Anti-Inflammation Activity and Chemical Composition of Flower Essential Oil from Hedychium Coronarium
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African Journal of Biotechnology Vol. 8 (20), pp. 5373-5377, 19 October, 2009 Available online at http://www.academicjournals.org/AJB ISSN 1684–5315 © 2009 Academic Journals Full Length Research Paper Anti-inflammation activity and chemical composition of flower essential oil from Hedychium coronarium Y. Lu*, C. X. Zhong, L. Wang, C. Lu, X. L. Li and P. J. Wang College of Biological and Environmental Engineering of Zhejiang Shuren University, Hangzhou, Zhejiang Province, China. Accepted 18 September, 2009 Hedychium coronarium Koen. (Family Zingiberaceae), popularly named butterfly ginger, is widely available in tropical and subtropical regions. It has been used in folk medicine for many conditions, such as contusion inflammation, anti-rheumatic and so on. In this study, chemical compositions and anti-inflammatory activity of this flowers’ essential oil were investigated for the first time. Followed by GC–MS analysis, a total of 29 components were identified and the main constituents included -trans- ocimenone (28.05%), linalool (18.52%), 1,8-cineole (11.35%), -terpineol (7.11%), 10-epi--eudesmol (6.06%), sabinene (4.59%) and terpinen-4-ol (3.17%). We measured the antioxidant activity of the essential oil in vitro (DPPH reduction assay and ferric reducing antioxidant power assay) and the anti- inflammatory activity in vivo (carrageenan-induced hind paw edema in rats). The oil (100 mg/kg p.o.) produced significant inhibition of paw oedema, but showed poor antioxidant activity (with the DPPH 2+ IC50 value of 1091.00 g/ml and FRAP value of 0.22 mol Fe /mg). The results reveal that there is no direct correlation between anti-inflammatory effect and antioxidant activity of this essential oil. Key words: Hedychium coronarium, essential oil, anti-inflammation, antioxidant activity. INTRODUCTION The Zingiberaceae plant Hedychium coronarium Koen., Muraoka et al., 2001; Morikawa et al., 2002a; Shrotriya et which has many common names including butterfly gin- al., 2007). Simultaneously, in the course of pharmaco- ger, butterfly lily, cinnamon jasmine, garland flower and logical studies of this natural medicine, it was reported to ginger lily is widely available in tropical and subtropical have analgesic and anti-inflammatory activities in animal regions, such as Japan, India, Brazil, South China, model (Seikou et al., 2008) and cytotoxic activity on Southeast Asian countries and so on. The rhizome of H. Chinese hamster V-79 cells (Itokawa et al., 1988a), coronarium (“Tuqianghuo”in Chinese) has been used for showing inhibitory effects on the vascular permeability the treatment of headache, diabetes, contusion inflam- induced by acetic acid (AcOH) in mice and nitric oxide mation and sharp pain due to rheumatism in Chinese (NO) production in lipopolysaccharide (LPS)-activated traditional medicine, while it is also used as a febrifuge, mouse peritoneal macrophages, as well as the inhibition tonic, excitant and anti-rheumatic in the Ayurvedic system of the release of b-hexosaminidase from rat basophilic of traditional Indian medicine (Jain et al., 1995). leukemia (RBL-2H3) cells (Matsuda et al., 2002b; Studies on the chemical composition of the rhizome Morikawa et al., 2002b) and representing a hepato- and flower of H. coronarium resulted in the isolation of protective effect on D-galactosamine (D-GalN)-induced several labdane-type diterpenes and farnesane-type cytotoxicity in primary cultured mouse hepatocytes sesquiterpenes (Itokawa et al., 1988a,b; Nakatani et al., (Shrotriya et al., 2007; Yoshikawa et al., 2008). In addi- 1994; Yoshikawa et al., 1994, 1998; Yamahara et al., tion, antimicrobial testing and gas chromatographic 1995; Matsuda et al., 1998, 2001a,b,c,d, 2002a,b; analysis of pure oxygenated monoterpenes and aroma chemicals with chiral and achiral columns were perfor- med and published (Jirovetz et al., 2005; Schmidt et al., 2005). However, the chemical constituents as well as the *Corresponding author. E-mail: [email protected]. Tel: +86-571- pharmacological properties of H. coronarium flowers 88298514. Fax: +86-571-88297097. essential oil remained uncertain. The objective of the 5374 Afr. J. Biotechnol. present study was to identify the main constituents of the from H. coronarium under consideration, carrageenan-induced paw essential oil from the flowers of H. coronarium and to edema test was examined according to the method of Winter et al. investigate the antioxidant and anti-inflammatory activi- (1962). Thirty minutes before application of the inflammatory agent, the animal groups were orally treated (p.o.) either with 1 ml of ties for the first time. essential oil (100 mg/kg) previously emulsified in 3% Tween 80, or with 1 ml of indomethacin (10 mg/kg; pos. control) or an equal volume of 