Summary of Study ST002026

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001287. The data can be accessed directly via it's Project DOI: 10.21228/M8BX22 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST002026
Study TitleMetabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women (Stool/GC)
Study SummarySamples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Institute
University of California, Davis
Last Namefolz
First Namejake
Address451 Health Science Drive
Emailjfolz@ucdavis.edu
Phone7155636311
Submit Date2021-12-21
Raw Data AvailableYes
Raw Data File Type(s)cdf
Analysis Type DetailGC-MS
Release Date2023-01-02
Release Version1
jake folz jake folz
https://dx.doi.org/10.21228/M8BX22
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR001287
Project DOI:doi: 10.21228/M8BX22
Project Title:Metabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women
Project Summary:Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Institute:University of California, Davis
Last Name:Folz
First Name:jake
Address:451 Health Science Drive
Email:jfolz@ucdavis.edu
Phone:7155636311

Subject:

Subject ID:SU002108
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Mammals

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Treatment
SA190097RS-01920609Post-intervention
SA190098RS-01916436Post-intervention
SA190099RS-02219083Post-intervention
SA190100RS-02217669Post-intervention
SA190101RS-01912456Post-intervention
SA190102RS-01923384Post-intervention
SA190103RS-01940439Post-intervention
SA190104RS-01933527Post-intervention
SA190105RS-02210756Post-intervention
SA190106RS-01909498Post-intervention
SA190107RS-02220151Post-intervention
SA190108RS-02254825Post-intervention
SA190109RS-02269483Post-intervention
SA190110RS-01865119Post-intervention
SA190111RS-01851707Post-intervention
SA190112RS-02248450Post-intervention
SA190113RS-01891737Post-intervention
SA190114RS-01950714Post-intervention
SA190115RS-02221392Post-intervention
SA190116RS-01899167Post-intervention
SA190117RS-01906962Post-intervention
SA190118RS-02209055Post-intervention
SA190119RS-02177788Post-intervention
SA190120RS-02099628Post-intervention
SA190121RS-02097213Post-intervention
SA190122RS-02179395Post-intervention
SA190123RS-02119956Post-intervention
SA190124RS-02129908Post-intervention
SA190125RS-02139810Post-intervention
SA190126RS-02136575Post-intervention
SA190127RS-02135615Post-intervention
SA190128RS-02186686Post-intervention
SA190129RS-02055701Post-intervention
SA190130RS-02207959Post-intervention
SA190131RS-02208413Post-intervention
SA190132RS-01975227Post-intervention
SA190133RS-02019533Post-intervention
SA190134RS-02193478Post-intervention
SA190135RS-02055250Post-intervention
SA190136RS-02045482Post-intervention
SA190137RS-02040176Post-intervention
SA190138RS-01838033Post-intervention
SA190139RS-01871547Post-intervention
SA190140RS-02622760Post-intervention
SA190141RS-01713712Post-intervention
SA190142RS-01707130Post-intervention
SA190143RS-01704567Post-intervention
SA190144RS-02309902Post-intervention
SA190145RS-02621442Post-intervention
SA190146RS-01735982Post-intervention
SA190147RS-01730343Post-intervention
SA190148RS-02583618Post-intervention
SA190149RS-02637716Post-intervention
SA190150RS-01694989Post-intervention
SA190151RS-01607340Post-intervention
SA190152RS-02756031Post-intervention
SA190153RS-01586115Post-intervention
SA190154RS-01574184Post-intervention
SA190155RS-01617214Post-intervention
SA190156RS-02750185Post-intervention
SA190157RS-02674498Post-intervention
SA190158RS-01686993Post-intervention
SA190159RS-01648850Post-intervention
SA190160RS-02354063Post-intervention
SA190161RS-01718537Post-intervention
SA190162RS-01776131Post-intervention
SA190163RS-01775777Post-intervention
SA190164RS-01747171Post-intervention
SA190165RS-01807614Post-intervention
SA190166RS-01830763Post-intervention
SA190167RS-01770486Post-intervention
SA190168RS-01803066Post-intervention
SA190169RS-01759730Post-intervention
SA190170RS-01764367Post-intervention
SA190171RS-02159560Pre-intervention
SA190172RS-02173570Pre-intervention
SA190173RS-02174019Pre-intervention
SA190174RS-02176985Pre-intervention
SA190175RS-02343753Pre-intervention
SA190176RS-02328916Pre-intervention
SA190177RS-02281190Pre-intervention
SA190178RS-02163124Pre-intervention
SA190179RS-02171917Pre-intervention
SA190180RS-02282708Pre-intervention
SA190181RS-02734868Pre-intervention
SA190182RS-02505149Pre-intervention
SA190183RS-02349304Pre-intervention
SA190184RS-02186695Pre-intervention
SA190185RS-02232959Pre-intervention
SA190186RS-02189596Pre-intervention
SA190187RS-02224703Pre-intervention
SA190188RS-02636728Pre-intervention
SA190189RS-02245717Pre-intervention
SA190190RS-02348498Pre-intervention
SA190191RS-02348451Pre-intervention
SA190192RS-02184855Pre-intervention
SA190193RS-02724295Pre-intervention
SA190194RS-02681370Pre-intervention
SA190195RS-02731752Pre-intervention
SA190196RS-01975879Pre-intervention
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Collection:

