Molecularly Imprinted Solid-Phase Extraction of Diazepam and Its Metabolites from Hair Samples

Abstract

An anti-diazepam, molecularly imprinted polymer (MIP) has been synthesized and used to extract diazepam and other benzodiazepines from hair samples via a molecularly imprinted solid-phase extraction (MISPE) protocol. Optimum retention of diazepam on the MIP columns was achieved using an apolar solvent, and the binding capacity of the polymer toward diazepam was found to be 110 ng of diazepam/mg of polymer. The recovery of a 50 ng diazepam standard spiked into blank hair was 93%, with good precision (RSD = 1.5%). The LOD and LOQ of diazepam in spiked hair samples were 0.09 and 0.14 ng/mg, respectively. The MISPE method was demonstrated to be applicable to the analysis of diazepam metabolites and other benzodiazepine drugs, in addition to diazepam itself. The application of the extraction method to postmortem hair samples yielded results that were in good agreement with the corresponding ELISA data (from blood samples) and data arising from the analysis of the same blood samples using a validated in-house SPE-LC-MS-MS method.