{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002994","ANALYSIS_ID":"AN004918","VERSION":"1","CREATED_ON":"01-26-2024"},

"PROJECT":{"PROJECT_TITLE":"Integrating uterine microbiome and metabolome to advance the understanding of the uterine environment in dairy cows with metritis","PROJECT_SUMMARY":"Background: Metritis is a prevalent uterine disease that affects the welfare, fertility, and survival of dairy cows. The uterine microbiome from cows that develop metritis and those that remain healthy do not differ from calving until 2 days after calving, after which there is a dysbiosis of the uterine microbiome characterized by a shift towards opportunistic pathogens such as Fusobacteriota and Bacteroidota. Whether these opportunistic pathogens proliferate and overtake the uterine commensals could be determined by the type of substrates present in the uterus. The objective of this study was to integrate uterine metabolome and microbiome data to advance the understanding of metritis development in dairy cows. Holstein cows (n = 104) had uterine fluid collected at calving and at the day of metritis diagnosis. Cows with metritis (n = 52) were paired with cows without metritis (n = 52) based on days after calving. First, the uterine metabolome and microbiome were evaluated individually, and then integrated using network analyses. Results: The uterine metabolome differed both at calving and on the day of metritis diagnosis between cows with and without metritis. The uterine microbiome did not differ at calving but differed on the day of metritis diagnosis between cows with and without metritis. Omics integration was performed between 153 significant metabolites and 6 significant bacteria genera on the day of metritis diagnosis. A total of 49 metabolites were correlated with 3 bacteria genera (i.e. Fusobacteria, Porphyromonas and Bacteroides) on the day of metritis diagnosis. The main metabolites have been associated with attenuation of biofilm formation by commensal bacteria, pathogenic bacterial overgrowth, defense mechanisms against the immune system, tissue damage and inflammation, and immune dysregulation. Conclusions: The data integration presented herein helps advance the understanding of metritis development in dairy cows. The identified metabolites may be promising targets for future interventions aiming to reduce pathogenic bacterial growth in the uterus, and therefore, reducing the incidence of metritis.","INSTITUTE":"University of Florida","DEPARTMENT":"Large Animal Clinical Sciences","LAST_NAME":"Segundo","FIRST_NAME":"Casaro","ADDRESS":"117 Deriso Hall, 2015 SW 16th Ave., Gainesville, FL 32610","EMAIL":"segundocasaro@ufl.edu","PHONE":"3522844016","DOI":"http://dx.doi.org/10.21228/M8S425"},

