Self-Classified Topic Area(s): Small Molecules / Tox / TDM
Integrated Toxicological Screening and Confirmation Analysis with Stable 24/7 Availability
Frank Streit (1), Gry Dihazi (2), Marcel Grapp (3), Tatjana Khromov (4), Lutz Binder (5) University Medical Center, Goettingen, Germany
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Frank Streit (Presenter) UMG Goettingen, Germany
Presenter Bio: Dr.rer.nat. Frank Streit from the university medical center in Goettingen, Germany.
As senior scientist in the interdisciplinary UMG-Laboratories his task area covers routine analysis, education, research and development. He is the head of their mass spec research unit. Next to TDM and Toxicology, their peptidomics, metabolomics and proteomics unit supports research groups in oncology cardiology and neurology.
Relevant Financial Disclosures
(within past 24 months)
No relevant financial relationship(s) to disclose.
Abstract
Introduction: Toxins or poisons are substances which already in small doses may lead to malfunction, damage to health or death. Prompt toxicological analysis may allow early identification of the toxic agent and give therapeutic guidance. In clinical laboratories, screening methods by immunoassays using polyclonal antibodies are commonly applied. Antibody-based screening, however, may cause a relevant number of false positive and false negative results. Detected drugs are not quantified. Furthermore, immunological screening requires a set of 5 to 10 different assays. Nonetheless, comprehensive drug screening and quantification of relevant substances by chromatography with mass spectrometry is often indispensable.
Objectives: To overcome these shortcomings, our aim was the development of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for a targeted screening of a broad toxicological spectrum with stable 24/7 availability and without need of special skilled personnel. To verify instrument stability, downtimes due do maintenance and service were investigated.
Methods: All relevant compounds from a list of the substances most frequently detected in patient samples addressed to the GIZ-Nord Poison Center at the University Medical Center Goettingen in the years 2014-2018 were selected and integrated into the targeted screening method.
We used a Clinical Laboratory Automated sample preparation Module (CLAM-2030, Shimadzu) connected to an LC-MS/MS system (LCMS-8050, Shimadzu). We reanalyzed routine patient samples and compared results of typical antibody-based screening and our targeted screening method. In addition all samples were analyzed with our untargeted LC-QToF screening method for forensic toxicological purposes. Downtimes, maintenance and service times of the CLAM-2030 and LCMS-8050 will be shown.
Results: We successfully developed a fast and fully automated LC-MS/MS method for the quantitation of 152 relevant toxicological compounds. 100 patient serum samples were investigated and results compared with our routine toxicological screening by LC-QToF. All relevant substances could be identified using the novel LC-MS/MS screening method (triple quad) in comparison with our well established comprehensive screening procedure with LC-QToF. Repeated measurements of quality control samples showed an inaccuracy of less than 20 % for most of the tested substances. Hence, quantification by one-point calibration is well suited for assessing the severity of an intoxication.
Conclusion: We established a fully automated, robust, quantitative screening method for the detection of more than 150 exogenous substances in serum. It allows a targeted toxicological emergency screening in clinical laboratories without LCMS-skilled personnel. To further refine the method, evaluation and interpretation of the results may be improved and automated.
= Discovery stage.
= Translation stage.
= Clinically available.
