Biochemistry & Pharmacology: Open Access
Open Access
ISSN: 2167-0501
+44-20-4587-4809
ISSN: 2167-0501
+44-20-4587-4809
Tamara Alvarez-Segura, Jose Ramon Torres-Lapasio and Maria Celia Garcia-Alvarez-Coque
University of Valencia, Spain
Posters & Accepted Abstracts: Biochem Pharmacol (Los Angel)
Liquid chromatography with single columns often does not succeed in the analysis of complex samples, in terms of resolution and
analysis time. A relatively simple solution to enhance chromatographic resolution is the modulation of the stationary phase through
the serial coupling of columns. This can be implemented with any type of column using compatible elution conditions and conventional
instruments. This review describes the key features of column coupling and published procedures, where two or more columns were
coupled in series to solve separation problems in chiral liquid chromatography. In most reports, two chiral columns are coupled.
However, coupling chiral and achiral stationary phases is also an active field, to analyze samples containing mixtures of chiral and
achiral compounds. In all reports, the authors could not resolve their samples with single columns, whereas significant enhancement in
chromatographic performance was obtained when the columns were combined. Particularly interesting is the reduction in the analysis
time in the isocratic mode, which alleviates the general elution problem of liquid chromatography, and may represent a stimulus for
the proposal of new procedures, especially in combination with mass spectrometric, electrochemical and refractometric detection.
Developments proposed to make the serial coupling of columns useful in routine and research laboratories are outlined, including
optimisation strategies that facilitate the selection of the appropriate column combination and elution conditions (solvent content, flow
rate or temperature) in both isocratic and gradient modes. The availability of zero dead volume couplers, able to connect standard
columns, and the commercialisation of short columns with multiple lengths, has expanded the possibilities of success.
Recent Publications
1. Alvarez-Segura T, Garc�?a-Alvarez-Coque MC, Ortiz-Bolsico C and Torres-Lapasi�?³ JR (2015) Interpretative approaches
to optimize serially-coupled columns in reversed-phase liquid chromatography. Current Chromatography 2:110-121.
2. Alvarez-Segura T, Torres-Lapasi�?³ JR, Ortiz-Bolsico C and Garc�?a-Alvarez-Coque C (2016) Stationary phase modulation
in liquid chromatography through the serial coupling of columns: A review. Analytica Chimica Acta 923:1-23.
3. Chu YQ and Wainer IW (1989) Determination of the enantiomers of verapamil and norverapamil in serum using
coupled achiral-chiral high-performance liquid. Journal of Chromatography B 497:191-200.
4. Ferretti R, Gallinella B, La Torre F and Zanitti L (1998) Direct resolution of a new antifungal agent, voriconazole
(UK-109,496) and its potential impurities, by use of coupled achiral-chiral high-performance liquid chromatography.
Chromatographia 47:649-654.
5. Johnson DV and Wainer IW (1996) Enantioselective separation of cyclic chiral ketones and their corresponding
diastereomeric alcohols by HPLC on chiral and chiral/chiral coupled stationary phases. Chirality 8:551-555.
Tamara Alvarez-Segura has completed her Degree in Chemistry in 2013 and Master degree in Experimental Techniques in Chemistry in 2014, both at the University of Valencia. She is now performing diverse research activities in the Department of Analytical Chemistry to get her PhD degree. She has received two pre-doctoral fellowships. During her Master studies, she began her collaboration with María Celia García-Alvarez-Coque and José Ramón Torres-Lapasió in the field of the modulation of the selectivity in HPLC, using serially coupled columns and other strategies to analyse complex samples. She has written 11 research articles and presented several communications in international meetings. The PhD period included a three-month stay (September to November 2017) in the Analytical Chemistry Department of the University of Barcelona (Spain), under the supervision of Prof Martí Rosés, working in the field of Hydrophilic Interaction Liquid Chromatography (HILIC).
E-mail: tamara.alvarez@uv.es