Merck
  • Home
  • Search Results
  • Highly sensitive GC/MS/MS method for quantitation of amino and nonamino organic acids.

Highly sensitive GC/MS/MS method for quantitation of amino and nonamino organic acids.

Analytical chemistry (2011-03-11)
Hans F N Kvitvang, Trygve Andreassen, Tomas Adam, Silas G Villas-Bôas, Per Bruheim
ABSTRACT

Metabolite profiling methods are important tools for measurement of metabolite pools in biological systems. While most metabolite profiling methods report relative intensities or depend on a few internal standards representing all metabolites, the ultimate requirement for a quantitative description of the metabolite pool in biological cells and fluids is absolute concentration determination. We report here a high-throughput and sensitive gas chromatography/tandem mass spectrometry (GC/MS/MS) targeted metabolite profiling method enabling absolute quantification of all detected metabolites. The method is based on methyl chloroformate derivatization and quantification by spiking samples with metabolite standards separately derivatized with deuterated derivatization reagents. The traditional electron impact ionization is replaced with positive chemical ionization since the latter to a much larger extent preserve the molecular ion and other high molecular weight fragments. This made it easier to select unique MS/MS transitions among the many coeluting metabolites. Currently, the novel GC/MS/MS method comprises 67 common primary metabolites of which most belong to the groups of amino and nonamino organic acids. We show the applicability of the method on urine and serum samples. The method is a significant improvement of present methodology for quantitative GC/MS metabolite profiling of amino acids and nonamino organic acids.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Threonine, reagent grade, ≥98% (HPLC)
Supelco
Glycine, analytical standard, for nitrogen determination according to Kjeldahl method
Supelco
L-Proline, certified reference material, TraceCERT®
Supelco
L-Tyrosine, certified reference material, TraceCERT®
Sigma-Aldrich
Glycine, BioXtra, ≥99% (titration)
Sigma-Aldrich
L-Proline, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
β-Alanine, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
L-Aspartic acid, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Aspartic acid, BioXtra, ≥99% (HPLC)
Sigma-Aldrich
L-Aspartic acid, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
Glycine, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
Gly-Pro
Sigma-Aldrich
Salicylic acid, BioXtra, ≥99.0%
Sigma-Aldrich
L-Proline, from non-animal source, meets EP, USP testing specifications, suitable for cell culture
Sigma-Aldrich
Glycine, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Glycine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, ≥98.5%
Supelco
L-Alanine, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Salicylic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
L-Aspartic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Glycine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
trans-4-Hydroxy-L-proline, BioReagent, suitable for cell culture, ≥98.5%
Sigma-Aldrich
L-Alanine, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Alanine, ≥98% (TLC)
Sigma-Aldrich
L-Valine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Valine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Threonine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 99.0-101.0%
Supelco
L-Valine, certified reference material, TraceCERT®
Supelco
L-Threonine, certified reference material, TraceCERT®
Sigma-Aldrich
L-Alanine, ≥99%
Sigma-Aldrich
Glycine, ACS reagent, ≥98.5%