{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003069","ANALYSIS_ID":"AN005027","VERSION":"1","CREATED_ON":"February 8, 2024, 1:30 am"},

"PROJECT":{"PROJECT_TITLE":"Plasma instead of serum avoids critical confounding of clinical metabolomics studies by platelets","PROJECT_SUMMARY":"Metabolomics is an emerging and powerful molecular profiling method supporting clinical investigations. Serum and plasma are commonly used without rational prioritization. Serum is collected after blood coagulation, a complex biochemical process involving active platelet metabolism. This may affect the metabolome and increase the variance as platelet counts and function may vary substantially in individuals. A multi-omics approach systematically investigating the suitability of serum and plasma for clinical studies demonstrated that metabolites correlated well (n=461, R2=0.991), whereas lipid mediators (n=104, R2=0.906) and proteins (n=322, R2=0.860) differed substantially between specimen. Independently, analysis of platelet releasates identified most biomolecules significantly enriched in serum when compared to plasma. A prospective, randomized, controlled parallel group metabolomics trial with acetylsalicylic acid administered for 7 days demonstrated that the apparent drug effects significantly differ depending on analyzed specimen. Only serum analyses of healthy individuals suggested a significant downregulation of TXB2 and 12-HETE, which were specifically formed during coagulation in vitro. Plasma analyses reliably identified acetylsalicylic acid effects on metabolites and lipids occurring in vivo such as a decrease in polyunsaturated fatty acids. The present data suggests that plasma should be preferred above serum for clinical metabolomics studies as the serum metabolome may be substantially confounded by platelets.","INSTITUTE":"University of Vienna","DEPARTMENT":"Department of Analytical Chemistry","LABORATORY":"Gerner lab","LAST_NAME":"Meier-Menches","FIRST_NAME":"Samuel","ADDRESS":"Währingerstraße 38, 1090 Vienna, Austria","EMAIL":"samuel.meier-menches@univie.ac.at","PHONE":"+43-1-4277-52373"},

"STUDY":{"STUDY_TITLE":"Plasma instead of serum avoids critical confounding of clinical metabolomics studies by platelets (Part 3/3 - Plasma and serum metabolomics)","STUDY_SUMMARY":"Metabolomics is an emerging and powerful molecular profiling method supporting clinical investigations. Serum and plasma are commonly used without rational prioritization. Serum is collected after blood coagulation, a complex biochemical process involving active platelet metabolism. This may affect the metabolome and increase the variance as platelet counts and function may vary substantially in individuals. A multi-omics approach systematically investigating the suitability of serum and plasma for clinical studies demonstrated that metabolites correlated well (n=461, R2=0.991), whereas lipid mediators (n=104, R2=0.906) and proteins (n=322, R2=0.860) differed substantially between specimen. Independently, analysis of platelet releasates identified most biomolecules significantly enriched in serum when compared to plasma. A prospective, randomized, controlled parallel group metabolomics trial with acetylsalicylic acid administered for 7 days demonstrated that the apparent drug effects significantly differ depending on analyzed specimen. Only serum analyses of healthy individuals suggested a significant downregulation of TXB2 and 12-HETE, which were specifically formed during coagulation in vitro. Plasma analyses reliably identified acetylsalicylic acid effects on metabolites and lipids occurring in vivo such as a decrease in polyunsaturated fatty acids. The present data suggests that plasma should be preferred above serum for clinical metabolomics studies as the serum metabolome may be substantially confounded by platelets.","INSTITUTE":"University of Vienna","DEPARTMENT":"Department of Analytical Chemistry","LABORATORY":"Gerner lab","LAST_NAME":"Meier-Menches","FIRST_NAME":"Samuel","ADDRESS":"Währingerstraße 38, 1090 Vienna, Austria","EMAIL":"samuel.meier-menches@univie.ac.at","PHONE":"+43-1-4277-52373"},

