#METABOLOMICS WORKBENCH Lamichhane_20210208_064812_mwtab.txt DATATRACK_ID:2460 STUDY_ID:ST001685 ANALYSIS_ID:AN002752 PROJECT_ID:PR001084 VERSION 1 CREATED_ON February 9, 2021, 2:47 pm #PROJECT PR:PROJECT_TITLE Exposure to per- and polyfluoroalkyl substances and lipid profile of breast milk PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY The objective of this study was to investigate whether the maternal levels of PR:PROJECT_SUMMARY PFAS are associated with lipid composition of human breast milk and further, if PR:PROJECT_SUMMARY the changes in composition have impact on the growth of the infants. PR:INSTITUTE University of Turku PR:DEPARTMENT Turku Bioscience PR:LABORATORY Turku Metabolomics Center PR:LAST_NAME Lamichhane PR:FIRST_NAME Santosh PR:ADDRESS Tykistökatu 6 , Turku PR:EMAIL santosh.lamichhane@utu.fi PR:PHONE 0452299070 PR:FUNDING_SOURCE This study was supported by the Swedish Research Council (grant no. 2016-05176 PR:FUNDING_SOURCE to T.H and M.O), Formas (grant no. 2019-00869 to T.H and M.O), and the PR:FUNDING_SOURCE NovoNordiskFoundation (xxx). The EDIA study was supported by the National PR:FUNDING_SOURCE Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National PR:FUNDING_SOURCE Institutes of Health (No. 1DP3DK094338-01 to M.K.), the Academy of Finland PR:FUNDING_SOURCE Centre of Excellence in Molecular Systems Immunology and Physiology Research PR:FUNDING_SOURCE 2012-17, No. 250114 to M.K. and M.O.). Further support was received by the PR:FUNDING_SOURCE Academy of Finland postdoctoral grant (No. 323171 to S.L.) and the Medical PR:FUNDING_SOURCE Research Funds, Tampere and Helsinki University Hospitals (to M.K.). PR:PROJECT_COMMENTS Part 2 PR:CONTRIBUTORS Santosh Lamichhane, Heli Siljander, Daniel Duberg, Jorma Ilonen, Suvi M. PR:CONTRIBUTORS Virtanen, Matej Orešič, Mikael Knip, Tuulia Hyötyläinen #STUDY ST:STUDY_TITLE Exposure to per- and polyfluoroalkyl substances associates with altered lipid ST:STUDY_TITLE profile of breast milk (Part 2) ST:STUDY_SUMMARY In this mother-infant study (n=44) we investigated the levels of PFAS in ST:STUDY_SUMMARY maternal serum and detailed lipidomic profile in breast milk at birth and at ST:STUDY_SUMMARY three months using ultra high performance liquid chromatography combined with ST:STUDY_SUMMARY quadrupole-time-of-flight mass spectrometry. ST:INSTITUTE University of Turku ST:LAST_NAME Lamichhane ST:FIRST_NAME Santosh ST:ADDRESS Tykistökatu 6, FI-20520 Turku, Finland ST:EMAIL santosh.lamichhane@utu.fi ST:PHONE 0452299070 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Not applicable #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 18 18 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0017.mzdata.xml SUBJECT_SAMPLE_FACTORS 38 38 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0011.mzdata.xml SUBJECT_SAMPLE_FACTORS 59 59 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0029.mzdata.xml SUBJECT_SAMPLE_FACTORS 72 72 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0023.mzdata.xml SUBJECT_SAMPLE_FACTORS 74 74 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0048.mzdata.xml SUBJECT_SAMPLE_FACTORS 84 84 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0012.mzdata.xml SUBJECT_SAMPLE_FACTORS 201 201 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0052.mzdata.xml SUBJECT_SAMPLE_FACTORS 226 226 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0004.mzdata.xml SUBJECT_SAMPLE_FACTORS 259 259 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0038.mzdata.xml SUBJECT_SAMPLE_FACTORS 261 261 