Quantitative HPLC determination and extraction of anthraquinones in Senna alata leaves.
A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of four anthraquinones: rhein, aloe-emodin, emodin, and chrysophanol in Senna alata leaves. The method involves the use of a TSK-gel ODS-80Tm column (5 microm, 4.6 x 150 mm) at 25 degrees C with the mixture of methanol and 2% aqueous acetic acid (70:30, v/v) as the mobile phase and detection at 254 nm. The parameters of linearity, precision, accuracy, and specificity of the method were evaluated. The recovery of the method is 100.3-100.5%, and linearity (r(2) > 0.9998) was obtained for all anthraquinones. A high degree of specificity as well as repeatability and reproducibility (relative standard deviation values less than 5%) were also achieved. The solvent for extraction of anthraquinones from S. alata leaves was examined in order to increase the anthraquinone content of the leaf extract. It was found that a solution of 5% hydrochloric acid (v/v), 5% ferric chloride (w/v), and 15% water in methanol (v/v) was capable of increasing the anthraquinone content in the leaf extract up to 1.67% (w/w).
Reduction of Candida tropicalis biofilm by photoactivation of a Heterophyllaea pustulata extract
Abstract Context: Biofilm formation is an important problem, since this growth mode confers resistance to drugs usually used in therapeutics. Objective: In vitro antifungal activity of extracts obtained from Heterophyllaea pustulata Hook f. (Rubiaceae) were studied against Candida tropicalis biofilms, evaluating the effect of irradiation and the oxidative and nitrosative stresses as possible mechanisms of action. Materials and methods: Hexane, benzene, ethyl acetate and ethanol extracts were evaluated at three concentrations (0.2, 0.1 and 0.05 mg/mL) over mature biofilm, under darkness and irradiation. After 48 h of incubation, biofilm quantitation was performed by the O'Toole and Kolter method. Reactive oxygen species (ROS) was measured by nitro-blue tetrazolium (NBT) reaction and reactive nitrogen intermediates (RNI) by the Griess reagent. Superoxide dismutase activation (SOD, NBT assay) and total antioxidant system (FRAP test) were studied. Results: Only the benzene extract at 0.2 mg/mL reduced the biofilms formation. The slight decrease achieved in darkness (17.06 ± 2.80% reduction) was increased by light action (39.31 ± 3.50% reduction), clearly observing a photostimulation. This great reduction was confirmed by confocal microscopy. In darkness, biofilm reduction was mediated by an increase in RNI, whereas under irradiation, the ROS action was most important. Although no SOD activation was observed, a strong stimulation of the total antioxidant system was detected. HPLC analysis established a high content of several anthraquinones in this extract. Discussion and conclusion: Biofilm reduction by benzene extract was mainly mediated by oxidative stress triggered under light action, confirming a photodynamic sensitization, which could be attributed to its high content of photosensitizing anthraquinones.
Microstructure of the solder-casting zone in bridges of dental gold alloys.
The composition and structure were studied in the cast and in the solder of bridges which had failed clinically. Both materials were gold alloys but with different contents of Pt, Ag and Zn. The metallographic investigation revealed defects mostly in the solder, situated in the subsurface layer. SEM studied of the fracture surface revealed large porosites and a structure of dense parallel lines--striations--, indicating that the material had failed from fatigue. The materials in the fractured bridges were identified by micro-probe measurements. It was stated that cast material and solder material used together, were of different composition.