The Analysis of Antidepressants Mixture using Chromatographic Methods

The object of the study – human plasma samples with the mixture of four antidepressant drugs (fluoxetine hydrochloride, paroxetine hydrochloride, escitalopram oxalate and buspirone hydrochloride). The aim of the study was to select the best conditions for chromatographic analysis methods – High-Performance Thin-Layer Chromatography (HPTLC) and High-Performance Liquid Chromatography (HPLC) for fluoxetine, paroxetine, escitalopram and buspirone separation from their mixture, identification and quantification in human plasma. The tasks were to select the most suitable solid phase extraction conditions for extraction of the analytes from blood plasma. To select the best conditions for the separation and identification of antidepressants by HPTLC and validate. To select the most suitable conditions for the separation and identification of the antidepressant mixture components by HPLC and validate. To select the most suitable conditions for quantification of extracted antidepressants from blood plasma. Used methods: Solid-Phase extraction, High-Performance Liquid Chromatography, Thin Layer Chromatography. The mobile phase (acetonitrile:methanol:ammonium hydroxide 85:10:10) of HPTLC was selected and validated during the experiment. This mobile phase allows to separate and identify all four components of antidepressants mixture, extracted from blood plasma. The specifity and repeatability were determined. The HPLC method was applied for quantitative and qualitative analysis of all four analytes. Validation of HPLC qualitative method was carried out by determination of specifity, linearity, limits of detection (fluoxetine–0,048 µg/ml, paroxetine–0,1 µg/ml, escitalopram–0,1 µg/ml, buspirone–0,088 µg/ml), and limits of quantification (fluoxetine–0,105 µg/ml, paroxetine–0,29 µg/ml, escitalopram–0,27 µg/ml, buspirone–0,201 µg/ml). For extraction from blood plasma by SPE, the best recovery of analytes was carried out when the chosen eluent was pure methanol