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- Labdanum resin from Cistus ladanifer L. : evaluation of residual water vs. extraction yieldPublication . Frazão, David F.; Mesquita, Maria da Conceição; Silva, Helena; Silva, Amélia M.; Gonçalves, José Carlos; APH- Associação Portuguesa de HorticulturaCistus ladanifer L. (Cistaceae) is an endemic and abundant resource in the Iberian Peninsula and North Africa. This plant exudes an aromatic resin nowadays valued in the perfumery and fragrance industry. Traditional processes for the extraction and isolation of such resin use boiling water or alkaline water followed by acidic precipitation. However, a concern arises about the effluents resulting from these extraction processes. To overcome this concern, labdanum resin was extracted with Na2CO3 solution (25 g/L) at 60 oC and precipitated with sulphuric acid (5 M). The residual water was evaluated regarding total phenolic content, suspended solids, electric conductivity, and sulphate, sodium, magnesium, and calcium content. The effluent was characterized by a total phenolic content of 1245 ± 455 mgGAeq/L, 1338 ± 101 mg/L of suspended solids, pH of approximately 2, electric conductivity of 34.8 ± 0.7 mS/cm, 22284 ± 710 mg/L of sulphate, 9696 ± 1072 mg/L of sodium, 3.97 ± 0.24 mg/L of magnesium, 3.52 ± 0.80 mg/L of calcium, and a Sodium Adsorption Ratio of 876 ± 112. Because the values were far from the limit values set by Portugal decree-law 236/98 for residual waters discharged and irrigation waters, it was concluded that efforts should be made to optimize the extraction process. In that regard, a factorial designed experiment was done to evaluate the effect of Na2CO3 concentration (0; 2.5; and 25 g/L), extraction temperature (60 and 100 oC) and acidification extent (pH 2, neutralization, and no acidification) on the residual water quality and on the yield of labdanum resin extraction. Alkalinization and acidification are important to obtain high resin extraction yields (Andalusian vs. Zamorean process), but mostly alkalinization may be reduced to meet sulphate criteria for discharge without significantly affecting resin extraction yields. Despite that, to meet salinity criteria for irrigation waters a higher reduction in alkalinization is needed for Andalusian processes. Phenolic content, although lower for extractions done at 60 oC, was far from the limit values for discharge, regardless experimental conditions. Given the high phenolic content the residual water from labdanum extraction by both traditional processes must be treated before discharge. If separated, phenolic compounds may be valorized as a by-product.
- Cistus ladanifer L. tissue culture from leaf and stem explants.Publication . Frazão, David F.; Barroca, Celina; Silva, Amélia M.; Delgado, F.M.G.; Gonçalves, José CarlosCistus ladanifer L. exudes a phenolic and terpenoid resin with interesting bioactive and aromatic properties. Despite its high abundance in the wild, this plant can be cultivated to advantage on oligotrophic and trace-elements contaminated soils. Plant tissue culture may be used to produce specific metabolites or for clonal propagation of specific genotypes for plantation. From a biotechnological perspective this is the second study that has attempted in vitro propagation of C. ladanifer from adult plant material. Its goal was to evaluate the potential of leaf and internodal stem explants from C. ladanifer for in vitro tissue culture. Three plant growth regulators were tested: 2,4-Dichlorophenoxyacetic acid (2,4-D), 6-Benzylaminopurine (BAP), and 1-Naphthaleneacetic acid (NAA). From both explants, shoots were regenerated under the influence of BAP (38%) and two types of compact calli were induced: dark green calli were induced under the influence of BAP (above 70%) and light green calli were induced under the influence of 2,4-D with or without BAP (100%). Light green calli grew between 558 and 708% during subsequent subcultures and showed rhizogenic capacity when the amounts of BAP were lower than of 2.4-D, but they showed low potential for shoot organogenesis. Dark green calli were associated with shoot organogenesis. The suitability of the two calli lines to produce metabolites and their transposition to liquid cultures is worth further study in comparison to organ in vitro cultures.