Aims:  To perform an activity‐guided purification, identification and quantification of antibacterial compounds from Tripodanthus acutifolius infusion. To validate the antibacterial activity of purified substances. Methods and Results:  Bioautographic methods were employed as screening assays for purifying bioactive substances. Purification procedures included sephadex LH‐20 column chromatography and reverse phase HPLC. Identification was achieved by spectroscopic methods (UV‐Vis, MS, NMR and polarimetry) and chromatographic assays (paper chromatography and HPLC). Antibacterial activity was studied by microdilution, colony count and photometric assays, Sytox green stain and transmission electron microscopy (TEM). Four glycoflavonoids (rutin, nicotiflorin, hyperoside and isoquercitrin) and an unusual phenylbutanoid glycoside (tripodantoside) were purified and identified. Tripodantoside was found at 6·59 ± 0·82 g per 100 g of dry leaves. The flavonoids showed bactericidal effect at a concentration of 4 mg ml−1 against Staphylococcus aureus and Pseudomonas aeruginosa strains from American Type Culture Collection, while tripodantoside was almost four times more active than those compounds, with a minimum bactericidal concentration = 1·024 mg ml−1 against these strains. Tripodantoside aglycone showed bacteriolytic effects on the assayed strains, causing evident damages on cell wall and membrane, while tripodantoside did not exhibit those effects. Conclusions:  The antibacterial activity of T. acutifolius infusion would be partially attributed to the purified glycoflavonoids and mainly to tripodantoside. Significance and Impact:  The high extraction yield and the antibacterial activity exhibited by tripodantoside makes this chemical structure of interest to support further studies dealing with chemical modifications to increase the antibacterial activity or to seek another activities.

Introduction: Controlling Enterococcus faecalis is of vital importance in endodontics, as this pathogen is associated with endodontic failure. Experimental evidence has shown that copper has antibacterial activity against other pathogens with similar characteristics. The objective of this study was to determine the antimicrobial activity of copper (II) or cupric (SC-II) sulfate on strains of Enterococcus faecalis and to compare it with the most commonly used antimicrobials. Methodology: We used 33 strains of Enterococcus faecalis isolated from different clinical pictures in different Chilean hospitals. The minimum inhibitory concentration (MIC) of SC-II, chlorhexidine and calcium hydroxide was determined by the broth microdilution technique, following the recommendations given by the Clinical and Laboratory Standards Institute. Results: The MIC for CHX varied in the range of 5-10 µg/ml; SC-II from 1.5 to 12 µg/ml, and HC was >32 mg/ml. The geometric mean of SC-II was 6 µg/ml, lower than that of CHX, which was 7.29 µg/ml. Conclusions: SC-II showed antimicrobial activity at lower concentrations than CHX. HC (which could have been affected by the buffer effect of the broth microdilution technique) showed high values, not comparable to other compounds. We suggest carrying out further studies on the properties of SC-II, such assessing its biocompatibility and reaction with other materials to be used clinically in endodontic therapy. Evaluación antimicrobiana del sulfato de cobre (II) sobre cepas de Enterococcus faecalis. Estudio in vitro.Resumen: Introducción: El control de Enterococcus faecalis es de vital importancia en endodoncia, ya que este patógeno está asociado al fracaso endodóntico. Evidencias experimentales que han demostrado que el cobre presenta actividad antibacteriana en otros patógenos de similares características. El objetivo de este estudio es determinar la actividad antimicrobiana del sulfato de cobre (II) o cúprico (SC-II) sobre cepas de Enterococcus faecalis y compararla con los antimicrobianos más usados en la actualidad. Metodología: Estudio in vitro. Se utilizó la técnica de microdilución en caldo según lineamientos del Clinical and Laboratory Standards Institute, incluyendo 33 cepas de Enterococcus faecalis obtenidas desde hospitales chilenos, para cada una de las cuales se determinó las concentraciones mínimas inhibitorias (CMI) de: SC-II, Clorhexidina (CHX) e Hidróxido de calcio (HC). Resultados: La CMI para CHX tuvo un rango de 5-10 µg/ml, el SC-II de 1,5-12 mM y el HC fue >32 mg/ml. Estas diferencias fueron estadísticamente significativas entre los 3 antimicrobianos utilizados (p<0,001). Conclusiones: El SC-II mostró actividad antimicrobiana a bajas concentraciones, superiores a CHX, pero menores a HC (que pudo ser afectado por el efecto tampón de la técnica de microdilución en caldo). Se sugiere seguir con los estudios de las propiedades del SC-II, como evaluación de biocompatibilidad y reacción con otros materiales para ser utilizados clínicamente en la terapia endodóntica. Pal...

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