#METABOLOMICS WORKBENCH danielabispo_20250911_112320 DATATRACK_ID:6423 STUDY_ID:ST004296 ANALYSIS_ID:AN007143 PROJECT_ID:PR002702 VERSION 1 CREATED_ON October 13, 2025, 12:06 pm #PROJECT PR:PROJECT_TITLE Global metabolomics identifies new extracellular biomarkers of PR:PROJECT_TITLE nanovibration-driven mesenchymal stem cells osteodifferentiation PR:PROJECT_SUMMARY Bone-related conditions are a leading cause of disability and rising healthcare PR:PROJECT_SUMMARY costs, prompting interest in tissue engineering solutions using mesenchymal stem PR:PROJECT_SUMMARY cells (MSC). As part of an effort to eliminate synthetic osteogenic compounds, PR:PROJECT_SUMMARY this study characterizes the metabolic adaptations of MSC to chemical-free PR:PROJECT_SUMMARY nanovibration (or nanokicking, NK)-induced osteodifferentiation. Through PR:PROJECT_SUMMARY articulation of conventional gene markers and a global metabolomics/lipidomics PR:PROJECT_SUMMARY strategy, our findings indicate successful slow-paced osteodifferentiation, PR:PROJECT_SUMMARY expressed by subtle and partially reversible intracellular changes, and PR:PROJECT_SUMMARY pronounced, largely irreversible, extracellular alterations. PR:INSTITUTE University of Aveiro PR:DEPARTMENT Chemistry PR:LABORATORY CICECO - Aveiro Institute of Materials PR:LAST_NAME Gil PR:FIRST_NAME Ana M. PR:ADDRESS CICECO, Departamento de química, Campus de Santiago, Aveiro, Portugal PR:EMAIL agil@ua.pt PR:PHONE +351 234 370 707 PR:FUNDING_SOURCE This work was developed within the scope of the CICECO-Aveiro Institute of PR:FUNDING_SOURCE Materials, UIDB/50011/2020 project (doi: 10.54499/UIDB/50011/2020), PR:FUNDING_SOURCE UIDP/50011/2020 (doi: 10.54499/UIDP/ 50011/2020) and LA/P/0006/2020 PR:FUNDING_SOURCE (doi:10.54499/LA/P/0006/2020), financed by national funds through the FCT/MCTES PR:FUNDING_SOURCE (PIDDAC). We acknowledge funds from the Foundation for Science and Technology PR:FUNDING_SOURCE through the BetterBone project (2022.04286.PTDC, doi: 10.54499/2022.04286.PTDC), PR:FUNDING_SOURCE the Portuguese National NMR Network (RNRMN), supported by Infrastructure Project PR:FUNDING_SOURCE Nº 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT PR:FUNDING_SOURCE through PIDDAC); and FCT/SPQ PhD grant (DSCB) (SFRH/BD/150655/2020, doi: PR:FUNDING_SOURCE 10.54499/SFRH/BD/150655/2020). EPSRC grant EP/P001114/1. #STUDY ST:STUDY_TITLE (MS data) Global metabolomics identifies new extracellular biomarkers of ST:STUDY_TITLE nanovibration-driven mesenchymal stem cells osteodifferentiation ST:STUDY_SUMMARY This study characterizes the metabolic adaptations of mesenchymal stem cells ST:STUDY_SUMMARY (MSCs) to chemical-free nanovibration (nanokicking, NK)-induced ST:STUDY_SUMMARY osteodifferentiation. Upon reaching the cell numbers required for metabolomic ST:STUDY_SUMMARY analysis (500,000 cells per sample for LC–MS), day 0 samples were collected in ST:STUDY_SUMMARY triplicate prior to stimulation. The remaining 24-well plates were divided into ST:STUDY_SUMMARY unstimulated controls (CTR) and NK-stimulated groups. The NK bioreactor uses the ST:STUDY_SUMMARY reverse piezoelectric effect to generate 30 nm vertical displacements at a ST:STUDY_SUMMARY frequency of 1000 