Pharmaceutical analysis of drugs and other biologically active substances

Analytical Chemistry Group

The activity of Analytical Chemistry group led by Prof. RNDr. Dalibor Šatínský, Ph.D. is divided into four research workgroups according to the analytical methods – 1. Liquid Chromatography and related separation techniques (Supercritical Fluid Chromatography and Gas Chromatography), 2. Mass Spectrometry hyphenated with separation methods for the analysis of small molecules and large biomolecules, 3. Flow techniques based on Flow Injection Analysis, Sequential Injection Analysis, and Lab-In-Syringe, and 4. Capillary Electrophoresis. The group benefits from both its own excellent scientists and international cooperation with renowned workgroups and scientists.

The first workgroup led by Prof. PharmDr. Lucie Nováková, Ph.D. develops methods for clinical analysis of biologically active substances in various biological materials (urine, blood serum, breast milk, lipoproteins, etc.), for analysis of selected natural substances (antioxidants) in biological samples, plant extracts, and nutraceuticals, and for analysis of drugs and markers in samples from in vitro and in vivo studies. Another significant aim is the analysis of biopharmaceuticals, both manufactured substances and clones of antibodies produced by means of biotechnology. The work includes original procedures for sample pre-treatment, chromatographic separation, and mass spectrometric detection. Original miniaturized and automated methods and processes are developed to process samples before analysis. New trends (modern stationary phases, Ultra-High Performance Liquid Chromatography, Supercritical Fluid Chromatography, 2D Chromatography,…) are followed in the development of new methods too. Supercritical Fluid Chromatography is rapidly developed these years, especially in the area of chiral separations. New methods are often developed in cooperation with Czech partners (the Palacký University Olomouc, the University Hospital Hradec Králové) and foreign institutions (the University of Geneva, the University of Oslo, the University of Tasmania,...).

The second workgroup led Prof. RNDr. Dalibor Šatínský, Ph.D. deals with automated methods for sample pre-treatment using High Performance Liquid Chromatography aiming the analysis of antioxidants, contaminants, and mycotoxins in food and environmental samples, the drug analysis, and analysis of nutraceuticals of plant origin. The methods use current trends – 2D Chromatography, Solid Phase Extraction using nanofibrous polymers, and molecularly imprinted polymers. The development of unique nanofibrous polymers is enabled thanks to close cooperation with the Technical University Liberec. Another cooperation with the Research and Breeding Institute of Pomology in Holovousy focuses on profiling of phenolic acids, flavonoids, and other antioxidants in fruits (apples, cherries, sour cherries, blueberries, currant,…) for evaluation of new cultivars.

The third workgroup led by Assoc. Prof. PharmDr. Hana Sklenářová, Ph.D. develops methods based on the flow methods for automated and miniaturized analysis, sample pre-treatment before further analysis, and monitoring of long-time processes. The methods utilize variability and properties of both continuous flow methods (Flow Injection Analysis and Sequential Injection Analysis for Solid Phase Extraction of organic and inorganic substances, monitoring of kinetics of long-time processes, separations in Sequential Injection Chromatography and Luminescence detection) and non-continual flow methods (Lab-In-Syringe – for miniaturized liquid-liquid extraction, dispersion micro-extraction,…). Due to the expansion of flow methods rather abroad, new methods are often developed in collaboration with the University of Balearic Islands, the University of Porto, the University of Melbourne, the University of Hawaii at Manoa, the Universidad Nacional del Sur, The Saint Petersburg State University and others. The close collaboration on new flow methods and components is linked to the two biggest flow systems manufacturers - FIAlab® (Seattle, USA) and GlobalFIA® (Fox Island, USA), which led to several significant innovations in flow techniques.

The fourth workgroup led by PharmDr. Pavel Jáč, Ph.D. focuses on the use of Capillary Electrophoresis (CE) in pharmaceutical analysis. Key activities of the workgroup include the development of novel CE methods for the analysis of the content of pharmaceuticals and dietary supplements, including analysis of impurities. Methods of chemometrics are broadly used for the optimization of the analysis conditions and for tests of the robustness of the developed methods. Another research topic is chiral separations using CE, wherein the cooperation with the Department of Organic and Bioorganic Chemistry, Faculty of Pharmacy, new commercial and lab-made chiral selectors based on ion liquids are tested. In the area of chiral separations, the workgroup cooperates with Prof. Gerhard Scriba (the Friedrich-Schiller-University Jena). The latest focus of the workgroup is on the use of CE hyphenated with Mass Spectrometry for analysis of the authenticity of dietary supplements, adulteration of food, and determination of residues of pesticides, drugs, and mycotoxins in food.

Pharmaceutical Analysis Group

The Pharmaceutical Analysis Group led by Assoc. Prof. PharmDr. Radim Kučera, Ph.D. focuses the research activities on the use of separation methods in pharmaceutical and analytical studies. The attention is paid especially to three main areas:

Bioanalytical studies of drugs - the main goal is the development of modern bioanalytical methods (especially UHPLC-MS / MS) for the determination of new drugs and potential drugs in biological materials. In cooperation within the faulty and with the Department of Pharmacology, Faculty of Medicine in Hradec Kralove the developed methods are applied in the study of bioactivation, transport mechanisms, pharmacokinetics and metabolism of selected drugs (Šesták et al., 2015; Jirkovský et al., 2018). Modern bioanalytical approach will serve as a basic methodological tool for studying the mechanism of action of the various substances as well as for the development of new drugs. Additionally, the attention is paid to sample preparation techniques prior to the analysis employing promising materials such as modified zirconia or polymers. The aspects of chirality in drug metabolism are addressed as well (Nobilis et al., 2013);

Alternative and classical materials in pharmaceutical analysis - in this field the experimental work is focused on the possibilities of using stationary phases especially on the basis of metal oxides in the analysis of pharmaceutically important substances and the study of retention mechanisms in chromatography on these materials (Kučera et al., 2014; et al., 2012). Alternative stationary phases are tested in development and validation of new HPLC and UHPLC methods for quality testing of active substances and medicinal products. These materials are also tested for use in high temperature chromatography. Newly the use of graphene in capillary electrochromatography is studied.

Stability study of drugs, analytical evaluation of API and excipients - the third area of ​​interest covers the issues of stability of API, potential drugs, and pharmaceutical preparations.  The special attention focuses on the identification of major degradation products, the investigation of degradation mechanism and the calculation of kinetic characteristics of the compound decay. The development of separation methods for the evaluation of related substances in pharmaceuticals, excipients, natural products and also in molecular probes is the subject of interest as well.

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