Collection ID:CO002101
Collection Summary:Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Sample Type:Feces

Treatment:

Treatment ID:TR002120
Treatment Summary:Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.

Sample Preparation:

Sampleprep ID:SP002114
Sampleprep Summary:Equipment: Centrifuge Eppendorf 5415 D Calibrated pipettes 20-200µL and 100-1000µL Multi-Tube Vortexer (VWR VX-2500) Orbital Mixing Chilling/Heating Plate (Torrey Pines Scientific Instruments) Speed vacuum concentration system (Labconco Centrivap cold trap) Chemicals: Product Manufacturer & Part Number Eppendorf tubes 1.5 mL, uncolored Eppendorf 022363204 Eppendorf tubes 2 mL, uncolored Eppendorf 022363352 Crushed ice UC Davis Water, LC/MS Grade Fisher Optima W6-4 MTBE, HPLC Grade Acros Organics 389050010 Methanol, LC/MS Grade Fisher A456-4 Bioreclamation human plasma (disodium EDTA) Bioreclamation HMPLEDTA Acetonitrile, HPLC Grade Fisher Optima A955-4 Iso-Propanol, HPLC Grade Fisher A461-4 Sample Preparation: Preparation of extraction solvent Combine 120 mL of chilled MeOH/QC mix with 400 mL of chilled MTBE/Cholesterol Ester 22:1 in a clean 500 mL stock bottle. Mix thoroughly by swirling or stir plate and store at -20°C until use. *See SOP “QC mix for LC-MS lipid analysis” for preparation of MeOH/QC mix and MTBE/Cholesterol Ester 22:1. Preparation of Clean Up solvent For 1 L of extraction solvent, combine 375 mL of acetonitrile, 375 mL of isopropanol, and 250 mL water in a 1 L bottle conditioned with the aforementioned chemicals. If a different total volume of extraction solvent is needed, simply mix acetonitrile, isopropanol, and water in volumes in proportion 3:3:2. Purge the extraction solution mix for 5 min with nitrogen with small bubbles. Make sure that the nitrogen line is flushed out of air before using it for degassing the extraction solvent solution. Store at -20°C until use. Note: if solvent freezes, sonicate until thawed and mix before use. Extraction Thaw raw samples/controls at room temperature (or in the refrigerator at 4˚C) and either invert the tube or vortex 10 sec at low speed to homogenize. Aliquot 20 μL of plasma sample into a 1.5 mL Eppendorf tube. Keep all samples on ice. Add 975 µL ice-cold 3:10 (v/v) MeOH/MTBE + QC mix/CE 22:1 extraction solvent mixture to each aliquot, keeping the extraction solvent on ice during the procedure. Vortex samples for 10 seconds, then shake for 5 minutes at 4°C on the orbital mixer. Add 188 µL room temperature LC/MS grade water to each tube. Vortex tubes for 20 seconds and then centrifuge for 2 min at 14,000 rcf. Transfer the upper organic phase to two separate tubes (350 µL/each tube) for lipidomics analysis. Transfer 75 µL of the remaining organic phase to a 2, 15, or 50 mL tube for pools, depending on number of samples in the study. Transfer the bottom aqueous phase to two separate tubes (110 µL/each tube) for HILIC/GC-TOF analysis. Dry down one tube from each phase by centrivap, keeping the undried tubes as backups. Store all tubes at -20˚C until ready for analysis. Clean up step for GC only (and pooling) Resuspend the dried aliquot with 500 μL 3:3:2 (v/v/v) ACN:IPA:H2O (degassed as given above) and vortex for about 10 sec. Centrifuge for 2 min at 14000 rcf. Remove 450 uL supernatant to a clean 1.5 mL eppendorf tube. Tranfering remainder to a 2, 15, 50 mL Tube, dependent on number of samples. Aliquot out 1.9 mL uL of supernatant to new 2ml eppendorf tubes. Centrifuge for 2 min at 14000 rcf Aliquot out 4x450 uL of supernatant into clean 1.5 mL Eppendorf tubes. Evaporate to comeplete dryness in the Labconco Centruvap cold trap concentrator. Submit to derivatization . Pooling (CSH platform only) Transfer multiple 350 µL aliquots of pooled samples to 1.5 mL Eppendorf tubes, one aliquot for every 10 samples in the study. If there is still pool remaining, prepare additional aliquots for backup. Evaporate to complete dryness in the Labconco Centrivap cold trap concentrator. Store all tubes at -20°C until ready for analysis. Quality assurance For every 10 samples, extract a method blank (20 µL of H2O) and a sample control (20 µL human Bioreclamation or analogous species plasma) in addition to samples. For large studies (>100 samples), for every 100 samples a NIST plasma extract should be prepared in the same manner as positive controls. Disposal of waste Collect all chemicals in appropriate bottles and follow the disposal rules. Collect residual plasma/serum samples in specifically designed red ‘biohazard’ waste bags. Extraction of Mammalian Tissue Samples: Liver 1. References: Fiehn O, Kind T (2006) Metabolite profiling in blood plasma. In: Metabolomics: Methods and Protocols. Weckwerth W (ed.), Humana Press, Totowa NJ (in press) 2.Starting material: Liver sample: weigh 4mg per sample into 2mL Eppendorf tubes. 