 MiaPaCa2_CON_siFADS2_p3 MiaPaCa2_CON_siNEG_p1 MiaPaCa2_CON_siNEG_p2 MiaPaCa2_CON_siNEG_p3 MiaPaCa2_CON_siSCD_p1 MiaPaCa2_CON_siSCD_p2 MiaPaCa2_CON_siSCD_p3 MiaPaCa2_GEMR_siFADS2_p1 MiaPaCa2_GEMR_siFADS2_p2 MiaPaCa2_GEMR_siFADS2_p3 MiaPaCa2_GEMR_siNEG_p1 MiaPaCa2_GEMR_siNEG_p2 MiaPaCa2_GEMR_siNEG_p3 MiaPaCa2_GEMR_siSCD_p1 MiaPaCa2_GEMR_siSCD_p2 MiaPaCa2_GEMR_siSCD_p3 Panc1_CON_siFADS2_p1 Panc1_CON_siFADS2_p2 Panc1_CON_siFADS2_p3 Panc1_CON_siFADS2_p4 Panc1_CON_siNEG_p1 Panc1_CON_siNEG_p2 Panc1_CON_siNEG_p3 Panc1_CON_siNEG_p4 Panc1_CON_siSCD_p1 Panc1_CON_siSCD_p2 Panc1_CON_siSCD_p3 Panc1_CON_siSCD_p4 Panc1_GEMR_siFADS2_p1 Panc1_GEMR_siFADS2_p2 Panc1_GEMR_siFADS2_p4 Panc1_GEMR_siNEG_p1 Panc1_GEMR_siNEG_p2 Panc1_GEMR_siNEG_p4 Panc1_GEMR_siSCD_p1 Panc1_GEMR_siSCD_p2 Panc1_GEMR_siSCD_p4 Factors Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:CON | batch:1 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siFADS2 | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siNEG | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:GEMR | batch:1 Sample source:MiaPaCa2 cells | group1:siSCD | group2:GEMR | batch:1 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:1 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:1 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:1 Sample source:Panc1 cells | group1:siFADS2 | group2:CON | batch:2 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:1 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:1 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:1 Sample source:Panc1 cells | group1:siNEG | group2:CON | batch:2 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:1 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:1 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:1 Sample source:Panc1 cells | group1:siSCD | group2:CON | batch:2 Sample source:Panc1 cells | group1:siFADS2 | group2:GEMR | batch:1 Sample source:Panc1 cells | group1:siFADS2 | group2:GEMR | batch:1 Sample source:Panc1 cells | group1:siFADS2 | group2:GEMR | batch:2 Sample source:Panc1 cells | group1:siNEG | group2:GEMR | batch:1 Sample source:Panc1 cells | group1:siNEG | group2:GEMR | batch:1 Sample source:Panc1 cells | group1:siNEG | group2:GEMR | batch:2 Sample source:Panc1 cells | group1:siSCD | group2:GEMR | batch:1 Sample source:Panc1 cells | group1:siSCD | group2:GEMR | batch:1 Sample source:Panc1 cells | group1:siSCD | group2:GEMR | batch:2 14:0-AMP 9.213018327 6.257385408 6.753414012 6.443082361 5.551923835 6.401136109 12.94137848 0 6.797071772 4.372373789 4.632275842 0 3.15317508 2.559920645 1.132167386 0 4.422496914 1.40169026 16.60546404 10.92586818 16.21843195 4.399323927 7.346311419 23.593961 6.534312029 7.694008138 7.777915004 10.08667111 9.648115799 7.195801768 11.39303268 17.44571419 11.41213291 9.613562815 9.342916787 9.288413499 0 20.21989191 6.19822701 15:0-AMP 0 0 0 0 0 0 0 0 0 0.579452565 0.460569862 0 0 0 0 0 0 0 1.162770664 1.0918609 0.979657082 0 0.29426026 3.912496884 0.001780816 0 0.516027659 0.533028061 0.79724263 0 1.156692394 1.749053267 0 0.771354253 1.199964388 0 0 0.586829641 0 15:1-AMP 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.037149697 0 0 0.028784493 0 0 0 0 0 0 0 0 0 0 0 0 0 13.51639679 0 0 16:0-AMP 220.3892299 154.3737586 195.1375687 147.706217 154.4804286 143.1814232 303.4112 0 186.63581 140.4303831 128.2286271 