Effects of alkylamines on electroosmotic flow and protein migration behaviour in capillary electrophoresis

Corradini, Danilo; Cannarsa, Gianfranco; Fabbri, Emanuela; Corradini, Claudio

doi:10.1016/0021-9673(95)00160-o

Cited by 57 publications

(28 citation statements)

References 24 publications

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“…To overcome these difficulties, several approaches have been suggested, including coating of the capillary wall [7,19.-22] and use of additives to the electrolyte (cf. [18]). Addition of ethylene glycol to samples (20-30% (v/v)) improves resolution of polypeptides in capillary electrophoresis [2,23].…”

Section: Resultsmentioning

confidence: 99%

“…A problem with fused silica capillaries is sample adsorption to the negatively charged wall with its ionized silanol groups which may result in broad peaks, loss of resolution and a 'memory effect' (carry over) [14,18]. To overcome these difficulties, several approaches have been suggested, including coating of the capillary wall [7,19.-22] and use of additives to the electrolyte (cf.…”

Section: Resultsmentioning

confidence: 99%

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Micropreparation of peptides by capillary electrophoresis for matrix assisted laser desorption mass spectrometry

Bergman

1996

FEBS Letters

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In the separation of peptides by capillary electrophoresis and analysis by matrix assisted laser desorption mass spectrometry, strong suppression of the mass spectrometric signals is a problem with many common electrolytes used in the separation step such as sodium phosphate. We describe an approach employing individual electrolytes selected for highest performance in each process. Suppression with samples collected into phosphate buffers is avoided when citrate, trifluoroacetic acid or hydrochloric acid is used for collection, while phosphate still provides excellent resolution in the capillary. Low concentrations of hydrochloric acid added to the sample/matrix mixture generate essentially adduct-free mass spectra with better signalto-noise ratios and detection limits (fmol range) than those obtained with citrate or trifluoroacetic acid. Addition of 0.25% ethylene glycol to both the phosphate electrolyte and the sample improves peak shape and resolution, and is crucial for preparative separations in large diameter capillaries.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Micropreparation of peptides by capillary electrophoresis for matrix assisted laser desorption mass spectrometry

Bergman

1996

FEBS Letters

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“…Depending on the ammonium salt concentration, a pH dependence has either been reported [397] or not been found [244]. High concentrations are necessary to obtain the maximum effect [400]. Low concentrations of these salts are sufficient to reduce or even reverse the EOF [397,464].…”

Section: Determination Of the Eof And Mobility Pmentioning

confidence: 99%

Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications

1998

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This review is in support of the development of selective, reproducible and validated capillary electrophoretis (CE) methods. Focusing on pharmaceutical and biological applications, the successful use of CE is demonstrated by more than 800 references, mainly from 1994 until 1998. Approximately 80 recent reviews have been catalogued. These articles sum up the existing strategies for method development in CE, especially in the search for generally accepted concepts, but also looking for new, promising reagents and ideas. General strategies for method development were derived not only with regard to selectivity and efficiency, but also with regard to precision, short analysis time, limit of detection, sample pretreatment requirements and validation. Standard buffer recipes, surfactants used in micellar electrokinetic capillary chromatography (MEKC), chiral selectors, useful buffer additives, polymeric separation media, electroosmotic flow (EOF) modifiers, dynamic and permanent coatings, actions to deal with complex matrices and aspects of validation are collected in 20 tables. Detailed schemes for the development of MEKC methods and chiral separations, for optimizing separation efficiency, means of troubleshooting, and other important information for key decisions during method development are given in 19 diagrams. Method development for peptide and protein separations, possibilities to influence the EOF and how to stabilize it, as well as indirect detection are considered in special sections.

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“…Logical ways for literature. It is widely accepted that the main reason circumventing these effects are either to oppress the for this phenomenon results from the charged silanol dissociation of the silanol groups by running the separation at a very low pH (|2.5, see [2,3]) or, on the contrary, to depress the dissociation of the [5][6][7][8][9][10][11][12][13].…”

Section: Introductionmentioning

confidence: 99%

Binding of proline- and hydroxyproline-containing peptides and proteins to the capillary wall

Hamrnı́ková

Mikšı́k

Deyl

et al. 1999

Journal of Chromatography A

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Sticking of peptides (as demonstrated by peak distortion) containing a high proportion of glycine and proline residues to the capillary wall was investigated. At acid pH where the carboxy terminal proline (e.g. Gly-Pro) is protonated, distinct sticking of these peptides occurred. On the contrary, the Pro-Gly peptide yielded a well shaped peak. At alkaline pH, where the situation was reversed, the peptide sticking most to the wall was Gly-Gly, while the behaviour of the dipeptides possessing N-or C-terminal proline did not suffer from sticking to the wall.It was concluded that the separation of simple proline-and glycine-containing dipeptides can be partly optimized by manipulating the pH of the background electrolyte; particularly if cetyltrimethylammonium bromide, methanol or acetonitrile is added to the run buffer (reversed polarity mode with cetyltrimethylammonium bromide). However, all attempts to resolve more complex mixtures of proline-(and hydroxyproline-) containing di-and tripeptides failed; addition of an organic modifier, i.e. methanol and acetonitrile, hexylamine, triethylamine and cetyltrimethylammonium bromide was unsuccessful. Such separations can be materialized by using simple 25 mmol / l phosphate buffer at pH 10.5 provided that the sample is dissolved in (aqueous) 17.5 mmol / l Brij or 33 mmol / l sodium dodecyl sulfate. These systems are also applicable to large polypeptides rich in proline, hydroxyproline and glycine: typically parent collagen a-chains, their dimers, trimers and higher polymers were successfully separated.

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Effects of alkylamines on electroosmotic flow and protein migration behaviour in capillary electrophoresis

Cited by 57 publications

References 24 publications

Micropreparation of peptides by capillary electrophoresis for matrix assisted laser desorption mass spectrometry

Micropreparation of peptides by capillary electrophoresis for matrix assisted laser desorption mass spectrometry

Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications

Binding of proline- and hydroxyproline-containing peptides and proteins to the capillary wall

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