Statistical optimization of two-phase hollow fiber liquid-phase microextraction for preconcentration of strychnine in the clinical and postmortem biological fluids prior to gas chromatography-mass spectroscopy detection

Strychnine is a herbal alkaloid whose role in military and doping applications as well as the nervous system stimulation and cancer therapy has been confirmed. Because of its lethal nature, even at low dosages, developing a rapid, sensitive, and accurate detection method for determination of this alkaloid in the clinical and postmortem samples is of great importance. In this study, a green analytical method based on the two-phase hollow fiber liquid-phase microextraction (HF-LPME) combined with gas chromatography-mass spectrometry (GC–MS) was presented for the preconcentration and determination of strychnine in biological fluids. A Box-Behnken design (BBD) with three variables, including extraction time (30–90 min), solution pH (9.0–11.0), and ionic strength (1.0–3.0 mol/L) was employed to maximize the extraction efficiency of strychnine by 1-octanol. The results were satisfactorily fitted to a quadratic response surface model (R2 = 0.979, F = 34.4) which predicted the optimum conditions of operation (extraction time of 89 min, pH = 11.0, and ionic strength of 3.0 mol/L). Based on the results, the studied variables and the interaction between pH and extraction time showed highly significant effects on the process efficiency. The calibration curve was linear in the concentration range of 2.7–1000.0 µg/L (R2 = 0.9978) and the limits of detection and quantitation were found to be 0.8 and 2.7 µg/L, respectively. Moreover, the method was successfully applied to different real samples, including urine (clinical sample), bile and stomach contents, blood, and vitreous humor (postmortem samples), and relative recoveries were obtained in the range of 93.8 % to 99.8 %. The findings of this study suggest that the environmentally friendly HF-LPME method which uses a small amount of a chlorine-free solvent, is a sensitive, cost-effective, simple, and efficient preconcentration method for trace determination of strychnine in biological fluids.