Summary of Study ST000434

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 PR000330. The data can be accessed directly via it's Project DOI: 10.21228/M8W02Q This work is supported by NIH grant, U2C- DK119886.

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Study IDST000434
Study TitleQuantitative measurements of free fatty acid in T1D poor control, good control, and controls.
Study TypeQuantitative measurements of free fatty acid
Study SummaryThe objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Institute
Mayo Clinic
DepartmentEndocrinology
LaboratoryMayo Clinic Metabolomics Resource Core
Last NameNair
First NameSreekumaran
Address200 First Street SW, Rochester, MN 55905
EmailNair.K@mayo.edu
Phone507-285-2415
Submit Date2016-07-20
Analysis Type DetailLC-MS
Release Date2016-09-23
Release Version1
Sreekumaran Nair Sreekumaran Nair
https://dx.doi.org/10.21228/M8W02Q
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000330
Project DOI:doi: 10.21228/M8W02Q
Project Title:Impact of Long-Term Poor and Good Glycemic Control on Metabolomics Alterations in Type 1 Diabetic People.
Project Type:Untargeted LC-MS Metabolomics
Project Summary:The objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Institute:Mayo Clinic
Department:Endocrinology
Laboratory:Mayo Clinic Metabolomics Resource Core
Last Name:Nair
First Name:Sreekumaran
Address:200 First Street SW, Rochester, MN 55905
Email:Nair.K@mayo.edu
Phone:507-285-2415

Subject:

Subject ID:SU000455
Subject Type:Animal
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human

Factors:

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

mb_sample_id local_sample_id treatment
SA021813sample33ND
SA021814sample34ND
SA021815sample35ND
SA021816sample32ND
SA021817sample28ND
SA021818sample24ND
SA021819sample26ND
SA021820sample36ND
SA021821sample31ND
SA021822sample37ND
SA021823sample42ND
SA021824sample43ND
SA021825sample44ND
SA021826sample41ND
SA021827sample40ND
SA021828sample38ND
SA021829sample39ND
SA021830sample22ND
SA021831sample30ND
SA021832sample16ND
SA021833sample14ND
SA021834sample10ND
SA021835sample12ND
SA021836sample8ND
SA021837sample6ND
SA021838sample2ND
SA021839sample20ND
SA021840sample18ND
SA021841sample4ND
SA021842sample11T1D good glycemic control
SA021843sample21T1D good glycemic control
SA021844sample3T1D good glycemic control
SA021845sample9T1D good glycemic control
SA021846sample5T1D good glycemic control
SA021847sample7T1D good glycemic control
SA021848sample25T1D good glycemic control
SA021849sample23T1D good glycemic control
SA021850sample13T1D good glycemic control
SA021851sample27T1D good glycemic control
SA021852sample19T1D good glycemic control
SA021853sample15T1D good glycemic control
SA021854sample17T1D good glycemic control
SA021855sample1T1D good glycemic control
SA021856sample29T1D good glycemic control
SA021857sample55T1D poor glycemic control
SA021858sample54T1D poor glycemic control
SA021859sample56T1D poor glycemic control
SA021860sample58T1D poor glycemic control
SA021861sample53T1D poor glycemic control
SA021862sample57T1D poor glycemic control
SA021863sample48T1D poor glycemic control
SA021864sample46T1D poor glycemic control
SA021865sample45T1D poor glycemic control
SA021866sample47T1D poor glycemic control
SA021867sample49T1D poor glycemic control
SA021868sample51T1D poor glycemic control
SA021869sample50T1D poor glycemic control
SA021870sample52T1D poor glycemic control
Showing results 1 to 58 of 58

Collection:

Collection ID:CO000449
Collection Summary:At 5:00 AM after an overnight fast, baseline blood samples were collected from study participants. Plasma samples were stored at 80°C until analysis.
Sample Type:Blood. Plasma was isolated for MS analysis.

Treatment:

Treatment ID:TR000469
Treatment Summary:Participants were admitted to the Clinical Research Unit at St Mary’s Hospital (Rochester, Minnesota) the evening before the study and spent overnight in the Clinical Research Unit. The participants were given a standard meal on the evening of the admission after which they fasted overnight. Participants with T1D were treated with insulin as per their usual individual programs. At 5:00 AM after an overnight fast, baseline blood samples were collected from study participants. Plasma samples were stored at 80°C until analysis.

