A fluorescence toolbox: A review of investigation of electrophoretic separations, process, and interfaces

The review provides a comprehensive overview of developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) for analysis, microscale isolation, and physicochemical characterization of peptides from 2019 up to approximately the middle of 2021. Advances in the investigation of electromigration properties of peptides and in the methodology of their analysis, such as sample preparation, sorption suppression, EOF control, and detection, are presented. New developments in the individual CE and CEC methods are demonstrated and several types of their applications are shown. They include qualitative and quantitative analysis, determination in complex biomatrices, monitoring of chemical and enzymatic reactions and physicochemical changes, amino acid, sequence, and chiral analyses, and peptide mapping of proteins. In addition, micropreparative separations and determination of significant physicochemical parameters of peptides by CE and CEC methods are described.

Section: Detectionmentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2019–mid 2021)

Kašička

2021

“…Different aspects of LIF detection in CE (light sources, detection elements, optimization of excitation conditions, stability of the fluorophores, spectral filtering, and fluorescent labels) including the possibilities to further decrease the LODs of on-capillary LIF detectors were in detail analyzed in the previously published tutorial [191]. Applications of the standard and advanced fluorescence spectroscopy techniques (fluorescence microscopy, fluorescence recovery after photobleaching, fluorescence correlation spectroscopy, and fluorescence anisotropy) for detection as well as for investigation of transport, interfacial phenomena, and intermolecular interactions in CE separations of peptides and other biomolecules are discussed in recent review [192].…”

Section: Fluorescencementioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2021–mid‐2023)

Kašička

2023

This review article brings a comprehensive survey of developments and applications of high‐performance capillary and microchip electromigration methods (zone electrophoresis in a free solution or in sieving media, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) for analysis, micropreparation, and physicochemical characterization of peptides in the period from 2021 up to ca. the middle of 2023. Progress in the study of electromigration properties of peptides and various aspects of their analysis, such as sample preparation, adsorption suppression, electroosmotic flow regulation, and detection, are presented. New developments in the particular capillary electromigration methods are demonstrated, and several types of their applications are reported. They cover qualitative and quantitative analysis of synthetic or isolated peptides and determination of peptides in complex biomatrices, peptide profiling of biofluids and tissues, and monitoring of chemical and enzymatic reactions and physicochemical changes of peptides. They include also amino acid and sequence analysis of peptides, peptide mapping of proteins, separation of stereoisomers of peptides, and their chiral analyses. In addition, micropreparative separations and physicochemical characterization of peptides and their interactions with other (bio)molecules by the above CE methods are described.

“…FD is an extremely sensitive and selective detection technique, especially its LIF variant. In this variant, a laser or LED is used as the excitation source [80].…”

Section: O‐phthaldialdehydementioning

confidence: 99%

Homocyclic o‐dicarboxaldehydes: Derivatization reagents for sensitive analysis of amino acids and related compounds by capillary and microchip electrophoresis

Celá

Glatz

2020

Amino acids are essential compounds for living organisms, and their determination in biological fluids is crucial for the clinical analysis and diagnosis of many diseases. However, the detection of most amino acids is hindered by the lack of a strong chromophore/fluorophore or electrochemically active group in their chemical structures. The highly sensitive determination of amino acids often requires derivatization. Capillary electrophoresis is a separation technique with excellent characteristics for the analysis of amino acids in biological fluids. Moreover, it offers the possibility of precapillary, on‐capillary, or postcapillary derivatization. Each derivatization approach has specific demands in terms of the chemistry involved in the derivatization, which is discussed in this review. The family of homocyclic o‐dicarboxaldehyde compounds, namely o‐phthalaldehyde, naphthalene‐2,3‐dicarboxaldehyde, and anthracene‐2,3‐dicarboxaldehyde, are powerful derivatization reagents for the determination of amino acids and related compounds. In the presence of suitable nucleophiles they react with the primary amino group to form both fluorescent and electroactive derivatives. Moreover, the reaction rate enables all of the derivatization approaches mentioned above. This review focuses on articles that deal with using these reagents for the derivatization of amino acids and related compounds for ultraviolet‐visible spectrometry, fluorescence, or electrochemical detection. Applications in capillary and microchip electrophoresis are summarized and discussed.