Chromatographic methods development for clinical practice: requirements and limitations

• Published in: Clinical chemistry and laboratory medicine Vol. 58; no. 11; pp. 1785 - 1793
• Main Authors: Krčmová, Lenka Kujovská, Melichar, Bohuslav, Švec, František
• Format: Journal Article
• Language: English
• Published: Germany De Gruyter 01.11.2020; Walter De Gruyter & Company
• Subjects: Biological materials; Chromatography; clinical research; clinical study; Data interpretation; laboratory medicine; liquid chromatography; Mathematical analysis; Optimization; Sample preparation; validation; clinical study; laboratory medicine; liquid chromatography; clinical research; validation
• ISSN: 1434-6621; 1437-4331
• DOI: 10.1515/cclm-2020-0517

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.