0.9% aq. NaCl (neg. control). Then male Wistar rats were MATERIALS AND METHODS briefly anesthetized with ethyl ether and injected subplantarly into the right hind paw with 0.1 mL of 1.0% carrageenan in isotonic Plant materials and essential oil preparation saline. The left hind paw was injected with 0.1 mL of saline to be used as a control. Measurement of paw size was carried out by The fragrant white flowers held in dense spikes of H. coronarium, gently wrapping a piece of white cotton thread around the paw and appeared in late spring through summer were collected in August measuring the circumference on a meter rule (Bamgbose and 2008 from plants growing in natural stands in different locations of Noamesi, 1981; Olajide et al., 2000) at hourly intervals for 5 h after Hangzhou city, Zhejiang Province, People’s Republic of China. The the stimulus. plants were selected at random. The samples were immediately placed in plastic bags and carried to the laboratory within a maxi- mum of 6 h prior to the experiment. The botanical identity was con- Antioxidant activity assay firmed by the Lab of Plant Systematic Evolution and Biodiversity, Zhejiang University. Voucher specimens were deposited in the The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging assay Herbarium of Zhejiang University (ZJUH, H2008026), China. was estimated according to the procedure described by Cardador- Flowers (0.2 kg) were cut into pieces and subjected to conven- Martinez et al. (2002) with a slight modification. The essential oil of tional hydrodistillation in a modified Clevenger-type apparatus for 3 H. coronarium was dissolved in EtOH at different concentrations. h, cooled, centrifuged at 15,000 rpm for 10 min and the super- Different dilutions of the oil (4 ml) were added to 1 ml of 0.5 Mm natants were extracted with diethyl ether. The ether phase was then DPPH (in EtOH), resulting in a final concentration of 0.1 mm DPPH. dried over anhydrous sodium sulphate (Na SO ) overnight, eva- 2 4 The mixture was shaken vigorously and left for 30 min in the dark at porated in vacuum at low temperature and reduced pressure by room temperature. The absorbance was measured at 517 nm. The rotary evaporator and kept in a labeled, sterile screw-capped bottle scavenging capacity was calculated as follow. at −20°C until use. Scavenging capacity % = [1 − (A /A )] × 100 test control GC–MS analyses of essential oil Where A was the absorbance of the test sample (DPPH solution test plus antioxidant) and A was the absorbance of the control The GC-MS analysis was done in a HP-6890 Series GC System / control (DPPH solution without test sample). BHT (butylated hydroxyl- HP-5973 Mass Selective Detector equipped with a HP-5ms fused toluene) and ascorbic acids were used as positive references while silica capillary column with a (5% phenyl)-methylpolysiloxane EtOH as negative and each assay was performed in triplicates. IC stationary phase (30 m long × 0.25 mm i.d., film thickness 0.25 m). 50 value, which represented the concentration of the essential oil that The temperature of the column was programmed at 50°C for the caused 50% scavenging was calculated by linear regression first 2 min and was then increased 3°C per minute up to the isother- module of Prism 5.0. mal at 250°C (being held for 30 min). The injector and detector The reducing power of the oil was investigated using Ferric temperatures were maintained at 250°C. The carrier gas was Reducing Antioxidant Power (FRAP) assay according to the helium at a flow rate of 1.0 ml/min. The quadrupole mass spectro- -1 method of Yen and Chen (1995) with a modification. FRAP reagent meter was scanned over the range 30–350 amu at 3 scan s , with was freshly prepared by mixing 2.5 ml sodium phosphate buffer an ionizing voltage of 70 eV, ionic source temperature of 230°C, (0.2 M, pH 6.6) and 2.5 ml 1% potassium ferricyanide (K Fe(CN) ). split ratio of 1:20 and an ionization current of 150 A. The oil com- 3 6 The oil (50-400 mg/l) in EtOH (1 ml) was mixed with FRAP reagent ponents were identified by retention times (Davies, 1990; Adams, stirred thoroughly and incubated at 50°C for 30 min. Afterwards, 2001), retention indices, co-injection of the standard and compare- 10% trichloroacetic acid (2.5 ml) was added followed by centrifu- son with a homologous series of National Institute of Standards and gating at 3000 rpm for 10 min. Finally, the supernatant solution (2.5 Technology (NIST) V.2.0 GC–MS library. Relative amount of the ml) was mixed with 2.5 ml of distilled water and 0.5 ml of 0.1% individual component was obtained electronically by FID peak-area FeCl . Then the absorbance was measured at 700 nm. Higher normalization without the use of correction factors. 3 absorbance of the reaction mixture indicated greater reducing power. The reducing power of the essential oil was compared with Experimental animals those of BHT and ascorbic acid as positive controls.