Development and validation of an HPLC method for the determination of benzodiazepines and tricyclic antidepressants in biological fluids after sequential SPE

Abstract

A novel method for the simultaneous determination of six benzodiazepines (BZDs) and four tricyclic antidepressants (TCAs) in biological fluids by HPLC with UV detection at 240 nm has been developed. After a deproteinization step biological fluids were analyzed by direct injection. SPE on Nexus cartridges was also applied. Since two compounds, namely imipramine and diazepam, were coeluting, a sequential SPE protocol has been developed. BZDs were eluted by a mixture of methanol/ACN(1:1), followed by the elution of TCAs with methanol. Separation was performed on a Kromasil C8 column (250 x 64 mm(2) id, 5 microm) using a mobile phase of 0.05 MCH3COONH4/ACN/methanol (initial composition 55:15:30 v/v/v) at a flow rate of 1.0 mL/min delivered by a gradient program within 15 min. Colchicine was used as the internal standard (4 ng/microL). The method was linear for all analytes up to 20 ng/lL, with coefficients of regression between 0.996 and 0.99996. LODs and LOQs were 0.08-1.17 and 0.28-3.91 ng/lL, respectively. Recovery was in the range of 92.8-108.7% for within-day and 91.9-109.9% for between-day assays, with RSD values lower than 10.0% for all matrices.

Improvement and validation the method using dispersive liquid–liquid microextraction with in situ derivatization followed by gas chromatography–mass spectrometry for determination of tricyclic antidepressants in human urine samples

“…Separation techniques, including high-performance liquid chromatography (HPLC) [3][4][5][6][7][8][9] and gas chromatography (GC) [10], have been employed for the analysis of TCAs. As regards sample preconcentration procedures, conventional extraction techniques, such as liquid-liquid extraction (LLE) [7][8][9] and solid phase extraction (SPE) [4][5][6], have been applied to the analysis of TCAs, but they are time-consuming and require large volumes of organic solvents. Microextraction techniques (extraction techniques that require small volumes of organic solvents), such as solid-phase microextraction (SPME) [11] and liquid-phase microextraction (LPME) [12], have also been applied to TCA analysis, but they have some drawbacks.…”

A Novel Electrochemical Sensor for Probing Doxepin Created on a Glassy Carbon Electrode Modified with Poly(4-Amino- benzoic Acid)/Multi-Walled Carbon Nanotubes Composite Film

“…Doxepin (Figure 1) has been widely used as an effective tricyclic antidepressant in the treatment of psychiatric disorders over the past decades [1,2]. Due to its importance, many analytical methods have been developed for its determination.…”

Validation of SPE‐HPLC determination of 1,4‐benzodiazepines and metabolites in blood plasma, urine, and saliva

“…However, SPE is still being exploited by the researchers for attaining objectives, like reduced solvent consumption and disposal, reduced time, cost, and labor providing higher concentration factors than LLE. It also provides greater opportunity for selective isolation, such as fractionation of the sample into different compounds or groups of compounds [38]. To the best of our knowledge only one paper has reported [39] applying SPE for the isolation of BDZs from saliva.…”