Hz, providing a purely mechanical stimulus without chemical ST:STUDY_SUMMARY induction. For intracellular lipidomics, CTR and NK cells were collected in ST:STUDY_SUMMARY triplicate on days 0, 7, and 21. Through the integration of conventional ST:STUDY_SUMMARY osteogenic gene markers with global metabolomics and lipidomics analyses, our ST:STUDY_SUMMARY findings reveal that NK stimulation promotes a slow-paced osteodifferentiation ST:STUDY_SUMMARY process characterized by subtle, partially reversible intracellular changes and ST:STUDY_SUMMARY pronounced, largely irreversible extracellular alterations, highlighting the ST:STUDY_SUMMARY metabolic reprogramming underlying mechanically induced osteogenesis. ST:INSTITUTE University of Aveiro ST:DEPARTMENT Chemistry ST:LABORATORY CICECO - Aveiro Institute of Materials ST:LAST_NAME Gil ST:FIRST_NAME Ana M. ST:ADDRESS CICECO, Departamento de química, Campus de Santiago, Aveiro, Portugal ST:EMAIL agil@ua.pt ST:PHONE +351234370707 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE 29 SU:WEIGHT_OR_WEIGHT_RANGE Healthy SU:GENDER Female SU:CELL_STRAIN_DETAILS Human adipose tissue-derived stem cells SU:CELL_PASSAGE_NUMBER 3 #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 - CTR_D0_S1 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D0_S2 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D0_S3 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D7_S1 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D7_S2 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D7_S3 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D21_S1 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D21_S2 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - CTR_D21_S3 Sample source:hASCs | Treatment:Proliferation conditions SUBJECT_SAMPLE_FACTORS - NK_D7_S1 Sample source:hASCs | Treatment:Osteogenic conditions SUBJECT_SAMPLE_FACTORS - NK_D7_S2 Sample source:hASCs | Treatment:Osteogenic conditions SUBJECT_SAMPLE_FACTORS - NK_D7_S3 Sample source:hASCs | Treatment:Osteogenic conditions SUBJECT_SAMPLE_FACTORS - NK_D21_S1 Sample source:hASCs | Treatment:Osteogenic conditions SUBJECT_SAMPLE_FACTORS - NK_D21_S2 Sample source:hASCs | Treatment:Osteogenic conditions SUBJECT_SAMPLE_FACTORS - NK_D21_S3 Sample source:hASCs | Treatment:Osteogenic conditions #COLLECTION CO:COLLECTION_SUMMARY hAMSC were donated from Histocell (Bilbao, Spain), derived from a healthy CO:COLLECTION_SUMMARY 29-year-old female donor undergoing liposuction with written informed consent. CO:COLLECTION_PROTOCOL_ID Protocol no. E08-30 CO:SAMPLE_TYPE Stem cells CO:STORAGE_CONDITIONS -80℃ CO:TISSUE_CELL_IDENTIFICATION Adipose Tissue #TREATMENT TR:TREATMENT_SUMMARY Upon reaching the cell numbers necessary for metabolomics (> 1 million TR:TREATMENT_SUMMARY cells/sample for NMR, and 500 k cells/sample for LC-MS), day 0 cell samples were TR:TREATMENT_SUMMARY collected in triplicate prior to stimulation (Figure S2). The remaining cell TR:TREATMENT_SUMMARY culture T150 flasks (for NMR metabolomics) and 24-well plates (for MS TR:TREATMENT_SUMMARY lipidomics, Alamar blue (AB) assay and qRT-PCR) were split into unstimulated TR:TREATMENT_SUMMARY (control, CTR) and NK-stimulated groups. As previously