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0015.mzdata.xml SUBJECT_SAMPLE_FACTORS 297 297 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0039.mzdata.xml SUBJECT_SAMPLE_FACTORS 305 305 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0020.mzdata.xml SUBJECT_SAMPLE_FACTORS 327 327 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0059.mzdata.xml SUBJECT_SAMPLE_FACTORS 333 333 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0034.mzdata.xml SUBJECT_SAMPLE_FACTORS 346 346 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0047.mzdata.xml SUBJECT_SAMPLE_FACTORS 353 353 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0049.mzdata.xml SUBJECT_SAMPLE_FACTORS 369 369 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0031.mzdata.xml SUBJECT_SAMPLE_FACTORS 399 399 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0065.mzdata.xml SUBJECT_SAMPLE_FACTORS 427 427 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0057.mzdata.xml SUBJECT_SAMPLE_FACTORS 450 450 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0025.mzdata.xml SUBJECT_SAMPLE_FACTORS 471 471 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0013.mzdata.xml SUBJECT_SAMPLE_FACTORS 487 487 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0018.mzdata.xml SUBJECT_SAMPLE_FACTORS 510 510 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0040.mzdata.xml SUBJECT_SAMPLE_FACTORS 514 514 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0032.mzdata.xml SUBJECT_SAMPLE_FACTORS 568 568 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0060.mzdata.xml SUBJECT_SAMPLE_FACTORS 673 673 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0002.mzdata.xml SUBJECT_SAMPLE_FACTORS 739 739 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0024.mzdata.xml SUBJECT_SAMPLE_FACTORS 740 740 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0050.mzdata.xml SUBJECT_SAMPLE_FACTORS 744 744 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0028.mzdata.xml SUBJECT_SAMPLE_FACTORS 757 757 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0010.mzdata.xml SUBJECT_SAMPLE_FACTORS 764 764 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0053.mzdata.xml SUBJECT_SAMPLE_FACTORS 776 776 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0041.mzdata.xml SUBJECT_SAMPLE_FACTORS 792 792 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0061.mzdata.xml SUBJECT_SAMPLE_FACTORS 806 806 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0036.mzdata.xml SUBJECT_SAMPLE_FACTORS 808 808 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0001.mzdata.xml SUBJECT_SAMPLE_FACTORS 820 820 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0022.mzdata.xml SUBJECT_SAMPLE_FACTORS 901 901 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0045.mzdata.xml SUBJECT_SAMPLE_FACTORS 969 969 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0006.mzdata.xml SUBJECT_SAMPLE_FACTORS 1098 1098 _2 Sample Code:3 months | QTL:Q3 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0063.mzdata.xml SUBJECT_SAMPLE_FACTORS 1172 1172 _2 Sample Code:3 months | QTL:Q4 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0044.mzdata.xml SUBJECT_SAMPLE_FACTORS 1294 1294 _2 Sample Code:3 months | QTL:Q2 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0026.mzdata.xml SUBJECT_SAMPLE_FACTORS 1300 1300 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0043.mzdata.xml SUBJECT_SAMPLE_FACTORS 1348 1348 _2 Sample Code:3 months | QTL:Q1 RAW_FILE_NAME=0045_LC_20200911_PFASBA_sample_0062.mzdata.xml #COLLECTION CO:COLLECTION_SUMMARY Blood samples were collected for this study. Plasma was separated within CO:COLLECTION_SUMMARY 30 minutes after the blood collection by centrifugation at room temperature. CO:COLLECTION_SUMMARY The plasma samples were stored at −80 °C until analyzed. CO:SAMPLE_TYPE Blood (plasma) CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Sample were stored at -80 as it is, no treatment done. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The serum samples were extracted as follows after randomization of the samples. SP:SAMPLEPREP_SUMMARY 90 µl of ecetonitrile (containing the BA+PFAS internal standard mixture (c=200 SP:SAMPLEPREP_SUMMARY ng/mL PFASs and 1000 ng/mL Bile acids in acetonitrile) was added to the 40 µl SP:SAMPLEPREP_SUMMARY of serum samples. The samples were then vortexted and centrifuged (9600 RCF, 10 SP:SAMPLEPREP_SUMMARY minutes). 90 µl of the supernatant was collected and evaporated to dryness. The SP:SAMPLEPREP_SUMMARY samples were reconstituted with 40 µl of MeOH/H2O (40%/60%). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Column name, BEH C18 (2.1 x 100 mm, particle size 1.7 µm) (Waters Corporation, CH:CHROMATOGRAPHY_SUMMARY Milford, MA, USA) CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity UPLC CH:COLUMN_NAME BEH C18 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6210 TOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS PFASs were analysed on a UHPLC-qTOF/MS (Agilent Technologies, Santa Clara, CA, MS:MS_COMMENTS The United States of America) with Acquity UPLC®, BEH C18 (2.1 x 100 mm, MS:MS_COMMENTS particle size 1.7 µm) (Waters Corporation, Milford, MA, The United States of MS:MS_COMMENTS America) column at 50°C with a C18 pre-column for column protection (Waters MS:MS_COMMENTS Corporation, Wexford, Ireland). Mobile phases used for the sample analysis were MS:MS_COMMENTS A: 2mM NH4Ac in H2O:MeOH (70:30) and B: 2mM NH4Ac in MeOH. NH4Ac was used as MS:MS_COMMENTS ionization agent. The samples were kept at 10°C during the whole sample MS:MS_COMMENTS acquisition and 1 µl of the sample volume was injected. The flow rate was set MS:MS_COMMENTS to 0.4 ml/min and the gradient started with 95%A and 5%B with a change after 1.5 MS:MS_COMMENTS minute to 70%A and 30%B, which followed a change after 4.5 minutes to 30%A and MS:MS_COMMENTS 70%B, the last change was after 7.5 minutes with 100%B until the end of run. MS:MS_COMMENTS Gradient program was 18 minutes long. 13 minutes for sample analysis and 5 MS:MS_COMMENTS minutes for clean-up, including equilibration in the end of the run. Maximum MS:MS_COMMENTS pressure limit in the binary pump was set on 850 bar. Dual jet stream MS:MS_COMMENTS electrospray (dual ESI) ion source was used and the ion polarity was on negative MS:MS_COMMENTS mode. The capillary voltage and the nozzle voltage were kept at 4500 V and 1500 MS:MS_COMMENTS V. The N2 pressure was set on 21 psi, with the sheath gas flow as 11 L/min and MS:MS_COMMENTS temperature at 379°C for the nebulizer. The data was acquired with MassHunter MS:MS_COMMENTS B.06.01 software (Agilent Technologies, Santa Clara, CA, The United States of MS:MS_COMMENTS America). MS data processing was performed using open source software MZmine MS:MS_COMMENTS 2.52 (Pluskal et al. 2010). