Fully automated sample preparation procedure to measure drugs of abuse in plasma by liquid chromatography tandem mass spectrometry
For the analysis of drugs and pharmaceutical compounds in biological matrices, extraction procedures are typically used for LC-MS/MS analysis often requiring manual steps in sample preparation. In this study, we report a fully automated extraction method carried out by a programable liquid handler directly coupled to an LC-MS/MS system for the determination of 42 components (illicit drugs and/or metabolites) (plus 20 deuterated internal standards). The acquisition was performed in positive ionization mode with up to 15 MRM transitions per compound, each with optimized collision energy (MRM spectrum mode) to enable qualitative library searching in addition to quantitation. After placing the sample tube into the system, no further intervention was necessary: automated preparation used 50 μL of blood or plasma with 3 μL of extracted sample injected for analysis. The method was validated according to the requirements of ISO 15189. The limit of detection and quantification was 1–5 ng/mL depending on the compound. Stability experiments found that historic calibration curve data files could accurately quantify for up to 1 month with less than 20% uncertainty. Comparison to a QuEChERS method was made using patient samples providing a regression correlation R2 = 0.98 between the two methods. This approach was successfully designed to support parallel sample preparation and analysis therefore significantly increasing sample throughput and reduced cycle times.
KeywordsLiquid chromatography Mass spectrometry Automated sample preparation Drugs of abuse
Drugs of abuse
Lower limit of detection
Lower limit of quantitation
Multiple reaction monitoring
Upper limit of quantitation
Compliance with ethical standards
The study has been carried out using biological samples of the biobank “CRBioLim” of the CHU of Limoges, authorized by regulatory authorities (French Ministry of Research and Regional Health Agency) and ethical committee “CPP SOOM 4” with the reference AC 2016-2748, according to the French regulation. All the samples stored in “CRBioLim” have been given by patients who did not oppose to the use of these samples for the development of an analytical method, as allowed by the French regulation (mainly the law on bioethics n° 2004-800 of August 6, 2004).
Conflict of interest
Author, Tiphaine Robin, is funded 50% by CIFRE and 50% by Shimadzu Corporation. All authors declare that they have no conflict of interest.
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