Development of a validated HPLC method for the determination of four 1,4-benzodiazepines in human biological fluids

A simple and sensitive HPLC method was developed and validated for the determination of four frequently prescribed 1,4‐benzodiazepines: alprazolam (ALP), bromazepam (BRZ), diazepam (DZP), and flunitrazepam (FNZ). Separation was achieved on an Inertsil C8 analytical (250 mm×4 mm, 5 μm) column, after...

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Bibliographic Details
Published in:Journal of separation science 2007-03, Vol.30 (5), p.679-687
Main Authors: Samanidou, Victoria F., Pechlivanidou, Anastasia P., Papadoyannis, Ioannis N.
Format: Article
Language:English
Subjects:
1,4‐Benzodiazepines
4-Benzodiazepines
Administration, Oral
Alprazolam
Analysis
Benzodiazepines - blood
Benzodiazepines - chemistry
Benzodiazepines - pharmacology
Benzodiazepines - urine
Biological and medical sciences
Biological fluids
Bromazepam
Calibration
Chromatography, High Pressure Liquid - methods
Diazepam
Flunitrazepam
General pharmacology
HPLC
Humans
Medical sciences
Molecular Structure
Pharmacology. Drug treatments
Plasma
Reproducibility of Results
Sensitivity and Specificity
SPE
Urine
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Summary:A simple and sensitive HPLC method was developed and validated for the determination of four frequently prescribed 1,4‐benzodiazepines: alprazolam (ALP), bromazepam (BRZ), diazepam (DZP), and flunitrazepam (FNZ). Separation was achieved on an Inertsil C8 analytical (250 mm×4 mm, 5 μm) column, after selective extraction of benzodiazepine drugs from biological matrices by means of SPE. Isocratic elution was performed with a mobile phase consisting of CH3COONH4, 0.05 M CH3OH, and CH3CN (33:57:10 by volume). Quantification was performed at 240 nm with mefenamic acid (6 ng/μL) as the internal standard. DSC‐18 Supelco cartridges provided high absolute recoveries (81–115%). The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability, and sensitivity. Repeatability (n = 8) and between‐day precision (n = 8) revealed RSD
ISSN:1615-9306
1615-9314
DOI:10.1002/jssc.200600365