Determination of Articaine in Human Blood by Gas and Liquid Chromatography and its Application in a Preliminary Pharmacokinetic Study

Abstract

Two simple, sensitive and specific gas and liquid chromatographic methods are developed and validated for quantification of articaine in human whole blood. Liquid-liquid extraction with n-hexane-isoamyl alcohol 90:10 (v/v) is used for sample preparation in both methods. GC-FID (gas chromatographic-flame ionization detection) analysis is performed with a gas chromatograph equipped with a split-splitless injector, flame-ionization detector and HP INNOwax capillary column. Reverse phase LC-PDA (liquid chromatographic-photo diode array detection) analysis is carried out using a C18 Hypersil GOLD column with two mobile phases: acetonitrile-water 10 mM ammonium acetate 50:50 (v/v), pH 7, and acetonitrile-buffer solution sodium acetate 10 mM and acetic acid 10 mM 50:50 (v/v), pH 4.7, respectively. A comparison between GC-FID and LC-PDA is performed, as well as between internal and external standard as quantification methods in LC. The best results in terms of accuracy (as recovery) and precision (as relative standard deviation) are obtained in GC (95-98% and 5.5-8.2%, respectively) with lidocaine internal standard and in LC (96-102% and 4.2-6.1%, respectively) with external standard and mobile phase pH 4.7. This method is applied in a preliminary pharmacokinetic study to three healthy volunteers to whom anesthesia with articaine is performed.