#METABOLOMICS WORKBENCH xpw877923714_20250606_185457 DATATRACK_ID:6009 STUDY_ID:ST003954 ANALYSIS_ID:AN006500 PROJECT_ID:PR002477 VERSION 1 CREATED_ON June 8, 2025, 3:41 am #PROJECT PR:PROJECT_TITLE Integrated analysis of irritable bowel syndrome comorbid with psychological PR:PROJECT_TITLE disorders based on microbiomics and metabolomics PR:PROJECT_SUMMARY Background: Irritable Bowel Syndrome (IBS), a chronic functional PR:PROJECT_SUMMARY gastrointestinal disorder, shows high comorbidity rates with psychiatric PR:PROJECT_SUMMARY disorders. However, comprehensive evaluations of psychiatric symptoms in IBS and PR:PROJECT_SUMMARY their interactions with gut microbiota and metabolic disorders are lacking. This PR:PROJECT_SUMMARY study aims to investigated mechanistic links among gut microbiota, serum amino PR:PROJECT_SUMMARY acid metabolites, and differently psychological comorbidities. Methods: Clinical PR:PROJECT_SUMMARY data from 267 IBS patients and 91 healthy controls (HCs) were collected using PR:PROJECT_SUMMARY validated questionnaires: IBS Severity Scoring System (IBS-SSS), Hospital PR:PROJECT_SUMMARY Anxiety and Depression Scale, Patient Health Questionnaire-15 (PHQ-15), Somatic PR:PROJECT_SUMMARY Symptom Disorder-12 (SSD-12), and Symptom Checklist-90 (SCL-90). IBS patients PR:PROJECT_SUMMARY were stratified into psychiatric comorbidity and non-comorbidity subgroups based PR:PROJECT_SUMMARY on SCL-90 scores. Serum amino acid metabolites and gut microbiota were analyzed PR:PROJECT_SUMMARY using LC-MS/MS targeted metabolomics and 16S rRNA gene sequencing, respectively. PR:PROJECT_SUMMARY followed by integrative correlation analyses to explore interactions among PR:PROJECT_SUMMARY microbiota, metabolites, and psychological comorbidities. Results: IBS patients PR:PROJECT_SUMMARY showed significantly higher prevalence rates of all assessed psychiatric PR:PROJECT_SUMMARY disorders compared to HCs, with somatization, anxiety, obsessive-compulsive PR:PROJECT_SUMMARY traits, and depression as predominant manifestations. IBS patients with PR:PROJECT_SUMMARY different psychiatric comorbidities exhibit distinct gut microbiota PR:PROJECT_SUMMARY compositions, including altered abundances of the class Clostridia (e.g., PR:PROJECT_SUMMARY Ruminococcus, Faecalibacterium, Monoglobus), the class Coriobacteriia (e.g., PR:PROJECT_SUMMARY Collinsella), and the class Bacilli (e.g., Lactococcus, Erysipelatoclostridium). PR:PROJECT_SUMMARY Metabolomic profiling revealed elevated arginine and its derivatives PR:PROJECT_SUMMARY (Nα-acetyl-L-arginine, argininosuccinate, succinate) and reductions in aromatic PR:PROJECT_SUMMARY amino acids (L-tryptophan, L-phenylalanine, L-tyrosine), branched-chain amino PR:PROJECT_SUMMARY acids (L-valine, L-leucine, L-isoleucine), and neuroactive amino acids PR:PROJECT_SUMMARY (L-glutamate, L-aspartate). Multi-omics integration implicated Clostridia and PR:PROJECT_SUMMARY Coriobacteriia might respectively participate in the pathophysiology of PR:PROJECT_SUMMARY somatization and anxiety comorbidities by modulating the ornithine cycle and PR:PROJECT_SUMMARY glutamatergic metabolism. Conclusions: IBS patients exhibit multidimensional PR:PROJECT_SUMMARY psychiatric disorders, with distinct gut microbiota and serum amino acids PR:PROJECT_SUMMARY profiles corresponding to specific psychiatric comorbidities. The interaction