Simultaneous determination of benzodiazepines in whole blood or serum by HPLC/DAD with a semi-micro column

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 23; no. 2; pp. 447 - 458
• Main Authors: El Mahjoub, Anissa, Staub, Christian
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.08.2000; Elsevier Science
• Subjects: Analysis; benzodiazepine; Benzodiazepines; Benzodiazepines - blood; Biological and medical sciences; Blood; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; Diode array detection; General pharmacology; High performance liquid chromatography; Medical sciences; Pharmacology. Drug treatments; Reproducibility of Results; Semi-micro HPLC column; Sensitivity and Specificity; Serum; Spectrophotometry, Ultraviolet - methods; Serum; Diode array detection; Semi-micro HPLC column; Benzodiazepines; Blood; High performance liquid chromatography; Validation; Reversed phase chromatography; HPLC chromatography; Benzodiazepine derivatives; Diode array; Quantitative analysis; Microbore column
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(00)00325-3

A simple and sensitive high performance liquid chromatographic (HPLC) method using a semi-micro column, C 8 reversed-phase column (3 mm i.d.) and a low flow rate 0.3 ml/min was developed and validated for the determination of five frequently prescribed benzodiazepines: clonazepam, diazepam, flunitrazepam, midazolam and oxazepam. Quantification was performed at 220 nm with methylclonazepam as internal standard. The method involved a simple extraction from alkalinized blood (1 ml) into 1-chlorobutane and provided excellent sensitivity, recovery, accuracy and reproducibility for benzodiazepines in therapeutic or toxic concentrations.