#METABOLOMICS WORKBENCH Emile_Callemeyn_20250127_071924 DATATRACK_ID:5574 STUDY_ID:ST003709 ANALYSIS_ID:AN006086 PROJECT_ID:PR002287 VERSION 1 CREATED_ON 02-06-2025 #PROJECT PR:PROJECT_TITLE The childhood salivary metabolome reflects inter-individual variation in weight PR:PROJECT_TITLE status, lifestyle, and mental wellbeing PR:PROJECT_TYPE (Un)targeted MS PR:PROJECT_SUMMARY Saliva is an emerging biofluid in metabolomics as it provides a similar window PR:PROJECT_SUMMARY into biological information as blood, while enabling repetitive and non-invasive PR:PROJECT_SUMMARY sampling. Here, the salivary metabolome of 965 children (7-17y) from 2 European PR:PROJECT_SUMMARY countries was comprehensively analyzed using ultra-high performance liquid PR:PROJECT_SUMMARY chromatography coupled to high-resolution mass spectrometry providing >3,500 PR:PROJECT_SUMMARY untargeted features and 188 consistently detected metabolites. Correlation PR:PROJECT_SUMMARY analysis with anthropometric, mental wellbeing, lifestyle and dietary data PR:PROJECT_SUMMARY revealed that weight status and age reflected the salivary metabolome best, with PR:PROJECT_SUMMARY functional analysis demonstrating alterations in multiple pathways. These PR:PROJECT_SUMMARY findings were replicated in 2 validation cohorts involving 471 children (8-18y) PR:PROJECT_SUMMARY confirming previously reported blood biomarkers and new molecules. For example, PR:PROJECT_SUMMARY acesulfame K, an artificial sweetener detected in all cohorts, increased with PR:PROJECT_SUMMARY weight and mental distress, while correlating to specific food/beverage PR:PROJECT_SUMMARY consumption. Our data offer a high-quality repository of the pediatric salivary PR:PROJECT_SUMMARY metabolome for providing guidance in exploring its association with children's PR:PROJECT_SUMMARY health. PR:INSTITUTE Ghent University PR:DEPARTMENT Department of Translational Physiology, Infectiology and Public Health PR:LABORATORY Laboratory of Integrative Metabolomics PR:LAST_NAME callemeyn PR:FIRST_NAME emile PR:ADDRESS Salisburylaan 133, 9820, Merelbeke, Belgium PR:EMAIL emile.callemeyn@ugent.be PR:PHONE 0032473271636 PR:FUNDING_SOURCE Fonds Wetenschappelijk Onderzoek PR:DOI http://dx.doi.org/10.21228/M80Z5F #STUDY ST:STUDY_TITLE Cross-sectional salivary metabolomics of the PANIC cohort (children 11-17 years) ST:STUDY_SUMMARY This study is part of a multi-part study, including salivary metabolomics of 4 ST:STUDY_SUMMARY patient cohorts, both negative and positive ionization mode. ST:INSTITUTE Ghent University ST:LAST_NAME callemeyn ST:FIRST_NAME emile ST:ADDRESS Salisburylaan 133, 9820 Merelbeke, Belgium ST:EMAIL emile.callemeyn@ugent.be ST:PHONE 0473271636 ST:SUBMIT_DATE 2025-01-27 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Male and female #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 220502s040_QC1_3 Sample source:saliva | Weight:- RAW_FILE_NAME(Raw file name)=220502s040_QC1_3.raw SUBJECT_SAMPLE_FACTORS - 220502s041_QC2_3 Sample source:saliva | Weight:- RAW_FILE_NAME(Raw file name)=220502s041_QC2_3.raw SUBJECT_SAMPLE_FACTORS - 220502s052_QC1_4 Sample source:saliva | Weight:- RAW_FILE_NAME(Raw file name)=220502s052_QC1_4.raw SUBJECT_SAMPLE_FACTORS - 220502s053_QC2_4 Sample source:saliva | Weight:- RAW_FILE_NAME(Raw file name)=220502s053_QC2_4.raw SUBJECT_SAMPLE_FACTORS - 220502s064_QC1_5 Sample source:saliva | Weight:- RAW_FILE_NAME(Raw file name)=220502s064_QC1_5.raw SUBJECT_SAMPLE_FACTORS - 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220502s389_A453 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s389_A453.raw SUBJECT_SAMPLE_FACTORS - 220502s390_A454 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s390_A454.raw SUBJECT_SAMPLE_FACTORS - 220502s391_A455 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s391_A455.raw SUBJECT_SAMPLE_FACTORS - 220502s394_A456 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s394_A456.raw SUBJECT_SAMPLE_FACTORS - 220502s395_A457 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s395_A457.raw SUBJECT_SAMPLE_FACTORS - 220502s396_A459 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s396_A459.raw SUBJECT_SAMPLE_FACTORS - 220502s399_A462 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s399_A462.raw SUBJECT_SAMPLE_FACTORS - 220502s400_A463 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s400_A463.raw SUBJECT_SAMPLE_FACTORS - 220502s401_A464 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s401_A464.raw SUBJECT_SAMPLE_FACTORS - 220502s402_A465 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s402_A465.raw SUBJECT_SAMPLE_FACTORS - 220502s406_A468 