PR:PROJECT_SUMMARY between gut microbiota and amino acids may be involved in the pathogenesis of PR:PROJECT_SUMMARY IBS with comorbid mental disorders. This study provide novel targets for early PR:PROJECT_SUMMARY stratification and precision interventions in IBS with psychological PR:PROJECT_SUMMARY comorbidities. PR:INSTITUTE Sun Yat-sen University PR:LAST_NAME Peiwei PR:FIRST_NAME Xie PR:ADDRESS Department of Gastroenterology and Hepatology, The First Affiliated Hospital, PR:ADDRESS Sun Yat-sen University, No. 58, Zhongshan Er Road, Guangzhou, China PR:EMAIL 877923714@qq.com PR:PHONE 020-87755766 #STUDY ST:STUDY_TITLE Integrated analysis of irritable bowel syndrome comorbid with psychological ST:STUDY_TITLE disorders based on microbiomics and metabolomics ST:STUDY_SUMMARY Background: Irritable Bowel Syndrome (IBS), a chronic functional ST:STUDY_SUMMARY gastrointestinal disorder, shows high comorbidity rates with psychiatric ST:STUDY_SUMMARY disorders. However, comprehensive evaluations of psychiatric symptoms in IBS and ST:STUDY_SUMMARY their interactions with gut microbiota and metabolic disorders are lacking. This ST:STUDY_SUMMARY study aims to investigated mechanistic links among gut microbiota, serum amino ST:STUDY_SUMMARY acid metabolites, and differently psychological comorbidities. Methods: Clinical ST:STUDY_SUMMARY data from 267 IBS patients and 91 healthy controls (HCs) were collected using ST:STUDY_SUMMARY validated questionnaires: IBS Severity Scoring System (IBS-SSS), Hospital ST:STUDY_SUMMARY Anxiety and Depression Scale, Patient Health Questionnaire-15 (PHQ-15), Somatic ST:STUDY_SUMMARY Symptom Disorder-12 (SSD-12), and Symptom Checklist-90 (SCL-90). IBS patients ST:STUDY_SUMMARY were stratified into psychiatric comorbidity and non-comorbidity subgroups based ST:STUDY_SUMMARY on SCL-90 scores. Serum amino acid metabolites and gut microbiota were analyzed ST:STUDY_SUMMARY using LC-MS/MS targeted metabolomics and 16S rRNA gene sequencing, respectively. ST:STUDY_SUMMARY followed by integrative correlation analyses to explore interactions among ST:STUDY_SUMMARY microbiota, metabolites, and psychological comorbidities. Results: IBS patients ST:STUDY_SUMMARY showed significantly higher prevalence rates of all assessed psychiatric ST:STUDY_SUMMARY disorders compared to HCs, with somatization, anxiety, obsessive-compulsive ST:STUDY_SUMMARY traits, and depression as predominant manifestations. IBS patients with ST:STUDY_SUMMARY different psychiatric comorbidities exhibit distinct gut microbiota ST:STUDY_SUMMARY compositions, including altered abundances of the class Clostridia (e.g., ST:STUDY_SUMMARY Ruminococcus, Faecalibacterium, Monoglobus), the class Coriobacteriia (e.g., ST:STUDY_SUMMARY Collinsella), and the class Bacilli (e.g., Lactococcus, Erysipelatoclostridium). ST:STUDY_SUMMARY Metabolomic profiling revealed elevated arginine and its derivatives ST:STUDY_SUMMARY (Nα-acetyl-L-arginine, argininosuccinate, succinate) and reductions in aromatic ST:STUDY_SUMMARY amino acids (L-tryptophan, L-phenylalanine, L-tyrosine), branched-chain amino ST:STUDY_SUMMARY acids (L-valine, L-leucine, L-isoleucine), and neuroactive amino acids ST:STUDY_SUMMARY (L-glutamate, L-aspartate). Multi-omics integration implicated Clostridia and ST:STUDY_SUMMARY Coriobacteriia might respectively participate in the pathophysiology of ST:STUDY_SUMMARY