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Optimization of the SPME Parameters and Its Online Coupling with HPLC for the Analysis of Tricyclic Antidepressants in Plasma Samples

2006; Oxford University Press; Volume: 44; Issue: 6 Linguagem: Inglês

10.1093/chromsci/44.6.340

1945-239X

Carlos Alves, Christian Fernandes, Álvaro J. Santos‐Neto, Juliana Chagas Rodrigues, Maria Eugênia Costa Queiroz, Fernando Mauro Lanças,

Pharmacological Effects and Assays

Solid-phase microextraction (SPME)-liquid chromatography (LC) is used to analyze tricyclic antidepressant drugs desipramine, imipramine, nortriptyline, amitriptyline, and clomipramine (internal standard) in plasma samples. Extraction conditions are optimized using a 23 factorial design plus a central point to evaluate the influence of the time, temperature, and matrix pH. A Polydimethylsiloxane-divinylbenzene (60-µm film thickness) fiber is selected after the assessment of different types of coating. The chromatographic separation is realized using a C18 column (150 × 4.6 mm, 5-µm particles), ammonium acetate buffer (0.05 mol/L, pH 5.50)-acetonitrile (55:45 v/v) with 0.1% of triethylamine as mobile phase and UV-vis detection at 214 nm. Among the factorial design conditions evaluated, the best results are obtained at a pH 11.0, temperature of 30°C, and extraction time of 45 min. The proposed method, using a lab-made SPME-LC interface, allowed the determination of tricyclic antidepressants in in plasma at therapeutic concentration levels.