described,[30,32] the NK TR:TREATMENT_SUMMARY bioreactor employs the reverse piezoelectric effect to induce mechanical TR:TREATMENT_SUMMARY expansions from applied voltages, enabling 30 nm vertical displacements to cell TR:TREATMENT_SUMMARY cultures at a frequency of 1000 Hz (Figure 1). To ensure consistent amplitudes TR:TREATMENT_SUMMARY across the growth surfaces while allowing for easy removal and maintenance, TR:TREATMENT_SUMMARY culture flasks/plates were firmly attached to the bioreactor using magnetic TR:TREATMENT_SUMMARY sheets. For intracellular NMR metabolomics (endometabolomics) and MS lipidomics, TR:TREATMENT_SUMMARY CTR and NK cells were trypsinized and collected in triplicate on days 0, 7 and TR:TREATMENT_SUMMARY 21 (Figure S2). The resulting cell suspensions were filtered through 100 μm TR:TREATMENT_SUMMARY pore strainers, centrifuged (300 g, 5 min, 4 °C) and rinsed twice with TR:TREATMENT_SUMMARY phosphate-buffered saline (PBS) solution. For extracellular NMR metabolomics TR:TREATMENT_SUMMARY (exometabolomics), media samples were collected on days 3, 6, 7, 10, 13, 17, 20 TR:TREATMENT_SUMMARY and 21. Blank media (not cell-exposed) was also obtained. Media and cell samples TR:TREATMENT_SUMMARY were stored (− 80 °C) until analysis. TR:CELL_MEDIA α-MEM (Gibco™ 12000063, Waltham, MA, USA) supplemented with 10% fetal bovine TR:CELL_MEDIA serum (FBS) and 1% antibiotics TR:CELL_HARVESTING Days 0, 7 and 21 TR:CELL_PCT_CONFLUENCE 100% #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Lipids were analysed by LC-MS using a Thermo Q-Exactive Orbitrap mass SP:SAMPLEPREP_SUMMARY spectrometer equipped with a heated electrospray ionization (HESI) probe and SP:SAMPLEPREP_SUMMARY interfaced with a Dionex UltiMate 3000 RSLC system (Thermo Fisher Scientific, SP:SAMPLEPREP_SUMMARY Hemel Hempstead, UK). Samples (10 µL) were injected onto a Thermo Hypersil Gold SP:SAMPLEPREP_SUMMARY C18 column (2.1 mm × 100 mm; 1.9 μm) maintained at 50 °C. Mobile phase A SP:SAMPLEPREP_SUMMARY consisted of water containing 10 mM ammonium formate and 0.1% (v/v) formic acid. SP:SAMPLEPREP_SUMMARY Mobile phase B consisted of a 90/10 (v/v) mixture of isopropanol/acetonitrile SP:SAMPLEPREP_SUMMARY containing 10 mM ammonium formate and 0.1% (v/v) formic acid. The initial SP:SAMPLEPREP_SUMMARY conditions for analysis were 65% mobile phase A, 35% mobile phase B and the SP:SAMPLEPREP_SUMMARY percentage of mobile phase B was increased from 35 to 65% over 4 min, followed SP:SAMPLEPREP_SUMMARY by 65% to 100% over 15 min, with a hold for 2 min before re-equilibration to the SP:SAMPLEPREP_SUMMARY starting conditions over 6 min. The flow rate was 400 μL/min. All samples were SP:SAMPLEPREP_SUMMARY analysed in positive and negative ionization modes over the mass-to-charge ratio SP:SAMPLEPREP_SUMMARY (m/z) range of 250 to 2,000 at a resolution of 60,000. SP:EXTRACTION_METHOD methanol-chloroform-water extraction method SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Q-Exactive Orbitrap mass spectrometer (Thermo Fisher Scientific) CH:COLUMN_NAME Thermo Hypersil Gold C18 (2.1 mm × 100 mm; 1.9 μm) CH:SOLVENT_A Water + 10 mM ammonium formate + 0.1% formic acid (v/v) CH:SOLVENT_B Isopropanol/acetonitrile (90/10, v/v) + 