Vinicius HV Fagundes1,
Rilson J Pinho1,
Luiz AM Wiirzler1,
Elza Kimura2,
Ciomar A Bersani-Amado1,
Roberto KN Cuman1
1Department of Pharmacology;
2Department of Pharmacy, University of Maringá, Avenue Colombo, 5790, Maringa- Brazil.
For correspondence:- Roberto Cuman
Email: rkncuman@uem.br Tel:+554430114923
Received: 12 May 2013
Accepted: 13 March 2014
Published: 23 May 2014
Citation:
Fagundes VH, Pinho RJ, Wiirzler LA, Kimura E, Bersani-Amado CA, Cuman RK.
High Performance Liquid Chromatography Method for the Determination of Anethole in Rat Plasma. Trop J Pharm Res 2014; 13(5):793-799
doi:
10.4314/tjpr.v13i5.21
© 2014 The authors.
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Abstract
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 ´ 2.1 mm ´ 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