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Separation of weak acids, neutral compounds, and permanent anions using sequential elution liquid chromatography with tandem columns.

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Title: Separation of weak acids, neutral compounds, and permanent anions using sequential elution liquid chromatography with tandem columns.
Authors: Lovejoy, Lauren Kline1,2 (AUTHOR), Foley, Joe P.1,2 (AUTHOR) jfoley@drexel.edu
Source: Journal of Chromatography A. Aug2024, Vol. 1731, pN.PAG-N.PAG. 1p.
Subjects: Liquid chromatography, Column chromatography, RF values (Chromatography), Chromatographic analysis, High performance liquid chromatography
Abstract: • An alternative elution mode to conventional HPLC to separate compounds by class. • Sequential elution was utilized to separate classes of compounds in under 15 min. • A tandem-column stationary phase combination (SAX and C18) was utilized. • SAX+C18 and C18+SAX column orientations yielded nearly identical separations. • Retention time and peak area precision were below 1.5 % RSD. This paper discusses the development of an analytical method by an alternative separation approach, sequential elution liquid chromatography (SE-LC), to separate permanently charged ions (anions), weak acids, and neutral compounds using anion exchange and reversed-phase columns in tandem. SE-LC separates classes of compounds by group by employing two or more elution modes. Advantages to using SE-LC over conventional HPLC are a greater peak capacity and a reduced separation disorder. Importantly, the same HPLC as used for a conventional HPLC separation may be used to afford a successful SE-LC separation. Mobile phase selection and gradient optimization are integral for a successful SE-LC class separation of permanent anions, weak acids, and neutral compounds and will be discussed in detail in this paper. The most successful (best resolution and repeatability) SE-LC separation was achieved by applying isocratic elution at low pH to elute the weak acids, followed by an acetonitrile gradient to elute the neutral compounds, and last a sodium methanesulfonate gradient to elute the anionic compounds using a superficially porous C18 column coupled with a strong anion exchange (SAX) column. Repeatability (RSD) in the retention times and peak areas of the analytes was less than 0.25 % and 1.5 %, respectively. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
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Abstract:• An alternative elution mode to conventional HPLC to separate compounds by class. • Sequential elution was utilized to separate classes of compounds in under 15 min. • A tandem-column stationary phase combination (SAX and C18) was utilized. • SAX+C18 and C18+SAX column orientations yielded nearly identical separations. • Retention time and peak area precision were below 1.5 % RSD. This paper discusses the development of an analytical method by an alternative separation approach, sequential elution liquid chromatography (SE-LC), to separate permanently charged ions (anions), weak acids, and neutral compounds using anion exchange and reversed-phase columns in tandem. SE-LC separates classes of compounds by group by employing two or more elution modes. Advantages to using SE-LC over conventional HPLC are a greater peak capacity and a reduced separation disorder. Importantly, the same HPLC as used for a conventional HPLC separation may be used to afford a successful SE-LC separation. Mobile phase selection and gradient optimization are integral for a successful SE-LC class separation of permanent anions, weak acids, and neutral compounds and will be discussed in detail in this paper. The most successful (best resolution and repeatability) SE-LC separation was achieved by applying isocratic elution at low pH to elute the weak acids, followed by an acetonitrile gradient to elute the neutral compounds, and last a sodium methanesulfonate gradient to elute the anionic compounds using a superficially porous C18 column coupled with a strong anion exchange (SAX) column. Repeatability (RSD) in the retention times and peak areas of the analytes was less than 0.25 % and 1.5 %, respectively. [ABSTRACT FROM AUTHOR]
ISSN:00219673
DOI:10.1016/j.chroma.2024.465178