10 mM ammonium formate + 0.1% formic CH:SOLVENT_B acid (v/v) CH:FLOW_GRADIENT 0–4 min: 35% → 65% B; 4–19 min: 65% → 100% B; 19–21 min: 100% B CH:FLOW_GRADIENT (hold); 21–27 min: re-equilibrate to 35% B CH:FLOW_RATE 400 µL/min CH:COLUMN_TEMPERATURE 50 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Lipids were analyzed by LC-MS using a Thermo Q-Exactive Orbitrap mass MS:MS_COMMENTS spectrometer equipped with a heated electrospray ionization (HESI) source and MS:MS_COMMENTS interfaced with a Dionex UltiMate 3000 RSLC system. Samples (10 µL) were MS:MS_COMMENTS injected onto a Thermo Hypersil Gold C18 column (2.1 mm × 100 mm; 1.9 µm) MS:MS_COMMENTS maintained at 50 °C. Mobile phase A consisted of water containing 10 mM MS:MS_COMMENTS ammonium formate and 0.1% (v/v) formic acid, and mobile phase B consisted of MS:MS_COMMENTS 90:10 (v/v) isopropanol/acetonitrile containing the same additives. The gradient MS:MS_COMMENTS was 35% B to 65% B over 4 min, 65% B to 100% B over 15 min, held for 2 min, then MS:MS_COMMENTS re-equilibrated over 6 min. The flow rate was 400 µL/min. Data were acquired MS:MS_COMMENTS over the m/z range 250–2000 at a resolution of 60 000 in negative ion mode. MS:MS_COMMENTS Raw LC-MS data were converted to mzML format using msConvert (ProteoWizard). MS:MS_COMMENTS Progenesis QI v2.4 (Non-linear Dynamics) was used for data processing and MS:MS_COMMENTS feature assignment. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Normalized abundance MS_METABOLITE_DATA_START Samples CTR_D0_S1 CTR_D0_S2 CTR_D0_S3 CTR_D7_S1 CTR_D7_S2 CTR_D7_S3 NK_D7_S1 NK_D7_S2 NK_D7_S3 CTR_D21_S1 CTR_D21_S2 CTR_D21_S3 NK_D21_S1 NK_D21_S2 NK_D7_S3 Factors Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Osteogenic conditions Sample source:hASCs | Treatment:Osteogenic conditions Sample source:hASCs | Treatment:Osteogenic conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Proliferation conditions Sample source:hASCs | Treatment:Osteogenic conditions Sample source:hASCs | Treatment:Osteogenic conditions Sample source:hASCs | Treatment:Osteogenic conditions Cer(d16:1/16:0)_negative_mode 246774 192563 270233 491243 467308 546083 528939 522009 607420 257825 291439 284248 339428 408463 487113 Cer(d18:1/14:0)_negative_mode 3614 1965 1072 89199 58165 56344 82413 16820 36968 1121 6016 6572 7017 11142 11397 Cer(d18:1/16:0)_negative_mode 1604777 1661772 1683293 2121138 2123695 2232808 2250456 2333422 2347384 1888903 1902859 1712473 2350008 1971977 1894775 Cer(d18:1/23:0)_negative_mode 277906 312654 336607 261850 307402 278374 281120 232130 268340 244683 242371 232135 189400 215659 214788 GlcCer(d18:1/16:0)_negative_mode 537178 521834 568437 493919 489313 466707 586145 556088 510088 441451 398597 463738 494992 559328 432787 LysoPC(18:1(9Z)/0:0)_negative_mode 50097 149522 144415 12661 18097 15386 8861 2460 13886 12960 57016 30464 5572 1500 6764 PC(14:0/15:0)_negative_mode 28522 18035 18186 80590 61407 69541 77927 79941 55725 36385 19101 7658 42059 34886 37227 PC(14:1(9Z)/18:1(9Z))_negative_mode 283398 212430 231522 443744 453323 460239 404938 413518 411432 377676 422038 362307 441541 442199 435292 PC(20:3(5Z,8Z,11Z)/P-18:1(11Z))_negative_mode 11331 7616 10163 32077 31548 32181 39539 35182 