Using reaction flow chromatography for the analysis of amino acid: Derivatisation with fluorescamine reagent

Pravadali-Cekic, Sercan; Jones, Andrew; Kazarian, Artaches A.; Paull, Brett; Soliven, Arianne; Ritchie, Harald; Camenzuli, Michelle; Dennis, Gary R.; Shalliker, R. Andrew

doi:10.1016/j.microc.2015.02.010

Cited by 14 publications

(2 citation statements)

References 23 publications

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“…The multiport design of the RF end-fitting allows for dual derivatization reagent analysis. Selim et al 23 investigated the performance of two reagents (4-aminoantipyrene and potassium ferricyanide) using RF-PCD conditions for the analysis of phenolic compounds compared to the conventional technique of PCD using 4-aminoantipyrene and potassium ferricyanide. This type of PCD technique requires two pumps and reaction loops for each derivatization reagent as opposed to one pump and reaction loop for DPPH…”

Section: Discussionmentioning

confidence: 99%

“…Reaction flow can be used for either a single reagent derivatization (1 port for the derivatization reagent, 1 port to pass the column eluent to the detector and 1 port blocked) or a dual reagent system (2 ports for the derivatization reagents and 1 port to pass the column eluent to the detector). The flow from the central stream can either be used to detect the underivatized column eluent, effectively multiplexing detection 23 , or passed to waste.…”

Section: Introductionmentioning

confidence: 99%

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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

Jones

Pravadali-Cekic

Hua

et al. 2016

JoVE

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A protocol for the use of reaction flow high performance liquid chromatography columns for methods employing post column derivatization (PCD) is presented. A major difficulty in adapting PCD to modern HPLC systems and columns is the need for large volume reaction coils that enable reagent mixing and then the derivatization reaction to take place. This large post column dead volume leads to band broadening, which results in a loss of observed separation efficiency and indeed detection in sensitivity. In reaction flow post column derivatization (RF-PCD) the derivatization reagent(s) are pumped against the flow of mobile phase into either one or two of the outer ports of the reaction flow column where it is mixed with column effluent inside a frit housed within the column end fitting. This technique allows for more efficient mixing of the column effluent and derivatization reagent(s) meaning that the volume of the reaction loops can be minimized or even eliminated altogether. It has been found that RF-PCD methods perform better than conventional PCD methods in terms of observed separation efficiency and signal to noise ratio. A further advantage of RF-PCD techniques is the ability to monitor effluent coming from the central port in its underivatized state. RF-PCD has currently been trialed on a relatively small range of post column reactions, however, there is currently no reason to suggest that RF-PCD could not be adapted to any existing one or two component (as long as both reagents are added at the same time) post column derivatization reaction.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%