Summary of Study ST000091

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, where it has been assigned Project ID PR000083. The data can be accessed directly via it's Project DOI: 10.21228/M8T884 This work is supported by NIH grant, U2C- DK119886.


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Study IDST000091
Study TitleQuantitative Metabolomics by 1H-NMR and LC-MS/MS Confirms Altered Metabolic Pathways in Diabetes
Study TypePre- and Post- insulin study with matched controls
Study SummaryWe obtained plasma samples from 7 c-peptide negative type 1 diabetic individuals (T1D) and 7 non-diabetic controls (Con) that were matched for age (T1D = 31.1±2.9 yrs, Con = 30.2±3.4 yrs), body mass (T1D = 80.2±4.7kg, Con = 81.9±7.4 kg) and BMI (T1D = 26.5±1.2 kg/m2, Con = 25.2±1.3 kg/m2). Type 1 diabetic people were studied while treated with insulin and also after 8 hours of insulin deprivation
Mayo Clinic
Last NameNair
First NameSreekumaran
Submit Date2014-07-15
Num Groups2
Raw Data AvailableNo
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2014-07-26
Release Version1
Sreekumaran Nair Sreekumaran Nair application/zip

Select appropriate tab below to view additional metadata details:


Project ID:PR000083
Project DOI:doi: 10.21228/M8T884
Project Title:Quantitative metabolomics by H-NMR and LC-MS/MS confirms altered metabolic pathways in diabetes
Project Type:Quantitative Metabolomics (Uploaded LC-MS data only)
Project Summary:Insulin is as a major postprandial hormone with profound effects on carbohydrate, fat, and protein metabolism. In the absence of exogenous insulin, patients with type 1 diabetes exhibit a variety of metabolic abnormalities including hyperglycemia, glycosurea, accelerated ketogenesis, and muscle wasting due to increased proteolysis. We analyzed plasma from type 1 diabetic (T1D) humans during insulin treatment (I+) and acute insulin deprivation (I-) and non-diabetic participants (ND) by 1H nuclear magnetic resonance spectroscopy and liquid chromatography-tandem mass spectrometry. The aim was to determine if this combination of analytical methods could provide information on metabolic pathways known to be altered by insulin deficiency. Multivariate statistics differentiated proton spectra from I- and I+ based on several derived plasma metabolites that were elevated during insulin deprivation (lactate, acetate, allantoin, ketones). Mass spectrometry revealed significant perturbations in levels of plasma amino acids and amino acid metabolites during insulin deprivation. Further analysis of metabolite levels measured by the two analytical techniques indicates several known metabolic pathways that are perturbed in T1D (I-) (protein synthesis and breakdown, gluconeogenesis, ketogenesis, amino acid oxidation, mitochondrial bioenergetics, and oxidative stress). This work demonstrates the promise of combining multiple analytical methods with advanced statistical methods in quantitative metabolomics research, which we have applied to the clinical situation of acute insulin deprivation in T1D to reflect the numerous metabolic pathways known to be affected by insulin deficiency.
Institute:Mayo Clinic
Last Name:Nair
First Name:Sreekumaran
Address:200 First Street SW, Rochester, MN 55905
Funding Source:UL1-RR-024150-01, R01-DK-41973
Project Comments:20479934


Subject ID:SU000110
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human


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

mb_sample_id local_sample_id Treatment
SA00499424 JH - CControl
SA0049959 JM-CControl
SA00499626 AW - CControl
SA0049973 JV-CControl
SA0049986 SS-CControl
SA00499921 PB - CControl
SA00500018 JF - CControl
SA00500112 GO-CControl
SA00500215 TK - CControl
SA0050038 AZ-DInsulin Deprived
SA0050042 AP-DInsulin Deprived
SA00500514 MB - DInsulin Deprived
SA00500617 MQ - DInsulin Deprived
SA00500720 RM - DInsulin Deprived
SA0050085 EK-DInsulin Deprived
SA00500923 JS - DInsulin Deprived
SA00501011 TZ-DInsulin Deprived
SA00501110 TZ-TInsulin Treatment
SA0050127 AZ-TInsulin Treatment
SA00501325 LH - TInsulin Treatment
SA00501419 RM - TInsulin Treatment
SA00501516 MQ - TInsulin Treatment
SA00501622 JS - TInsulin Treatment
SA00501713 MB - TInsulin Treatment
SA0050181 AP-TInsulin Treatment
SA0050194 EK-TInsulin Treatment
Showing results 1 to 26 of 26