35514 0 2812 0 0 0 3530 PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/16:1(9Z))_negative_mode 49227 19954 16071 97204 106892 80688 90556 90813 126639 15355 42358 12481 45295 28582 41715 PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/22:5(4Z,7Z,10Z,13Z,16Z))_negative_mode 0 0 0 12470 7858 7728 15459 15274 16021 0 0 0 2668 0 0 PC(O-18:0/18:2(9Z,12Z))_negative_mode 31147 40803 51211 50947 53474 53703 58027 56855 54692 82197 92613 71268 83827 78873 99889 PE(14:0/18:1(9Z))_negative_mode 183091 153351 202249 269103 263719 276600 276855 307595 258724 305912 336064 299583 381320 396281 343902 PE(20:1(11Z)/22:0)_negative_mode 11080 12583 19750 22153 37569 28475 28284 30036 31186 13885 40721 43551 15762 17552 23852 SM(d18:1/14:0)_negative_mode 105562 109374 132994 172589 182173 194693 200542 185922 202452 211684 231849 220483 284637 248946 270630 SM(d18:1/16:0(2OH))_negative_mode 11899 22760 16160 77236 83275 91155 73337 92800 91417 86057 85945 76972 125201 99910 97704 SM(d18:1/16:0)_negative_mode 2921390 2819261 3169451 5952971 6131760 6336655 6642916 6343439 6552708 5703707 5290660 5079210 6876010 6971100 6300386 SM(d18:1/16:1(9Z))_negative_mode 137949 146235 177006 316279 324673 317407 345430 312757 332788 185995 220884 191856 283754 251470 229326 SM(d18:1/17:0)_negative_mode 44084 38007 47400 239390 238493 246351 260921 247850 266244 106636 129260 80376 157508 133460 117666 SM(d18:1/20:0)_negative_mode 18880 26605 26530 78234 68511 75373 85367 97054 85795 90419 112177 103730 115331 93578 76195 Cer(d18:1/25:0)_negative_mode 112213 86242 66029 70551 101811 84848 115416 109099 92220 44176 67798 24334 23292 17244 25196 CL(16:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z)/18:0/16:0)_negative_mode 94996 188619 159522 306202 324199 357247 222771 179981 268317 0 199097 0 0 20872 159604 CL(18:2(9Z,12Z)/18:2(9Z,11Z)/16:0/16:0)_negative_mode 311700 194108 271967 41651 70203 0 44317 7609 17733 35627 333978 0 340319 221719 229161 LacCer(d18:1/24:1(15Z))_negative_mode 195922 176191 229062 182721 216215 199918 325162 202356 197004 104999 71902 35066 103063 112959 137071 PC(20:3(5Z,8Z,11Z)/20:0)_negative_mode 2032 0 7070 33945 30806 38695 41022 40975 41382 15315 60767 4777 47903 34052 50527 PC(20:3(5Z,8Z,11Z)/P-18:0)_negative_mode 49892 35554 31454 45419 45413 65319 48697 50667 47294 33967 40364 31468 44391 44761 55140 PC(22:1(13Z)/15:0)_negative_mode 10595 5204 12585 44608 45686 46139 53117 43406 55299 15945 34582 10728 37469 23628 32802 PE(20:3(5Z,8Z,11Z)/P-18:0)_negative_mode 74496 78511 55994 82401 85768 93192 107113 93502 108191 222360 253009 199208 224080 223145 246000 PE-NMe(16:1(9Z)/22:2(13Z,16Z))_negative_mode 2962 5359 0 8577 11531 11330 17557 16265 12811 21919 27111 8785 26530 30816 28238 PI(16:0/20:4(8Z,11Z,14Z,17Z))_negative_mode 298747 310368 315429 698618 660579 703848 791281 721241 753732 450841 450812 455463 477720 499338 510522 PI(16:0/22:3(10Z,13Z,16Z))_negative_mode 448951 483777 522560 565476 584506 608982 655867 663448 664278 892852 781001 795389 962286 925593 1004765 PI(20:1(11Z)/15:0)_negative_mode 18826 12343 15005 90948 76550 90003 96637 93087 107345 59137 78033 57517 119476 100741 103753 PI(20:3(6,8,11)-OH(5)/20:0)_negative_mode 894280 601246 729878 