The extractions of the samples were realized using Toxivial A® (Interchim, France) and 1 μL of each extracted sample was injected on to Adriamycin DB-5 ms UI column (30 mm × 0.25 mm internal diameter; 0.25 μm film thickness) using pulsed splitless injection with an injector temperature of 250 °C . Temperature conditions were as follows: initial temperature of 70 °C for 1 min, increased to 100 °C at 10 °C/min, then increased to 300 °C at 20 °C/min and held for 12 min. The flow of the carrier gas (helium) was maintained at 1 mL/min in constant flow mode. The gas chromatograph interface temperature was held at 315 °C. Electron impact ionization was performed at 70 eV, with an ion source temperature of 230 °C and mass spectra collected from 40 to 600 m/z.
3.1.2. Liquid chromatography-diode array detection analysis
A liquid–liquid extraction of the sample was performed. Internal standard solution (30 μL of glafenine at 10 mg/L), 2 mL of carbonate buffer pH 9.2 and 4 mL of dicholoromethane/hexane solution (3/4, v/v) were added to 1 mL of plasma in a glass tube. Then sample was shaken for 10 min and centrifuged 10 min at 2500 rpm, the organic layer was evaporated to dryness under nitrogen. Each extract was reconstituted with 100 μL of phosphate buffer and CH3CN/H2O (70/30, v/v) and injected into the chromatograph.