"STUDY":{"STUDY_TITLE":"Integrating uterine microbiome and metabolome to advance the understanding of the uterine environment in dairy cows with metritis","STUDY_SUMMARY":"Background: Metritis is a prevalent uterine disease that affects the welfare, fertility, and survival of dairy cows. The uterine microbiome from cows that develop metritis and those that remain healthy do not differ from calving until 2 days after calving, after which there is a dysbiosis of the uterine microbiome characterized by a shift towards opportunistic pathogens such as Fusobacteriota and Bacteroidota. Whether these opportunistic pathogens proliferate and overtake the uterine commensals could be determined by the type of substrates present in the uterus. The objective of this study was to integrate uterine metabolome and microbiome data to advance the understanding of metritis development in dairy cows. Holstein cows (n = 104) had uterine fluid collected at calving and at the day of metritis diagnosis. Cows with metritis (n = 52) were paired with cows without metritis (n = 52) based on days after calving. First, the uterine metabolome and microbiome were evaluated individually, and then integrated using network analyses. Results: The uterine metabolome differed both at calving and on the day of metritis diagnosis between cows with and without metritis. The uterine microbiome did not differ at calving but differed on the day of metritis diagnosis between cows with and without metritis. Omics integration was performed between 153 significant metabolites and 6 significant bacteria genera on the day of metritis diagnosis. A total of 49 metabolites were correlated with 3 bacteria genera (i.e. Fusobacteria, Porphyromonas and Bacteroides) on the day of metritis diagnosis. The main metabolites have been associated with attenuation of biofilm formation by commensal bacteria, pathogenic bacterial overgrowth, defense mechanisms against the immune system, tissue damage and inflammation, and immune dysregulation. Conclusions: The data integration presented herein helps advance the understanding of metritis development in dairy cows. The identified metabolites may be promising targets for future interventions aiming to reduce pathogenic bacterial growth in the uterus, and therefore, reducing the incidence of metritis.","INSTITUTE":"University of Florida","LAST_NAME":"Casaro","FIRST_NAME":"Segundo","ADDRESS":"117 Deriso Hall, 2015 SW 16th Ave., Gainesville, FL 32610","EMAIL":"segundocasaro@ufl.edu","PHONE":"3522844016","SUBMIT_DATE":"2023-11-30"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Bos taurus","TAXONOMY_ID":"9913","GENDER":"Female","ANIMAL_HOUSING":"Free-stalls","ANIMAL_FEED":"TMR"},
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"Sample ID":"10246_Calving_Con",
"Factors":{"Group":"Con","Time":"Calving","Parity":"Prim"}
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"Sample ID":"10254_Calving_Con",
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"Factors":{"Group":"Con","Time":"Calving","Parity":"Prim"}
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{
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"Factors":{"Group":"Con","Time":"Calving","Parity":"Prim"}
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{
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"Factors":{"Group":"Con","Time":"Calving","Parity":"Prim"}
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{
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"Factors":{"Group":"Con","Time":"Diagnosis","Parity":"Mult"}
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{
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"Factors":{"Group":"Con","Time":"Diagnosis","Parity":"Mult"}
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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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"Factors":{"Group":"Met","Time":"Calving","Parity":"Mult"}
},
{
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"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
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{
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"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
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},
{
"Subject ID":"10193",
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"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10198",
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"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10206",
"Sample ID":"10206_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10207",
"Sample ID":"10207_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10208",
"Sample ID":"10208_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10213",
"Sample ID":"10213_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10215",
"Sample ID":"10215_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10219",
"Sample ID":"10219_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10222",
"Sample ID":"10222_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10223",
"Sample ID":"10223_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10230",
"Sample ID":"10230_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10231",
"Sample ID":"10231_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10239",
"Sample ID":"10239_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10249",
"Sample ID":"10249_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10251",
"Sample ID":"10251_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10253",
"Sample ID":"10253_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10256",
"Sample ID":"10256_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10257",
"Sample ID":"10257_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10258",
"Sample ID":"10258_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10259",
"Sample ID":"10259_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10260",
"Sample ID":"10260_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10263",
"Sample ID":"10263_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10267",
"Sample ID":"10267_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10278",
"Sample ID":"10278_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10293",
"Sample ID":"10293_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10294",
"Sample ID":"10294_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10298",
"Sample ID":"10298_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10304",
"Sample ID":"10304_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10308",
"Sample ID":"10308_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10318",
"Sample ID":"10318_Calving_Met",
"Factors":{"Group":"Met","Time":"Calving","Parity":"Prim"}
},
{
"Subject ID":"10004",
"Sample ID":"10004_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"8790",
"Sample ID":"8790_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9193",
"Sample ID":"9193_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9206",
"Sample ID":"9206_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9221",
"Sample ID":"9221_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9232",
"Sample ID":"9232_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9292",
"Sample ID":"9292_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9481",
"Sample ID":"9481_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9490",
"Sample ID":"9490_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9505",
"Sample ID":"9505_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9669",
"Sample ID":"9669_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9796",
"Sample ID":"9796_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9810",
"Sample ID":"9810_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9832",
"Sample ID":"9832_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9848",
"Sample ID":"9848_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9908",
"Sample ID":"9908_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9910",
"Sample ID":"9910_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9977",
"Sample ID":"9977_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"9980",
"Sample ID":"9980_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Mult"}
},
{
"Subject ID":"10156",
"Sample ID":"10156_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10158",
"Sample ID":"10158_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10175",
"Sample ID":"10175_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10193",
"Sample ID":"10193_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10198",
"Sample ID":"10198_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10206",
"Sample ID":"10206_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10207",
"Sample ID":"10207_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10208",
"Sample ID":"10208_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10213",
"Sample ID":"10213_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10215",
"Sample ID":"10215_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10219",
"Sample ID":"10219_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10222",
"Sample ID":"10222_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10223",
"Sample ID":"10223_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10230",
"Sample ID":"10230_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10231",
"Sample ID":"10231_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10239",
"Sample ID":"10239_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10249",
"Sample ID":"10249_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10251",
"Sample ID":"10251_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10253",
"Sample ID":"10253_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10256",
"Sample ID":"10256_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10257",
"Sample ID":"10257_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10258",
"Sample ID":"10258_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10259",
"Sample ID":"10259_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10260",
"Sample ID":"10260_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10263",
"Sample ID":"10263_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10267",
"Sample ID":"10267_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10278",
"Sample ID":"10278_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10293",
"Sample ID":"10293_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10294",
"Sample ID":"10294_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10298",
"Sample ID":"10298_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10304",
"Sample ID":"10304_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10308",
"Sample ID":"10308_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
},
{
"Subject ID":"10318",
"Sample ID":"10318_Diagnosis_Met",
"Factors":{"Group":"Met","Time":"Diagnosis","Parity":"Prim"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"All cows had uterine fluid collected at calving (first 24h after calving), and at diagnosis (day of metritis diagnosis). Briefly, cows’ cervix was stabilized by rectal palpation, the vulva was rinsed with alcohol 70% and dried with paper towels. Subsequently, a single-use plastic round-tip pipette (UterFlush pipettes, Van Beek) was introduced into the vagina at a 45° angle and manipulated through the cervix. A total of 50 mL of sterile saline solution (0.9% sodium chloride irrigation, Baxter) was infused into the uterine lumen using a 60-mL syringe (Covidien) attached to the end of the pipette. Uterine contents were homogenized, retrieved into the same 60-mL syringe, and transferred to a sterile 15-mL conical tube (VWR). After collection, tubes were placed on ice and transported to the laboratory within 2 hours. Once in the laboratory, uterine fluid samples were aliquoted into 2-mL microcentrifuge tubes (Eppendorf) and stored at -80 oC until essayed. One frozen uterine fluid aliquot was submitted to the University of California’s West Coast Metabolomics Center in Davis, CA for metabolome analysis.","SAMPLE_TYPE":"Uterine fluid"},