72204 57620 39327 84258 84308 92425 49444 37248 21312 64089 73594 71207 SM(d17:2(4E,8Z)/20:3(8Z,11Z,14Z)-2OH(5,6))_negative_mode 14592 22608 12618 50810 49530 56553 63009 59001 71159 39753 36381 31870 51463 15541 35743 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Compound m/z Retention time (min) Accepted Compound ID Cer(d16:1/16:0)_negative_mode 10.17_554.4787m/z 554.4787 10.17 HMDB0240681 Cer(d18:1/14:0)_negative_mode 10.17_508.4741m/z 508.4741 10.17 HMDB0011773 Cer(d18:1/16:0)_negative_mode 11.10_537.5119n 582.5101 11.1 HMDB0004949 Cer(d18:1/23:0)_negative_mode 14.06_680.6221m/z 680.6221 14.06 HMDB0000950 GlcCer(d18:1/16:0)_negative_mode 10.42_699.5657n 744.5639 10.42 HMDB0004971 LysoPC(18:1(9Z)/0:0)_negative_mode 5.78_566.3448m/z 566.3448 5.78 HMDB0002815 PC(14:0/15:0)_negative_mode 9.70_736.5141m/z 736.5141 9.7 HMDB0007868 PC(14:1(9Z)/18:1(9Z))_negative_mode 10.12_774.5313m/z 774.5313 10.12 HMDB0007906 PC(20:3(5Z,8Z,11Z)/P-18:1(11Z))_negative_mode 11.74_838.5966m/z 838.5966 11.74 HMDB0008392 PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/16:1(9Z))_negative_mode 9.76_848.5452m/z 848.5452 9.76 HMDB0008726 PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/22:5(4Z,7Z,10Z,13Z,16Z))_negative_mode 10.06_924.5766m/z 924.5766 10.06 HMDB0008746 PC(O-18:0/18:2(9Z,12Z))_negative_mode 12.00_816.6134m/z 816.6134 12 HMDB0013418 PE(14:0/18:1(9Z))_negative_mode 10.46_688.4919m/z 688.4919 10.46 HMDB0008828 PE(20:1(11Z)/22:0)_negative_mode 14.58_828.6489m/z 828.6489 14.58 HMDB0009270 SM(d18:1/14:0)_negative_mode 9.20_719.5344m/z 719.5344 9.2 HMDB0012097 SM(d18:1/16:0(2OH))_negative_mode 9.81_763.5622m/z 763.5622 9.81 LMSP03010101 SM(d18:1/16:0)_negative_mode 10.15_747.5663m/z 747.5663 10.15 HMDB0010169 SM(d18:1/16:1(9Z))_negative_mode 9.40_745.5516m/z 745.5516 9.4 HMDB0240613 SM(d18:1/17:0)_negative_mode 10.61_761.5822m/z 761.5822 10.61 HMDB0240609 SM(d18:1/20:0)_negative_mode 11.98_803.6278m/z 803.6278 11.98 HMDB0012102 Cer(d18:1/25:0)_negative_mode 14.78_708.6522m/z 708.6522 14.78 HMDB0004957 CL(16:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z)/18:0/16:0)_negative_mode 16.63_1451.9947m/z 1451.9947 16.63 HMDB0056884 CL(18:2(9Z,12Z)/18:2(9Z,11Z)/16:0/16:0)_negative_mode 16.43_1399.9633m/z 1399.9633 16.43 HMDB0111446 LacCer(d18:1/24:1(15Z))_negative_mode 12.87_1016.7268m/z 1016.7268 12.87 HMDB0004872 PC(20:3(5Z,8Z,11Z)/20:0)_negative_mode 12.57_884.6404m/z 884.6404 12.57 HMDB0008373 PC(20:3(5Z,8Z,11Z)/P-18:0)_negative_mode 12.09_840.6133m/z 840.6133 12.09 HMDB0008391 PC(22:1(13Z)/15:0)_negative_mode 12.57_846.6248m/z 846.6248 12.57 HMDB0008560 PE(20:3(5Z,8Z,11Z)/P-18:0)_negative_mode 12.43_752.5601m/z 752.5601 12.43 HMDB0009346 PE-NMe(16:1(9Z)/22:2(13Z,16Z))_negative_mode 12.41_782.5707m/z 782.5707 12.41 HMDB0113077 PI(16:0/20:4(8Z,11Z,14Z,17Z))_negative_mode 9.79_857.5193m/z 857.5193 9.79 HMDB0009790 PI(16:0/22:3(10Z,13Z,16Z))_negative_mode 10.95_887.5673m/z 887.5673 10.95 HMDB0009792 PI(20:1(11Z)/15:0)_negative_mode 10.81_849.5480m/z 849.548 10.81 LMGP06010500 PI(20:3(6,8,11)-OH(5)/20:0)_negative_mode 8.35_932.6028n 931.5956 8.35 HMDB0277062 SM(d17:2(4E,8Z)/20:3(8Z,11Z,14Z)-2OH(5,6))_negative_mode 9.40_813.5416m/z 813.5416 9.4 HMDB0290421 METABOLITES_END #END