52.81379298 127.4820401 96.37272574 80.2214895 0 159.7755388 82.48297775 260.1609007 193.9317521 243.163318 57.56145568 0 215.3087302 137.3963173 84.89767167 139.3951578 182.2952968 200.7225281 77.38689148 165.7258748 252.8195503 113.5202554 134.7761333 155.4063587 80.97524899 0 265.4337706 77.32033418 16:1n10-AMP 3.750107527 3.138867605 3.4279086 1.425044746 3.111977123 3.52413906 4.451274191 2.094216973 3.131223325 2.609630195 3.093534298 3.291942467 2.504882326 1.670590165 2.759758841 3.600999993 2.853568562 2.754208389 5.302307171 3.888485164 6.061483647 2.581650525 7.707789888 23.03151929 7.62350039 10.2255859 16.56787503 22.71929419 20.80677423 16.84779936 2.856385856 5.929125418 3.829083923 5.155123026 4.546522046 7.884622016 11.61969608 21.55892431 19.0488486 16:1n7-AMP 50.69090973 36.49500839 43.39466048 33.38125138 34.63842982 39.28082302 19.4138944 11.44365908 15.79557281 23.44801315 29.61886803 15.58876402 20.23168693 19.16889623 8.812627504 14.11077668 12.95444439 5.226998882 44.95499465 31.51564838 39.16490252 16.54558671 12.67949932 40.16288466 13.05541911 17.63809068 6.625095651 8.918197908 9.13770902 7.103026219 35.50971714 45.93662404 43.65374296 17.9503833 17.85572599 25.50569729 9.389328821 14.58435915 10.53037857 16:1n9-AMP 7.709109394 7.252144151 8.72276712 5.58867675 6.425109318 6.746516653 5.06880781 4.394395501 4.346300226 4.539449579 7.938451509 7.126817429 4.131913246 3.993761778 3.944605233 4.933019826 4.688865248 3.202074787 9.380370328 4.83249772 7.367503262 2.483283818 2.584404695 8.322972804 2.878175181 3.05670132 2.353517613 3.250521435 3.157564107 2.206046639 5.041849015 9.119454036 8.080656667 3.099333743 3.961443696 2.849403304 4.70239176 5.782262472 2.654637442 17:0-AMP 0 0 0 0 0 0 0 0 0 1.191564486 0.858786315 0 0 0 0 0 0 0 0.194047021 2.725325051 0 0 1.247629431 10.05369114 0 0 0 1.462726065 0 0 0 0 0 2.188611396 1.968446228 0 0 0 0 17:1-AMP 8.101359449 6.058617823 6.203800157 5.374178566 6.119586894 5.293479164 9.053986309 5.545667406 6.08968679 6.076619261 7.01288422 6.964494829 5.732514537 4.530837424 5.208643545 5.90186859 7.10735702 5.096758767 7.20900648 5.435305783 6.680839617 2.863745618 3.228501421 7.379485836 3.608887237 3.337614694 2.753567716 4.296087059 4.817538212 2.921877525 4.678417556 7.923217093 4.749277739 3.134685989 4.6090929 3.212313381 1.031272886 5.911678271 3.363476866 18:0-AMP 75.51656682 55.08814449 66.42775189 61.84181296 43.81954518 38.85823819 237.2532601 0 157.3084836 120.3700622 115.1838083 65.90770552 82.46667296 86.19426764 96.00841691 0 221.2376735 191.7039578 145.9490248 142.828632 116.5402633 51.02533386 105.8022377 195.8884334 109.7587602 62.25455465 193.7534959 208.9681087 227.106782 126.7917376 106.579172 124.1053245 45.9696692 104.7795143 112.2258573 47.36005281 0 203.4494358 103.5488691 18:1n10-AMP 54.93727663 54.18796828 69.58772627 56.96377333 51.79922595 60.0256008 90.61587302 64.31372264 87.14323817 60.10969535 65.84397015 96.68551487 79.07798419 39.03783458 74.14814618 145.2380174 71.65728889 89.68970184 88.79684739 69.59843598 115.9966256 38.80418657 66.30668362 97.1692666 90.63867608 88.98864341 93.85963816 122.1174166 120.5209528 83.39422577 32.85196288 73.57046651 60.89757813 48.95655363 69.32749122 54.87784939 64.73970397 130.4502163 97.67526993 18:1n7-AMP 138.563483 88.93916884 135.1517446 132.3099015 