<abstract xmlns="http://www.w3.org/1999/xhtml"><p>A rapid high performance liquid chromatography method, using a monolithic column, was developed for quantitative determinations of benzodiazepines (diazepam, clonazepam, lorazepam, midazolam) in whole blood. A liquid-liquid extraction step with n-chlorobutane isolates the drugs from alkalinized blood. The separation was carried out in reversed phase conditions using a Chromolith Performance (RP-18 100x4.6 mm) column. For the mobile phase, a mixture of a phosphate buffer (pH= 2.5)/acetonitrile (65/35 v/v), in isocratic mode at 2 mL/min. An ultraviolet spectrophotometer was used as the detector at the wavelength of 220 nm. The total run time of the analytical method is less than 4-6 minutes. The calibration curves showed linearity and the correlation coefficient of each individual curve was greater than 0.995. The method was linear over a concentration range of 0.03-0.6 μg/mL for clonazepam, lorazepam and midazolam. For diazepam of linearity was over the range 0.04-5.0μg/mL. Quantification limits ranged from 0.03-0.04μg/mL and the accuracy were from 80% to 105% for the recovery test. </p><p>The results indicate that this analytical method is simple, specific, accuracy, sensitive, demonstrating from the validation data and a higher robustness. The proposed method is applied routinely in forensic toxicological analysis involving blood.</p></abstract>

A rapid high performance liquid chromatography method, using a monolithic column, was developed for quantitative determinations of benzodiazepines (diazepam, clonazepam, lorazepam, midazolam) in whole blood. A liquid-liquid extraction step with n-chlorobutane isolates the drugs from alkalinized blood. The separation was carried out in reversed phase conditions using a Chromolith Performance (RP-18 100x4.6 mm) column. For the mobile phase, a mixture of a phosphate buffer (pH= 2.5)/acetonitrile (65/35 v/v), in isocratic mode at 2 mL/min. An ultraviolet spectrophotometer was used as the detector at the wavelength of 220 nm. The total run time of the analytical method is less than 4-6 minutes. The calibration curves showed linearity and the correlation coefficient of each individual curve was greater than 0.995. The method was linear over a concentration range of 0.03-0.6 μg/mL for clonazepam, lorazepam and midazolam. For diazepam of linearity was over the range 0.04-5.0μg/mL. Quantification limits ranged from 0.03-0.04μg/mL and the accuracy were from 80% to 105% for the recovery test. The results indicate that this analytical method is simple, specific, accuracy, sensitive, demonstrating from the validation data and a higher robustness. The proposed method is applied routinely in forensic toxicological analysis involving blood.

Simultaneous quantification of four benzodiazepines from whole blood by highperformance liquid chromatography in forensic toxicological analysis

Ovidius University, Faculty of Pharmacy, University Street No.1, Constanta, Romania

Ovidius University, Faculty of Applied Science and Engineering, 124 Mamaia Blvd., Constanta, Romania

UMF ”Carol Davila”, Faculty of Pharmacy, Traian Vuia Street No.6, Bucharest, Romania

Ovidius University Annals of Chemistry

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

A rapid high performance liquid chromatography method, using a monolithic column, was developed for quantitative determinations of benzodiazepines (diazepam,...

Simultaneous quantification of four benzodiazepines from whole blood by highperformance liquid chromatography in forensic toxicological analysis

Published Online: Sep 12, 2014

Page range: 24 - 27

Received: May 28, 2013

Accepted: May 10, 2014

DOI: https://doi.org/10.2478/auoc-2014-0004

Keywords
benzodiazepines, forensic toxicological analysis, HPLC, monolithic column

© Ovidius University Press

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Simultaneous quantification of four benzodiazepines from whole blood by highperformance liquid chromatography in forensic toxicological analysis

Published Online: Sep 12, 2014

Page range: 24 - 27

Received: May 28, 2013

Accepted: May 10, 2014

DOI: https://doi.org/10.2478/auoc-2014-0004

Keywordsbenzodiazepines, forensic toxicological analysis, HPLC, monolithic column

© Ovidius University Press

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
benzodiazepines, forensic toxicological analysis, HPLC, monolithic column