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β-sitosterol and two triterpenoids: ursolic acid acetate and platanic acid have been isolated from ethanolic extract of Vitex trifola leaves. β-sitosterol was previously isolated from the leaves, stem and seeds of Vitex trifolia. Ursolic acid acetate has been isolated for the first time in this plant species. Platanic acid has been reported for the first time in Vitex trifolia and even in the family of this plant: Verbenaceae. These compounds were characterized using spectroscopic methods including 1D-1HNMR, 13CNMR, ESIMS and 2D-NMR (HSQC, HMBC, COSY) experiments and confirmed by comparison of their NMR data with those from the literature. A preliminary molluscicidal test for ethanol, chloroform and n-hexane extracts of leaves of Vitex trifolia against Biomphalaria alexandrina adult snails showed that ethanol extract of leaves with LC50 value 26.42 mg/l (27.92 mg/l – 24.99 mg/l) was more effective than n-hexane extract with LC50 value 35.48 mg/l (43.81 mg/l – 28.72mg/l) and chloroform extract with LC50 value 46.77 mg/l (53.59 mg/l – 43.81 mg/l) after 24 h exposure.
The molluscicidal activity of six monoterpenes and two phenylpropenes against Theba pisana adults was evaluated using fumigation and direct contact methods. In the fumigant toxicity assay, (-)-citronellal showed the highest toxicity with LC50 value of 7.79 µl · l−1 air after 24 h of treatment, followed by (-)-terpinen-4-ol (LC50 = 12.06 µl · l−1), (-)-menthone (LC50 = 12.28 µl · l−1 air) and p-cymene (LC50 = 16.07 µl · l−1 air). Eugenol and trans-cinnamaldehyde were the most potent contact toxicants against T. pisana. Their LD50 values were 0.18 and 0.29 mg · snail−1 after 24 h of treatment, respectively. These two compounds were more toxic than a reference molluscicide, methomyl. In contrast, α-terpinene and (-)-citronellal were the least toxic compounds. In another experiment, the synergistic effect of piperonyl butoxide (PBO) on tested monoterpenes and phenylpropenes by topical application was examined. The results showed that the toxicity of the tested compounds was increased when mixed with PBO at a ratio [compound/PBO (1 : 2)] except for α-pinene and (-)-terpinen-4-ol in which the toxicity of binary mixtures was less than for single compounds. The synergistic effect of PBO improved with increased exposure time. The highest synergistic effect was observed with (-)-menthone and α-terpinene with synergistic ratios of 9.25 and 4.37, respectively. Monoterpenes and phenylpropenes and their mixtures with PBO described herein merit further studies as potential T. pisana control agents.
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