Souleiman El Balkhi (Presenter)
CHU de LIMOGES
Authorship: Lauranne Poncelet(1), Maxime Chaslot(1), Sylvain Dulaurent(1), Souleiman El Bakhi(1), Levi Mikaël(2), Franck Saint-Marcoux(1)
(1) CHU Limoges, France. Department of pharmacology and toxicology (2) Shimadzu, France
| Short Abstract A method for routine analysis of 43 benodiazepines by UHPLC-MS/MS was developed. An original sample preparation method was used and validated to simplify workflow in the laboratory and accelarate result reporting. The method was validated in blood, serum and urine and performances were tested in 50 pateint samples. |
Long Abstract
Objectives: The aim of this study was to develop an analytical procedure for the measurement of benzodiazepines, where the extraction step is based on QuEChERS salts, for application in clinical toxicology and forensic science.
Methods: 43 benzodiazepines were considered: 24 commercialized compounds, 10 of their metabolites, 8 designer benzodiazepines and 2-amino-5 chloropyridine (ACP ; degradation product of zopiclone). The LC-MS/MS system consisted in a Pinnacle DB PFPP (50 mm x 2.1 mm with particle size of 1.9 µm ; Restek) column coupled with a 8050 MS system (Shimadzu®, France). The whole procedure was validated according to ISO 15189 standard (except for the 8 designer benzodiazepines and ACP) and its performance was tested in the biological samples of 50 patients.
Results: The procedure was validated from 1 or 5 µg/L to 2000 µg/L, for the 24 benzodiazepines and 10 metabolites, when measured on a 100 µL biological sample (whole blood, serum or urine). 123 molecules were detected in the 50 samples; among them, 19 at concentrations below 5 µg/L. In the routine, about 30 to 45min were needed for the preparation of 10 patient samples, 2 internal quality controls and 6 points of calibration standards. This produced results in less than 3 hours, which reduces to 1 hour for a single patient.
Conclusion: We developed and validated a procedure with a one-step extraction using QuEChERS salts for the determination of multiple benzodiazepines and their metabolites.
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