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Benzodiazepines: sample preparation and HPLC methods for their determination in biological samples
Victoria F Samanidou1, Mohammad N Uddin, Ioannis N Papadoyannis
1Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki, GR-541 24, Greece. samanidu@chem.auth.gr
Bioanalysis
|November 19, 2010
Summary
Benzodiazepines (BDZs) analysis in biological samples requires effective sample preparation and HPLC methods. This review details techniques for accurate and sensitive BDZ determination in clinical and forensic toxicology.
Area of Science:
- Analytical Chemistry
- Forensic Toxicology
- Pharmacology
Background:
- Benzodiazepines (BDZs) possess sedative, hypnotic, and anticonvulsant properties, necessitating their accurate determination in biological fluids.
- Clinical assays, forensic investigations, and toxicological studies rely on precise quantification of BDZs and their metabolites.
- Current analytical methods for BDZ determination face challenges due to trace levels in complex biological matrices.
Purpose of the Study:
- To provide a comprehensive review of sample preparation techniques for benzodiazepine analysis.
- To critically evaluate High-Performance Liquid Chromatography (HPLC) conditions used in various studies for BDZ determination.
- To highlight advancements in achieving sensitive and accurate detection of benzodiazepines in biological samples.
Main Methods:
- Review of extraction techniques including solid-phase extraction (SPE) and liquid-liquid extraction (LLE).
- Analysis of various HPLC conditions, column types, and mobile phases.
- Comparison of detection methods such as UV/diode array detection (DAD), electron capture detection (ECD), nitrogen phosphorus detection (NPD), and mass spectrometry (MS).
Main Results:
- Sample preparation is crucial for isolating trace levels of BDZs from interfering compounds in biological matrices.
- HPLC methods coupled with sensitive detectors allow for detection limits down to 1-2 ng/ml (UV/DAD) and below 1 ng/ml (ECD, NPD).
- Mass spectrometry, particularly with negative chemical ionization or sonic spray ionization, offers the highest sensitivity, reaching 0.1 ng/ml.
Conclusions:
- Effective sample preparation and optimized HPLC conditions are vital for accurate benzodiazepine quantification.
- Advanced detection techniques significantly enhance sensitivity, enabling reliable trace-level analysis in toxicological contexts.
- The review provides a valuable resource for bioanalysts developing and validating methods for benzodiazepine determination.
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