Rapid and simple multi-analyte LC–MS/MS method for the determination of benzodiazepines and Z-hypnotic drugs in blood samples: Development, validation and application based on three years of toxicological analyses
Section snippets
INTRODUCTION
Benzodiazepines (BZDs) are a large class of substances that are amongst the most frequently prescribed psychotropic pharmaceuticals worldwide [1]. They were introduced to medical practice in the 1960s and have led to the phase out of the previously used meprobamate and compounds from the barbiturate group due to relatively low toxicity and high efficacy [2,3]. It is estimated that approximately 10 - 20% of adults take BZDs regularly. [2]. Recently, a number of new benzodiazepines (named
Chemicals and reagents
Certified standards were purchased from the following manufacturers: 7-aminoflunitrazepam, bromazepam, chlordiazepoxide, estazolam, etizolam, flubromazepam, lormetazepam, medazepam, phenazepam, tetrazepam, triazolam, diazepam-D5, zaleplon and zopiclone were from LGC Standards (London, UK); benzodiazepine multi-component mixture-8 (alprazolam, clonazepam, diazepam, flunitrazepam, lorazepam, nitrazepam, oxazepam, temazepam), 7-aminoclonazepam (7-AC), midazolam, and prazepam were from Cerilliant
Results and discussion
The analysis of BZDs is still a challenge for both clinical and forensic laboratories, mainly because the number of new compounds is constantly growing. Therefore, it is incredibly important to develop a new method that will combine all the desired characteristics, such as, simplicity, specificity, sensitivity, low cost and the ability to detect a large number of compounds in one analytical run.
Conclusion
Currently, LLE has become an increasingly popular sample preparation method in forensic and clinical applications due to its many advantages compared to SPE-based procedures. In this paper, a fast, selective, sensitive, and robust method utilizing LLE and LC–MS/MS for the determination of wide range of BZD, their metabolites and Z-Drugs most often used in medicine was developed. The validation was performed for 26 compounds because bromazepam is included only for the detection.. A few DBZDs,
Ethical approval
All blood collections from living suspects and deceased were made by judicial authorities and the samples were sent to Forensic Science Department (Gdańsk) for routine toxicology analysis. All analyses were made according to the request of judicial authorities.
CRediT authorship contribution statement
Laura Banaszkiewicz: Conceptualization, Methodology, Validation, Formal analysis, Investigations, Writing-original draft. Mateusz Kacper Woźniak: Conceptualization, Data analysis, Methodology, Funding acquisition, Writing-original draft, Writing-review & editing, Supervision. Marzena Kata: Formal analysis, Investigations, Writing-review & editing. Ewa Domagalska: Investigations, Data curation, Writing-review & editing. Marek Wiergowski: Supervision, Writing-review & editing. Beata Szpiech:
Declaration of Competing Interest
The authors report no declarations of interest.
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Overview of the bioanalytical methods used for the determination of benzodiazepines in biological samples and their suitability for emergency toxicological analysis
2023, Journal of Pharmacological and Toxicological MethodsA multi-analyte LC-MS/MS method for the determination of 57 pharmaceuticals and illicit drugs in plasma, and its application to poisoning cases
2023, Journal of Pharmaceutical and Biomedical AnalysisCitation Excerpt :Compared to other protocols using PP, the developed method required smaller amounts of sample (50 µL) and solvent (240 µL), in addition to dispensing with the solvent drying step [20,21,25,26]. Besides that, the chromatographic run time was 7 min, shorter than other multi-analyte studies using LC-MS/MS already published [14,21,26,27]. For optimization of precipitation solvent, the geometric mean of the absolute areas of the peaks of all analytes was calculated.
Liquid chromatography in forensic toxicology
2023, Liquid Chromatography: ApplicationsInterpol review of toxicology 2019–2022
2023, Forensic Science International: SynergyCitation Excerpt :Method validations using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for detection of NPS had been done on various biological matrices including blood, urine and oral fluid [3–9]. The use of LC-MS/MS for analyzing benzodiazepines and hypnotic drugs in blood and/or urine was validated for toxicology analyses [10,11]. The determination of opiates in blood [12] and hair [13] by derivatization of opiates prior to gas chromatography-tandem mass spectrometry (GC-MS/MS) was validated and demonstrated for quantifying analytes at low levels.