Alternative Extraction Methods of Essential Oil From the Flowers of Citrus aurantium L. Var Daidai Tanaka: Evaluation of Oil Quality and Sedative-Hypnotic Activity

Abstract

The aim of this study was to analyze the content of hypnotic components in the essential oil from Citrus aurantium flowers (EDD), extracted by different methods, and to characterize its sedative-hypnotic effects. The sedative-hypnotic capacity of EDD was evaluated using pentobarbital-induced sleeping assays, locomotor activity tests and GABAA receptor antagonists. The results showed that EDD extracted by steam and water distillation (SWD), hydrodistillation (HD), and ultrasound-assisted hydrodistillation (UHD) had as their main components linalool, linalyl acetate, and limonene, comprising more than 55% of the total peak area. Compared with EDD extracted by HD and UHD, the total content of linalool and linalyl acetate in EDD obtained by SWD was highest, whereas the content of limonene in EDD extracted by the 3 different methods was not different. Oral and intraperitoneal administration of EDD resulted in reduced sleep latency and increased sleep duration of mice, as well as reduced locomotor activity, which was proven by decreases in the total distance travelled, average velocity, number of activities, and central distance. Interestingly, intraperitoneal injection of EDD had better sedative and hypnotic effects than oral ingestion. In vitro assays using SH-SY5Y cells showed that EDD dose-dependently increased Cl− influx, which could be blocked by the GABAA receptor antagonists, picrotoxin, bicuculline, and flumazenil, suggesting that EDD promoted sedative-hypnotic activity by potentiating GABAA receptor-mediated Cl− current responses. Altogether, these results suggest that the important hypnotic-sedative activity of EDD appears to be due to the effects of limonene, and particularly the high contents of linalool and linalyl acetate, which were effectively extracted by SWD.