somatization and anxiety comorbidities by modulating the ornithine cycle and ST:STUDY_SUMMARY glutamatergic metabolism. Conclusions: IBS patients exhibit multidimensional ST:STUDY_SUMMARY psychiatric disorders, with distinct gut microbiota and serum amino acids ST:STUDY_SUMMARY profiles corresponding to specific psychiatric comorbidities. The interaction ST:STUDY_SUMMARY between gut microbiota and amino acids may be involved in the pathogenesis of ST:STUDY_SUMMARY IBS with comorbid mental disorders. This study provide novel targets for early ST:STUDY_SUMMARY stratification and precision interventions in IBS with psychological ST:STUDY_SUMMARY comorbidities. ST:INSTITUTE Sun Yat-sen University ST:LAST_NAME Peiwei ST:FIRST_NAME Xie ST:ADDRESS Department of Gastroenterology and Hepatology, The First Affiliated Hospital, ST:ADDRESS Sun Yat-sen University, No. 58, Zhongshan Er Road, Guangzhou, China ST:EMAIL 877923714@qq.com ST:PHONE 020-87755766 #SUBJECT SU:SUBJECT_TYPE Human 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 - A01 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A01.mzML SUBJECT_SAMPLE_FACTORS - A02 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A02.mzML SUBJECT_SAMPLE_FACTORS - A03 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A03.mzML SUBJECT_SAMPLE_FACTORS - A04 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A04.mzML SUBJECT_SAMPLE_FACTORS - A05 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A05.mzML SUBJECT_SAMPLE_FACTORS - A06 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A06.mzML SUBJECT_SAMPLE_FACTORS - A07 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A07.mzML SUBJECT_SAMPLE_FACTORS - A08 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A08.mzML SUBJECT_SAMPLE_FACTORS - A09 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A09.mzML SUBJECT_SAMPLE_FACTORS - A10 Sample source:serum | Group1:HC | Group2:1 RAW_FILE_NAME(Raw file name)=A10.mzML SUBJECT_SAMPLE_FACTORS - A11 Sample source:serum | Group1:IBS-C-1 | Group2:1 RAW_FILE_NAME(Raw file name)=A11.mzML SUBJECT_SAMPLE_FACTORS - A12 Sample source:serum | Group1:IBS-C-2 | Group2:1 RAW_FILE_NAME(Raw file name)=A12.mzML SUBJECT_SAMPLE_FACTORS - A13 Sample source:serum | Group1:IBS-C-3 | Group2:1 RAW_FILE_NAME(Raw file name)=A13.mzML SUBJECT_SAMPLE_FACTORS - A14 Sample source:serum | Group1:IBS-C-4 | Group2:1 RAW_FILE_NAME(Raw file name)=A14.mzML SUBJECT_SAMPLE_FACTORS - A15 Sample source:serum | Group1:IBS-C-5 | Group2:1 RAW_FILE_NAME(Raw file name)=A15.mzML SUBJECT_SAMPLE_FACTORS - A16 Sample source:serum | Group1:IBS-C-6 | Group2:1 RAW_FILE_NAME(Raw file name)=A16.mzML SUBJECT_SAMPLE_FACTORS - A17 Sample source:serum | Group1:IBS-C-7 | Group2:1 RAW_FILE_NAME(Raw file name)=A17.mzML SUBJECT_SAMPLE_FACTORS - A18 Sample source:serum | Group1:IBS-C-8 | Group2:1 RAW_FILE_NAME(Raw file name)=A18.mzML SUBJECT_SAMPLE_FACTORS - A19 Sample source:serum | Group1:IBS-C-9 | Group2:1 RAW_FILE_NAME(Raw file name)=A19.mzML SUBJECT_SAMPLE_FACTORS - A20 Sample source:serum | Group1:IBS-C-10 | Group2:1 RAW_FILE_NAME(Raw file name)=A20.mzML SUBJECT_SAMPLE_FACTORS - A21 Sample source:serum | Group1:IBS-C-11 | Group2:1 RAW_FILE_NAME(Raw file name)=A21.mzML SUBJECT_SAMPLE_FACTORS - A22 Sample source:serum | Group1:IBS-C-12 | Group2:1 RAW_FILE_NAME(Raw file name)=A22.mzML SUBJECT_SAMPLE_FACTORS - A23 Sample source:serum | Group1:IBS-C-13 | Group2:1 RAW_FILE_NAME(Raw file name)=A23.mzML SUBJECT_SAMPLE_FACTORS - A24 Sample source:serum | Group1:IBS-C-14 | Group2:1 RAW_FILE_NAME(Raw file name)=A24.mzML SUBJECT_SAMPLE_FACTORS - A25 Sample source:serum | Group1:IBS-C-15 | Group2:1 RAW_FILE_NAME(Raw file name)=A25.mzML SUBJECT_SAMPLE_FACTORS - A26 Sample source:serum | Group1:IBS-C-16 | Group2:1 RAW_FILE_NAME(Raw file name)=A26.mzML SUBJECT_SAMPLE_FACTORS - A27 Sample source:serum | Group1:IBS-D-1 | Group2:1 RAW_FILE_NAME(Raw file name)=A27.mzML SUBJECT_SAMPLE_FACTORS - A28 Sample source:serum | Group1:IBS-D-2 | Group2:1 RAW_FILE_NAME(Raw file name)=A28.mzML SUBJECT_SAMPLE_FACTORS - A29 Sample source:serum | Group1:IBS-D-3 | Group2:1 RAW_FILE_NAME(Raw file name)=A29.mzML SUBJECT_SAMPLE_FACTORS - A30 Sample source:serum | Group1:IBS-D-4 | Group2:1 RAW_FILE_NAME(Raw file name)=A30.mzML SUBJECT_SAMPLE_FACTORS - A31 Sample source:serum | Group1:IBS-D-5 | Group2:1 RAW_FILE_NAME(Raw file name)=A31.mzML SUBJECT_SAMPLE_FACTORS - A32 Sample source:serum | Group1:IBS-D-6 | Group2:1 RAW_FILE_NAME(Raw file name)=A32.mzML SUBJECT_SAMPLE_FACTORS - A33 Sample source:serum | Group1:IBS-D-7 | Group2:1 RAW_FILE_NAME(Raw file name)=A33.mzML SUBJECT_SAMPLE_FACTORS - A34 Sample source:serum | Group1:IBS-D-8 | Group2:1 RAW_FILE_NAME(Raw file name)=A34.mzML SUBJECT_SAMPLE_FACTORS - A35 Sample source:serum | Group1:IBS-D-9 | Group2:1 RAW_FILE_NAME(Raw file name)=A35.mzML SUBJECT_SAMPLE_FACTORS - A36 Sample source:serum | Group1:IBS-D-10 | Group2:1 RAW_FILE_NAME(Raw file name)=A36.mzML SUBJECT_SAMPLE_FACTORS - A37 Sample source:serum | Group1:IBS-D-11 | Group2:1 RAW_FILE_NAME(Raw file name)=A37.mzML SUBJECT_SAMPLE_FACTORS - A38 Sample source:serum | Group1:IBS-D-12 | Group2:1 RAW_FILE_NAME(Raw file name)=A38.mzML SUBJECT_SAMPLE_FACTORS - A39 Sample source:serum | Group1:IBS-D-13 | Group2:1 RAW_FILE_NAME(Raw file name)=A39.mzML SUBJECT_SAMPLE_FACTORS - A40 Sample source:serum | Group1:IBS-D-14 | Group2:1 RAW_FILE_NAME(Raw file name)=A40.mzML SUBJECT_SAMPLE_FACTORS - A41 Sample source:serum | Group1:IBS-D-15 | Group2:1 RAW_FILE_NAME(Raw file name)=A41.mzML SUBJECT_SAMPLE_FACTORS - A42 Sample source:serum | Group1:IBS-D-16 | Group2:1 RAW_FILE_NAME(Raw file name)=A42.mzML SUBJECT_SAMPLE_FACTORS - A43 Sample source:serum | Group1:IBS-D-17 | Group2:1 RAW_FILE_NAME(Raw file name)=A43.mzML SUBJECT_SAMPLE_FACTORS - A44 Sample source:serum | Group1:IBS-D-18 | Group2:1 RAW_FILE_NAME(Raw file name)=A44.mzML SUBJECT_SAMPLE_FACTORS - A45 Sample source:serum | Group1:IBS-D-19 | Group2:1 RAW_FILE_NAME(Raw file name)=A45.mzML SUBJECT_SAMPLE_FACTORS - A46 Sample source:serum | Group1:IBS-D-20 | Group2:1 RAW_FILE_NAME(Raw file name)=A46.mzML SUBJECT_SAMPLE_FACTORS - A47 Sample source:serum | Group1:IBS-D-21 | Group2:1 RAW_FILE_NAME(Raw file name)=A47.mzML SUBJECT_SAMPLE_FACTORS - A48 Sample source:serum | Group1:IBS-D-22 | Group2:1 RAW_FILE_NAME(Raw file name)=A48.mzML SUBJECT_SAMPLE_FACTORS - A49 Sample source:serum | Group1:IBS-D-23 | Group2:1 RAW_FILE_NAME(Raw file name)=A49.mzML SUBJECT_SAMPLE_FACTORS - A50 Sample source:serum | Group1:IBS-D-24 | Group2:1 RAW_FILE_NAME(Raw file name)=A50.mzML SUBJECT_SAMPLE_FACTORS - A51 Sample source:serum | Group1:IBS-D-25 | Group2:1 RAW_FILE_NAME(Raw file name)=A51.mzML SUBJECT_SAMPLE_FACTORS - A52 Sample source:serum | Group1:IBS-D-26 | Group2:1 RAW_FILE_NAME(Raw file