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Capillary Electrophoresis Analysis: Trends and Recent Advances
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Capillary Electrophoresis Analysis: Trends and Recent Advances

A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Analytical Chemistry".

Deadline for manuscript submissions: closed (31 December 2022) | Viewed by 8867

Special Issue Editor

Prof. Dr. Catherine Perrin

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Guest Editor
Institut des Biomolécules Max Mousseron (IBMM), Université de Montpellier– CNRS, UMR 5247, 34095 Montpellier, France
Interests: electrophoretic methodologies; miniaturized analytical techniques; pharmaceutical analysis; biomolecule analysis
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Capillary electrophoretic methodologies have become an essential, highly efficient separation technique for quality control, characterization, purification and quantification purposes. The wide operating conditions in an open separation support free from a stationary phase provide the unique versatility of capillary electrophoresis ranging from small to large, hydrophilic to hydrophobic molecules in various matrices, cells, particles, etc. Recent developments have shown that capillary electrophoresis is more than a separation technique as it allows the development of fully automated integrated analytical methodologies from sample preparation/derivatization to separation and detection. The potential of electrophoretic methods for miniaturization as well as its recent hyphenation to mass spectrometry has dramatically expanded its application domains and, in particular, proteins and low concentration samples.

The target of this Special Issue is to present the state of the art in electrophoretic methodologies in capillary and microchip formats. Recent, original and efficient methodologies are reported for all analysis domains.

Prof. Dr. Catherine Perrin
Guest Editor

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Keywords

  • capillary electrophoresis
  • electrophoretic methodologies
  • electrophoretic µ-chip
  • electrophoretic techniques and hyphenation
  • free solution electrophoresis
  • electrophoretic µ-reactor
  • isoelectric focusing (IEF)
  • capillary polyacrylamide gel electrophoresis (PAGE)—sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE)
  • protein analysis
  • pharmaceutical analysis
  • food analysis
  • environmental analysis

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Published Papers (4 papers)