Sample Preparation:

Sampleprep ID:SP000462
Sampleprep Summary:Plasma quality-control samples used in the study were prepared from pooled plasma spiked with a selection of metabolites to mimic elevated levels of metabolites during I− (insulin withdrawn) condition. Plasma was spiked with a standard mixture (3:1 ratio of plasma to spiking solution) containing 100 μg/mL niacin, hypoxanthine, leucine, isoleucine, phenylalanine, tryptophan, citric acid, glucose, hippuric acid, and taurocholic acid dissolved in 1:1 acetonitrile/water. All plasma samples (200 μL) were thawed on ice at 4°C followed by deproteinization with methanol (1:4 ratio of plasma to methanol) and vortexed for 10 s, followed by incubation at −20°C for 2 h. The samples were then centrifuged at 15,871g for 30 min at 4°C. The supernatants were lyophilized (Savant, Holbrook, NY) and stored at −20°C prior to analysis. The samples were reconstituted in 50% H2O/acetonitrile and passed through a Microcon YM3 filter (Millipore Corporation). The supernatants were transferred to analytical vials, stored in the autosampler at 4°C, and analyzed within 48 h of reconstitution in buffer.

Combined analysis:

Analysis ID AN000684
Analysis type MS
Chromatography type Reversed phase
Chromatography system Cohesive TX2 liquid chromatography system
Column Ascentis C18 (150 x 2.1mm,2.7um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name ABI Sciex API 5000 QQQ
Ion Mode NEGATIVE
Units micromolar

Chromatography:

Chromatography ID:CH000496
Chromatography Summary:Concentration and isotopic enrichment of palmitic acid from extracted plasmas were simultaneously measured against an extracted concentration standard curve as well as an enrichment standard curve on the Applied Biosystem API5000 mass spectrometer-MS (Foster City, CA) coupled with a Cohesive TX2 liquid chromatography system-LC (Franklin, MA). This system provides two LCs connected in parallel to the mass spectrometer, allowing twice the number of samples to be analyzed by using tandem sample injection. The LCs were controlled by Aria software (Cohesive Technology). Palmitic and heptadecanoic acids were separated on the LCs using an Ascentis C18, 2.7 μm, 2.1 × 150 mm column using two buffers. Buffer A was 80% acetonitrile, 0.5 mM ammonium acetate; buffer B was 99% acetonitrile, 1% 0.5 mM ammonium acetate. The flow rate was 0.4 ml/min, and the gradient conditions were as follows: 0–8 min isocratic at 55% B, 8–8.5 min 55⇒95% B, 8.5–10 min isocratic at 95% B, 10–10.5 min 95⇒55% B, and 10.5–12 min isocratic at 55% B. One tenth of the volume of each concentration standard and each plasma sample were resuspended in 400 μl of buffer A prior to injecting 10 μl onto the LC/MS. The chromographic separation of linoleic acid, palmitic acid, oleic acid, eladic acid, and heptadecanoic acid (internal standard) are depicted in Fig. 1A. Although palmitic and oleic acid peaks overlap, they are distinguishable by MS, whereas eladic acid, which is indistinguishable from oleic acid by MS, is easily separated by chromatography. Fig. 1B depicts the M+2 peaks for palmitate and heptadecanoate and the M+16 peak for palmitate of a plasma sample from a volunteer receiving a [U-13C]palmitate infusion. Palmitate elutes at 6.2 min and heptadecanoate acid at 7.1 min; however, in Fig. 1B the retention times are shown as 1.2 and 2.9 min because of the 3 min delay set in the LC method before diverting the flow to the MS for acquisition. The MS was set to acquire for only 5 min, allowing MS data acquisition from the second LC to begin while the first LC finishes its 12 min gradient. The MS was equipped with a turbo ion spray interface with the heater set at 60°C, spray voltage at 5500 V, sheath gas at 50, sweep gas at 40, and transfer capillary at 270°C. Palmitate, [13C16]palmitate and heptadecanoate were selectively monitored at m/z 257, m/z 271 in negative mode. Palmitate was monitored as [M+2−H]− and [M+16−H]−. Heptadecanoic was also monitor as [M+2−H]− . Therefore m/z 271 was either [13C16]palmitate or heptadecanoate, depending on where it eluted in the LC gradient.
Instrument Name:Cohesive TX2 liquid chromatography system
Column Name:Ascentis C18 (150 x 2.1mm,2.7um)
Flow Gradient: 0–8 min isocratic at 55% B, 8–8.5 min 55⇒95% B, 8.5–10 min isocratic at 95% B, 10–10.5 min 95⇒55% B, and 10.5–12 min isocratic at 55% B.
Flow Rate:0.4 ml/min
Solvent A:80% acetonitrile/20% water; 0.5 mM ammonium acetate
Solvent B:99% acetonitrile/1% water; 0.5 mM ammonium acetate
Chromatography Type:Reversed phase

MS:

MS ID:MS000610
Analysis ID:AN000684
Instrument Name:ABI Sciex API 5000 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
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