"TREATMENT":{"TREATMENT_SUMMARY":"This was a case-control study, hence, there were not treatments applied. Cows were self-assigned to the groups. Cows that developed metritis were paired with healthy cows by days in milk."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Uterine fluid samples were aliquoted into 2-mL microcentrifuge tubes (Eppendorf) and stored at -80 oC until essayed."},

"CHROMATOGRAPHY":{"INSTRUMENT_NAME":"Leco Pegasus IV GC","COLUMN_NAME":"Restek Rtx-5Sil MS (30m x 0.25mm, 0.25um)","COLUMN_TEMPERATURE":"50-330","FLOW_GRADIENT":".","FLOW_RATE":"1 mL min-1","SOLVENT_A":".","SOLVENT_B":".","CHROMATOGRAPHY_TYPE":"GC"},

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

"MS":{"INSTRUMENT_NAME":"Leco Pegasus IV TOF","INSTRUMENT_TYPE":"GC-TOF","MS_TYPE":"EI","MS_COMMENTS":"Data are acquired using the following chromatographic parameters, with more details to be found in Fiehn O. et al. Plant J. 53 (2008) 691–704. Column: Restek corporation Rtx-5Sil MS (30 m length x 0.25 mm internal diameter with 0.25 μm film made of 95% dimethyl/5%diphenylpolysiloxane) Mobile phase: Helium Column temperature: 50-330°C Flow- rate: 1 mL min-1 Injection volume: 0.5 μL Injection: 25 splitless time into a multi-baffled glass liner Injection temperature: 50°C ramped to 250°C by 12°C s-1 Oven temperature program: 50°C for 1 min, then ramped at 20°C min-1 to 330°C, held constant for 5 min. The analytical GC column is protected by a 10 m long empty guard column which is cut by 20 cm intervals whenever the reference mixture QC samples indicate problems caused by column contaminations. We have validated that at this sequence of column cuts, no detrimental effects are detected with respect to peak shapes, absolute or relative metabolite retention times or reproducibility of quantifications. This chromatography method yields excellent retention and separation of primary metabolite classes (amino acids, hydroxyl acids, carbohydrates, sugar acids, sterols, aromatics, nucleosides, amines and miscellaneous compounds) with narrow peak widths of 2–3 s and very good within-series retention time reproducibility of better than 0.2 s absolute deviation of retention times. We use automatic liner exchanges after each set of 10 injections which we could show to reduce sample carryover for highly lipophilic compounds such as free fatty acids. Mass spectrometry parameters are used as follows: a Leco Pegasus IV mass spectrometer is used with unit mass resolution at 17 spectra s-1 from 80-500 Da at - 70 eV ionization energy and 1800 V detector voltage with a 230°C transfer line and a 250°C ion source.","ION_MODE":"POSITIVE"},

"MS_METABOLITE_DATA":{
"Units":"peak heights",

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