Abstract
Saliva is far less popular as a diagnostic material than blood. This has resulted in a lack of procedures for the sampling and handling of saliva, e.g., effective ways to purify endogenous compounds from saliva to enable a simultaneous determination of xenobiotics such as neuroleptics. Therefore, the aim of this study was to develop an analytical procedure to purify saliva samples so that it is then possible to simultaneously determine five neuroleptics (aripiprazole, clozapine, olanzapine, quetiapine and risperidone) and the antiepileptic drug carbamazepine, and their respective metabolites (dehydroaripiprazole, N-desmethylclozapine, N-demethylolanzapine, norquetiapine, 9-OH-risperidone and carbamazepine-10,11-epoxide). A study of three types of solid-phase extraction (SPE) columns showed that the purest eluates were obtained using columns containing ion exchange sorbent. The sorbents were first washed with water then with a mixture of water and methanol (1:1), and the adsorbed residue was eluted with a 5% ammonia solution in methanol. Saliva samples for SPE were diluted with 2% formic acid and a mixture of methanol and water (1:1). This procedure was developed to purify a saliva sample spiked with a mixture of neuroleptics and carbamazepine, and their respective metabolites. A chromatographic analysis confirmed the isolation of all compounds, indicating that this procedure can be used in further development and validation for a method designed to monitor the levels of neuroleptic drugs in saliva and to monitor their uptake by patients.
Highlights
Solid phase extraction (SPE) is one of the most frequently used methods of sample purification and isolation of selected substances
Taking the above into account, this study developed a method for purifying endogenous substances from saliva so that it was possible to quickly and reliably determine the most commonly used neuroleptics and carbamazepine, along with their respective metabolites in this diagnostic material
This study was undertaken to select the best procedure for saliva purification with SPE, with the intention of developing a method allowing the determination of five neuroleptics, carbamazepine and their respective metabolites
Summary
Solid phase extraction (SPE) is one of the most frequently used methods of sample purification and isolation of selected substances. Blood is one of the most frequently tested biological matrices, inconveniences associated with blood sampling such as the possibility of infection and the required presence of qualified medical staff, are reasons why other alternative biological matrices are being considered. The presence of qualified medical personnel is not required during saliva sampling. Sampling is not sterile, which means that the compounds to be determined may be degraded by microorganisms [1]; the concentration of the test substance(s) is usually lower than in blood and depends on many factors, including the molecular weight of the analyte, lipid solubility, pKa of the compound, the degree of binding to blood proteins or salivary flow; the concentration of a given substance is an individual feature of the tested person; saliva contains only the unbound forms of substances, i.e., active forms [2]
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