RESUMEN
The dielectric properties of Cymbopogon nardus, Eucalyptus sp., Piper aduncum and Piper hispidinervum were investigated as a function of frequency and temperature, using dry plant matter and its mixtures with water at different concentrations. This was followed by the extraction of essential oils performed with microwave heating in single-mode and multimode cavities with a variable power 6.0 kW generator operating at 2.45 GHz. The dielectric properties of the dry plant matter changed markedly with increasing water content, exhibiting high loss factors and small penetration depths. Due to the high level of absorption, even with low water contents, microwave-assisted extraction (MAE) showed better green performance employing lower plant matter/water ratios (1 : 2 or 1 : 4) and applying shorter extraction times compared with conventional hydrodistillation (HD). Using the single-mode MAE reactor, in the case of Cymbopogon nardus, for a plant matter/water ratio of 1 : 4 the energy efficiency was 1.78 g kW-1 h-1, applying 0.3 kW for 16.7 min. By way of comparison, for the same extraction time using HD, the corresponding efficiency was only 0.50 g kW-1 h-1. In experiments with citronella using multimode MAE, the best energy efficiency of 2.53 g kW-1 h-1 was obtained with a plant matter/water ratio of 1 : 2 applying 1.8 kW of power for 30 min. Single and multimode MAE experiments showed optimum conditions with lower water content. Thus, greater amounts of material can be processed in a shorter time, in accordance with the ideals of a green chemistry. The resulting extractions showed an energy efficiency up to 27 times greater compared with conventional HD, applying the same extraction time.
RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Toads known as "cururú" (Rhinella schneideri) have been used in the Brazilian Pantanal and Paraguayan Chaco wetlands to treat erysipelas and cancer. The aim of the study was to assess the antiproliferative effect of the venom obtained from Rhinella schneideri and to identify its constituents by spectroscopic and spectrometric methods. MATERIALS AND METHODS: The venom was obtained by gentle pressing the parotid glands of the toads. The dry crude drug was analyzed by HPLC-MS-MS and chromatographed on Sephadex LH-20 to obtain purified compounds and fractions for spectroscopic analysis. The venom and fractions were evaluated for antiproliferative activity towards normal human lung fibroblasts (MRC-5) and four human cancer cell lines: gastric epithelial adenocarcinoma (AGS), lung cancer (SK-MES-1), bladder carcinoma (J82) and promyelocytic leukemia (HL-60). RESULTS: From the Rhinella schneideri venom, 29 compounds were isolated and/or identified by spectroscopic and spectrometric means. Three known alkaloids and five argininyl diacids were identified in the complex mixture by HPLC-MS-MS. Nine out of fifteen argininyl diacid derivatives of the bufadienolides bufalin, marinobufagin and telocinobufagin are reported for the first time and four argininyl diacids are described for the first time as natural products. The venom and the fractions 9-13 showed a remarkable antiproliferative effect, with IC50 values in the range 0.019-0.022, 0.035-0.040, 0.028-0.064, 0.042-0.056 and 0.044-0.052 µg/mL for MRC-5, AGS, SK-MES-1, J82 and HL-60 cell lines, respectively. Under the same experimental conditions, IC50 values of the reference compound etoposide were 2.296, 0.277, 1.295, 1.884 and 1.059 µg/mL towards MRC-5, AGS, SK-MES-1, J82 and HL-60 cells, respectively. CONCLUSIONS: The venom showed a strong antiproliferative effect towards human cancer cells and presented a high chemical diversity in its constituents, supporting its use as anticancer agent. These findings encourage further work on the chemistry and bioactivity of South American toad venoms.