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Postcolumn derivatization of amino acids using reaction flow chromatography columns with fluorescence detection: A fast new approach to selective derivatization techniques |
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| Authors: | Sercan Pravadali-Cekic Andrew Jones Artaches A Kazarian Brett Paull Arianne Soliven Harald Ritchie |
| Institution: | 1. Australian Centre for Research on Separation Sciences, Western Sydney University, Parramatta, NSW, Australia;2. School of Science and Health, Western Sydney University, Parramatta, NSW, Australiaspcekic@hotmail.com;4. School of Science and Health, Western Sydney University, Parramatta, NSW, Australia;5. Australian Centre for Research on Separation Sciences, University of Tasmania, Hobart, TAS, Australia;6. School of Physical Sciences (Chemistry), University of Tasmania, Tasmania, Australia;7. Faculty of Chemistry, Department of Natural Products and Pharmacognosy, Universidad de la República, Montevideo, Uruguay;8. Trajan Scientific Europe Ltd, Crownhill, Milton Keynes, United Kingdom |
| Abstract: | Reaction flow (RF) chromatography with fluorescamine reagent and fluorescence detection (FLD) was used for the analysis of amino acids. The performance of RF chromatography was tested against several optimized conventional postcolumn derivatization (PCD) methods. RF columns achieved greater sensitivity compared to conventional PCD methods, without the need for reaction loops, which resulted in more efficient separations. The RF-PCD method also achieved limits of detection (LOD) from the low picomole to subnanomole range. The calibration data of the RF-PCD technique yielded R2?≥?0.99 and % relative standard deviation in peak areas ranging from 0.34% to 5%. Through reaction flow chromatography, multiplexed detection was also achieved allowing the monitoring and analysis of derivatized and nonderivatized flow streams simultaneously. |
| Keywords: | Amino acids fluorescamine fluorescence detection multiplexed detection postcolumn derivatization reaction flow chromatography |