Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1218, Issue 46, Pages 8379-8387Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2011.09.046
Keywords
Enantioselective LC MS MS; Amino acid derivatization; Enantioselective amino acid analysis
Funding
- MSMT, Czech Republic [CZ.1.05/2.1.00/03.0058]
- Palacky University [PrF_2011_025]
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D-Amino acid analysis in biological samples still poses a challenge to analytical chemists. In higher developed species trace amounts of D-amino acids have to be detected in vast excesses of the corresponding L-enantiomers. This method utilizes an easy-to-carry-out derivatization step on the amino group with an iron ferrocenyl propionate hydroxy succinimide ester followed by one-dimensional enantioselective anion exchange chromatography with cinchona alkaloid based chiral stationary phases (CSPs). MS detection is carried out in the highly sensitive SRM (selected reaction monitoring) mode, which allows a chemoselective differentiation of amino acid derivatives as well as their enantioselective separation in one step. Application of this method allows LOD (limits of detection) in the low mu mol L-1 range and baseline enantioseparation for all proteinogenic amino acids except for Pro, Arg and His. The D-enantiomers of isomeric Leu and Ile were separated chromatographically and pose an example for the complementary selectivities of LC and MS. A successful application of this procedure to unprocessed human urine indicated the eligibility to analyse biological samples. (C) 2011 Elsevier B.V. All rights reserved.
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