Validation of a High‐Performance Liquid Chromatography Separation of Weak Acids, Neutral Compounds, and Permanent Ions Using Sequential Elution with Tandem‐Column Liquid Chromatography
Year: 2025
Type: article
Abstract: ABSTRACT Sequential elution‐liquid chromatography (SE‐LC) employs conventional high‐performance LC instrumentation to separate compounds by class and within each class using two or more sequential elution modes. The aim of this work was to validate a novel SE‐LC method used to separate weak acids, permanently charged ions (anions), and neutral compounds by applying first a low pH isocratic elution to elute the weak acids, then an acetonitrile gradient to elute the neutral compounds, and lastly a sodium methanesulfonate gradient to elute the anionic compounds using a tandem‐column arrangement comprised of a C18 column and a strong anion exchange column. The effects of flow rate, starting mobile phase organic content, column temperature, mobile phase composition, and gradient start time were investigated using a 25‐1 fractional factorial design with center points for robustness. The method was demonstrated to be robust, specific, linear, accurate, and precise (repeatable) over the range of 6%–120% of the target analyte concentrations.
Source: Journal of Separation Science
DOI: 10.1002/jssc.70124
Institutions: GlaxoSmithKline (United States), Drexel University
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: bronze
OpenAlex ID: W4408724965