Evaluation of the antibacterial synergism of two plant extracts belonging to Bignoniaceae family and development of a topical formulation

Abstract Fridericia caudigera and Cuspidaria convoluta (Bignoniaceae) species, which grow in the northwest of Argentina, have shown antibacterial effect against strains isolated from skin infections, and each one displayed synergism with commercial antibiotics. The aims of this work were to evaluate the antibacterial activity and toxicity of the combination of these two plant species, and to design a stable gel for topical use including the blend of extracts. The combination of extracts was evaluated for synergistic effects (chequerboard assay), genotoxicity (Ames test) and cytotoxicity (Artemia salina test). A gel was subsequently formulated with the combination of extracts using carboxymethylcellulose as a polymer. The following physico- chemical characteristics of the gel formulation: pH, viscosity, spreadability and total phenol content, as well as resistance to severe temperature changes, biological activity (diffusion in agar), in vitro permeation (Franz cells) and primary dermal irritation (Draize test) were analyzed. The combination of extracts showed a synergistic effect on pathogenic bacteria and was not toxic in the in vitro tests. The gel was stable and retained the antimicrobial activity of the original extracts. The formulation proposed in this work could constitute an alternative for primary skin infections since it proved to be safe for topical administration.

Actividad antioxidante de cuatros especies vegetales del nordeste argentino / Antioxidant activity of four plant species in the Argentine Northeast / Atividade antioxidante de quatro espécies de plantas no Nordeste argentino

RESUMEN Objetivos: Determinar la composición fitoquímica y evaluar la actividad antioxidante de los extractos hidroalcohólicos de S. haematospermum; P. niruri; P. tenellus; E. serpens. Metodología: La composición fitoquímica se analizó por cromatografía en capa fina y la actividad antioxidante se observó de modo cualitativo por ensayo autográfico. Los fenoles totales se cuantificaron con el método de Folin-Ciocalteu y flavonoides totales con la reacción de complejación de aluminio. Para determinar la actividad antioxidante se utilizaron las técnicas de decoloración del radical libre DPPH• (2,2- difenil-1-picrilhidracilo) y del radical catión ABTS•+ (ácido 2,2'-azino-bis (3-etilbenzotiazolin)-6-sulfónico) y del poder reductor del catión hierro (III). Resultados: El estudio fitoquímico evidenció en todas las especies la presencia de metabolitos bioactivos. El contenido de fenoles totales osciló entre: 627,71 ± 1,45 y 369,8±52,6 mg EAG/g muestra y de flavonoides totales fue de 12,2 ± 0,1 hasta 4,5 ± 0,3 mg Q/g muestra. Las especies S. haematospermum y P. tenellus presentaron mayor contenido de polifenoles. Los ensayos de actividad antioxidante permitieron reconocer que S. haematospermum y P. tenellus tienen capacidad de mayor inhibición de los radicales libres probados y mostraron mayor poder reductor los extractos hidroalcohólicos S. haematospermum y P. niruri.

SUMMARY Objectives: To determine the phytochemical composition and evaluate the anti-oxidant activity of the hydroalcoholic extracts of S. haematospermum; P. niruri; P. tenellus; E. serpens. Methodology: The phytochemical composition was analyzed by thin layer chromatography and the antioxidant activity was observed qualitatively by autographic assay. Total phenols were quantified with the Folin-Ciocalteu method and total flavonoids with the aluminum complexation reaction. To determine the antioxidant activity, the discoloration techniques of the free radical DPPH• (2,2-diphenyl-1-picrilhydracil) and of the cation radical ABTS•+ (2,2-Azino-bis-3-ethylbenzothiazolin-6-sulfonic acid) and the reducing power of the iron (III) cation. Results: The phytochemical study showed the presence of bioactive metabolites in all species. The total phenol content ranged from: 627.71 ± 1.45 to 369.8 ± 52.6 mg EAG / g sample and total flavonoids was 12.2 ± 0.1 to 4.5 ± 0.3 mg Q / g sample. The species S. haematospermum and P. tenellus presented higher content of polyphenols. The antioxidant activity tests allowed recognizing that S. haematospermum and P. tenellus have a capacity for greater inhibition of the tested free radicals and the hydroalcoholic extracts S. haematospermum and P. niruri showed greater reducing power.

RESUMO Objetivos: Determinar a composição fitoquímica e avaliar a atividade antioxidante dos extratos hidroalcoólicos de S. haematospermum; P. niruri; P. tenellus; E. serpens. Metodologia: A composição fitoquímica foi analisada por cromatografía em camada delgada e a atividade antioxidante foi observada qualitativamente por ensaio autográ-fico. Fenóis totais foram quantificados pelo método de Folin-Ciocalteu e flavonóides totais pela reação de complexação de alumínio. Para determinar a atividade antioxidante, as técnicas de descoloração do radical livre DPPH• (2,2-difenil-1-picrilhidracil) e do radical catiônico ABTS•+ (2,2-azino-bis-3-etilbenzotiazolin-6-sulfônico ácido) e o poder redutor do cátion ferro (III). Resultados: O estudo fitoquímico mostrou a presença de metabólitos bioativos em todas as espécies. O conteúdo total de fenol variou de: 627,71 ± 1,45 a 369,8 ± 52,6 mg EAG / g amostra e flavonóides totais foi de 12,2 ± 0,1 a 4,5 ± 0,3 mg Q / g amostra. As espécies S. haematospermum e P. tenellus apresentaram maior teor de polifenóis. Os testes de atividade antioxidante permitiram reconhecer que S. haematospermum e P. tenellus apresentam maior capacidade de inibição dos radicais livres testados e os extratos hidroalcoólicos S. haematospermum e P. niruri apresentaram maior poder redutor.