126.2654883 109.2566617 111.1181041 56.29555679 97.3526033 80.1020033 89.23663212 54.17812621 93.94685197 88.26838755 36.19666509 73.88485681 62.69535268 51.89842641 108.4359442 93.40730976 127.3942864 59.46395534 58.26975329 125.1503174 79.53295538 104.2449733 52.78386703 64.13477203 71.55530838 39.0064576 132.9200938 142.5234096 98.09549478 80.17102238 62.24499541 101.258607 17.96479692 63.61152528 63.10940528 18:1n9-AMP 360.9585282 333.4583749 327.0285857 283.4040343 325.2306023 318.5081251 433.2737877 300.6301173 339.0945064 286.2827733 285.8573672 367.7278978 341.8888305 278.8701584 320.0039188 336.648614 335.9650288 302.5749009 371.662369 290.2169879 391.2927689 173.4376928 210.7449224 279.7795491 284.7386832 250.0472231 192.3423345 274.8027343 310.7475982 166.1383177 199.3194669 346.4599754 241.4463612 204.8146567 267.2028355 185.7048355 130.0286667 343.6463928 226.2981462 18:1n9t-AMP 7.02064311 6.541787751 6.254042334 2.824364825 5.996573959 5.737297043 8.419542387 6.919326202 6.690654318 5.85484285 6.057042065 8.408577111 5.618104945 4.575549504 5.387283464 8.909231154 7.082920216 6.035151214 8.624049931 6.276397486 8.430002738 3.549950587 6.760593241 16.15111699 7.64399607 8.241898034 15.71043445 22.45862905 20.69316847 17.02995295 4.549572109 7.76294168 4.734403742 4.58156304 6.145471827 4.776727536 13.88268151 14.29110819 15.93456759 18:2n6-AMP 14.66957465 12.21351792 10.83855054 9.176557224 12.5778319 9.701860909 20.50100246 13.70456218 13.60117997 10.42018919 11.50799718 12.05641318 9.648135856 7.849183171 8.942863077 14.15238337 14.98278165 10.02514035 11.07155386 9.53803191 10.02803402 5.035719176 5.535877985 12.52360567 7.3490142 6.802156277 5.938270318 9.207800822 10.4312856 6.342983897 8.059580551 13.77670311 7.603397274 5.591509599 7.77188049 6.501493858 8.807929235 16.45118066 6.928164629 18:2n6t-AMP 0.62910841 0.297406745 0.48701888 0.206200629 0.541499847 0.59315199 0.81364595 0.340839117 0.747074283 0.331060723 0.502862789 0.686649789 0.306778243 0.289685046 0.420582073 0.267142266 0.656825523 0.35985283 1.393756765 1.073703386 1.418539437 0.578836896 1.053925087 2.931938705 1.31595831 1.957922019 1.695157752 2.464619188 2.219740596 1.949157954 0.464894513 0.946124869 1.087938141 0.782872158 0.844847974 1.280345307 4.991333345 1.68041882 1.907970291 18:3n3-AMP 0.352691909 0.219962387 0.203617048 0.201073124 0.229108715 0.216428129 0.270303027 0.092232225 0.186063621 0.148427758 0.282550854 0.383849527 0.215169853 0.146866922 0.110006258 0 0.221853673 0.064957144 0.024315129 0.121499391 0.080168606 0.060347789 0 0.086827633 0.031268245 0.038675909 0.000679381 0.067716118 0.022772804 0.050376909 0.160571111 0.210895647 0.081493415 0.037188938 0.060389828 0.015100537 10.14108875 0.077908912 0.100369161 18:3n6-AMP 0.230019461 0.034130442 0.053401066 0.046651582 0.080080513 0.112296396 0.212133852 0 0 0.111942607 0.135058541 0.156775282 0.042900761 0.021060694 0.128088067 0 0.089131864 0.159940831 0.055816359 0.088484958 0.030356788 0.015436877 0.072416925 0.285511217 0.037420424 0.095058376 0.049244501 0.099638223 0.15855528 0.103221814 0.034521316 0.057197149 0.039573987 0.039739504 0.033955408 0.078864417 9.096487839 0.141692049 0.192612793 19:0-AMP 145.0199089 155.7837324 106.72493 102.1558925 118.6265273 90.11431954 220.286971 198.252676 158.9634205 139.3133432 