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"Sample ID":"MXP500L-0-5711_1036867631_06_1_1_1_00_1036866545_27_R26V3",
"Factors":{"Sample material":"Serum","Treatment":"after_Aspirin","Sex":"m","Age":"26"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_06_1_1_1_00_1036866545_27_R26V3"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_93_1_1_1_00_1036866564_29_R28V2",
"Factors":{"Sample material":"Serum","Treatment":"before_Aspirin","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_93_1_1_1_00_1036866564_29_R28V2"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_45_1_1_1_00_1036865818_30_R28V3",
"Factors":{"Sample material":"Serum","Treatment":"after_Aspirin","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_45_1_1_1_00_1036865818_30_R28V3"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_24_1_1_1_00_1036865822_31_R29V2",
"Factors":{"Sample material":"Serum","Treatment":"before_Aspirin","Sex":"m","Age":"24"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_24_1_1_1_00_1036865822_31_R29V2"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_33_1_1_1_00_1036865837_32_R29V3",
"Factors":{"Sample material":"Serum","Treatment":"after_Aspirin","Sex":"m","Age":"24"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_33_1_1_1_00_1036865837_32_R29V3"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_05_1_1_1_00_1036865841_33_R30V2",
"Factors":{"Sample material":"Serum","Treatment":"before_Aspirin","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_05_1_1_1_00_1036865841_33_R30V2"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_17_1_1_1_00_1036865856_34_R30V3",
"Factors":{"Sample material":"Serum","Treatment":"after_Aspirin","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_17_1_1_1_00_1036865856_34_R30V3"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_66_1_1_1_00_1036867045_2_D1",
"Factors":{"Sample material":"Plasma","Treatment":"Plasma","Sex":"w","Age":"20"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_66_1_1_1_00_1036867045_2_D1"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_42_1_1_1_00_1036867079_5_D2",
"Factors":{"Sample material":"Plasma","Treatment":"Plasma","Sex":"w","Age":"26"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_42_1_1_1_00_1036867079_5_D2"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_67_1_1_1_00_1036867128_10_D3",
"Factors":{"Sample material":"Plasma","Treatment":"Plasma","Sex":"m","Age":"28"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_67_1_1_1_00_1036867128_10_D3"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_72_1_1_1_00_1036866380_11_D4",
"Factors":{"Sample material":"Plasma","Treatment":"Plasma","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_72_1_1_1_00_1036866380_11_D4"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_46_1_1_1_00_1036866405_13_D5",
"Factors":{"Sample material":"Plasma","Treatment":"Plasma","Sex":"m","Age":"24"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_46_1_1_1_00_1036866405_13_D5"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_52_1_1_1_00_1036866424_15_D6",
"Factors":{"Sample material":"Plasma","Treatment":"Plasma","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_52_1_1_1_00_1036866424_15_D6"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_82_1_1_1_00_1036866477_20_D1",
"Factors":{"Sample material":"Serum","Treatment":"Serum","Sex":"w","Age":"20"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_82_1_1_1_00_1036866477_20_D1"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_48_1_1_1_00_1036866507_23_D2",
"Factors":{"Sample material":"Serum","Treatment":"Serum","Sex":"w","Age":"26"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_48_1_1_1_00_1036866507_23_D2"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_07_1_1_1_00_1036866550_28_D3",
"Factors":{"Sample material":"Serum","Treatment":"Serum","Sex":"m","Age":"28"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_07_1_1_1_00_1036866550_28_D3"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_93_1_1_1_00_1036866564_29_D4",
"Factors":{"Sample material":"Serum","Treatment":"Serum","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_93_1_1_1_00_1036866564_29_D4"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_24_1_1_1_00_1036865822_31_D5",
"Factors":{"Sample material":"Serum","Treatment":"Serum","Sex":"m","Age":"24"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_24_1_1_1_00_1036865822_31_D5"}
},
{
"Subject ID":"-",
"Sample ID":"MXP500L-0-5711_1036867631_05_1_1_1_00_1036865841_33_D6",
"Factors":{"Sample material":"Serum","Treatment":"Serum","Sex":"w","Age":"23"},
"Additional sample data":{"RAW_FILE_NAME":"MXP500L-0-5711_1036867631_05_1_1_1_00_1036865841_33_D6"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"SERUM/PLASMA: Blood samples were obtained at baseline and after 7 days intake of the study medication. On both study days two blood samples using 6 mL K3EDTA and serum collection tubes (both Vacuette, Greiner Bio-One GmbH, Kremsmünster, Austria) were obtained from each subject. EDTA-anticoagulated tubes were carefully inverted two times after blood draw and centrifuged immediately at room temperature at 2000 g for 10 min. In contrast, filled serum tubes were carefully inverted after blood draw and placed to sit upright for 15 to 30 minutes to allow clot formation. Then, tubes were centrifuged at room temperature at 2000 g for 10 min. Directly after centrifugation, 500 µL of plasma or serum, respectively, were transferred into pre-labelled Eppendorf safe-lock tubes and stored at -80°C until analysis.","SAMPLE_TYPE":"Blood (serum) and blood (plasma)"},

