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Issue 15, 2014
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Reliable and simple analytical methods for determination of citrulline and metabolically related amino acids by liquid chromatography after derivatization: comparison between monolithic and core–shell columns

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Abstract

We describe the development of a high-performance liquid chromatography (HPLC) method for the determination of citrulline and other amino acids relevant to intestinal diseases. The amino acids were derivatized with 9-fluorenylmethylchloroformate (FMOC-Cl) and their derivatives were separated on two different columns, a core–shell column (Halo C18) and a silica-based monolith (Chromolith Performance RP-18). The derivatization reaction was optimized with respect to pH, buffer concentration and reproducibility. The optimal derivatization conditions were achieved with 0.4 M borate buffer at pH 9.20, a constant ratio of FMOC-Cl/total amino acids (10 : 1) and 75 mM tyramine after 1 min (quenching reaction). The separation conditions with both chromatographic supports were also optimized. The chromatographic performance (peak capacity and global resolution) of these two columns was compared. This proposed HPLC-UV method was satisfactorily applied to the analysis of a real plasma sample.

Graphical abstract: Reliable and simple analytical methods for determination of citrulline and metabolically related amino acids by liquid chromatography after derivatization: comparison between monolithic and core–shell columns

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Article information


Submitted
27 Feb 2014
Accepted
19 May 2014
First published
20 May 2014

Anal. Methods, 2014,6, 5830-5837
Article type
Paper

Reliable and simple analytical methods for determination of citrulline and metabolically related amino acids by liquid chromatography after derivatization: comparison between monolithic and core–shell columns

A. Acquaviva, L. Romero, C. Castells, G. Ramis-Ramos and J. M. Herrero-Martinez, Anal. Methods, 2014, 6, 5830
DOI: 10.1039/C4AY00496E

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