name)=A52.mzML SUBJECT_SAMPLE_FACTORS - A53 Sample source:serum | Group1:IBS-D-27 | Group2:1 RAW_FILE_NAME(Raw file name)=A53.mzML SUBJECT_SAMPLE_FACTORS - A54 Sample source:serum | Group1:IBS-D-28 | Group2:1 RAW_FILE_NAME(Raw file name)=A54.mzML SUBJECT_SAMPLE_FACTORS - A55 Sample source:serum | Group1:IBS-D-29 | Group2:1 RAW_FILE_NAME(Raw file name)=A55.mzML SUBJECT_SAMPLE_FACTORS - A56 Sample source:serum | Group1:IBS-D-30 | Group2:1 RAW_FILE_NAME(Raw file name)=A56.mzML SUBJECT_SAMPLE_FACTORS - A57 Sample source:serum | Group1:IBS-D-31 | Group2:1 RAW_FILE_NAME(Raw file name)=A57.mzML SUBJECT_SAMPLE_FACTORS - A58 Sample source:serum | Group1:IBS-M-1 | Group2:1 RAW_FILE_NAME(Raw file name)=A58.mzML SUBJECT_SAMPLE_FACTORS - A59 Sample source:serum | Group1:IBS-M-2 | Group2:1 RAW_FILE_NAME(Raw file name)=A59.mzML SUBJECT_SAMPLE_FACTORS - A60 Sample source:serum | Group1:IBS-M-3 | Group2:1 RAW_FILE_NAME(Raw file name)=A60.mzML SUBJECT_SAMPLE_FACTORS - A61 Sample source:serum | Group1:IBS-M-4 | Group2:1 RAW_FILE_NAME(Raw file name)=A61.mzML SUBJECT_SAMPLE_FACTORS - A62 Sample source:serum | Group1:IBS-M-5 | Group2:1 RAW_FILE_NAME(Raw file name)=A62.mzML SUBJECT_SAMPLE_FACTORS - A63 Sample source:serum | Group1:IBS-M-6 | Group2:1 RAW_FILE_NAME(Raw file name)=A63.mzML SUBJECT_SAMPLE_FACTORS - A64 Sample source:serum | Group1:IBS-M-7 | Group2:1 RAW_FILE_NAME(Raw file name)=A64.mzML SUBJECT_SAMPLE_FACTORS - A65 Sample source:serum | Group1:IBS-M-8 | Group2:1 RAW_FILE_NAME(Raw file name)=A65.mzML SUBJECT_SAMPLE_FACTORS - A66 Sample source:serum | Group1:IBS-M-9 | Group2:1 RAW_FILE_NAME(Raw file name)=A66.mzML SUBJECT_SAMPLE_FACTORS - A67 Sample source:serum | Group1:IBS-M-10 | Group2:1 RAW_FILE_NAME(Raw file name)=A67.mzML SUBJECT_SAMPLE_FACTORS - A68 Sample source:serum | Group1:IBS-M-11 | Group2:1 RAW_FILE_NAME(Raw file name)=A68.mzML SUBJECT_SAMPLE_FACTORS - A69 Sample source:serum | Group1:IBS-M-12 | Group2:1 RAW_FILE_NAME(Raw file name)=A69.mzML SUBJECT_SAMPLE_FACTORS - A70 Sample source:serum | Group1:IBS-M-13 | Group2:1 RAW_FILE_NAME(Raw file name)=A70.mzML SUBJECT_SAMPLE_FACTORS - A71 Sample source:serum | Group1:IBS-M-14 | Group2:1 RAW_FILE_NAME(Raw file name)=A71.mzML #COLLECTION CO:COLLECTION_SUMMARY Venous blood samples were obtained after overnight fasting, followed by serum CO:COLLECTION_SUMMARY separation through aseptic processing (centrifugation at 3,000 × g for 10 min CO:COLLECTION_SUMMARY at 4℃) and subsequent storage at -80℃.In this study, we collected serum CO:COLLECTION_SUMMARY samples from 61 IBS patients and 10 healthy controls. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY Diagnosed with IBS based on the Rome IV criteria. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Venous blood samples were obtained after overnight fasting, followed by serum SP:SAMPLEPREP_SUMMARY separation through aseptic processing (centrifugation at 3,000 × g for 10 min SP:SAMPLEPREP_SUMMARY at 4 °C) and subsequent storage at -80 °C. