Cristiane Pimentel Victório

Therapeutic value of the genus Alpinia, Zingiberaceae

Plants containing bioactive substances have increasingly become the object of research studies, particularly those plants with therapeutic value. Many species of the genus Alpinia provide a variety of medicinal properties, such as, Alpinia zerumbet (Pers.) Burtt et Smith and A. purpurata (Vieill) K. Schum, which have a significant presence in Brazil. These species have been commercialized in the food and cosmetic industries. However, their greatest importance arises from the medicinal properties of their essential oils containing flavonoids, terpenoids and kavalactones which have been used in folk medicine to treat, for example, arterial hypertension and inflammatory processes. In addition, such species are also used in multidisciplinary studies, including phytochemistry, ethnobotany and biology, indicating the key pharmacological role of this genus in everyday life. Therefore, this work aims to present a bibliographic review of the genus Alpinia and its significance in therapeutic applications.

Chemical Composition of the Leaf Oils of Alpinia zerumbet (Pers.) Burtt et Smith and A. purpurata (Vieill) K. Schum. From Rio de Janeiro, Brazil

Abstract Firesh leaves of Alpinia zerumbet and A. purpurata (Zingiberaceae) were collected in Rio de Janeiro city and subjected to hydrodistillation; their chemical composition was analyzed by GC/MS. These species are used as ornamental plants and in folk medicine. Variations in the leaf oils content and composition of A. zerumbet were found between April and August 2005. In April, the main constituents identiified in leaf oil of A. zerumbet were terpinen-7-al (40.5%) and sabinene hydrate (15.4%); in August, the major components identiified were terpinen-4-ol (29.4%) and 1,8-cineole (23.1%). Leaf oil of A. purpurata was rich in P-pinene (34.7%) and α-pinene (11.8%).

Glandular trichomes of Rosmarinus officinalis L.: Anatomical and phytochemical analyses of leaf volatiles

Abstract Rosmarinus officinalis is known for the production of volatile compounds used in medicinal and food preparations. Leaves of R. officinalis are densely covered with capitate and peltate glandular trichomes where biosynthesis of volatiles mainly occurs. This study aims to conduct a morphological assessment to identify anatomical characteristics of both leaves and trichomes, as well as a chemical analysis of leaf volatile compounds, using histochemistry and stem distillation extraction. Specifically, anatomical and chemical constituents of the secretory structures of R. officinalis leaves were investigated using light and scanning electron microscopy, in addition to gas chromatography. One peltate and three types of capitate glandular trichomes were observed on the leaves of R. officinalis. Histochemical tests showed positive reactions to lipophilic compounds for both capitate and peltate trichomes, with only a slight detection of terpenoids with carbonyl group in peltate glands. Gas chromatography revealed camphor (23.2%) as the main volatile compound, mostly accumulating in peltate glandular trichomes. This phytochemical study of volatile compounds, together with anatomical and histochemical analyses of R. officinalis leaves, demonstrated the importance of leaves as a center of volatile production in peltate and capitate trichomes, as well as the nature of volatile composition, which is involved in species survival.

Effects of Supplemental UV-A on the Development, Anatomy and Metabolite Production of Phyllanthus tenellus Cultured In Vitro

Phyllanthus tenellus is widely used for its antiviral, analgesic and hepatoprotective properties. Although the production of several chemical classes of secondary metabolites is influenced by UV radiation, particularly phenolic compounds, we also know that UV radiation can result in anatomical and developmental damage. However, the morphological, anatomical and phytochemical changes in response to UV‐A exposure are generally understudied in the Phyllanthaceae. Therefore, we evaluated the effects of UV‐A radiation on plant development and leaf anatomy, as well as the production of secondary metabolites and the contents of carotenoids and chlorophylls a and b, in P. tenellus. To accomplish this, in vitro cultures of P. tenellus were maintained for 60 days under white light (WL) and WL plus UV‐A radiation. Results showed different phenotypic responses under additional UV‐A, such as high phenolic metabolite production, increasing dimensions of abaxial epidermis and thickness of palisade parenchyma. Compared to plants cultured under WL, UV‐A radiation caused damage to plant morphogenesis, including a reduced number of branches and shoots, consequently reducing the rate of proliferation. On the other hand, geraniin, ellagic acid and carotenoid contents increased after UV‐A exposure, indicating that this light source is an important resource for inducing phenolic compounds.

VOLATILE COMPOUNDS FROM Rosmarinus officinalis L. AND Baccharis dracunculifolia DC. GROWING IN SOUTHEAST COAST OF BRAZIL

Chemical composition of leaf volatiles of Rosmarinus officinalis and Baccharis dracunculifolia cultured in Southeast of Brazil has been characterized by GC/MS after simultaneous distillation-extraction. The main components in volatiles of these species showed in common α-pinene, myrcene, 1,8 cineole and camphor. Camphor was the major component among volatiles of B. dracunculifolia and R. officinalis with concentrations exceeding 25%. B. dracunculifolia volatiles possessed more sesquiterpenes (21.4%) than R. officinalis (16.7%), such as caryophyllene (1.9%) and α-humulene (0.4%). Lower concentrations of nerolidol and spathulenol were achieved in volatiles of B. dracunculifolia. Considering both species, there was a predominance of monoterpenes.

Tissue culture techniques in the proliferation of shoots and roots of Calendula officinalis.

