An ethanolic extract of cloves was analyzed by gas chromatography directly to identify eugenol and other major phenolic compounds without previous separation of other components. Separation was performed on a fused-silica capillary column of 30 m x 0.53 mm I.D., 0.53 microns film thickness. The detector was a flame ionization detector. Helium gas at a flow-rate of 3 ml/min was used as a carrier gas. The analysis were performed with linear temperature programming. Nine components were detected and special attention was given to the major phenolic compound, eugenol.
Pharmacophagy of methyl eugenol (ME)--a highly potent male attractant, by Bactrocera papayae results in the hydroxylation of ME to sex pheromonal components, 2-ally-4,5-dimethoxyphenol (DMP) and (E)-coniferyl alcohol (CF). These compounds, which are also male attractants, are then sequestered and stored in the rectal gland prior to their release during courtship at dusk. Chemical analyses of the digestive tract (excluding the crop and rectal gland) showed the absence of the sex pheromonal components and their precursor, ME. However, B. papayae males were attracted to and fed on the ME-fed male hemolymph extracts but not on hemolymph extracts of ME-deprived males. After thin layer chromatography in a hexane:ethyl acetate solvent system, flies were attracted to and fed on the original point on the TLC plate where the hemolymph extract had been spotted, suggesting that the pheromone components were bound in polar complexes. Chemical analyses of the ME-fed male hemolymph and crop extracts revealed the presence of the sex pheromonal components. The presence of the ME-derived pheromonal components and the absence of ME in the hemolymph suggest that the hemolymph is involved in the transportation of sex pheromonal components from the crop to the rectal gland.
Candida species have been associated with the emergence of strains resistant to selected antifungal agents. Plant products have been used traditionally as alternative medicine to ease mucosal fungal infections. This study aimed to investigate the effects of Piper betle extract on the growth profile and the ultrastructure of commonly isolated oral candidal cells. The major component of P. betle was identified using liquid chromatography-mass spectrophotometry (LC-MS/MS). Seven ATCC control strains of Candida species were cultured in yeast peptone dextrose broth under four different growth environments: (i) in the absence of P. betle extract; and in the presence of P. betle extract at respective concentrations of (ii) 1 mg⋅mL(-1); (iii) 3 mg⋅mL(-1); and (iv) 6 mg⋅mL(-1). The growth inhibitory responses of the candidal cells were determined based on changes in the specific growth rates (µ). Scanning electron microscopy (SEM) was used to observe any ultrastructural alterations in the candida colonies. LC-MS/MS was performed to validate the presence of bioactive compounds in the extract. Following treatment, it was observed that the µ-values of the treated cells were significantly different than those of the untreated cells (P<0.05), indicating the fungistatic properties of the P. betle extract. The candidal population was also reduced from an average of 13.44×10(6) to 1.78×10(6) viable cell counts (CFU)⋅mL(-1). SEM examination exhibited physical damage and considerable morphological alterations of the treated cells. The compound profile from LC-MS/MS indicated the presence of hydroxybenzoic acid, chavibetol and hydroxychavicol in P. betle extract. The effects of P. betle on candida cells could potentiate its antifungal activity.