Micellar electrokinetic chromatography separations of dynorphin peptide analogs

Fürtös‐Matei, Alexandra; Day, Robert; St‐Pierre, Serge; St-Pierre, Louis G.; Waldron, Karen C.

doi:10.1002/(sici)1522-2683(20000301)21:4<715::aid-elps715>3.0.co;2-5

Cited by 17 publications

(5 citation statements)

References 27 publications

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“…Resolution of closely related species can also be problematic in these applications. MEKC has been used to improve the resolution of opioid peptide analogs providing an additional mode of separation within a bare fused silica capillary 12. In addition, CDs have been exploited as run buffer additives and have been shown to improve resolution for peptide metabolites and enantiomers of relevant pharmaceuticals 43, 44.…”

Section: Discussionmentioning

confidence: 99%

“…Another disadvantage of RIA is the use of expensive radioactive reagents and assay kits, as well as the costs associated with the disposal of radioactive waste. A variety of other methods have been used to quantify opioid peptides such as the enkephalins, endomorphins, and smaller Dyn peptides including, HPLC with UV detection 10, LC‐MS 3, 11, CE‐UV 12, 13, CE‐LIF 13, 14, and CE‐MS 15–19. Qualitative and semi‐quantitative MALDI methods have also demonstrated potential for monitoring Dyn peptides 20–23.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of an on‐capillary copper complexation methodology for the investigation of in vitro metabolism of dynorphin A 1–17

Kuhnline

Lunte

2010

J of Separation Science

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Dynorphin A 1–17 is an endogenous neuropeptide implicated in a variety of neurological disorders including Alzheimer’s and Parkinson’s diseases and neuropathic pain. Metabolites of this peptide can exhibit their own unique effects in vivo, and it is possible that one of these metabolites is responsible for the neurotoxicity. In this article, the use of CE for the separation of dynorphin A 1–17 from four of its metabolites is described. Buffer additives were investigated to eliminate peptide adsorption to the capillary wall and to improve resolution between closely related metabolites. On-capillary copper complexation was employed and was shown to improve separation efficiency as compared with the separation of native peptides. The method was then applied to in vitro dynorphin metabolism in human plasma as well as rat brain and rat spinal cord slices.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Evaluation of an on‐capillary copper complexation methodology for the investigation of in vitro metabolism of dynorphin A 1–17

Kuhnline

Lunte

2010

J of Separation Science

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“…It allows the partitioning of analytes between the aqueous phase and the micellar pseudostationary phase, which allows the neutrally charged compounds to be separated as long as they differ in hydrophobicity [18]. Several other examples of the use of MEKC for the analysis of hydrophobic peptides were provided [19][20][21][22].…”

Section: Introductionmentioning

confidence: 99%

Rapid methods for the separation of natural mixtures of beauverolides, cholesterol acyltransferase inhibitors, isolated from the fungus Isaria fumosorosea

Šimčíková

Tůma

Jegorov

et al. 2020

J of Separation Science

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Beauverolides (beauveriolides) are abundant, biologically active cyclodepsipeptides produced by many entomopathogenic fungi, including those that are used as biopesticides. Beauverolides act as cholesterol acyltransferase inhibitors in humans; thus, their mode of action has been the subject of pharmacological and clinical research. The cost‐effective analytical methods are needed for fast, routine laboratory analysis of beauverolides. We isolated beauverolides from the fungal strain Isaria fumosorosea PFR 97‐Apopka and opened the rings of the isolated beauverolides using a pyridine alkaline medium. We separated fractions of cyclic and linearized beauverolides by thin‐layer chromatography, and found the chloroform–acetate (9:1, v/v) and chloroform–acetonitrile–acetate (8:1:1, v/v/v) mobile phases, respectively, to be the most efficient. We examined all the fractions by liquid chromatography–mass spectrometry using ion trap and Orbitrap high resolution mass spectrometry. For rapid screening of the contents of cyclic, and, particularly, linearized beauverolides, we developed a novel analytical method that consisted of using capillary electrophoresis coupled with contactless conductivity detection. Furthermore, we improved the separation of the peptides by applying capillary micellar electrokinetic chromatography with the N‐cyclohexyl‐2‐aminoethanesulfonic acid:SDS:NaOH buffer, pH 9.8 as the background electrolyte. The described novel methods allow fast and cost‐effective separation of chemically related groups of beauverolides.

