Summary of Study ST000091
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 PR000083. The data can be accessed directly via it's Project DOI: 10.21228/M8T884 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000091 |
| Study Title | Quantitative Metabolomics by 1H-NMR and LC-MS/MS Confirms Altered Metabolic Pathways in Diabetes |
| Study Type | Pre- and Post- insulin study with matched controls |
| Study Summary | We 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 |
| Institute | Mayo Clinic |
| Department | Endocrinology |
| Last Name | Nair |
| First Name | Sreekumaran |
| Address | -- |
| nair@mayo.edu | |
| Phone | -- |
| Submit Date | 2014-07-15 |
| Num Groups | 2 |
| Raw Data Available | No |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2014-07-26 |
| Release Version | 1 |
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Project:
| 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 |
| Department: | Endocrinology |
| Last Name: | Nair |
| First Name: | Sreekumaran |
| Address: | 200 First Street SW, Rochester, MN 55905 |
| Email: | Dasari.Surendra@mayo.edu |
| Phone: | -- |
| Funding Source: | UL1-RR-024150-01, R01-DK-41973 |
| Project Comments: | 20479934 |
Subject:
| Subject ID: | SU000110 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Species Group: | Human |
Factors:
Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA004994 | 24 JH - C | Control |
| SA004995 | 9 JM-C | Control |
| SA004996 | 26 AW - C | Control |
| SA004997 | 3 JV-C | Control |
| SA004998 | 6 SS-C | Control |
| SA004999 | 21 PB - C | Control |
| SA005000 | 18 JF - C | Control |
| SA005001 | 12 GO-C | Control |
| SA005002 | 15 TK - C | Control |
| SA005003 | 8 AZ-D | Insulin Deprived |
| SA005004 | 2 AP-D | Insulin Deprived |
| SA005005 | 14 MB - D | Insulin Deprived |
| SA005006 | 17 MQ - D | Insulin Deprived |
| SA005007 | 20 RM - D | Insulin Deprived |
| SA005008 | 5 EK-D | Insulin Deprived |
| SA005009 | 23 JS - D | Insulin Deprived |
| SA005010 | 11 TZ-D | Insulin Deprived |
| SA005011 | 10 TZ-T | Insulin Treatment |
| SA005012 | 7 AZ-T | Insulin Treatment |
| SA005013 | 25 LH - T | Insulin Treatment |
| SA005014 | 19 RM - T | Insulin Treatment |
| SA005015 | 16 MQ - T | Insulin Treatment |
| SA005016 | 22 JS - T | Insulin Treatment |
| SA005017 | 13 MB - T | Insulin Treatment |
| SA005018 | 1 AP-T | Insulin Treatment |
| SA005019 | 4 EK-T | Insulin Treatment |
| Showing results 1 to 26 of 26 |
Collection:
| 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:
| 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 Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Thermo Quantum Ultra |
| Ion Mode | POSITIVE |
| Units | uM |
Chromatography:
| 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:
| MS ID: | MS000121 |
| Analysis ID: | AN000145 |
| Instrument Name: | Thermo Quantum Ultra |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| 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. |
| Ion Mode: | POSITIVE |
