Post-column reactor systems in liquid chromatography

Deelder, R.S.; Kroll, Moritz; Beeren, A.J.B.; Berg, J. H. M. van den

doi:10.1016/s0021-9673(00)81019-5

Cited by 110 publications

(21 citation statements)

References 38 publications

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“…An alternative to fraction collection for analysis is postcolumn derivatization in a continuous flow reactor . In continuous flow postcolumn reactors, it is important to keep the reaction fast and mixing efficient so that not much resolution will be sacrificed.…”

Section: Resultsmentioning

confidence: 99%

Capillary liquid chromatography fraction collection and postcolumn reaction using segmented flow microfluidics

Nie

Kennedy

2013

J. Sep. Science

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A challenge for capillary liquid chromatography (cLC) is fraction collection and manipulation of fractions from microscale columns. An emerging approach is use of segmented flow or droplet technology to perform such tasks. In this work, a fraction collection and post-column reaction system based on segmented flow was developed for gradient cLC of proteins. In the system, column effluent and immiscible oil are pumped into separate arms of a tee resulting in regular fractions of effluent segmented by oil. Fractions were generated at 1 Hz corresponding to 5 nL volumes. Fraction collection rate was high enough to generate over 30 fractions per peak and preserve chromatographic resolution achieved for a 5 protein test mixture. The resulting fractions could be stored and subsequently derivatized for fluorescence detection by pumping them into a second tee where naphthalene dicarboxyaldehyde, a fluorogenic reagent, was pumped into a second arm and added to each fraction. Proteins were derivatized within the droplets enabling post-column fluorescence detection of the proteins. The experiments demonstrate that fraction collection from cLC by segmented flow can be extended to proteins. Further they illustrate a potential workflow for protein analysis based on post-column derivatization for fluorescence detection.

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Section: Resultsmentioning

confidence: 99%

Capillary liquid chromatography fraction collection and postcolumn reaction using segmented flow microfluidics

Nie

Kennedy

2013

J. Sep. Science

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“…Several studies have dealt with the theoretical aspects of band broadening in post-column reactor systems [17][18][19][20]; however, such in-depth discussions are beyond the scope of this chapter. Briefly, to describe band broadening in open tubes, Deelder [20] has proposed the equation…”

Section: Reactorsmentioning

confidence: 99%

Post-Column Derivatization Techniques

Woolf

1993

A Practical Guide to HPLC Detection

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“…Moreover, most of the derivatization reactions are carried out under strict conditions. Post-column chemical derivatization (PCD) is one of chemical derivatization modes, 24 which has been used for LC−MS analysis (LC−PCD−MS). 25 The derivatization reactions of chromatographically separated analytes in PCD mode are usually performed in a post-column derivatization device between LC and MS. PCD has some advantages over the pre-column derivatization.…”

Section: ■ Introductionmentioning

confidence: 99%

Rapid Analysis of Monosaccharides in Sub-milligram Plant Samples Using Liquid Chromatography–Mass Spectrometry Assisted by Post-column Derivatization

Cai

Wang

et al. 2020

J. Agric. Food Chem.

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Monosaccharides play important roles in plant growth and development, and their biofunctions are closely related to their endogenous contents. Therefore, the determination of monosaccharides is beneficial for the further study of monosaccharide biofunction. In this work, we developed a liquid chromatography–mass spectrometry analytical method assisted by a post-column derivatization technique (LC–PCD–MS) for the fast and automatic determination of 16 monosaccharides in samples. Post-column chemical derivatization of monosaccharides was performed by a reaction of monosaccharides with 4-benzylaminobenzeneboronic acid (4-PAMBA) through boronate ester formation in a three-way connector. 4-PAMBA worked as a derivatization reagent to improve the selectivity and sensitivity of monosaccharide detection by MS. The developed LC–PCD–MS method integrates LC separation, chemical derivatization, and MS detection in one run, thus greatly reducing the analysis time for each sample. The limits of detection and limits of quantification for 16 monosaccharides were in the range of 0.002–0.1 and 0.007–0.5 ng/mL, respectively. Good linearity was obtained from the linear regression, with a determination coefficient (R 2) ranging from 0.9928 to 1.0000. The relative recoveries were in the range of 80.7–117.8%, with the intra- and interday relative standard deviations less than 19.7 and 16.5%, respectively, indicating good accuracy and acceptable reproducibility of the method. Finally, the method was successfully applied to investigate the spatial and temporal distribution of 16 monosaccharides in the developing flower and germinating seed of Arabidopsis thaliana.

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Post-column reactor systems in liquid chromatography

Cited by 110 publications

References 38 publications

Capillary liquid chromatography fraction collection and postcolumn reaction using segmented flow microfluidics

Capillary liquid chromatography fraction collection and postcolumn reaction using segmented flow microfluidics

Post-Column Derivatization Techniques

Rapid Analysis of Monosaccharides in Sub-milligram Plant Samples Using Liquid Chromatography–Mass Spectrometry Assisted by Post-column Derivatization

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