100.3318789 210.737048 127.049857 81.98916117 192.6043117 170.1090097 124.0619887 243.8038577 135.1778035 124.8652969 148.993582 73.9123785 102.5671134 118.6469714 147.483956 102.4882025 136.3191385 147.0915498 174.820651 98.7931333 126.782328 172.1326153 84.5851603 127.7918905 169.5232231 93.24832591 159.9905909 213.6607575 116.5520243 19:1-AMP 3.238559182 3.237625717 2.47705057 2.948707815 3.348168464 2.989787028 3.690075542 2.237019755 2.648222771 2.815527309 2.530864509 1.659032187 3.273306957 2.834455932 2.602182275 1.471782328 3.511104912 2.5080314 3.335739161 3.283065792 2.933958792 1.892174084 2.273739653 4.499258494 2.862003451 2.871176529 1.984796409 2.592670024 3.405995793 2.044848995 2.834076784 3.347682581 2.221392149 2.434012144 2.788395038 2.443915639 3.474679072 2.361764519 1.8886734 20:0-AMP 1.288631598 1.032489749 0.894947797 0.764971412 0 0.384580357 3.520661346 0 2.900046468 3.027360907 2.980785199 1.159933871 1.767076245 2.33569955 1.984317173 0 6.507836486 3.405644229 0.95580892 2.26877724 0.856571381 1.12352352 1.449094042 2.876510953 1.133842044 1.186558587 4.025653985 3.43034615 2.657160865 3.385394071 1.85457341 0.820612698 0.714128397 1.79204113 1.442318231 0.990233856 0 2.281518746 2.87952072 20:1-AMP 18.65744342 14.70505552 19.01900986 17.7034881 19.38457196 21.04932812 20.43625189 12.8893029 17.966844 16.89630358 18.16871336 9.398604647 20.84546249 22.95021185 12.5047451 12.66896041 24.70244448 11.66964947 26.13502472 29.08710667 23.40211629 17.79173543 19.76435486 58.88392033 23.08586088 28.56139705 19.55368359 25.49436422 24.84734243 22.58016204 22.9655354 19.40133154 23.22698892 19.98835248 19.80613573 24.29998982 10.76431682 15.8155679 22.06750501 20:2-AMP 3.904858121 2.894366041 3.202852653 3.17554797 3.861826431 3.060122919 5.593044081 3.852480472 3.414218893 2.942913376 2.757086181 2.423864397 3.2197045 2.688998866 2.335141085 2.609845319 4.060978009 3.066284053 8.194764563 8.225266762 6.971224772 4.251070067 8.38788302 25.73387652 8.669154266 12.45284607 7.104566224 10.43419193 10.62264464 7.112704882 5.490335936 5.775210392 8.105114295 7.173223454 6.386327899 14.50955621 2.190331763 6.130795834 8.782104122 20:3-AMP 17.87682953 14.29426808 11.13743675 9.470160731 14.59925829 10.16697822 25.00336558 18.99969507 16.28999215 7.847424799 7.732909534 13.3603701 8.226686199 5.248325712 8.250184027 10.38967807 11.77081759 11.96252135 13.26284885 11.98250155 13.58646364 6.454125811 8.699024559 15.72813813 11.10997644 12.12543634 11.26222967 17.8530342 20.7502416 11.31395916 8.078398299 15.76220544 7.524664602 6.770889693 9.295148523 8.12413013 5.41387826 17.60352587 10.13728519 20:4-AMP 48.7974485 35.83493455 36.28204537 26.9818278 41.34982803 31.85642768 69.64724978 50.08200154 48.98668592 34.17958907 36.44448525 42.44639075 31.81226124 23.87883669 30.78178353 58.78463333 48.63792587 41.96501585 55.27520434 40.5253087 48.87176657 21.64979414 28.63316318 0.246682617 32.16371895 32.654008 33.42802421 46.86768251 57.18639778 29.88297331 25.88469127 50.93734733 27.2252807 21.07554185 30.64179466 23.05412441 16.2390812 64.80032862 30.98829061 20:5-AMP 2.903919194 1.935502887 2.102758908 1.61579121 2.320778991 1.729878168 4.010502743 2.415766534 2.988608674 1.550103076 1.901243799 2.335002287 1.059328887 0.829792664 1.029889434 2.585312004 2.180447574 1.780860596 