3. Equipment: Centrifuge (Eppendorf 5415 D) Calibrated pipettes 1-200μl and 100-1000μl Eppendorf tubes 2mL, clear (Cat. No. 022363204) Centrifuge tubes 50mL, polypropylene Eppendorff Tabletop Centrifuge (Proteomics core Lab.) ThermoElectron Neslab RTE 740 cooling bath at –20°C MiniVortexer (VWR) Orbital Mixing Chilling/Heating Plate (Torrey Pines Scientific Instruments) Speed vacuum concentration system (Labconco Centrivap cold trap) Turex mini homogenizer 4. Chemicals Acetonitrile, LCMS grade (JT Baker; Cat. No.9829-02) Isopropanol, HPLC grade (JT Baker; Cat. No. 9095-02) Methanol Acetone Crushed ice 18 MΩ pure water (Millipore) Nitrogen line with pipette tip pH paper 5-10 (EMD Chem. Inc.) 5. Procedure Preparation of extraction mix and material before experiment: Switch on bath to pre-cool at –20°C (±2°C validity temperature range) Check pH of acetonitrile and isopropanol (pH7) using wetted pH paper Make the extraction solution by mixing acetonitrile, isopropanol and water in proportions 3 : 3 : 2 De-gas the extraction solution for 5 min with nitrogen. Make sure that the nitrogen line was flushed out of air before using it for degassing the extraction solvent solution Sample Preparation Weigh 4mg tissue sample in to a 2mL Eppendorf tube. Add 1mL extraction solvent to the tissue sample and homogenize for 45 seconds ensuring that sample resembles a powder. In between samples, clean the homogenizer in solutions of methanol, acetone, water, and the extraction solvent in the order listed. Vortex samples for 10 seconds, then 5 minutes on 4°C shaker. Centrifuge the samples for 2 minutes at 14,000 rcf. Aliquot 500µL supernatant for analysis, and 500µL for a backup. Store backup aliquots in the -20°C freezer. Evaporate one 500µl analysis aliquot in the Labconco Centrivap cold trap concentrator to complete dryness (typically overnight). The dried aliquot is then re-suspended with 500μl 50% acetonitrile (degassed as given) Centrifuge for 2 minutes at 14,000 rcf using the centrifuge Eppendorf 5415. Remove supernatant to a new Eppendorf tube. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. Submit to derivatization. The residue should contain membrane lipids because these are supposedly not soluble enough to be found in the 50% acetonitrile solution. Therefore, this ‘membrane residue’ is now taken for membrane lipidomic fingerprinting using the nanomate LTQ ion trap mass spectrometer. Likely, a good solvent to redissolve the membrane lipids is e.g. 75% isopropanol (degassed as given above). If the ‘analysis’ aliquot is to be used for semi lipophilic compounds such as tyrosine pathway intermediates (incl. dopamine, serotonine etc, i.e. polar aromatic compounds), then these are supposedly to be found together with the ‘GCTOF’ aliquot. We can assume that this mixture is still too complex for Agilent chipLCMS. Therefore, in order to develop and validate target analysis for such aromatic compounds, we should use some sort of Solid Phase purification. We re-suspend the dried ‘GCTOF’ aliquot in 300 l water (degassed as before) to take out sugars, aliphatic amino acids, hydroxyl acids and similar logP compounds. The residue should contain our target aromatics .We could also try to adjust pH by using low concentration acetate or phosphate buffer. The residue could then be taken up in 50% acetonitrile and used for GCTOF and Agilent chipMS experiments. The other aliquot should be checked how much of our target compounds would actually be found in the ‘sugar’ fraction. 6. Problems To prevent contamination disposable material is used. Control pH from extraction mix. 7. Quality assurance For each sequence of sample extractions, perform one blank negative control extraction by applying the total procedure (i.e. all materials and plastic ware) without biological sample. 8. Disposal of waste Collect all chemicals in appropriate bottles and follow the disposal rules.

