Capillary electrophoretic separations of peptides using micelle-forming compounds and cyclodextrins as additives

Liu, Jinping; Cobb, Kelly A.; Novotný, Miloš V.

doi:10.1016/0021-9673(90)85147-n

Cited by 111 publications

(20 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The hydrophobicity-based mechanism is located at an interface between the BGE and the hydrophobic capillary wall surface in OT-CEC and between the BGE and micelle surface in MEKC in an analogous manner to the chromatographic hydrophobic mechanism located at the interface of the aqueous mobile phase and chemical bonded phase of RP-HPLC. It should be stressed that, when employing micelles or cyclodextrins in MEKC [14], while this approach does involve peptide and micelle or cyclodextrin interactions within the BGE, such interactions are specifically with the surface of the micelles or cyclodextrins and not with the bulk ions of the BGE. In each case, the selected combination of a chargebased mechanism and an hydrophobicity-based mechanism proved to be compatible.…”

Section: Introductionmentioning

confidence: 99%

Ion-interaction–capillary zone electrophoresis of cationic proteomic peptide standards

Popa¹,

Mant

Hodges

2006

Journal of Chromatography A

View full text Add to dashboard Cite

We have employed a novel capillary electrophoresis (CE) approach recently developed in our laboratory, termed ion-interaction-capillary zone electrophoresis (II-CZE), to the resolution of a mixture of 27 synthetic cationic proteomic peptide standards. These peptides were comprised of three groups of nine peptides (with net charges of +1, +2 and +3 for all nine peptides within a group), the hydrophobicity of the nine peptides within a group varying only subtly between adjacent peptides. This bidimensional CE approach achieved excellent resolution of the peptides with high peak capacity by combining the powerful CZE mechanism located in the background electrolyte (BGE) with an hydrophobicity-based mechanism also located in the BGE, the latter consisting of high concentrations (up to 0.4 M) of aqueous perfluorinated acids ( trifluoroacetic acid, pentafluoropropionic acid and heptafluorobutyric acid). Thus, concomitant with a CZE separation of the three differently charged groups of peptides, there is an hydrophobically-mediated separation of the peptides within these groups effected through interaction of the hydrophobic anions of the perfluorinated acids with hydrophobic amino acid side-chains in the peptides. This methodology is dramatically different from other CE methods that have used complexing agents such as micelles or cyclodextrins in MEKC. Overall, the results presented here demonstrate the value of CE as a peptide separative tool in its own right, including its use for proteomic applications, and not merely as a complementary technique to reversed-phase high-performance liquid chromatography (RP-HPLC).

show abstract

Section: Introductionmentioning

confidence: 99%

Ion-interaction–capillary zone electrophoresis of cationic proteomic peptide standards

Popa¹,

Mant

Hodges

2006

Journal of Chromatography A

View full text Add to dashboard Cite

show abstract

“…3 [93] and OPA solution as sheath fluid with a reaction time of 1-2 s, it was possible to achieve LODs of 1.8 for carbonic anhydrase to 23 amol for b-lactoglobulin B (b-LGB), representing, respectively, five-and twofold improvements in sensitivity and a ten-fold improvement in separation efficiency by comparison to previous results obtained with a conventional fluorescence detector [101]. In peptide analysis, the behaviour of similar model peptides and peptides from the enzymatic digestion of proteins derivatized with OPA was evaluated using oncolumn LIF detection [102]. It was shown that addition of anionic (SDS) or cationic (alkylammonium bromides) surfactants and CDs to the buffer has beneficial effects on the separation of peptides having similar net charges but different hydrophobicities.…”

Section: Ortho-phthalaldehyde (Opa)mentioning

confidence: 99%

“…Neuropeptide b-endorphine and neuropeptide Y were detected at a 10 25 ng/mL concentration and luteinizing hormone-releasing hormone at 10 26 ng/mL. CDs added in the electrolyte not only enhanced the resolution of FC-labeled derivatives, but also the fluorescence intensity [102].…”