The high demand for plant material from Calendula officinalis in the production of herbal medicines and cosmetics, turns the technique of plant-tissue culture into one of the alternatives for the improvement of crops over a short period of time. A protocol for tissue culture was developed from segments of seedlings of C. officinalis, in order to improve the proliferation of shoots and roots. We used a Murashige and Skoog (MS½N) medium, reduced to half the concentrations of NH4NO3 and KNO3 to verify the effect of different types of explants (basal, intermediate, and apical), a medium containing beach sand as support instead of agar, and the effect of auxins and cytokinins (TDZ tidiazuron; BAP, 6-benzylaminopurine, IAA, indol-3-acetic acid, IBA, indol-3-butyric acid, NAA, naphthalene-acetic acid) on plant development in vitro. The results showed pronounced rooting from the apical explants, as well as a greater elongation of shoots and number of leaves. The solid medium was more suitable for the C. officinalis cultures. Shoot proliferation was dependent on cytokinins with better results from the influence of TDZ or BAP compared to the other treatments. Plants regenerated from the medium containing TDZ displayed a glazed appearance and morphogenetic deformations. The highest rate for rooting (80%) was obtained using IAA 0.1mgL-1. Through in-vitro propagation, healthy C. officinalis plants were obtained with roots which can acclimatise, allowing the continuous supplement of raw material.

Standardized production of Phyllanthus tenellus Roxb. by plant tissue culture

Exigencies as ethic plant raw material are part of the needs of modern phytotherapy. Micropropagation offers opportunities to obtain mass propagation of superior genotypes in short time. This study aimed to develop a protocol of direct and indirect organogenesis of Phyllanthus tenellus Roxb. Nodal segments from plantlets obtained by in vitro germination were subcultured in modified Murashige and Skoog medium added with different plant growth regulators: IAA (indole-3-acetic acid), IBA (indole-3-butyric acid), GA 3 (3-giberelic acid) and KIN (kinetin). The highest proliferation rate was obtained using the combinations: IBA, KIN + GA 3 (3.5 mg L -1 ) and IBA + KIN (2.4 mg L -1 ). Rooting was intensified after 40 days, reaching 100% for all media with indole-3-butyric acid. Addition of 2,4 dichlorophenoxyacetic acid (2,4D) provided the best results for production of friable calli. Acclimatization was 100% effective for plantlets cultured in control medium, with decrease in survival rate in grown plantlets from media added with growth regulators.

Detection of flavonoids in Alpinia purpurata (Vieill.) K. Schum. leaves using high-performance liquid chromatography

The species Alpinia purpurata is scarcely cited as to ethnopharmacology and phytochemistry. This study aimed to analyze bioactive compounds through high-performance liquid chromatography (HPLC). Hydroalcoholic crude extract was obtained from A. purpurata dried leaves. Folin-Ciocalteau method was used to quantify total phenols, using gallic acid as standard. The obtained result was 15.6 mg GAE g-1. The crude extract was partitioned with the solvents ethyl acetate and butanol, followed by thin-layer chromatography (TLC) and HPLC. The flavonoids kaempferol-3-O-glucuronide and rutin were detected at a higher concentration in ethyl acetate and butanolic extracts. The butanolic extract contains the highest flavonoid percentage (94.3%). A. purpurata presents important flavonoids of therapeutic use, already verified for A. zerumbet. This is the first study verifying the presence of flavonoids in A. purpurata extracts.

Chemical composition of the fractions of leaf oil of Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm. and antimicrobial activity.

Leaf oil prepared by hydrodistillation of Alpinia zerumbet Pers.) B.L. Burtt & R.M. Sm. was analyzed by GC/FID and GC/MS to determine the major compounds and it was also evaluated for antimicrobial activity. The oil presented a high content of oxygenated monoterpenes (52.5%), terpinen-4-ol, 1,8 cineole and γ-terpinene as the major constituents. The antimicrobial activity of leaf oil was tested by drop diffusion and bioauthography methods. Through drop test, all bacteria and fungi tested were inhibited by leaf oil. Preparative TLC (thin-layer chromatography) plates were developed using the mobile phase hexane: ethyl acetate (70:20, v/v) and fractions of the leaf oil were separated in three zones, scraped, extracted from silica and identified by GC/MS. The bioauthography method permitted to verify pronounced inhibition of Cryptococcus neoformans by fractions F2 and F3, both rich in oxygenated monoterpenes. The fraction F2 comprised 1,8 cineole (9.6%), linalool (3.7%) and caryophyllene oxide (5.4%), while fraction F3 showed mainly terpinen-4-ol (43.6%) and an amount of 32.7% oxygenated sesquiterpenes. The obtained data may be used to suggest the constituents of A. zerumbet leaf oil involved in antimicrobial activity.

Simultaneous Distillation-Extraction, Hydrodistillation and Static Headspace Methods for the Analysis of Volatile Secondary Metabolites of Alpinia zerumbet (Pers.) Burtt et Smith. from Southeast Brazil

Abstract Alpinia zerumbet (Zingiberaceae) is cultivated in several tropical areas. It has a high value all over the world as a spice in culinary preparations and for its medicinal properties. The composition of the essential oil obtained by simultaneous distillation-extraction (SDE) and hydrodistillation (HD) from fresh leaves of A. zerumbet was analyzed by GC-MS. In both samples the main constituents were terpinen-4-ol, 1,8-cinole, sabinene and λ-terpinene together with caryophyllene and caryophyllene oxide sesquiterpenes. However, individual percentage values of compounds varied between the methods. The components camphene (0.3 %), p-2-mentha-4(8)-diene (1.4 %) and trans-sabinene hydrate (1.0 %) were obtained only by SDE. Static-headspace (S-HS) was for the first time used to analyze the aroma from fresh leaves of A. zerumbet. Sabinene, 1,8 cineole and λ- terpinene were the main terpenes detected through S-HS. Flower oil was extracted by HD method, and the major components identified were 1,8 cineole (15.5 %), λ-terpinene (13.1 %) and terpinen-4- ol (42.3 %). There was a predominance of monoterpenes.