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The data acquisition instrument system primarily includes Ultra Performance CH:CHROMATOGRAPHY_SUMMARY Liquid Chromatography (UPLC) (ExionLC™ AD, https://sciex.com.cn/) and Tandem CH:CHROMATOGRAPHY_SUMMARY Mass Spectrometry (MS/MS) (QTRAP® 6500+, https://sciex.com.cn/). The liquid CH:CHROMATOGRAPHY_SUMMARY chromatography conditions mainly comprise: 1) Chromatographic column: ACQUITY CH:CHROMATOGRAPHY_SUMMARY BEH Amide column (1.7 µm, 100 mm×2.1 mm i.d.); 2) Mobile phase: Phase A, CH:CHROMATOGRAPHY_SUMMARY ultrapure water (containing 2 mM ammonium acetate, 0.04% formic acid); Phase B, CH:CHROMATOGRAPHY_SUMMARY acetonitrile (containing 2 mM ammonium acetate, 0.04% formic acid); 3) Gradient CH:CHROMATOGRAPHY_SUMMARY elution program: 0-1.2 min A/B at 10:90 (V/V), 9 min A/B at 40:60 (V/V), 10-11 CH:CHROMATOGRAPHY_SUMMARY min at 60:40 (V/V), 11.01-15 min at 10:90 (V/V); 4) Flow rate 0.4 mL/min; column CH:CHROMATOGRAPHY_SUMMARY temperature 40°C; injection volume 2 μL. The mass spectrometry conditions CH:CHROMATOGRAPHY_SUMMARY mainly include: Electrospray Ionization (ESI) temperature at 550°C, mass CH:CHROMATOGRAPHY_SUMMARY spectrometry voltage at 5500 V in positive ion mode, and -4500 V in negative ion CH:CHROMATOGRAPHY_SUMMARY mode, with curtain gas (CUR) at 35 psi. In the Q-Trap 6500+, each ion pair is CH:CHROMATOGRAPHY_SUMMARY scanned and detected based on optimized declustering potential (DP) and CH:CHROMATOGRAPHY_SUMMARY collision energy (CE). CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME SCIEX QTRAP 6500+ CH:COLUMN_NAME ACQUITY BEH Amide (1.7 µm，100 mm×2.1 mm i.d.) CH:SOLVENT_A ultrapure water containing 2 mM ammonium acetate and 0.04% formic acid CH:SOLVENT_B acetonitrile containing 2 mM ammonium acetate and 0.04% formic acid CH:FLOW_GRADIENT 0-1.2 minutes A/B is 10:90 (V/V), 9 minutes A/B is 40:60 (V/V), 10-11 minutes is CH:FLOW_GRADIENT 60:40 (V/V), 11.01-15 minutes is 10:90 (V/V) CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 40°C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 6500+ QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS The Metware Database (MWDB) is constructed based on standard samples to perform MS:MS_COMMENTS qualitative analysis on mass spectrometry data. Quantitative analysis is MS:MS_COMMENTS accomplished using the Multiple Reaction Monitoring (MRM) mode of triple MS:MS_COMMENTS quadrupole mass spectrometry. In the MRM mode, the quadrupole first selects the MS:MS_COMMENTS precursor ions (parent ions) of the target substances, eliminating ions MS:MS_COMMENTS corresponding to other molecular weights to preliminarily exclude interference. MS:MS_COMMENTS The precursor ions undergo collision-induced ionization in the collision cell, MS:MS_COMMENTS resulting in the formation of multiple fragment ions. These fragment ions are MS:MS_COMMENTS then filtered through the triple quadrupole to select the required MS:MS_COMMENTS characteristic fragment ions, excluding interference from non-target ions, MS:MS_COMMENTS thereby enhancing the accuracy and reproducibility of the quantification. After MS:MS_COMMENTS obtaining the mass spectrometry analysis data of different samples, the MS:MS_COMMENTS chromatographic peaks of all target substances are integrated, and quantitative MS:MS_COMMENTS analysis is conducted using a standard curve. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS ng/ml MS_METABOLITE_DATA_START Samples A01 A02 A03 A04 A05 A06 A07 A08 A09 A10 A11 A12 A13 A14 A15 A16 A17 A18 A19 A20 A21 A22 A23 A24 A25 A26 A27 A28 A29 A30 A31 A32 A33 A34 A35 A36 A37 A38 A39 A40 A41 A42 A43 A44 A45 A46 A47 A48 A49 A50 A51 A52 A53 A54 A55 A56 A57 A58 A59 A60 A61 A62 A63 A64 A65 A66 A67 A68 A69 A70 A71 Factors Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:HC | Group2:1 Sample source:serum | Group1:IBS-C-1 | Group2:1 Sample source:serum | Group1:IBS-C-2 | Group2:1 Sample source:serum | Group1:IBS-C-3 | Group2:1 Sample source:serum | Group1:IBS-C-4 | Group2:1 Sample source:serum | Group1:IBS-C-5 | Group2:1 Sample source:serum | Group1:IBS-C-6 | Group2:1 Sample source:serum | Group1:IBS-C-7 | Group2:1 Sample source:serum | Group1:IBS-C-8 | Group2:1 Sample source:serum | Group1:IBS-C-9 | Group2:1 Sample source:serum | Group1:IBS-C-10 | Group2:1 Sample source:serum | Group1:IBS-C-11 | Group2:1 Sample source:serum | Group1:IBS-C-12 | Group2:1 Sample source:serum | Group1:IBS-C-13 | Group2:1 Sample source:serum | Group1:IBS-C-14 | Group2:1 Sample source:serum | Group1:IBS-C-15 | Group2:1 Sample source:serum | Group1:IBS-C-16 | Group2:1 Sample source:serum | Group1:IBS-D-1 | Group2:1 Sample source:serum | Group1:IBS-D-2 | Group2:1 Sample source:serum | Group1:IBS-D-3 | Group2:1 Sample source:serum | Group1:IBS-D-4 | Group2:1 Sample source:serum | Group1:IBS-D-5 | Group2:1 Sample source:serum | Group1:IBS-D-6 | Group2:1 Sample source:serum | Group1:IBS-D-7 | Group2:1 Sample source:serum | Group1:IBS-D-8 | Group2:1 Sample source:serum | Group1:IBS-D-9 | Group2:1 Sample source:serum | Group1:IBS-D-10 | Group2:1 Sample source:serum | Group1:IBS-D-11 | Group2:1 Sample source:serum | Group1:IBS-D-12 | Group2:1 Sample source:serum | Group1:IBS-D-13 | Group2:1 Sample source:serum | Group1:IBS-D-14 | Group2:1 Sample source:serum | Group1:IBS-D-15 | Group2:1 Sample source:serum | Group1:IBS-D-16 | Group2:1 Sample source:serum | Group1:IBS-D-17 | Group2:1 Sample source:serum | Group1:IBS-D-18 | Group2:1 Sample source:serum | Group1:IBS-D-19 | Group2:1 Sample source:serum | Group1:IBS-D-20 | Group2:1 Sample source:serum | Group1:IBS-D-21 | Group2:1 Sample source:serum | Group1:IBS-D-22 | Group2:1 Sample source:serum | Group1:IBS-D-23 | Group2:1 Sample source:serum | Group1:IBS-D-24 | Group2:1 Sample source:serum | Group1:IBS-D-25 | Group2:1 Sample source:serum | Group1:IBS-D-26 | Group2:1 Sample source:serum | Group1:IBS-D-27 | Group2:1 Sample source:serum | Group1:IBS-D-28 | Group2:1 Sample source:serum | Group1:IBS-D-29 | Group2:1 Sample source:serum | Group1:IBS-D-30 | Group2:1 Sample source:serum | Group1:IBS-D-31 | Group2:1 Sample source:serum | Group1:IBS-M-1 | Group2:1 Sample source:serum | Group1:IBS-M-2 | Group2:1 Sample source:serum | Group1:IBS-M-3 | Group2:1 Sample source:serum | Group1:IBS-M-4 | Group2:1 Sample source:serum | Group1:IBS-M-5 | Group2:1 Sample source:serum | Group1:IBS-M-6 | Group2:1 Sample source:serum | Group1:IBS-M-7 | Group2:1 Sample source:serum | Group1:IBS-M-8 | Group2:1 Sample source:serum | Group1:IBS-M-9 | Group2:1 Sample source:serum | Group1:IBS-M-10 | Group2:1 Sample source:serum | Group1:IBS-M-11 | Group2:1 Sample source:serum | Group1:IBS-M-12 | Group2:1 Sample source:serum | Group1:IBS-M-13 | Group2:1 Sample source:serum | Group1:IBS-M-14 | Group2:1 1,3-Dimethyluric-Acid 1429.446 3276.78 18346.98 0 1746.09 1648.662 0 0 3654.678 2568.168 0 0 0 0 143.925 0 8156.76 0 2637.378 0 12716.4 0 1414.752 162.9996 12721.86 374.829 0 3734.142 0 1714.224 1817.016 0 6783.54 0 5741.166 0 1185.372 0 2320.116 0 95.5662 360.0558 2346.864 8621.04 0 262.5696 0 2923.302 10410.96 1535.652 0 514.29 0 1101.33 1.348158 572.6256 1449.222 0 0 0 632.01 309.1986 3.79131 