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s406_A468.raw SUBJECT_SAMPLE_FACTORS - 220502s407_A469 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s407_A469.raw SUBJECT_SAMPLE_FACTORS - 220502s410_A473 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s410_A473.raw SUBJECT_SAMPLE_FACTORS - 220502s411_A474 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s411_A474.raw SUBJECT_SAMPLE_FACTORS - 220502s412_A475 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s412_A475.raw SUBJECT_SAMPLE_FACTORS - 220502s414_A480 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s414_A480.raw SUBJECT_SAMPLE_FACTORS - 220502s419_A483 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s419_A483.raw SUBJECT_SAMPLE_FACTORS - 220502s422_A491 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s422_A491.raw SUBJECT_SAMPLE_FACTORS - 220502s423_A493 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s423_A493.raw SUBJECT_SAMPLE_FACTORS - 220502s424_A494 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s424_A494.raw SUBJECT_SAMPLE_FACTORS - 220502s425_A495 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s425_A495.raw SUBJECT_SAMPLE_FACTORS - 220502s426_A496 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s426_A496.raw SUBJECT_SAMPLE_FACTORS - 220502s430_A499 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s430_A499.raw SUBJECT_SAMPLE_FACTORS - 220502s431_A500 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s431_A500.raw SUBJECT_SAMPLE_FACTORS - 220502s432_A502 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s432_A502.raw SUBJECT_SAMPLE_FACTORS - 220502s433_A505 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s433_A505.raw SUBJECT_SAMPLE_FACTORS - 220502s434_A507 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s434_A507.raw SUBJECT_SAMPLE_FACTORS - 220502s435_A508 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s435_A508.raw SUBJECT_SAMPLE_FACTORS - 220502s436_A510 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s436_A510.raw SUBJECT_SAMPLE_FACTORS - 220502s437_A511 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=220502s437_A511.raw SUBJECT_SAMPLE_FACTORS - 230502s018_A001 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=230502s018_A001.raw SUBJECT_SAMPLE_FACTORS - 230502s020_A004 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=230502s020_A004.raw SUBJECT_SAMPLE_FACTORS - 230502s021_A007 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=230502s021_A007.raw SUBJECT_SAMPLE_FACTORS - 230502s022_A008 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=230502s022_A008.raw SUBJECT_SAMPLE_FACTORS - 230502s024_A011 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=230502s024_A011.raw SUBJECT_SAMPLE_FACTORS - 230502s025_A013 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=230502s025_A013.raw SUBJECT_SAMPLE_FACTORS - 230502s026_A014 Sample source:saliva | Weight:Healthy weight RAW_FILE_NAME(Raw file name)=230502s026_A014.raw SUBJECT_SAMPLE_FACTORS - 220502s145_A159 Sample source:saliva | Weight:Obesity RAW_FILE_NAME(Raw file name)=220502s145_A159.raw SUBJECT_SAMPLE_FACTORS - 220502s162_A179 Sample source:saliva | Weight:Obesity RAW_FILE_NAME(Raw file name)=220502s162_A179.raw SUBJECT_SAMPLE_FACTORS - 220502s202_A222 Sample source:saliva | Weight:Obesity RAW_FILE_NAME(Raw file name)=220502s202_A222.raw SUBJECT_SAMPLE_FACTORS - 220502s243_A273 Sample source:saliva | Weight:Obesity RAW_FILE_NAME(Raw file name)=220502s243_A273.raw SUBJECT_SAMPLE_FACTORS - 220502s244_A274 Sample source:saliva | Weight:Obesity RAW_FILE_NAME(Raw file name)=220502s244_A274.raw SUBJECT_SAMPLE_FACTORS - 220502s427_A498 Sample source:saliva | Weight:Obesity RAW_FILE_NAME(Raw file name)=220502s427_A498.raw SUBJECT_SAMPLE_FACTORS - 220502s068_A071 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s068_A071.raw SUBJECT_SAMPLE_FACTORS - 220502s072_A075 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s072_A075.raw SUBJECT_SAMPLE_FACTORS - 220502s074_A079 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s074_A079.raw SUBJECT_SAMPLE_FACTORS - 220502s085_A095 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s085_A095.raw SUBJECT_SAMPLE_FACTORS - 220502s093_A103 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s093_A103.raw SUBJECT_SAMPLE_FACTORS - 220502s095_A106 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s095_A106.raw SUBJECT_SAMPLE_FACTORS - 220502s107_A122 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s107_A122.raw SUBJECT_SAMPLE_FACTORS - 220502s130_A140 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s130_A140.raw SUBJECT_SAMPLE_FACTORS - 220502s154_A168 