"TREATMENT":{"TREATMENT_SUMMARY":"Subjects received acetylsalicylic acid for 7 days. The study cohort was instructed to take 500 mg acetylsalicylic acid (Aspirin® 500 mg acetylsalicylic acid, Cellulose powder, maize starche) per day in the evening."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Targeted metabolomics experiments were conducted by applying the MxP® Quant 500 Kit (Biocrates Life Sciences AG, Innsbruck, Austria). Therefore, 10 µL of sample was used and the kit was performed according to the manufacturer’s instructions. Phenyl isothiocyanate (Sigma-Aldrich, St. Louis, USA) was purchased separately and was used for derivatization of amino acids and biogenic amines according to the kit manual."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Measurements were carried out using LC-MS/MS and flow injection (FIA)-MS/MS analyses on a Sciex 6500+ series mass spectrometer coupled to an ExionLC AD chromatography system (AB Sciex, Framingham, MA, USA), utilizing the Analyst 1.7.1 software with hotfix 1 (also AB SCIEX). All required standards, quality controls and eluents were included in the kit, as well as the chromatographic column for the LC-MS/MS analysis part. LC1_pos: The UHPLC autosampler and column oven are held at 10 °C and 50 °C, respectively. The injection volume was 5 µL. Eluent A is water (0.2% formic acid) and eluent B is acetonitrile (0.2% formic acid). Gradient run time of 5.8 mins. Remain at 0% B for 0.25 min, linear gradient to 12% B at 1.5 min and 17.5% B at 2.7 min. Further linear increase to 50% B at 4 mins and 100% B at 4.5 min, where it remains until 5 min. Reduction to 0% at 5.1 min and remaining at 0% until 5.8 min. Flow rate of 0.8 mL/min until 4.5 min, then increase to 1 mL/min until 4.7 min and remaining until 5.1 min. Finally, reduction to 0.8 mL/min until 5.8 min.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"ExionLC AD chromatography system (AB Sciex, Framingham, MA, USA)","COLUMN_NAME":"MxP Quant 500 Column System (Biocrates Part No: 21117)","SOLVENT_A":"100% Water; 0.2% FA","SOLVENT_B":"100% ACN; 0.2% FA","FLOW_GRADIENT":"Gradient run time of 5.8 mins. Remain at 0% B for 0.25 min, linear gradient to 12% B at 1.5 min and 17.5% B at 2.7 min. Further linear increase to 50% B at 4 mins and 100% B at 4.5 min, where it remains until 5 min. Reduction to 0% at 5.1 min and remaining at 0% until 5.8 min. Flow rate of 0.8 mL/min until 4.5 min, then increase to 1 mL/min until 4.7 min and remaining until 5.1 min. Finally, reduction to 0.8 mL/min until 5.8 min.","FLOW_RATE":"0.8 mL/min","COLUMN_TEMPERATURE":"50°C"},

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

"MS":{"INSTRUMENT_NAME":"ABI Sciex 6500+","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Measurements were carried out using LC-MS/MS and flow injection (FIA)-MS/MS analyses on a Sciex 6500+ series mass spectrometer coupled to an ExionLC AD chromatography system (AB Sciex, Framingham, MA, USA), utilizing the Analyst 1.7.1 software with hotfix 1 (also AB SCIEX). All required standards, quality controls and eluents were included in the kit, as well as the chromatographic column for the LC-MS/MS analysis part. Preparation of the measurement worklist as well as data validation and evaluation were performed with the software supplied with the kit (MetIDQ-Oxygen-DB110-3005, Biocrates Life Sciences). MS(LC1_pos): Scheduled MRM experiment in positive polarity. Detection window of 30 s, target scan time of 0.15 s and 3 ms pause between mass ranges. Q1 and Q3 were held at unit resolution. Source parameters were as follows: CUR 45, CAD 9, voltage 5.5 kV, temperature 500 °C, ion source gas at 60 and 70."},