Section: -Phenylspiro[furan-2(3h) 1'-phthalan]-33'-dione (Fc)mentioning

confidence: 99%

“…The reaction takes place at room temperature for 1 h. Novotny and co-workers [127] used CBCQA as a fluorogenic agent for primary amines in CE with LIF detection using 442 nm He-Cd laser. It has been used to study the role of the CD cavity in peptide separations [102]. Good resolution and peak shapes were achieved using a-CD as additive for peptides derivatized with CBQCA.…”

Section: -(4-carboxybenzoyl)-2-quinolinecarboxaldehyde (Cbqca)mentioning

confidence: 99%

See 1 more Smart Citation

LIF detection of peptides and proteins in CE

2007

View full text Add to dashboard Cite

CE- and microchip-based separations coupled with LIF are powerful tools for the separation, detection and determination of biomolecules. CE with certain configurations has the potential to detect a small number of molecules or even a single molecule, thanks to the high spatial coherence of the laser source which permits the excitation of very small sample volumes with high efficiency. This review article discusses the use of LIF detection for the analysis of peptides and proteins in CE. The most common laser sources, basic instrumentation, derivatization modes and set-ups are briefly presented and special attention is paid to the different fluorogenic agents used for pre-, on- and postcapillary derivatization of the functional groups of these compounds. A table summarizing major applications of these derivatization reactions to the analysis of peptides and proteins in CE-LIF and a bibliography with 184 references are provided which covers papers published to the end of 2005.

show abstract

“…When Terabe's group [13,14] demonstrated micellar electrokinetic capillary chromatography (MEKC) in the mid-1980s, it was expected to rival liquid chromatography (LC) for the analysis of amino acids and peptides. In reality, MEKC using charged micelles can be problematic for the separation of complex mixtures of peptides [15,16]; their zwitterionic nature leads to strong, yet nonspecific, electrostatic interactions with charged micelles masking weaker hydrophobic effects that make reversed-phase LC selective. This underlines the importance of solution pH.…”

Section: Introductionmentioning

confidence: 99%

A comparison between electrokinetic capillary chromatography and absorption spectroscopy for the analysis of peptide‐micelle association by weak hydrophobic interactions

2003

View full text Add to dashboard Cite

Micellar electrokinetic capillary chromatography (MEKC) was compared to absorption spectroscopy to estimate equilibrium association constans (K(as)) for peptide-micelle systems involving three peptides (leucine-enkephalin, methionine-enkephalin and leucine-phenylalanine (LF)) and two surfactant micelles (sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB)). Buffer pH was chosen to minimize purely electrostatic interactions between peptides and micelles that could not be interrogated by absorption spectroscopy. Viscosity-corrected MEKC mobilities gave reasonably similar estimates of K(as) between the two methods for all three peptide-SDS micelle systems, with K(as) values ranging from 13.7 +/- 0.3 to 49 +/- 1 M(-1). For CTAB, estimates of K(as) for LF-CTAB micelle association were of the same order of magnitude as the SDS micelle by the two methods of estimation. On the other hand, enkephalin-CTAB micelle binding was about 10 times stronger (K(as) = 122 +/- 3 M(-1) to 311 +/- 9 M(-1)) than the enkephalin-SDS micelle binding. In addition, MEKC underestimated the K(as) values relative to spectroscopy by a factor of 2-3 for the enkephalin-CTAB system.

show abstract

Capillary electrophoretic separations of peptides using micelle-forming compounds and cyclodextrins as additives

Cited by 111 publications

References 21 publications

Ion-interaction–capillary zone electrophoresis of cationic proteomic peptide standards

Ion-interaction–capillary zone electrophoresis of cationic proteomic peptide standards

LIF detection of peptides and proteins in CE

A comparison between electrokinetic capillary chromatography and absorption spectroscopy for the analysis of peptide‐micelle association by weak hydrophobic interactions

Contact Info

Product

Resources

About