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“…Of particular relevance to the present study, MEKC‐CE has been used in several studies to analyze hydrophobic peptides (Idei et al. 1998; Fürtös‐matei et al. 2000; Gong et al.…”

Section: Introductionmentioning

confidence: 99%

“…Analytes are partitioned between the aqueous phase and the micellar (pseudo-stationary) phases, and based on hydrophobic interaction with the micelles, neutral analytes or analytes with identical charge-to-mass ratios can be separated if they differ in hydrophobicity (Idei et al 1998). Of particular relevance to the present study, MEKC-CE has been used in several studies to analyze hydrophobic peptides (Idei et al 1998;Fürtös-matei et al 2000;Gong et al 2006).…”

Section: Introductionmentioning

confidence: 99%

QUANTITATIVE ANALYSIS OF THE BACILLUS CEREUS EMETIC TOXIN, CEREULIDE, USING MICELLAR ELECTROKINETIC CHROMATOGRAPHY‐CAPILLARY ELECTROPHORESIS

Cox

2010

Journal of Food Safety

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A micellar electrokinetic chromatography capillary electrophoresis (MEKC‐CE) method was developed for quantitative analysis of cereulide, the emetic toxin produced by Bacillus cereus. Valinomycin, a highly hydrophobic cyclic dodecadepsipeptide with a chemical structure similar to that of cereulide, was used as a surrogate for method development. A series of buffer systems and electrophoretic conditions were investigated and optimized. The final method, yielding good peak resolution and reproducibility of analysis, utilized 20 mM borate buffer, pH 8.5, containing 75 mM SDS. The capillary had a 50 µm internal diameter and 36.4 cm effective length, and was maintained at 25C throughout column preparation and analysis. The sample compartment was maintained at 22C. A constant voltage of 15 kV was applied, after hydrodynamic injection of the sample at 34.5 KPa for 2 s. Detection used ultraviolet light, at 195 nm. A calibration curve, using peak area rather than peak height, was obtained with purified cereulide, with linearity between 2 and 30 ng/µL. PRACTICAL APPLICATIONS Detection of the emetic toxin cereulide, as well as enumeration of B. cereus, is important for determining the risk of emetic food poisoning. The novel capillary electrophoresis method developed in this study, employing the principles of MEKC‐CE, can be used for the quantitative analysis of cereulide. MEKC‐CE gives results quickly and simply, and offers advantages over reversed phase high performance liquid chromatography (RP‐HPLC), through simple sample handling and preparation, less sample required and lower cost of operation. After calibration with purified cereulide was obtained, the applicability of the method to model and real food samples was evaluated with rice samples inoculated with an emetic B. cereus strain and samples from an outbreak of emetic food poisoning. The results showed that the method can be successfully applied to both semi‐purified toxin extracts of cultures and food samples.

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Micellar electrokinetic chromatography separations of dynorphin peptide analogs

Cited by 17 publications

References 27 publications

Evaluation of an on‐capillary copper complexation methodology for the investigation of in vitro metabolism of dynorphin A 1–17

Evaluation of an on‐capillary copper complexation methodology for the investigation of in vitro metabolism of dynorphin A 1–17

Rapid methods for the separation of natural mixtures of beauverolides, cholesterol acyltransferase inhibitors, isolated from the fungus Isaria fumosorosea

QUANTITATIVE ANALYSIS OF THE BACILLUS CEREUS EMETIC TOXIN, CEREULIDE, USING MICELLAR ELECTROKINETIC CHROMATOGRAPHY‐CAPILLARY ELECTROPHORESIS

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