{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003124","ANALYSIS_ID":"AN005122","VERSION":"1","CREATED_ON":"March 14, 2024, 10:32 pm"},

"PROJECT":{"PROJECT_TITLE":"Serum metabolites in inherited retinal degenerations","PROJECT_TYPE":"Untargeted meetabolomics analysis","PROJECT_SUMMARY":"Inherited retinal degeneration (IRD) contains a group of retinopathies characterized by high heterogeneity in phenotypes and a widely variable genetic background. The diagnosis of IRD is challenging owing to its phenotypic and genotypic complexity. Metabolomics, the emerging tool investigating comprehensive small molecule constitution of biological samples, provides instantaneous phenotypic information as metabolites reflect both genetic and environmental factors. In this project, we aimed to discover potential biomarkers and establish a diagnosis model for classifying specific IRDs.","INSTITUTE":"National Taiwan University","DEPARTMENT":"Department of Chemistry","LABORATORY":"Cheng-Chih Hsu''s lab","LAST_NAME":"Chung","FIRST_NAME":"Hsin-Hsiang","ADDRESS":"No. 1, Sec. 4, Roosevelt Rd., Taipei, Taipei, 106, Taiwan","EMAIL":"hhchung@ntu.edu.tw","PHONE":"+886-2-3366-1681"},

"STUDY":{"STUDY_TITLE":"Serum metabolites in inherited retinal degenerations","STUDY_SUMMARY":"The diagnosis of inherited retinal degeneration (IRD) is challenging owing to its phenotypic and genotypic complexity. Clinical information is important before a genetic diagnosis is made. Metabolomics studies the entire picture of bioproducts, which are determined using genetic codes and biological reactions. We demonstrated that the common diagnoses of IRD, including retinitis pigmentosa (RP), cone-rod dystrophy (CRD), Stargardt disease (STGD), and Bietti’s crystalline dystrophy (BCD), could be differentiated based on their metabolite heatmaps. Hundreds of metabolites were identified in the volcano plot compared with that of the control group in every IRD except BCD, considered as potential diagnosing markers. The phenotypes of CRD and STGD overlapped but could be differentiated by their metabolomic features with the assistance of a machine learning model with 100% accuracy. Moreover, EYS-, USH2A-associated, and other RP, sharing considerable similar characteristics in clinical findings, could also be diagnosed using the machine learning model with 85.7% accuracy. Further study would be needed to validate the results in the external dataset. By incorporating mass spectrometry and machine learning, a metabolomics-based diagnostic workflow for the clinical and molecular diagnoses of IRD was proposed in our study.","INSTITUTE":"National Taiwan University","DEPARTMENT":"Department of Chemistry","LABORATORY":"Cheng-Chih Hsu''s lab","LAST_NAME":"Chung","FIRST_NAME":"Hsin-Hsiang","ADDRESS":"No. 1, Sec. 4, Roosevelt Rd.","EMAIL":"hhchung@ntu.edu.tw","PHONE":"+886-2-3366-1681","TOTAL_SUBJECTS":"155","NUM_MALES":"90","NUM_FEMALES":"65"},