"MS_METABOLITE_DATA":{
"Units":"µM",

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"Metabolites":[{"Metabolite":"1-Met-His","Precursor Ion":"305.1","Fragment Ion":"124.1"},{"Metabolite":"3-IAA","Precursor Ion":"176","Fragment Ion":"130.1"},{"Metabolite":"3-IPA","Precursor Ion":"190.1","Fragment Ion":"130.1"},{"Metabolite":"3-Met-His","Precursor Ion":"305.1","Fragment Ion":"168.2"},{"Metabolite":"AABA","Precursor Ion":"239","Fragment Ion":"104.1"},{"Metabolite":"ADMA","Precursor Ion":"338.2","Fragment Ion":"46"},{"Metabolite":"Ala","Precursor Ion":"225.2","Fragment Ion":"44.2"},{"Metabolite":"alpha-AAA","Precursor Ion":"297.1","Fragment Ion":"144.2"},{"Metabolite":"Arg","Precursor Ion":"310","Fragment Ion":"217"},{"Metabolite":"Asn","Precursor Ion":"268.2","Fragment Ion":"77"},{"Metabolite":"Asp","Precursor Ion":"269.2","Fragment Ion":"116.2"},{"Metabolite":"beta-Ala","Precursor Ion":"225.1","Fragment Ion":"90.1"},{"Metabolite":"Betaine","Precursor Ion":"118.1","Fragment Ion":"59.1"},{"Metabolite":"Choline","Precursor Ion":"104","Fragment Ion":"58.1"},{"Metabolite":"Cit","Precursor Ion":"311.2","Fragment Ion":"113.1"},{"Metabolite":"Cortisol","Precursor Ion":"363.1","Fragment Ion":"121.1"},{"Metabolite":"Creatinine","Precursor Ion":"114.1","Fragment Ion":"44.1"},{"Metabolite":"Cys","Precursor Ion":"392","Fragment Ion":"177"},{"Metabolite":"GABA","Precursor Ion":"239","Fragment Ion":"104.1"},{"Metabolite":"Gln","Precursor Ion":"282.2","Fragment Ion":"130"},{"Metabolite":"Glu","Precursor Ion":"283.2","Fragment Ion":"130.2"},{"Metabolite":"Gly","Precursor Ion":"211.2","Fragment Ion":"75.9"},{"Metabolite":"HArg","Precursor Ion":"324.1","Fragment Ion":"189.1"},{"Metabolite":"HCys","Precursor Ion":"406","Fragment Ion":"236.9"},{"Metabolite":"HipAcid","Precursor Ion":"180.1","Fragment Ion":"105"},{"Metabolite":"His","Precursor Ion":"291.1","Fragment Ion":"110.2"},{"Metabolite":"Hypoxanthine","Precursor Ion":"137.1","Fragment Ion":"110.1"},{"Metabolite":"Ile","Precursor Ion":"267.3","Fragment Ion":"69"},{"Metabolite":"Kynurenine","Precursor Ion":"344.2","Fragment Ion":"146.2"},{"Metabolite":"Leu","Precursor Ion":"267.3","Fragment Ion":"43"},{"Metabolite":"Lys","Precursor Ion":"417.2","Fragment Ion":"324.2"},{"Metabolite":"Met","Precursor Ion":"285.1","Fragment Ion":"104.2"},{"Metabolite":"Met-SO","Precursor Ion":"301.2","Fragment Ion":"88.1"},{"Metabolite":"Orn","Precursor Ion":"403.2","Fragment Ion":"310.2"},{"Metabolite":"Phe","Precursor Ion":"301.2","Fragment Ion":"120.2"},{"Metabolite":"Pro","Precursor Ion":"251.2","Fragment Ion":"70.3"},{"Metabolite":"ProBetaine","Precursor Ion":"144.1","Fragment Ion":"84.1"},{"Metabolite":"Putrescine","Precursor Ion":"359.1","Fragment Ion":"266.1"},{"Metabolite":"Sarcosine","Precursor Ion":"225","Fragment Ion":"90.1"},{"Metabolite":"SDMA","Precursor Ion":"338.2","Fragment Ion":"70.1"},{"Metabolite":"Ser","Precursor Ion":"241.2","Fragment Ion":"60"},{"Metabolite":"Serotonin","Precursor Ion":"312.3","Fragment Ion":"160.2"},{"Metabolite":"t4-OH-Pro","Precursor Ion":"267.2","Fragment Ion":"68"},{"Metabolite":"Taurine","Precursor Ion":"261","Fragment Ion":"126.1"},{"Metabolite":"Thr","Precursor Ion":"255.2","Fragment Ion":"74.1"},{"Metabolite":"TMAO","Precursor Ion":"75.9","Fragment Ion":"59.1"},{"Metabolite":"Trp","Precursor Ion":"340.2","Fragment Ion":"188.2"},{"Metabolite":"TrpBetaine","Precursor Ion":"247","Fragment Ion":"188.1"},{"Metabolite":"Tyr","Precursor Ion":"317.2","Fragment Ion":"136.1"},{"Metabolite":"Val","Precursor Ion":"253.2","Fragment Ion":"72.2"},{"Metabolite":"Xanthine","Precursor Ion":"153","Fragment Ion":"110.1"}]
}

}