Secondary Metabolites

Plants are the main source for obtaining secondary metabolites that are used in the pharmaceutical, cosmetic and food industries. In nature, the performance of bioactive compounds is affected by biotic and abiotic factors, an alternative to overcome this adversity is in vitro plant cultures and particularly plant cell culture that has multiple advantages, highlighting the possibility of controlling variables to increase the content. of compounds of interest. The objective of this research was to determine and optimize the effect of some elicitors on the production of secondary flavonoid metabolites in suspension cell cultures of Thevetia peruviana on a shake flask scale. The experimental part was performed from cell cultures in suspension cells of T. peruviana maintained by the Bioconversion Laboratory of the Universidad Nacional de Colombia, sede Medellin. Firstly, methyl jasmonate (MeJa) 3 µM and salicylic acid (SA) 300 µM were added to two los of cell cultures, that circumstantially they differed in their time suspension state lot 1 (9 months) and lot 2 (3 months). Secondly, certain were evaluated, in order to be optimized; the operational parameters cosidered were: day of addition, concentration and hours of harvest. Finally, the combined effect of MeJa (0.3 µM) and SA (100 µM) was determined in different proportions, the day of elicitation and the hours of harvest were constant. Flavonoid content was quantified by UV-vis spectrophotometry usinfg the AlCl3 complexation method and was evaluated at extracellular and intracellular level. In the first part, at intracellular level, SA generated 14.6 % and 9.56 % more flavonoid content than MeJa in lots 1 and 2, respectively. In the second part, 4.14 mg EQ/g DW were generated with MeJa 0.3 µM, elicitation day 5 and harvest at 90 h; 3.75 mg EQ/g DW were generated with 100 µM SA, elicitation day 0 and harvests at 96 h. In the third part, 4.62 mg EQ / g DW were generated with the combination of MeJa (0.3 µM) – SA (100 µM) in a ratio of 20-80, elicitation on day 0 and harvest at 96 h. Eliciting cell cultures of Thevetia peruviana with MeJa and SA under optimal conditions of concentration, day of addition and hours of harvest increased the content of flavonoid compounds. The results obtained could serve as a basis for the development of investigations at the bioreactor scale.Las plantas son la principal fuente para la obtención de metabolitos secundarios que se usan en la industria farmacéutica, cosmética y alimentaria. En la naturaleza el rendimiento de compuestos bioactivos se ve afectado por factores bióticos y abióticos, una alternativa para superar esta adversidad son los cultivos vegetales in vitro y particularmente el cultivo de células en suspensión que presenta múltiples ventajas destacándose la posibilidad de controlar variables para aumentar el contenido de compuestos de interés. El objetivo del presente trabajo de investigación fue determinar y optimizar el efecto de algunos elicitores sobre la producción de metabolitos secundarios tipo flavonoides en cultivos de células en suspensión de Thevetia peruviana a escala de matraz agitado. La parte experimental se realizó a partir de cultivos de células en células en suspensión de T. peruviana que mantenía el laboratorio de bioconversiones de la Universidad Nacional de Colombia sede Medellín. Primeramente, metil jasmonato (MeJa) 3 μM y ácido salicílico (AS) 300 μM se adicionaron a dos lotes de cultivos de células; que circunstancialmente diferían en su tiempo en estado de suspensión lote 1 (9 meses) y lote 2 (3 meses). Segundamente, determinadas condiciones fueron evaluadas, con el fin de ser optimizadas; los parámetros operacionales que se consideraron fueron: día de adición, concentración y las horas de cosecha. Por último, el efecto combinado de MeJa (0,3 μM) – AS (100 μM) se determinó en diferentes proporciones, el día de elicitación y las horas de cosecha fueron constantes. El contenido de flavonoides se cuantificó por espectrofotometría UV – vis por el método de complejación de AlCl3 y se evaluaron a nivel extracelular e intracelular. En la primera parte, a nivel intracelular AS generaron 14,6 % y 9,56 % más contenido de flavonoides que MeJa en lotes 1 y 2, respectivamente. En la segunda parte, 4,14 mg EQ/g MS fueron generados con MeJa 0,3 μM, elicitación día 5 y cosecha a 90 h; por otro lado, 3,75 mg EQ/g MS fueron generados con AS 100 μM, elicitación día 0 y cosecha a 96 h. En la tercera parte, 4,62 mg EQ/g MS fueron generados con la combinación de MeJa (0,3 μM) – AS (100 μM) en proporción 20 – 80, elicitación el día 0 y cosecha a las 96 h. Elicitar cultivos celulares de Thevetia peruviana con MeJa y AS en condiciones óptimas de concentración, día de adición y horas de cosecha incrementaron el contenido de compuestos flavonoides. Los resultados obtenidos podrían servir como base para el desarrollo de investigaciones a escala de biorreactor.Maestrí