Combined analysis:

Analysis ID AN003296
Analysis type MS
Chromatography type GC
Chromatography system Leco Pegasus IV
Column Restek Rtx-5Sil MS (30 x 0.25mm,0.25um)
MS Type EI
MS instrument type GC-TOF
MS instrument name Leco Pegasus IV TOF
Ion Mode UNSPECIFIED
Units normalized peak height

Chromatography:

Chromatography ID:CH002435
Chromatography Summary:A 30 m long, 0.25 mm i.d. Rtx-5Sil MS column (0.25 μm 95% dimethyl 5% diphenyl polysiloxane film) with additional 10 m integrated guard column is used (Restek, Bellefonte PA). 99.9999% pure Helium with built-in purifier (Airgas, Radnor PA) is set at constant flow of 1 ml/min. The oven temperature is held constant at 50°C for 1 min and then ramped at 20°C/min to 330°C at which it is held constant for 5 min.
Instrument Name:Leco Pegasus IV
Column Name:Restek Rtx-5Sil MS (30 x 0.25mm,0.25um)
Chromatography Type:GC

MS:

MS ID:MS003066
Analysis ID:AN003296
Instrument Name:Leco Pegasus IV TOF
Instrument Type:GC-TOF
MS Type:EI
MS Comments:A Leco Pegasus IV time of flight mass spectrometer is controlled by the Leco ChromaTOF software vs. 2.32 (St. Joseph, MI). The transfer line temperature between gas chromatograph and mass spectrometer is set to 280°C. Electron impact ionization at 70V is employed with an ion source temperature of 250°C. Acquisition rate is 17 spectra/second, with a scan mass range of 85-500 Da.
Ion Mode:UNSPECIFIED
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