1.557193861 1.026789755 1.310494971 0.59109563 0.501886166 1.48431755 0.483529489 0.588095672 0.628105059 0.982631559 1.030819435 0.843491858 1.175556178 2.92900671 1.190442655 0.698618201 1.048988821 0.778182227 0.993367497 2.660701964 1.253618258 203:3n9-AMP 2.442954808 1.880754266 2.169276787 1.623810768 1.922976119 1.726884411 3.710340354 3.019353539 2.758383781 1.184833521 1.026346068 1.605706797 1.25359969 0.898218579 0.966152578 2.239742797 2.4284367 1.946194754 7.310938876 5.633748245 6.260622007 3.276621894 8.633061629 12.07218671 8.980090722 12.41076455 7.591912538 12.16937064 10.87803993 8.886812804 2.704421578 3.514756793 2.912729606 4.925750778 4.205519265 6.775478087 3.491354971 6.110738112 7.095876771 21:0-AMP 0 0 0 0 0 0 0 0 0 0.050584229 0 0 0 0 0 0 0 0 0 0 0 0.023794279 0 0.18771962 0 0 0.019900894 0.069101626 0 0.051303037 0.141551918 0 0 0.095419847 0 0.079176928 0 0 0 22:0-AMP 0.65474473 0.762944484 0.626434854 0.724894313 0.24500396 0.310459505 1.946456722 0 1.442837721 2.130263676 2.137897946 0.851965006 0.605342479 1.703508351 0.79646452 0 2.448747167 1.153974699 0 0.839596902 0 1.285880655 0.57113504 1.35562753 0.357939395 0.557366564 1.802939891 0.82511954 0 1.820065064 1.826828585 0 0.511497248 1.057586319 0.982551075 1.081850157 0 0 1.258530489 22:1-AMP 2.702659669 1.970613788 3.093345077 2.692550724 2.356217982 2.991415445 2.540108089 0 1.3174605 4.523600844 5.490761298 1.622518005 4.881247353 5.345962492 3.150351185 0 4.220689585 3.659514856 4.327228636 6.954719871 3.50017615 2.506431918 2.989958161 7.712749261 3.607934177 4.356423519 4.595773234 4.932233199 4.662065988 4.347163613 3.967183955 3.821694118 3.432270326 3.517653552 3.295709213 3.975815716 0 0.654454082 2.672620736 22:2-AMP 0.51469562 0.44643344 0.668529325 0.579531139 0.495245805 0.709995141 0.685909803 0.329492377 0.562283945 0.764533406 0.820583532 0.003235211 0.951576912 1.174178683 0.519752148 0 1.411008844 0.590931448 2.767075378 3.147929038 2.037459891 1.871353913 2.27342582 6.885873675 2.673585956 3.840587064 2.371502484 2.962887519 2.67498168 2.434821074 1.687590064 1.299494012 2.205774562 2.078769899 1.565041317 3.381756815 2.250349782 1.414976359 2.508025442 22:4-AMP 11.94088848 9.022906824 8.941524856 8.096482104 10.53002108 8.713838343 16.08744789 12.83857881 11.89039981 10.95341736 10.01422849 10.07656737 12.54421094 9.261763808 10.61117186 11.94170556 14.0157043 12.19995433 17.58515618 15.06217196 14.8136533 6.770852684 11.08491636 26.86405593 13.42308576 10.6458019 11.53915124 18.18158942 22.74849939 9.41684052 14.71947872 22.44431934 14.69250097 12.20354801 14.98250059 14.86379871 8.461937386 18.82419563 16.6534841 22:5n3-AMP 35.02254629 26.77642893 27.32167658 22.95621066 30.57331606 24.42115908 48.77928381 35.92564172 36.62327773 27.88059172 29.91560991 32.01273532 30.37184051 20.90623027 27.41184685 42.71161627 40.5710995 33.00037401 15.99111797 13.61073335 13.03909749 6.399014768 7.401770753 17.69584498 7.878582006 7.840725781 8.577889128 15.09091965 16.2274086 8.550265399 15.50810678 28.10631848 15.55286424 11.03051893 15.01679671 13.04487898 6.788016954 24.48646654 15.16613576 22:5n6-AMP 1.871260669 0.978894876 1.300683109 0.305279124 1.398745092 1.079276926 2.352668647 0.914789276 1.749574108 0.935562793 0.93944992 1.054674225 0.922851899 0.648640363 0.71798063 1.600767273 