Analytical and Bioanalytical Chemistry

, Volume 410, Issue 20, pp 4855–4866 | Cite as

Characterization of bonded stationary phase performance as a function of qualitative and quantitative chromatographic factors in chaotropic chromatography with risperidone and its impurities as model substances

  • Jelena Čolović
  • Milena Rmandić
  • Anđelija MalenovićEmail author
Research Paper


Numerous stationary phases have been developed with the aim to provide desired performances during chromatographic analysis of the basic solutes in their protonated form. In this work, the procedure for the characterization of bonded stationary phase performance, when both qualitative and quantitative chromatographic factors were varied in chaotropic chromatography, was proposed. Risperidone and its three impurities were selected as model substances, while acetonitrile content in the mobile phase (20–30%), the pH of the aqueous phase (3.00–5.00), the content of chaotropic agents in the aqueous phase (10–100 mM), type of chaotropic agent (NaClO4, CF3COONa), and stationary phase type (Zorbax Eclipse XDB, Zorbax Extend) were studied as chromatographic factors. The proposed procedure implies the combination of D-optimal experimental design, indirect modeling, and polynomial-modified Gaussian model, while grid point search method was selected for the final choice of the experimental conditions which lead to the best possible stationary phase performance for basic solutes. Good agreement between experimentally obtained chromatogram and simulated chromatogram for chosen experimental conditions (25% acetonitrile, 75 mM of NaClO4, pH 4.00 on Zorbax Eclipse XDB column) confirmed the applicability of the proposed procedure. The additional point was selected for the verification of proposed procedure ability to distinguish changes in solutes’ elution order. Simulated chromatogram for 21.5% acetonitrile, 85 mM of NaClO4, pH 5.00 on Zorbax Eclipse XDB column was in line with experimental data. Furthermore, the values of left and right peak half-widths obtained from indirect modeling were used in order to evaluate performances of differently modified stationary phases applying a half-width plots approach. The results from half-width plot approach as well as from the proposed procedure indicate higher efficiency and better separation performance of the stationary phase extra densely bonded and double end-capped with trimethylsilyl group than the stationary phase with the combination of end-capping and bidentate silane bonding for chromatographic analysis of basic solutes in RP–HPLC systems with chaotropic agents.

Graphical abstract


Indirect modeling Polynomial modified Gaussian model Chromatogram simulation Half-width plots approach 



The authors thank the Ministry of Education, Science and Technological Development of the Republic of Serbia for supporting this investigation through the Project 172052.

Compliance with ethical standards:

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

216_2018_1122_MOESM1_ESM.pdf (151 kb)
ESM 1 (PDF 151 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Jelena Čolović
    • 1
  • Milena Rmandić
    • 1
  • Anđelija Malenović
    • 1
    Email author
  1. 1.Faculty of Pharmacy, Department of Drug AnalysisUniversity of BelgradeBelgradeSerbia

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