US EPA

7426494 

Journal Article 

Multifunctional and highly sensitive precolumn reagents for amino acids in liquid chromatography/tandem mass spectrometry 

Shimbo, K; Yahashi, A; Hirayama, K; Nakazawa, M; Miyano, H; , 

2009 

Yes 

Analytical Chemistry
ISSN: 0003-2700
EISSN: 1520-6882 

AMER CHEMICAL SOC 

WASHINGTON 

81 

13 

5172-5179 

English 

19480430 

10.1021/ac900470w 

WOS:000267609500012 

https://pubs.acs.org/doi/10.1021/ac900470w
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We have developed novel precolumn derivatization reagent, p-N,N,N-trimethylammonioanilyl N'-hydroxysuccinimidyl carbamate iodide (TAHS), for sensitive analyses of amino acids using high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). TAHS, an activated carbamate, was reacted briefly with the amino group to form a ureide bond under mild condition. The derivatives provided selective cleavage at the binding site between the reagent and the amino acid in the collision cell of the mass spectrometer and produced a characteristic fragment derived from the reagent moiety. Using the precursor ion scan mode of the tandem mass spectrometry, amino acids derivatized with the reagents were simultaneously measured on the chromatogram. Selective cleavage also enabled the straightforward isotope ratio analysis of amino acids by the selected reaction monitoring mode, which was applicable in (13)C metabolic flux analysis. TAHS, which contains a cationic quaternary amine, achieved subfemtomole to attomole levels of amino acids detection by measurement in the selected reaction monitoring mode. We also synthesized trideuteriummethyl-substituted TAHS, TAHS-d(3), and demonstrated that the combination of TAHS and TAHS-d(3) is useful in comparing amino acid concentrations between two different samples using a single LC/MS/MS measurement. 

Acid concentrations; Amino group; Attomole; Collision cells; Highly sensitive; Isotope ratio; LC/ESI-MS/MS; LC/MS/MS; Liquid chromatography/tandem mass spectrometry; Metabolic flux analysis; Precolumn derivatization; Precursor-ion scan; Quaternary amines; Selected reaction monitoring; Sensitive analysis; Tandem mass spectrometry; Amination; Amino acids; Binding energy; Binding sites; Ionization of liquids; Liquids; Mass spectrometers; Mass spectrometry; Monitoring; Organic acids; Spectrum analysis; Amines; 4 n,n,n trimethylammonioanilyl n' hydroxysuccinimidylcarbamate; amine; amino acid; carbamic acid derivative; carbon 13; reagent; unclassified drug; article; binding site; derivatization; electrospray mass spectrometry; high performance liquid chromatography; liquid chromatography; measurement; monitoring; precursor; tandem mass spectrometry; Amino Acids; Animals; Carbamates; Chromatography, High Pressure Liquid; Indicators and Reagents; Rats; Rats, Sprague-Dawley; Spectrometry, Mass, Electrospray Ionization; Staining and Labeling; Succinimides; Tandem Mass Spectrometry