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s154_A168.raw SUBJECT_SAMPLE_FACTORS - 220502s156_A171 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s156_A171.raw SUBJECT_SAMPLE_FACTORS - 220502s171_A191 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s171_A191.raw SUBJECT_SAMPLE_FACTORS - 220502s174_A195 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s174_A195.raw SUBJECT_SAMPLE_FACTORS - 220502s186_A209 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s186_A209.raw SUBJECT_SAMPLE_FACTORS - 220502s206_A234 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s206_A234.raw SUBJECT_SAMPLE_FACTORS - 220502s208_A237 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s208_A237.raw SUBJECT_SAMPLE_FACTORS - 220502s260_A298 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s260_A298.raw SUBJECT_SAMPLE_FACTORS - 220502s321_A373 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s321_A373.raw SUBJECT_SAMPLE_FACTORS - 220502s333_A386 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s333_A386.raw SUBJECT_SAMPLE_FACTORS - 220502s350_A415 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s350_A415.raw SUBJECT_SAMPLE_FACTORS - 220502s367_A431 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s367_A431.raw SUBJECT_SAMPLE_FACTORS - 220502s372_A438 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s372_A438.raw SUBJECT_SAMPLE_FACTORS - 220502s375_A443 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s375_A443.raw SUBJECT_SAMPLE_FACTORS - 220502s408_A470 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s408_A470.raw SUBJECT_SAMPLE_FACTORS - 220502s409_A471 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s409_A471.raw SUBJECT_SAMPLE_FACTORS - 220502s418_A482 Sample source:saliva | Weight:Overweight RAW_FILE_NAME(Raw file name)=220502s418_A482.raw #COLLECTION CO:COLLECTION_SUMMARY In the PANIC cohort, saliva was collected by a research nurse from 7 to 10 a.m. CO:COLLECTION_SUMMARY after an overnight fast. Participants chewed paraffin to stimulate saliva CO:COLLECTION_SUMMARY excretion, with saliva excreted over the first 30 sec discarded, and the rest CO:COLLECTION_SUMMARY collected over the next three min. These samples were stored at -80 °C until CO:COLLECTION_SUMMARY further analysis. CO:COLLECTION_PROTOCOL_FILENAME wijnant-et-al-2020-validated-ultra-high-performance-liquid-chromatography-hybrid-high-resolution-mass-spectrometry-and_(1).pdf CO:SAMPLE_TYPE Saliva CO:STORAGE_CONDITIONS -80â #TREATMENT TR:TREATMENT_SUMMARY No treatment was applied #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Saliva samples were randomized, extracted and analyzed per cohort study using a SP:SAMPLEPREP_SUMMARY validated salivary UHPLC-Q-Orbitrap HRMS method, as previously described by SP:SAMPLEPREP_SUMMARY Wijnant et al. 2020â2â. In short, 450 mL of saliva was pipetted into a 1.5 SP:SAMPLEPREP_SUMMARY mL Eppendorf tube, followed by the addition of 10 mL of an internal standard SP:SAMPLEPREP_SUMMARY mixture (Table S9). The solution was vortexed briefly and centrifuged for 5 min SP:SAMPLEPREP_SUMMARY at 17,000 x g at room temperature. The supernatant was then collected with a 1 SP:SAMPLEPREP_SUMMARY mL syringe and passed over a polyamide membrane filter (25 mm diameter, 0.45 mm SP:SAMPLEPREP_SUMMARY pore size, Machery-Nagel, Germany). Finally, 150 mL of the undiluted extract was SP:SAMPLEPREP_SUMMARY transferred to a UHPLC-vial with a glass insert. SP:SAMPLEPREP_PROTOCOL_FILENAME wijnant-et-al-2020-validated-ultra-high-performance-liquid-chromatography-hybrid-high-resolution-mass-spectrometry-and_(1).pdf #CHROMATOGRAPHY CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) CH:COLUMN_TEMPERATURE 45°C CH:FLOW_GRADIENT A gradient profile with following proportions (v/v) of solvent A was applied: CH:FLOW_GRADIENT 0e1.5 min at 98%, 1.5e7.0 min from 98% to 75%, 7.0e8.0 min from 75% to 40%, CH:FLOW_GRADIENT 8.0e12.0 min from 40% to 5%, 12.0e14.0 min at 5%, 14.0e14.1 min from 5 to 98%, CH:FLOW_GRADIENT followed by 4.0 min of re-equilibration. CH:FLOW_RATE 0.4 mL/min CH:SOLVENT_A 100% Water; 0.1% Formic acid CH:SOLVENT_B 100% Acetonitrile; 0.1% Formic acid CH:CHROMATOGRAPHY_TYPE Reversed phase #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap Exploris 120 MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS MS acquisition: Xcalibur, Compound Discoverer, R studio. Polarity switching MS:MS_COMMENTS mode, thereby generating data for both positive and negative ionization at the MS:MS_COMMENTS same time. MS:ION_MODE NEGATIVE MS:MS_RESULTS_FILE ST003709_AN006086_Results.txt UNITS:a.u. Has m/z:Yes Has RT:Yes RT units:Minutes #END