Estudo etnofarmacológico e fitoquímico das espécies medicinais Cleome spinosa Jacq, Pavonia varians Moric e Croton cajucara Benth

In this present work an ethnographic research was performed with 84 native medicinal specimens from the Litoral Norte Riograndense, from which two plants Cleome spinosa Jacq e Pavonia varians Moric were submitted to ethnobotanic, phytochemistry and pharmacologic investigations. Additionally, a phytopharmacological research of the medicinal specimen Croton cajucara Benth ( native plant of the Amazon region of Brazil) was improved. The obtained phytochemical results of the C. spinosa and P. varians showed the presence of flavonoids constituents, among other components. The two flavonoids (2S)-5-hydroxy-7,4 -dimethoxy-flavanone and 5,4 -dihydroxy-3,7,3 -trimethoxy-flavone were isolated from C. spinosa. The antioxidant activity of the hydroalcoholic extracts of C. spinosa and P. varians solubilized in the microemulsion systems SME-1 and SME-4, was evaluated in the DPPHmethod. The used SME systems [obtained with Tween 80: Span 20 (3:1) and isopropyl myristate (IPM)] improved the dissolution of those tested polar extracts, with higher efficacy to the SME-1 system (in which ethanol was included as cosurfactant). The CE50 values evidenced for P. varians were 114 [g/mL (SME-1) and 246 [g/mL (SME-4); for C. spinosa it was 224 [g/mL (SME-1) and 248 [g/mL (SME-4), being the system SME-1 more effective for both tested extracts. The hydroalcoholic extracts of P. varians (HAE-PV) was also submitted to pharmacological screening for antinociceptive activity in animal models. The oral administration of this extract (100, 300 and 1000 mg/kg) inhibited the acetic acid-induced writhing in mice. The higher inhibition (74%) was evidenced to the 1000 mg/kg administered dose. Its effect on the central nervous system (CNS) was investigated by tail flick and formalin-method and reveled that it has negligible antinociceptive action on the CNS. After taking consideration of HAE-PV interaction, Pavonia varians Moric could be used as a potent analgesic agent in case of peripheral algesia, without affecting the CNS. The phytochemical study of the stem bark of Croton cajucara Benth lead to the isolation of 19-nor-clerodanetype diterpenes, as well as to the separation of its fixed oil FO-CC. This non polar oil material reveled to be rich in sesquiterpenes and 19-nor-clerodanes components. The biologic effect of OF-CC was evaluated in the development in vitro of the fungis phytopatogens such as Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii. Significant inhibitory effect of the tested fungis (at 0,2 mg.mL-1 dosage) were comproved. A Mass Spectrometry study of clerodane-type diterpenes was developed in order to identify characteristic fragments on mass spectrometra of both clerodane and 19-nor-clerodane presenting an α,β-insaturated carbonyl moiety at ring A of the decalin-system. For that study, mass spectroscopy data were analysed for 19-nor-clerodanes [trans-dehydrocrotonin (DCTN), trans-crotonin (CTN), cis-cajucarin B (c-CJC-B), and cajucarinolide (CJCR)] and for clerodanes [isosacacarin (ISCR) and transcajucarin A (t-CJC-A)] obtained from the stem bark of C. cajucara, and also clerodane-type from other species. The trans-junction of the enone-system clerodanes was clear correlated with the presence of the characteristic ions at m/z 95, 121 e 205. Meanwhile, the characteristics ions at m/z 122 e 124 were correlated to cis-junction. The trans-junction of the enone-system 19-nor-clerodanes showed characteristics ions at m/z 161, 134 e 121. This study could be successful employed for identification of clerodane constituents from other specimens without any additional spectroscopic analyses, as well as a previously phytochemical analyzes in clerodane project search

Purificação e caracterização de uma β-glucuronidase extraída do molusco mesogastropoda ampularidade Pomacea sp

This work studies the involved enzymatic way in the metabolism of glycosaminoglycans sulfateds in the mollusc Pomacea sp. Had been identified endoglycosidases and exoglycosidases in the enzymatic extract of the mollusc Pomacea sp by means of hydrolysis activity in condroitim sulphate of whale cartilage and of the p-Nitrofenil-β-glucuronide, respectively. The enzymatic extracts qere obtained of Pomacea sp. being used of 0.1 sodium acetate buffer, pH 5.0 and later centrifugated the 8,000 x g and the presents proteins in the sobrenadante were submitted to the fractionament with two crescents ammonium sulphate concentrations, the visualized activity biggest in the F2 fraction (50-80%). The β-glucuronidase (F3) was isolated in gel chromatography filtration Biogel 1.5m, the purification degree was ratified in Chromatography Liquid of high efficiency (HPLC). The enzyme was purificated 6.362,5 times with 35,6% yield. The β -glucuronidase isolated in this work showed a molecular mass of 100 kDa, determined for eletroforese in poliacrilamida gel . The determination of the ideal kinetic parameters for the catalysis of the p-nitrofenil- β -glucuronide for β-glucuronidase, showed excellent activity in pH 5,0 and temperature 65ºC for 6 hours and apparent Km of 72 x 10-2 mM. It is necessary for the total degradation of 3mM of p-N-β-glucoronide, the amount of 1,2μg of ss-glucuronidase. The BaCl2 increased the activity of ss-glucuronidase, and the activity was inhibited completely by the composites SDS and NaH2PO4

Extração e análise da fração lipídica da microalga monoraphidium sp., síntese e caracterização do seu biodiesel

The cultivation of microalgae biomass in order to produce biodiesel arises as an extremely promising aspect, in that the microalgae culture includes short cycle of reproduction, smaller areas for planting and residual biomass rich in protein content. The present dissertation evaluates the performance and features, through spectrometry in the region of infrared with transformed Fourier (FTIR) and spectrometry in the region of UVvisible (UV-Vis), of the extracted lipid material (LM) using different techniques of cell wall disruption (mechanical agitation at low and at high spin and agitation associated with cavitation). The technique of gas chromatography (GC) brought to light the success of alkaline transesterification in the conversion of oil into methyl monoesters (MME), which was also analyzed by spectroscopic techniques (FTIR, proton magnetic resonance (1H NMR) and carbon (13C NMR). Through thermogravimetric analysis (TGA) were analyzed the lipid material (LM), biodiesel and the microalgae biomass. The method which provided the best results concerning the efficiency in extraction of the LP of Monoraphidium sp. (12,51%) was by mechanical agitation at high spin (14 000 rpm), for 2 hours being the ideal time, as shown by the t test. The spectroscopic techniques (1H NMR, 13C NMR and FTIR) confirmed that the structure of methyl monoesters and the chromatographic data (CG) revealed a high content of saturated fatty acid esters (about 70%) being the major constituent eicosanoic acid (33,7%), which justifies the high thermal stability of microalgae biodiesel. The TGA also ratified the conversion rate (96%) of LM into MME, pointing out the quantitative results compatible with the values obtained through GC (about 98%) and confirmed the efficiency of the extraction methods used, showing that may be a good technique to confirm the extraction of these materials. The content of LM microalgae obtained (12,51%) indicates good potential for using such material as a raw material for biodiesel production, when compared to oil content which can be obtained from traditional oil for this use, since the productivity of microalgae per hectare is much larger and requires an extremely reduced period to renew its cultivation