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Research

19 pages, 1897 KiB  
Article
Reliable N-Glycan Analysis–Removal of Frequently Occurring Oligosaccharide Impurities by Enzymatic Degradation
by Robert Burock, Samanta Cajic, René Hennig, Falk F. R. Buettner, Udo Reichl and Erdmann Rapp
Molecules 2023, 28(4), 1843; https://doi.org/10.3390/molecules28041843 - 15 Feb 2023
Cited by 2 | Viewed by 1886
Abstract
Glycosylation, especially N-glycosylation, is one of the most common protein modifications, with immense importance at the molecular, cellular, and organismal level. Thus, accurate and reliable N-glycan analysis is essential in many areas of pharmaceutical and food industry, medicine, and science. However, [...] Read more.
Glycosylation, especially N-glycosylation, is one of the most common protein modifications, with immense importance at the molecular, cellular, and organismal level. Thus, accurate and reliable N-glycan analysis is essential in many areas of pharmaceutical and food industry, medicine, and science. However, due to the complexity of the cellular glycosylation process, in-depth glycoanalysis is still a highly challenging endeavor. Contamination of samples with oligosaccharide impurities (OSIs), typically linear glucose homo-oligomers, can cause further complications. Due to their physicochemical similarity to N-glycans, OSIs produce potentially overlapping signals, which can remain unnoticed. If recognized, suspected OSI signals are usually excluded in data evaluation. However, in both cases, interpretation of results can be impaired. Alternatively, sample preparation can be repeated to include an OSI removal step from samples. However, this significantly increases sample amount, time, and effort necessary. To overcome these issues, we investigated the option to enzymatically degrade and thereby remove interfering OSIs as a final sample preparation step. Therefore, we screened ten commercially available enzymes concerning their potential to efficiently degrade maltodextrins and dextrans as most frequently found OSIs. Of these enzymes, only dextranase from Chaetomium erraticum and glucoamylase P from Hormoconis resinae enabled a degradation of OSIs within only 30 min that is free of side reactions with N-glycans. Finally, we applied the straightforward enzymatic degradation of OSIs to N-glycan samples derived from different standard glycoproteins and various stem cell lysates. Full article
(This article belongs to the Special Issue Capillary Electrophoresis Analysis: Trends and Recent Advances)
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14 pages, 1949 KiB  
Article
Development of Capillary Zone Electrophoresis Method for the Simultaneous Separation and Quantification of Metformin and Pioglitazone in Dosage Forms; and Comparison with HPLC Method
by Maymonah K. I. AlThikrallah, Abubakr M. Idris, Abdalla Ahmed Elbashir, Rafea E. E. Elgorashe, Alyah Buzid and Ahmed O. Alnajjar
Molecules 2023, 28(3), 1184; https://doi.org/10.3390/molecules28031184 - 25 Jan 2023
Cited by 7 | Viewed by 2115
Abstract
A capillary zone electrophoretic (CZE) method was developed, validated, and applied for the assay of metformin (MET) and pioglitazone (PIO) in pharmaceutical formulations. The optimum running buffer composition was found to be 75 mmol/L phosphate buffer containing 30% acetonitrile (ACN) at pH 4.0. [...] Read more.
A capillary zone electrophoretic (CZE) method was developed, validated, and applied for the assay of metformin (MET) and pioglitazone (PIO) in pharmaceutical formulations. The optimum running buffer composition was found to be 75 mmol/L phosphate buffer containing 30% acetonitrile (ACN) at pH 4.0. The optimum instrumental conditions were found to be injection time, 10 s; applied voltage, 25 kV; hydrodynamic injection pressure, 0.5 psi for 10 s, capillary temperature, 25 °C; and the detection wavelength, 210 nm. The quantifications were calculated based on the ratio of the peak areas of analytes to atenolol as an internal standard. The CZE method was validated in terms of accuracy (98.21–104.81%), intra- and inter-day precision of migration time and peak area (relative standard deviation ≤ 5%), linearity (correlation coefficients ≥ 0.9985), limit of detection (≤0.277 μg/mL), and limit of quantitation (≤0.315 μg/mL). The proposed method was applied for the analysis of PIO and MET both individually and in a combined dosage tablet formulation. All electrophoretic parameters were calculated and evaluated. A previously reported high-performance liquid chromatographic (HPLC) method was also applied to the same samples. A comprehensive comparison was then carried out for the analytical features of both methods CZE and HPLC. Comparable results were obtained with the advantage of reagent consumption and separation efficiency of CZE over HPLC and shorter analysis time by HPLC compared with CZE. Full article
(This article belongs to the Special Issue Capillary Electrophoresis Analysis: Trends and Recent Advances)
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8 pages, 1389 KiB  
Article
The Effect of Sample Glucose Content on PNGase F-Mediated N-Glycan Release Analyzed by Capillary Electrophoresis
by Rebeka Torok, Felicia Auer, Robert Farsang, Eszter Jona, Gabor Jarvas and Andras Guttman
Molecules 2022, 27(23), 8192; https://doi.org/10.3390/molecules27238192 - 24 Nov 2022
Cited by 2 | Viewed by 1922
Abstract
Protein therapeutics have recently gained high importance in general health care along with applied clinical research. Therefore, it is important to understand the structure–function relationship of these new generation drugs. Asparagine-bound carbohydrates represent an important critical quality attribute of therapeutic glycoproteins, reportedly impacting [...] Read more.
Protein therapeutics have recently gained high importance in general health care along with applied clinical research. Therefore, it is important to understand the structure–function relationship of these new generation drugs. Asparagine-bound carbohydrates represent an important critical quality attribute of therapeutic glycoproteins, reportedly impacting the efficacy, immunogenicity, clearance rate, stability, solubility, pharmacokinetics and mode of action of the product. In most instances, these linked N-glycans are analyzed in their unconjugated form after endoglycosidase-mediated release, e.g., PNGase F-mediated liberation. In this paper, first, N-glycan release kinetics were evaluated using our previously reported in-house produced 6His-PNGase F enzyme. The resulting deglycosylation products were quantified by sodium dodecyl sulfate capillary gel electrophoresis to determine the optimal digestion time. Next, the effect of sample glucose content was investigated as a potential endoglycosidase activity modifier. A comparative Michaelis-Menten kinetics study was performed between the 6His-PNGase F and a frequently employed commercial PNGase F product with and without the presence of glucose in the digestion reaction mixture. It was found that 1 mg/mL glucose in the sample activated the 6His-PNGase F enzyme, while did not affect the release efficiency of the commercial PNGase F. Capillary isoelectric focusing revealed subtle charge heterogeneity differences between the two endoglycosidases, manifested by the lack of extra acidic charge variants in the cIEF trace of the 6His-PNGase F enzyme, which might have possibly influenced the glucose-mediated enzyme activity differences. Full article
(This article belongs to the Special Issue Capillary Electrophoresis Analysis: Trends and Recent Advances)
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13 pages, 1729 KiB  
Article
Comparison of the Performance of Different Bile Salts in Enantioselective Separation of Palonosetron Stereoisomers by Micellar Electrokinetic Chromatography
by Shaoqiang Hu, Tao Sun, Rui Li, Dongdong Zhang, Yonghua Zhang, Zhuo Yang, Ge Feng and Xuming Guo
Molecules 2022, 27(16), 5233; https://doi.org/10.3390/molecules27165233 - 16 Aug 2022
Cited by 4 | Viewed by 1818
Abstract
Bile salts are a category of natural chiral surfactants which have ever been used as the surfactant and chiral selector for the separation of many chiral compounds by micellar electrokinetic chromatography (MEKC). In our previous works, the application of sodium cholate (SC) in [...] Read more.
Bile salts are a category of natural chiral surfactants which have ever been used as the surfactant and chiral selector for the separation of many chiral compounds by micellar electrokinetic chromatography (MEKC). In our previous works, the application of sodium cholate (SC) in the separation of four stereoisomers of palonosetron (PALO) by MEKC has been studied systematically. In this work, the parameters of other bile salts, including sodium taurocholate (STC), sodium deoxycholate (SDC), and sodium taurodeoxycholate (STDC) in the separation of PALO stereoisomers by MEKC were measured and compared with SC. It was found that all of four bile salts provide chiral recognition for both pairs of enantiomers, as well as achiral selectivity for diastereomers of different degrees. The structure of steroidal ring of bile salts has a greater impact on the separation than the structure of the side chain. The varying separation results by different bile salts were elucidated based on the measured parameters. A model to describe the contributions of the mobility difference of solutes in the aqueous phase and the selectivity of micelles to the chiral and achiral separation of stereoisomers was introduced. Additionally, a new approach to measure the mobility of micelles without enough solubility for hydrophobic markers was proposed, which is necessary for the calculation of separation parameters in MEKC. Under the guidance of derived equations, the separation by SDC and STDC was significantly improved by using lower surfactant concentrations. The complete separation of four stereoisomers was achieved in less than 3.5 min by using 4.0 mM of SDC. In addition, 30.0 mM of STC also provided the complete resolution of four stereoisomers due to the balance of different separation mechanisms. Its applicability for the analysis of a small amount of enantiomeric impurities in the presence of a high concentration of the effective ingredient was validated by a real sample. Full article
(This article belongs to the Special Issue Capillary Electrophoresis Analysis: Trends and Recent Advances)
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