Journal
ANALYTICA CHIMICA ACTA
Volume 1221, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.aca.2022.340035
Keywords
Marker; Isoelectric point; Focusing; Capillary; Colored
Categories
Funding
- Czech Academy of Sciences [VI20172020069, BV III/1-VS]
- Safety Research Programme of the Czech Republic for 2015-2020 [VI20172020069, BV III/1-VS]
- Czech Academy of Sciences institutional [RVO:68081715]
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We present a set of novel low-molecular-mass compounds with ampholytic properties that can be used as markers of isoelectric point in different isoelectric focusing formats. These compounds have excellent focusing properties and stability, making them suitable for various isoelectric focusing methods.
We present a set of novel low-molecular-mass (LMM) compounds possessing ampholytic properties. The compounds were designed to perform as markers of isoelectric point (pI) in different isoelectric focusing (IEF) formats and feature direct detectability in UV and visible wavelength regions. Capillary isoelectric focusing (cIEF) was used to determine the purity of the focusing species and the compounds' pI values. Nitrophenol-based pI markers (NPIMs) published previously were used as standards for the pI value calibration. The presented compounds focused very well, but small portion of them contained focusing impurities, thus, we recommend them for use in other IEF formats like gel IEF and preparative IEF. Moreover, multi-wavelength detection enabled determination of individual markers based on their specific spectral profiles and different absorption at selected detection wavelengths in the electropherogram. The presented compounds compose a group of chemicals featuring excellent shelf stability and isoelectric focusing properties, inexpensive synthesis, universal/multimode detectability, and good solubility at pI. The presented results provide a solid ground for their use as reference standards in various isoelectric focusing methods.
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