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS ng/ml MS_METABOLITE_DATA_START Samples 18 _2 38 _2 59 _2 72 _2 74 _2 84 _2 201 _2 226 _2 259 _2 261 _2 297 _2 305 _2 327 _2 333 _2 346 _2 353 _2 369 _2 399 _2 427 _2 450 _2 471 _2 487 _2 510 _2 514 _2 568 _2 673 _2 739 _2 740 _2 744 _2 757 _2 764 _2 776 _2 792 _2 806 _2 808 _2 820 _2 901 _2 969 _2 1098 _2 1172 _2 1294 _2 1300 _2 1348 _2 Factors Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q3 Sample Code:3 months | QTL:Q4 Sample Code:3 months | QTL:Q2 Sample Code:3 months | QTL:Q1 Sample Code:3 months | QTL:Q1 PFDA 9.742988991 5.364438896 3.461182094 8.676159822 4.519607417 3.254017692 4.416603492 7.35831805 5.620181239 9.642088874 8.221322987 2.451176622 8.343957816 6.545109639 7.291944977 7.210584469 3.587319696 8.339470986 7.171623189 3.958866346 7.776845584 8.060720707 6.099545005 7.441880489 4.430602691 4.403917637 5.475031343 8.477094221 2.686980971 8.229451479 5.455342696 5.574588609 10.01636241 2.779062132 4.001054521 2.036433612 5.529742997 2.772660714 8.942583847 9.778749721 9.347070601 6.5887657 2.618737169 PFHxS 1.651559696 3.561216612 2.62829761 2.740022521 1.546544755 2.16026146 1.802299459 3.350735228 2.748531885 2.013170076 2.034313619 2.517269406 1.974037435 2.810524146 2.889855802 2.191348567 2.695356156 1.649363526 1.639370619 2.315435667 2.724057782 2.686988414 2.235487444 2.58873993 3.802765398 3.624071867 1.944602495 3.666420985 2.366533172 1.5382006 2.569760132 1.981503455 2.459422912 2.733756989 2.258967654 1.88046534 2.458946133 1.705588956 1.784582262 3.673450925 2.054801919 1.744944909 2.163119763 PFNA 7.912388525 14.95315612 8.585393112 20.33662436 15.79873242 16.65169525 12.8145425 29.39620198 15.77942343 13.36859687 12.21866997 6.137177906 18.5971537 27.96670727 19.6511242 24.45001273 14.87858559 10.83155805 8.684247828 19.76627861 23.20110464 10.11806901 22.48854007 26.75189572 32.20853861 53.13672577 10.53473938 25.62668682 36.22205305 17.46876677 17.69053408 2.448678212 11.15610756 18.56369362 15.38729129 4.759227196 23.34807577 7.846521375 15.68033607 26.85186091 11.72297062 8.794221082 10.14161425 PFOA 23.447431 35.93411556 63.47741838 68.94268047 46.90142073 35.04091831 26.58885758 102.1813494 52.84445946 34.41269402 81.70831985 34.3895696 33.61868393 67.03155384 55.72446882 80.59314366 59.75819534 21.1753003 38.28782586 50.82533167 50.44513263 51.36192679 34.80439984 39.48696433 87.01979285 49.8657037 29.10341052 55.9068401 113.3173646 36.7177813 82.34383518 22.92445104 56.95203386 61.58143311 12.16460733 24.34206435 83.58406282 19.1249388 23.99416769 53.26544964 47.35948495 38.89387151 18.44741334 PFOS1 31.40050767 54.49425535 73.44003295 68.96499552 58.05217763 49.31882276 26.27454442 127.0847774 45.56071357 41.00887531 33.86581285 65.31285291 46.78023412 108.3447 79.92592962 40.05902026 75.4465874 20.41806537 22.98776882 67.67345581 151.162346 64.43901669 58.0582161 66.40045905 84.10451061 86.38044872 48.49313397 85.07287031 62.68226911 22.77102443 74.10719368 34.8586746 53.92440679 117.1616341 82.09927364 21.31133326 71.46549631 22.41199155 85.31312057 111.8174822 59.29445397 26.37478326 25.23547131 PFOS2 41.41137931 85.84458661 73.41430574 110.7325442 79.6624526 92.55036454 61.91689848 112.3080361 63.33215423 49.85526485 39.95013137 72.06563257 66.88434261 178.0623644 116.9088566 51.48223561 91.77341335 47.51791572 38.77190085 91.5579036 173.9670504 73.26246714 87.01962515 155.9922222 95.34571152 103.3122981 61.2488302 128.257114 72.4262715 45.90971344 64.77772333 38.34225746 54.96886632 124.7080754 71.85082572 35.29271125 118.0372417 34.85474977 96.32102033 194.8289862 43.09235675 36.40448711 30.1686089 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name quantitated m/z retention time PFDA 512.979909 5.17 PFHxS 398.9369138 3.96 PFNA 462.9643268 4.91 PFOA 412.9669347 4.45 PFOS1 498.9301269 4.77 PFOS2 498.9301787 4.93 METABOLITES_END #END