Formulación de productos cosméticos con aceite de semillas de Cucurbita argyrosperma C. Huber / Formulation of cosmetic products from Cucurbita argyrosperma C. Huber seed oil

RESUMEN El presente trabajo tiene como objetivo aprovechar el aceite de las semillas de Cucurbita argyrosperma C. Huber (sin. Cucurbita mixta Pangalo), como fitoingrediente en la elaboración de productos cosméticos. El aceite empleado se obtuvo por extracción con soxhlet y se realizó su caracterización fisicoquímica. Se elaboraron emulgeles (O/W) variando el agente viscosante y emulgente. Las formulaciones logradas se valoraron mediante la evaluación de las propiedades organolépticas (color, olor, brillo), fisicoquímicas (pH, conductividad) y extensibilidad y fueron sometidas a estudios de estabilidad. Se obtuvieron productos tipo emulgel de color amarillento con ligero olor ceroso, con pH aceptable para la piel. Todas las formulaciones resultaron estables. Los resultados demostraron que las formulaciones propuestas permitieron incorporar el aceite de origen vegetal logrando un preparado cosmético de características y estabilidad aceptables.

SUMMARY The objective of this work is to take advantage of the oil of the seeds of Cucurbita argyrosperma C. Huber (sin. Cucurbita mixta Pangalo), as a phytoingredient in the elaboration of cosmetic products. The oil used was obtained by extraction with soxhlet and its physicochemical characterization was carried out. Emulgels (O/W) were prepared by varying the viscosifying and emulsifying agent. The formulations achieved were evaluated by evaluating the organoleptic properties (color, odor, brightness), physicochemical (pH, conductivity) and extensibility and were subjected to stability studies. Emulgel type products with yellowish color with slight waxy odor were obtained, with acceptable pH for the skin. All the formulations were stable. The results showed that the proposed formulations allowed incorporating the oil of vegetable origin achieving a cosmetic preparation of acceptable characteristics and stability.

In vitro antioxidant, antilipoxygenase and antimicrobial activities of extracts from seven climbing plants belonging to the Bignoniaceae.

OBJECTIVES: This study aimed to evaluate the in vitro antioxidant capacity, to determine the anti-inflammatory effect due to lipoxygenase inhibition and to test the antimicrobial activity of ethanolic extracts from leaves of seven climbing species belonging to the Bignoniaceae family. These species are Adenocalymma marginatum (Cham.) DC., Amphilophium vauthieri DC., Cuspidaria convoluta (Vell.) A. H. Gentry, Dolichandra dentata (K. Schum.) L. G. Lohmann, Fridericia caudigera (S. Moore) L. G. Lohmann, Fridericia chica (Bonpl.) L. G. Lohmann and Tanaecium selloi (Spreng.) L. G. Lohmann. METHODS: The antioxidant activity was evaluated using three methods, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power. Lipoxygenase-inhibiting activity was assayed spectrophotometrically; the result was expressed as percent inhibition. The antimicrobial activity was assessed using the agar disk diffusion method. Minimal inhibitory concentration (MIC) and minimal bactericidal/fungicidal concentration were also determined for each extract against 12 pathogenic bacterial strains of Staphylococcus aureus and seven fungal strains of the Candida genus. The identification of the major compounds present in the most promising extract was established by high-performance liquid chromatography-tandem mass spectrometry. RESULTS: C. convoluta, F. caudigera, and F. chica exhibited the best antioxidant activity by scavenging DPPH and ABTS+ radicals and reducing Fe3+ ion. These extracts showed a notable inhibition of lipoxygenase. F. caudigera was found to have the lower MIC value against S. aureus strains and six Candida species. The extracts of F. caudigera and C. convoluta were active even against methicillin-resistant S. aureus. C. convoluta had higher total phenol content, better antioxidant activity and superior anti-inflammatory and antimicrobial activity. The main phenolic compounds found in this extract were coumaric and hydroxybenzoic acid derivatives and glycosylated and nonglycosylated flavones. CONCLUSION: Most of the extracts exhibited antioxidant activity as well as in vitro inhibition of lipoxygenase. The excellent antimicrobial activity of T. selloi and F. chica supports their use in traditional medicine as antiseptic agents. The extracts of F. caudigera and C. convoluta, both with notable biological activities in this study, could be used as herbal remedies for skin care. In addition, this study provides, for the first time, information about phenolic compounds present in C. convoluta.

In vitro antimicrobial activity of 20 selected climber species from the Bignoniaceae family.

Hydroalcoholic and aqueous extracts of some climber species from the Bignoniaceae family that grow in the north of Argentina were evaluated for in vitro antibacterial activity against Gram-positive and Gram-negative strains. By means of bioautography and disc diffusion methods, it could be determined that all infusions were not active, whereas the hydroalcoholic extracts of seven species were able to inhibit bacterial growth. The minimum inhibitory concentration and minimum bactericidal concentration observed were between 62.5 and 1000 µg gallic acid equivalent (GAE)/mL and between 125 and 1000 µg GAE/mL, respectively. The tested extracts were more active against Gram-positive microorganisms. Time-kill experiments indicated that all extracts have bacteriostatic activity. Phytochemical screening showed the presence of terpenoids, phenols and flavonoids. The amount of phenolic compounds and flavonoids was higher in tinctures when compared with infusions. These results suggest the presence of antibacterial substances in the hydroalcoholic extracts, which could be used for the treatment of infections.