Collection ID:CO000093
Collection Summary:Arterialized venous blood was obtained from a catheterized hand vein maintained at 60°C using a hot box for the duration of the study. Plasma samples were stored at ?80°C until analysis.
Sample Type:Blood. Plasma was isolated for MS analysis.
Collection Location:Hand vein


Treatment ID:TR000111
Treatment Summary:Saline Infusion | Insulin Withdrawal | Insulin Infusion
Treatment Protocol ID:SI, IW, II
Treatment Protocol Comments:ND participants were kept on a saline infusion from the evening following their meal, Insulin was discontinued for 8.6 ± 0.6 h starting at 0400 h., Insulin was infused into a forearm vein to maintain blood glucose between 4.44 and 5.56 mmol/L overnight until 1200 h the next day.
Treatment Compound:Saline, Insulin

Sample Preparation:

Sampleprep ID:SP000106
Sampleprep Summary:Plasma samples and amino acid calibration standards were prepared with MassTrak Amino Acid Analysis Solution (AAA) kit from Waters according to instructions with slight modifications for detection on a mass spectrometer. A 10 point standard concentration curve was made from the calibration standard solution to calculate amino acid concentrations in plasma samples. A solution containing U-13C4-L-aspartic acid, U-13C3-L-alanine, U-13C4-L-threonine, U-13C5-L-proline, U-13C5-L-valine, U-13C6-leucine, U-13C6-phenylalanine all from Cambridge Isotope Laboratories, 13C6-tyrosine from Isotec, L-arginine (15N2, 2H2) from MassTrace, norvaline from Sigma dissolved in 0.01N HCl was used as the internal standard solution. Frozen plasma samples were thawed, spiked with internal standard then deproteinized with cold MeOH followed by centrifugation at 10,000 g for 5 minutes prior to derivatization according to MassTrak instructions. The amino acid derivatizing reagent used was 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate.

Combined analysis:

Analysis ID AN000145
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column Waters Acquity BEH C18 (150 x 2.1mm,1.7um)
MS instrument type Triple quadrupole
MS instrument name Thermo Quantum Ultra
Units uM


Chromatography ID:CH000103
Chromatography Summary:High resolution separation was done using an Acquity UPLC system, injecting 1 l of derviatized solution, with a UPLC BEH C18 1.7 micron 2.1150 mm column from Waters. Column flow was set to 400 l/min with a gradient from 99.9%A to 98%B where buffer A is 1% acetonitrile in 0.1% formic acid and buffer B is 100% acetonitrile. A column temp of 43 degrees Celsius and a sample tray temp of 6 degree Celsius.
Instrument Name:Waters Acquity
Column Name:Waters Acquity BEH C18 (150 x 2.1mm,1.7um)
Column Temperature:43
Flow Gradient:99.9% A to 98% B
Flow Rate:401 µl/min
Solvent A:99% water/1% acetonitrile; 0.1% formic acid
Solvent B:100% acetonitrile
Chromatography Type:Reversed phase


MS ID:MS000121
Analysis ID:AN000145
Instrument Name:Thermo Quantum Ultra
Instrument Type:Triple quadrupole
MS Comments:Mass detection was completed on a TSQ Ultra Quantum from Thermo Finnigan running in ESI positive mode. A scan width of 0.002, scan time of 0.04 seconds per transition mass, collision energy of 25, collision gas pressure of 1.5 mTorr, tube lens value set to 90, monitoring a signature ion of the derivitized amines at m/z 171.04 by selected reaction monitoring.