Improved detection and derivatization in capillary electrophoresis

Szulc, Michael; Krull, Ira S.

doi:10.1016/0021-9673(94)85064-x

Cited by 48 publications

(14 citation statements)

References 64 publications

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“…As many fluorescent amino acid derivatives are known [27][28][29][30], very sensitive determination, of these analytes using laser induced fluorescence detection is possible. …”

Section: Determination Of a M I N O Acidsmentioning

confidence: 99%

Determination of inorganic anions, carboxylic acids and amino acids in plant matrices by capillary zone electrophoresis

et al. 1997

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SummaryPhysiological investigations of solute transport in plants affords knowledge of solute distribution between different tissues. Capillary electrophoresis using indirect UV and laser induced fluorescence (LIF) detection is demonstrated as a useful technique for the simultaneous determination of inorganic anions, amino acids and carboxylic acids in plant samples. Cell sap obtained from plant tissues as well as simple ethanolic or aqueous plant extracts can be analysed directly without any pretreatment. This matrix stability and the very small volumes required allow fast determinations of various compounds in small plant tissue sections. In the case of carboxylic acids, resolution can be optimized using calcium for selective complexation of some of the compounds. Selective and sensitive determination of amino acids is possible using precolumn derivatisation with orthophthaldialdehyde (OPA) and laser induced fluorescence detection.

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“…As many fluorescent amino acid derivatives are known [27][28][29][30], very sensitive determination, of these analytes using laser induced fluorescence detection is possible. …”

Section: Determination Of a M I N O Acidsmentioning

confidence: 99%

Determination of inorganic anions, carboxylic acids and amino acids in plant matrices by capillary zone electrophoresis

et al. 1997

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“…will be changed, either positively or negatively [5]. The majority of the probing reactions in CE is performed in this mode because of (i) the large flexibility in optimizing the reaction conditions, (ii) the fact that the conditions during the derivatization do not have to be compatible with the electrophoretic buffer, (iii) the availability of a wide variety of labeling reagents and (iv) the fact that no complex instrumentation is needed [6,15]. Disadvantages of this approach are that in a number of cases the excess probe should be removed from the reaction mixture before the actual separation and that the derivatives are not always sufficiently hydrolytically or thermally stable [15].…”

Section: He-electrophoretic Derivatizations Reactionsmentioning

confidence: 99%

“…The result is that this type of labeling normally is applied when non-stable derivatives are formed or when the analyte contains multiple derivatization sites [6, 151. The inevitable disadvan-tages of post-electrophoretic reactions are, that (i) the choice of the reaction conditions is rather critical and should be compatible with the required separation conditions, (ii) the reaction times are relatively short, which means that detectability is normally lower in comparison with pre-electrophoretic separations, (iii) special equipment is necessary, and (iv) the detection properties of the derivatives should be different from the reagent [6]. Several post-electrophoretic reactor designs have been described [lo].…”

Section: Post-electrophoretic Derivatization Reactionsmentioning

confidence: 99%

Pre‐, on‐ and post‐column derivatization in capillary electrophoresis

et al. 1997

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This survey gives a short overview of the various reagents and procedures that can be used for pre-, post- and on-column derivatization in capillary electrophoresis. First there is an introduction about capillary electrophoresis as an analytical technique; this is followed by a discussion of the pros and cons of the various modes of derivatization and a comparison with liquid chromatography. In the following paragraphs the reagents for a number of functional groups are discussed. The emphasis is on derivatization of the amino group. Most of the information on the reagents and derivatization procedures is listed in tables together with information on the detection mode, analytes, sensitivity and samples. In addition to the amino group, information is given on labeling of aldehyde, keto, carboxyl, hydroxyl and sulfhydryl groups.

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“…As a consequence, this detection limit does not represent the true assay detection limit [23,24]. In addition, the incomplete labeling caused by multiple labeling sites on protein results to broad peak profiles [25,26]. Another approach is to avoid the complication of covalent labeling by detecting the intrinsic fluorescence of proteins using an UV laser.…”

Section: Introductionmentioning

confidence: 96%

On‐line concentration of proteins by SDS‐CGE with LIF detection

Chang

Tseng

2008

Electrophoresis

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We present a simple approach for on-line concentration of SDS-protein complexes by using poly(vinyl alcohol) (PVA) solution in CGE. In comparison to the coated capillary, the presence of EOF in CGE omitted the need to fill the capillaries with polymer solutions prior to the analysis. More importantly, we found that highly reproducible separation of eight proteins by 3.5% PVA was achieved between runs and without the regeneration of high bulk EOF; the RSD of migration times was less than 0.7%. To further improve the concentration sensitivity, neutral PVA was introduced into the capillary with the help of EOF to act as sieving matrix. The occurrence of stacking at the boundary between the PVA and the sample zone is mainly due to the retardation of proteins by PVA. As a result, the LODs at an S/N of 3 for SDS-protein complexes are of the order of sub-nM to several nM. For example, the LOD for BSA is 0.78 nM, which is a 91-fold sensitivity enhancement over the normal injection. In addition, our stacking method has been applied to the analyses of proteins in Escherichia coli cells. The peak for b-galactosidase (E. coli) was observed after 0.1 mM b-galactosidase was spiked into the E. coli samples.

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Improved detection and derivatization in capillary electrophoresis

Cited by 48 publications

References 64 publications

Determination of inorganic anions, carboxylic acids and amino acids in plant matrices by capillary zone electrophoresis

Determination of inorganic anions, carboxylic acids and amino acids in plant matrices by capillary zone electrophoresis

Pre‐, on‐ and post‐column derivatization in capillary electrophoresis

On‐line concentration of proteins by SDS‐CGE with LIF detection

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