Journal Article

Stability of Benzodiazepines and Cocaine in Blood Spots Stored on Filter Paper

Abdulkareem A. Alfazil and Robert A. Anderson

in Journal of Analytical Toxicology

Volume 32, issue 7, pages 511-515
Published in print September 2008 | ISSN: 0146-4760
Published online September 2008 | e-ISSN: 1945-2403 | DOI:
Stability of Benzodiazepines and Cocaine in Blood Spots Stored on Filter Paper

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Previous studies have shown that drug concentrations in blood can change during storage, especially at room temperature, but even labile drugs such as cocaine may be stable in dried blood spots (DBS). A new method has been developed for the analysis of hydrolytically labile drugs in blood spots on filter paper in order to assess their degradation during a storage period of one month. The drugs selected included flunitrazepam, temazepam, oxazepam, lorazepam, nitrazepam, diazepam, and cocaine. A Guthrie card 903 was spotted with 100 µL of blood containing the drugs at concentrations of 1000 ng/mL and left overnight to dry at room temperature. The filter paper was suspended in extraction buffer for 1 h with ultrasonication. Drugs were then extracted from the buffer by solid-phase extraction using Clean Screen® columns and analyzed by liquid chromatography-tandem mass spectrometry. Method validation showed that all calibration curves were linear over the concentration range 5–200 ng/spot with correlation coefficients of 0.994–0.999. Interday and intraday precisions at three concentrations (10, 50, and 100 ng/spot) were 1.6–18.3% and 2.8–14.7%, respectively. Limits of detection were 0.29–0.74 ng/spot, and lower limits of quantitation were 0.99–2.46 ng/spot. Recoveries of all analytes were in the range 81–106%. DBS were stored in duplicate at room temperature, 4°C, and −20°C for up to one month. Degradation of the drugs in DBS at all storage conditions was less than for the corresponding liquid blood samples stored under similar conditions and more than 80% of each analyte could be recovered from the samples.

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Subjects: Medical Toxicology ; Toxicology (Non-medical)

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