High Performance Liquid Chromatography Method for the Determination of Anethole in Rat Plasma

Purpose: To identify and quantify anethole in the essential oil of fruits of Illicium verum Hook (star anise) and in vivo in rat plasma using reverse-phase liquid chromatography.Methods: Anethole was identified in the essential oil of the fruits of Star anise and determined by gas chromatography-tandem mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), ultraviolet visible spectrophotometry (UV-VIS). A simple, sensitive and validated high performance liguid chromatography (HPLC) technique with UV-VIS detection method was developed for the determination of the compound in rat plasma using: methanol-water (85:15, v/v) as mobile phase at a flow rate of 0.2 ml/min Hypersil ODS Thermo (150 mm x 2.1 mm x 3.0 μM) as column with wavelength detection at 259 nm.Results: GC determination showed that anethole in the essential oil of star anise exhibited a retention time of 21.02 min. The validation results for anethole in plasma were satisfactory, with coefficient of determination (R2) of 0.9945 and relative standard deviation of < 3 %. HPLC run time of 4 min with a retention time of 2.73 min was the faster method to determine anethole when compared to a previously reported method which had a run time of 15 min.Conclusion: Anethole in the essential oil of Illicium verum Hook can be identified and determined by GC-MS, NMR and UV-VIS, and a superior HPLC method has been developed for the determination of the compound in rat plasma.Keywords: Anethole, High performance liguid chromatography, Star anise, Essential oil, Rat plasma, Illicium verum Hook

Effect of Camphor on the Behavior of Leukocytes In vitro and In vivo in Acute Inflammatory Response

Purpose: To evaluate the effect of camphor on acute inflammatory  response by leukocytes on chemotaxis, antiedematogenic and phagocytic activities.Methods: The effect of camphor in acute inflammatory response evaluated by neutrophil chemotaxis, ear edema induced by croton oil, activity of the enzyme myeloperoxidase (MPO) and phagocytic activity of macrophages in Swiss mice.Results: Camphor treatment did not show any cytotoxicity. Camphor at 3, 10, and 30 ìg/ml doses exhibited significant (p < 0.01) reduction on leukocyte migration toward N-formyl methionyl leucyl phenylalanine fMLP. Topical treatment with camphor did not reduce significant ear edema or MPO activity at any of the doses tested. However, in contrast, oral treatment with 100 and 200 mg/kg camphor significantly (p < 0.01) reduced ear edema and myeloperoxidase (MPO) activity. Additionally, the phagocytic activity of macrophages was not affected by camphor.Conclusions: These results indicate that the anti-inflammatory activity of camphor may be related to the inhibition of leukocyte migration and antiedematogenic activity.Keywords: Camphor, Inflammatory response, Chemotaxis, Macrophages, Phagocytic, Leukocytes, Edem