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Licensed Unlicensed Requires Authentication Published by De Gruyter July 1, 2020

Chromatographic methods development for clinical practice: requirements and limitations

Lenka Kujovská Krčmová ORCID logo, Bohuslav Melichar and František Švec ORCID logo

Abstract

Development of a chromatographic method in bioanalysis is a challenging and complex procedure with many pitfalls and often unexpected reversals that can require several months to accomplish. Even an experienced analytical team must contend many limitations mainly in connection with the strict requirements imposed on current clinical research. These restrictions typically persist throughout the whole development process, from clinical trial assignment, across optimization of extraction of biological materials and chromatographic separation, to validation and data interpretation. This paper describes questions and their possible answers raised during the pre-analytical phase such as use of modern sample preparation techniques in clinical methods, application of internal standards, as well as selection of stationary phases and detection techniques in the analytical phase. Validation problems and interpretation of results are demonstrated with three typical examples of characteristics to be considered, i.e. recovery, matrix effect, and limit of detection vs. lower limit of quantification.


Corresponding author: Assoc. Prof. Lenka Kujovská Krčmová, PhD, The Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Akademika Heyrovského 1203, 500 05 Hradec Králové, Czech Republic; The Department of Clinical Biochemistry and Diagnostics, University Hospital, Sokolská 581, 500 05 Hradec Králové, Czech Republic, Phone: +420 49583337, Fax: +420 495834841, E-mail:

Funding source: Charles University, Faculty of Pharmacy in Hradec Králové

Funding source: University Hospital in Hradec Králové

Funding source: Ministry of Health of the Czech Republic

Funding source: Ministry of Health of the Czech Republic

Funding source: Ministry of Health of the Czech Republic

Funding source: PERSONMED – Center for Development of Personalized Medicine in Age-Related Diseases

Funding source: STARSS project co-funded by ERDF

Acknowledgments

The authors gratefully acknowledge Filip Hátle for technical support. Thanks are also due for the financial support by Charles University, Faculty of Pharmacy in Hradec Králové (Project SVV 260 458), project MH CZ – DRO (University Hospital in Hradec Králové, 00179906), the Ministry of Health of the Czech Republic, grants numbers 18-03-00130, 17-28882A, and 17-29241A, PERSONMED – Center for Development of Personalized Medicine in Age-Related Diseases, Reg Nr. CZ.02.1.01/0.0/0.0/17_048/0007441 co-financed by the ERDF state budget of the Czech Republic. FS gratefully acknowledges the STARSS project (Reg. No. CZ.02.1.01/0.0/0.0/15_003/0000465) co-funded by ERDF.

  1. Research funding: Thanks are due for the financial support by Charles University, Faculty of Pharmacy in Hradec Králové (Project SVV 260 458), project MH CZ – DRO (University Hospital in Hradec Králové, 00179906), the Ministry of Health of the Czech Republic, grant numbers 18-03-00130, 17-28882A, and 17-29241A, PERSONMED – Center for Development of Personalized Medicine in Age-Related Diseases, Reg Nr. CZ.02.1.01/0.0/0.0/17_048/0007441 co-financed by the ERDF state budget of the Czech Republic. FS gratefully acknowledges the STARSS project (Reg. No. CZ.02.1.01/0.0/0.0/15_003/0000465) co-funded by ERDF.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

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Received: 2020-04-16
Accepted: 2020-05-25
Published Online: 2020-07-01
Published in Print: 2020-10-25

© 2020 Walter de Gruyter GmbH, Berlin/Boston

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