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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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"Sample ID":"RU102_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_102.mzXML"}
},
{
"Subject ID":"RU104",
"Sample ID":"RU104_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_104.mzXML"}
},
{
"Subject ID":"RU110",
"Sample ID":"RU110_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_110.mzXML"}
},
{
"Subject ID":"RU163",
"Sample ID":"RU163_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_163.mzXML"}
},
{
"Subject ID":"RU172",
"Sample ID":"RU172_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_172.mzXML"}
},
{
"Subject ID":"RU272",
"Sample ID":"RU272_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_272.mzXML"}
},
{
"Subject ID":"RU293",
"Sample ID":"RU293_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_293.mzXML"}
},
{
"Subject ID":"RU322",
"Sample ID":"RU322_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_322.mzXML"}
},
{
"Subject ID":"RU333",
"Sample ID":"RU333_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_333.mzXML"}
},
{
"Subject ID":"RU382",
"Sample ID":"RU382_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_382.mzXML"}
},
{
"Subject ID":"RU405",
"Sample ID":"RU405_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_405.mzXML"}
},
{
"Subject ID":"RU429",
"Sample ID":"RU429_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_429.mzXML"}
},
{
"Subject ID":"RU430",
"Sample ID":"RU430_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_430.mzXML"}
},
{
"Subject ID":"RU434",
"Sample ID":"RU434_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_434.mzXML"}
},
{
"Subject ID":"RU455",
"Sample ID":"RU455_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_455.mzXML"}
},
{
"Subject ID":"RU456",
"Sample ID":"RU456_MN",
"Factors":{"Sample source":"Serum","Disease":"RP","Causative gene":"USH2A"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_RP_USH2A_456.mzXML"}
},
{
"Subject ID":"S019",
"Sample ID":"S019_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_019.mzXML"}
},
{
"Subject ID":"S026",
"Sample ID":"S026_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_026.mzXML"}
},
{
"Subject ID":"S041",
"Sample ID":"S041_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_041.mzXML"}
},
{
"Subject ID":"S044",
"Sample ID":"S044_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_044.mzXML"}
},
{
"Subject ID":"S066",
"Sample ID":"S066_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_066.mzXML"}
},
{
"Subject ID":"S083",
"Sample ID":"S083_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_083.mzXML"}
},
{
"Subject ID":"S155",
"Sample ID":"S155_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_155.mzXML"}
},
{
"Subject ID":"S157",
"Sample ID":"S157_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_157.mzXML"}
},
{
"Subject ID":"S159",
"Sample ID":"S159_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_159.mzXML"}
},
{
"Subject ID":"S253",
"Sample ID":"S253_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_253.mzXML"}
},
{
"Subject ID":"S268",
"Sample ID":"S268_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_268.mzXML"}
},
{
"Subject ID":"S286",
"Sample ID":"S286_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_286.mzXML"}
},
{
"Subject ID":"S287",
"Sample ID":"S287_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_287.mzXML"}
},
{
"Subject ID":"S321",
"Sample ID":"S321_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_321.mzXML"}
},
{
"Subject ID":"S356",
"Sample ID":"S356_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_356.mzXML"}
},
{
"Subject ID":"S390",
"Sample ID":"S390_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_390.mzXML"}
},
{
"Subject ID":"S391",
"Sample ID":"S391_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_391.mzXML"}
},
{
"Subject ID":"S392",
"Sample ID":"S392_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_392.mzXML"}
},
{
"Subject ID":"S401",
"Sample ID":"S401_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_401.mzXML"}
},
{
"Subject ID":"S435",
"Sample ID":"S435_MN",
"Factors":{"Sample source":"Serum","Disease":"STGD","Causative gene":"ABCA4"},
"Additional sample data":{"RAW_FILE_NAME(Raw_FILE_NAME)":"20210216_Metabolites_neg_HRD_STGD_ABCA4_435.mzXML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Patients diagnosed with IRDs, including RP, STGD, CD/CRD, and BCD, at the National Taiwan University Hospital (NTUH) between 2015 and 2020 were prospectively enrolled in this cross-sectional observational study. The study protocol adhered to the tenets of the Declaration of Helsinki and was approved by the Institutional Review Board of NTUH. Serum samples of all participants were collected within 1 to 3 p.m. without fasting status. The serum samples were stored under -80°C before analysis.","SAMPLE_TYPE":"Blood (serum)"},

"TREATMENT":{"TREATMENT_SUMMARY":"Not applicable"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The MTBE extraction protocol, with laboratory modifications, was used to extract lipids and polar metabolites from the serum. The 10 μL internal standards (IS) mixture contains 15:0-18:1-d7-PC (2 ppm), 15:0-18:1-d7-PG (2 ppm), and L-tryptophan-(indole-d5) (10 ppm) were spiked into an aliquot of 50 μL serum. Then the sample was extracted by adding 600 μL MTBE and 150 μL MeOH and vortexed for 30 min at room temperature. Next, the sample was added with 200 μL water and centrifuged for 3 min at 13,697 g for phase separation. The upper portion containing serum lipids was transferred to another tube. The extraction was repeated by adding 100 μL water, 100 μL MeOH, and 300 μL MTBE. The sample was vortexed for an additional 10 min and centrifuged for 3 min at 13,697 g. The upper portion was mixed with the lower portion, and the combined solution was dried in a vacuum concentrator (Vacufuge plus Vacuum Concentrator, Eppendorf) for 3 h. The sample reconstitution was performed by adding 100 μL of reconstituted solution (ACN/IPA/water, v/v/v = 65/30/5). For the lower portion, 150 μL cold MeOH was added and stored under a -20°C environment for 2 h, followed by 10 min of 21,401 g centrifugation for protein precipitation. Next, the supernatant was dried using a vacuum concentrator overnight and then reconstituted by adding 100 μL reconstituted solution (ACN/water, v/v = 50/50). The protein precipitation was repeated by mixing 60 μL reconstituted sample with 120 μL cold ACN and then putting the mixtures in a -20°C freezer for an hour. After a 15 min centrifugation at 21,401 g under 4°C, super supernatants (120 μL) were collected and stored under -80°C before further analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Chromatography methods for metabolite extract, negative-ion mode.","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um)","SOLVENT_A":"100% Water; 5 mM ammonium acetate","SOLVENT_B":"100% Acetonitrile; 5 mM ammonium acetate","FLOW_GRADIENT":"0–1 min, 90% B; 1–9 min, 90-40% B; 9–11.5 min, 40% B; 11.5–12 min, 40–90% B; and 12–14 min, 90% B","FLOW_RATE":"0.25 mL/min","COLUMN_TEMPERATURE":"40℃"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"MS data were processed using Thermo Scientific Compound Discoverer v3.2 software.","CAPILLARY_TEMPERATURE":"280℃","ION_SOURCE_TEMPERATURE":"180℃","ION_SPRAY_VOLTAGE":"-3.5 kV","MS_RESULTS_FILE":"ST003124_AN005122_Results.txt UNITS:Peak area Has m/z:Neutral masses Has RT:Yes RT units:Minutes"}

}