3479.832 2417.676 1267.926 326.9958 1985.328 104.1324 0 3475.668 2-Aminoethanesulfonic-Acid 42459.06 90343.8 74934.6 43029.36 105417 69818.4 35596.38 46248.24 39812.1 37178.94 58297.2 66791.4 63188.4 56845.32 92026.8 76625.4 69597 50906.76 57471 48432.6 72181.8 58372.5 119636.4 55533.06 68254.8 74009.4 72256.2 82832.4 49490.22 55506.66 101937 69610.8 80280 80151 76392.6 81113.4 94869.6 76224 83818.8 31167.78 56806.32 73832.4 75841.2 67101 51245.16 60907.8 90714.6 73979.4 55670.64 52701.66 79803 55538.64 76918.2 71838 64962.6 76892.4 48070.62 79099.8 52423.86 51303.36 82933.8 76470 75961.2 59217.24 82429.8 54084.96 93566.4 60276.6 69957.6 75265.8 58186.14 3,7-Dimethyluric-Acid 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5-Hydroxy-tryptophan 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5300.598 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Cystine 32741.1 32964.12 30931.08 36178.2 29078.64 53090.52 30382.26 34266.9 43996.14 35055.54 31203.18 28316.94 50082.9 35507.88 32483.76 24152.04 26923.62 25310.76 50005.62 29307.06 38865.6 47068.32 39046.56 39371.82 36958.38 43901.34 27808.92 27535.56 19628.4 28760.28 26023.02 24283.56 27386.28 27776.46 29599.08 27066.6 31865.58 27393 29792.94 25073.94 33665.46 28199.22 33270.78 40713.6 25683.42 43483.86 41625.12 31437.48 41239.44 34568.58 47435.28 31274.16 27549.36 31988.82 48611.28 37902.96 32059.62 20898.54 41604.12 22194.9 31104.66 38651.88 29997.42 23398.14 32369.4 38716.86 28241.58 28323.18 36025.86 33638.58 47390.76 N-Isovaleroylglycine 0 0 0 0 0.4517028 0 2.512134 0 0 0 11.15376 15.13146 17.81934 0 0 35.424 0 0 0 69.5442 0 52.05198 0 0 8.46984 0 19.89576 7.0392 0 5.986338 0 25.64556 0 40.64322 48.38154 0 0 38.41074 51.83166 48.10584 0 0 8.0694 16.78626 4.84269 37.93878 0 0 0 6.25698 0 0 0 26.25084 0 0 0 0 0 0 0 0 21.58224 0 0.674544 1.08927 2.18616 24.7611 0 0 0 N-Propionylglycine 45.882 0 0 0 93.3474 78.1014 77.9466 0 41.02104 98.8314 140.8356 132.5586 132.1836 0 78.8118 102.6234 0 0 97.0656 116.226 39.94146 86.4954 142.71 118.0962 46.17132 0 69.3312 96.8706 123.8988 107.4156 79.2894 196.6644 69.0972 119.9016 175.077 119.8068 63.3528 109.497 181.7502 233.709 0 84.2748 113.7108 133.0032 116.9526 158.1822 104.9724 50.26878 72.1182 112.4022 46.06362 51.44334 47.36688 180.06 0 61.4238 61.4112 65.3208 0 77.9364 76.3572 64.0722 189.4572 46.75314 111.3432 95.769 64.7418 90.4002 78.1254 0 43.5024 argininosuccinic-acid 0 3265.71 3289.188 3385.806 4861.188 3773.232 3915.372 2544.87 3832.038 2433.138 0 3392.814 2872.908 2030.142 2696.16 2973.21 2238.534 2619.756 4467.042 3078.648 2807.358 3798.468 2897.478 2568.216 2611.758 3115.392 0 0 0 0 3134.976 2432.85 0 0 2999.61 2501.106 0 0 2289.75 0 2571.408 2355.132 2905.674 2446.164 2194.206 3624.27 2622.864 2347.764 2314.758 2610.768 2382.882 2572.65 2664.534 2859.264 4691.97 4427.37 2617.788 2297.166 2437.626 2091.33 3395.094 2707.266 2896.23 1966.374 2976.138 2594.172 1838.07 2842.632 3051.312 2278.962 2749.674 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID 1,3-Dimethyluric-Acid 1.54 169.18 HMDB01857 - 2-Aminoethanesulfonic-Acid 6.85 89.05 HMDB00251 C00245 3,7-Dimethyluric-Acid 1.68 174.11 HMDB01982 - 5-Hydroxy-tryptophan 6.77 280.11 - - argininosuccinic-acid 10.97 195.05 - - L-Cystine 10.88 204.09 HMDB00192 C00491 N-Isovaleroylglycine 1.43 240.26 HMDB00678 - N-Propionylglycine 2.45 174.1 HMDB00783 - METABOLITES_END #END