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United States Department of Agriculture

Agricultural Research Service

Newman - Methods & Techniques
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Methods and Techniques

 

Liquid chromatography with tandem mass spectrometry for the quantification of endogenous lipid mediators isolated from biological samples (e.g. plasma, tissues, lipoprotein particles).

 

Gas chromatography with mass spectroscopy for the quantification of fatty acids isolated or liberated from various biological samples.

 

Gel permeation chromatography for lipoprotein particle isolation.

 

Cell culture and molecular biology approaches with adipose cell types to investigate the impact of lipoprotein composition on cell growth, proliferation, and gene expression.

 

Animal and human feeding studies to investigate the impact of dietary intake on physiology and metabolism.

 

Instrumentation

 

 

Waters Acquity UPLC - Applied Biosystems/Sciex API 4000 Q TRAP® MS/MS

 

The combined technology of ultra high performance liquid chromatography with sub-two micron separations and hybrid triple quadrupole/linear ion trap mass spectrometer gives us highly resolved, high-sensitivity full-scan MS, MS/MS, and MS3 with high-selectivity in precursor and neutral loss modes. Our current applications include targeted quantitative Oxylipin Metabolite Profiling of over 80 endogenous analytes coupled with non-targeted peak collection. Methods in development are targeted/non-target bile acid, endocannabinoid/vanilloid, and selective peptide marker metabolomic profiling.

 

 

Agilent 6890/5973 GC-MSD

 

The Agilent Technologies gas chromatograph coupled to their single-quad mass selective detector allows for selective and sensitive quantitative analysis of fatty acid methyl esters (FAME analysis). Our current FAME multi-residue analysis consists of 52 fatty acids ranging from C10:0 to C24:6n3.

 

 

 

Waters 600 Series FPLC System

 

A Waters 626 HPLC pump, 717 autosampler and 996 PDA detector are coupled to a chilled FRAC 200 to perform lipoprotein particle separation with a Superose 6 gel permeation (size exclusion) column. Fresh plasma samples are fractionated according to Scheffer (1997) and frozen immediately at 80ºC for compositional analysis.

 

 

 

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Last Modified: 9/12/2011
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