1.381974202 1.014034666 8.1516938 6.7046975 6.452195545 3.844862957 8.512420535 18.71811746 10.14111141 10.13135369 7.853937028 11.14844909 13.36638144 7.310869564 2.990020565 3.576239236 2.417208498 3.40819238 3.571762877 3.824857009 1.849791277 4.181364725 3.522524349 22:6-AMP 38.15102173 27.52072103 29.17950057 23.55026086 34.92447215 25.68787838 49.73537416 38.97595976 38.06522487 25.63111394 27.12325497 26.52894383 27.30920073 20.05933069 21.40124465 50.06255288 37.93422741 23.82189968 59.64164012 42.86724915 51.29425979 23.83221142 37.02004928 61.61733909 39.66591136 41.90748668 38.90120382 61.28965317 66.09290128 40.44830413 25.58808617 44.44808268 24.79033497 22.82528382 30.02565431 25.38798256 18.12539898 53.29911103 32.42940286 23:0-AMP 0 0 0.286298472 0 0 0 0.74037622 0 0 0.269363321 0.157078003 0 0.124659082 0 0 0 0 0 0 0 0 0.250382125 0 0.376621261 0 0 0.280611306 0 0 0.268204747 0.491781529 0 0 0.419915638 0.368726686 0.313639456 0 0 0 24:0-AMP 0.538609507 0.775306932 1.418451242 0.370798226 0.070866643 0.737193796 2.157258042 0.177722146 1.534384364 2.753476737 3.191303781 0.377538764 0.269421944 2.712329889 0.478565918 0.789408028 2.235984806 1.932484038 0 1.700731032 0 1.365523986 1.697366203 1.708699236 1.084304407 0.299530509 3.514837064 1.648819548 0.174166873 1.964483829 1.977967167 0.321380821 0.00881901 1.296674754 0.981795898 0.838979692 0 0 0.342518827 24:1-AMP 2.022037198 2.08305474 2.256783512 1.792250172 1.614312189 1.818458754 1.541508246 1.143902951 1.296431805 2.291679864 3.209362039 0.610024744 1.531711095 3.582251486 1.11664871 1.070054693 1.733976915 1.179502744 1.079349499 1.759365662 0.68293094 1.441396827 1.475613879 2.232445459 1.507274556 1.265295755 1.235281596 1.095690856 0.896857316 1.00778934 2.141555707 0.862738589 0.900200787 1.337154768 1.250919167 1.632748922 1.567003778 0.64620599 0.444255706 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Formula Mass RT 14:0-AMP C26 H39 N2 O 395.3024 2.39 15:0-AMP C27 H41 N2 O 409.318 3.21 15:1-AMP C27 H39 N2 O 407.3024 2.12 16:0-AMP C28 H43 N2 O 423.3343 4.23 16:1n10-AMP C28 H41 N2 O 421.3182 3.02 16:1n7-AMP C28 H41 N2 O 421.3183 2.79 16:1n9-AMP C28 H41 N2 O 421.3181 2.87 17:0-AMP C29 H45 N2 O 437.3494 5.62 17:1-AMP C29 H43 N2 O 435.3338 3.69 18:0-AMP C30 H47 N2 O 451.3657 7.47 18:1n10-AMP C30 H45 N2 O 449.3499 4.86 18:1n7-AMP C30 H45 N2 O 449.35 4.69 18:1n9-AMP C30 H45 N2 O 449.35 4.78 18:1n9t-AMP C30 H45 N2 O 449.3496 5.25 18:2n6-AMP C30 H43 N2 O 447.3337 3.39 18:2n6t-AMP C30 H43 N2 O 447.3336 3.94 18:3n3-AMP C30 H41 N2 O 445.3185 2.53 18:3n6-AMP C30 H41 N2 O 445.3181 2.68 19:0-AMP C31 H49 N2 O 465.3814 8.37 19:1-AMP C31 H47 N2 O 463.3651 6.16 20:0-AMP C32 H51 N2 O 479.3966 8.68 20:1-AMP C32 H49 N2 O 477.381 7.92 20:2-AMP C32 H47 N2 O 475.365 5.58 20:3-AMP C32 H45 N2 O 473.3495 4.17 20:4-AMP C32 H43 N2 O 471.334 3.46 20:5-AMP C32 H41 N2 O 469.3179 2.6 203:3n9-AMP C32 H45 N2 O 473.3493 4.77 21:0-AMP C33 H53 N2 O 493.4125 8.92 22:0-AMP C34 H55 N2 O 507.4318 9.15 22:1-AMP C34 H53 N2 O 505.4123 8.69 22:2-AMP C34 H51 N2 O 503.396 8.33 22:4-AMP C34 H47 N2 O 499.3651 5.12 22:5n3-AMP C34 H45 N2 O 497.3494 3.9 22:5n6-AMP C34 H45 N2 O 497.3494 4.51 22:6-AMP C34 H43 N2 O 495.3338 3.43 23:0-AMP C35 H57 N2 O 521.4443 9.38 24:0-AMP C36 H59 N2 O 535.4637 9.6 24:1-AMP C36 H57 N2 O 533.4438 9.13 METABOLITES_END #END