Analysis of the chemical composition of the essential oils extracted from Lippia lacunosa Mart. & Schauer and Lippia rotundifolia Cham. (Verbenaceae) by gas chromatography and gas chromatography-mass spectrometry

Lippia lacunosa and L. rotundifolia (Verbenaceae) are two Brazilian species of complex taxonomic delimitation. The composition of the essential oils from leaves and flowers of these plants was investigated by gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS) analysis. The major components of the essential oils of flowers and leaves of L. lacunosa were: myrcene (14.7% and 11.9%), myrcenone (45.2% and 64.2%), Z-ocimenone (5.7% and 5.2%), and E-ocimenone (14.7% and 4.1%), respectively; whereas in L. rotundifolia (flowers and leaves) were a-pinene (8.7% and 1.8%), myrcene (5.1% and 3.6%), limonene (26.0% and 7.9 %), cis-pinocamphone (4.5% and 3.1%) and myrtenal (22.3% and 16.7%), respectively. The essential oils from L. lacunosa exhibited a strong and pleasant mango aroma, which was related to the presence of myrcene and myrcenone. The marked differences in the chemical composition of their essential oils may represent a powerful tool for the botanical classification.Lippia lacunosa e L. rotundifolia são duas espécies brasileiras que formam um complexo de difícil delimitação taxonômica. A composição química do óleo essencial das folhas e flores dessas plantas foi investigada por cromatografia com fase gasosa (CG) e por cromatografia com fase gasosa acoplada à espectrometria de massas (CG-EM). Principais constituintes dos óleos essenciais de L. lacunosa (flores e folhas): mirceno (14,7% e 11,9%), mircenona (45,2% e 64,2%), Z-ocimenona (5,7% e 5,2%), e E-ocimenona (14,7% e 4,1%), respectivamente; L. rotundifolia (flores e folhas): a-pineno (8,7% e 1,8%), mirceno (5,1% e 3,6%), limoneno (26,0% e 7,9 %), cis-pinocanfona (4,5% e 3,1%) e mirtenal (22,3% e 16,7%), respectivamente. Os óleos essenciais de L. lacunosa apresentaram um forte e agradável aroma de manga, que foi relacionado à presença de mirceno e mircenona. Diferenças fundamentais na composição química de seus óleos essenciais podem representar uma poderosa ferramenta na classificação botânica das espécies

Scents from the Brazilian Cerrado: The essential oil from Siparuna brasiliensis (Siparunaceae)

Cerrado is a term used to describe a savannah-like vegetation, occurring in Central Brazil. It is considered one of the 25 most important biodiversity hotspots in the world and has numerous herbs, including several aromatic plant families, many of which have never been subjected to chemical study [1]. Siparuna brasiliensis (Spreng.) A. DC. (family Siparunaceae) is an endemic Brazilian species, occurring in both the Cerrado and the Atlantic Forest [2]. Differently from other Siparuna species, very few chemical data are available about S. brasiliensis, and none so far regarding its essential oil. During a systematic investigation on the Cerrado flora, S. brasiliensis (CEN herbarium voucher 88294) was sampled in Brasilia, Brazil, and the essential oil obtained by hydrodistillation. According to Brazilian law, collection and access were authorized by the Ministry of Environment (process IBAMA 02001.003166/2013-26). The oil was analyzed by GC-FID and GC-MS on Agilent 7890A and 5975C systems, both with HP-5MS fused silica capillary columns (30 m x 0.25 mm x 0.25 μm). Oil components were identified by comparison of both mass spectra and linear retention indices with spectral libraries and literature. Oil yield was 0.7%. Only 11 compounds were detected, all but one identified by mass spectra and retention indices. Most of the constituents were closely related sesquiterpenes, with gurjunane and guaiane skeletons. The major compound was cyclocolorenone (75.5%). Other components present were 11-hydroxy-3,5-guaiadiene (tentative identification), 2-tridecanone (3.6%), α-cadinol (3.4%) and viridiflorol (3.4%)

Scents from the Brazilian Cerrado: The essential oil from Calea hymenolepis (Asteraceae)

The Brazilian Cerrado is a savannah-like biome with more than 12,000 botanical species in Central Brazil. It is an endangered biome, and considered to be a biodiversity hotspot [1]. Calea hymenolepis Baker is a shrub native from the Cerrado. Samples from a population (n>5) were collected from a rupestrian field in the Chapada dos Veadeiros National Park, in Goiás State, Brazil. According to Brazilian law, collection and access were authorized by the Ministry of Environment (process IBAMA 02001.003166/2013-26). A voucher was deposited at the Embrapa Genetic Resources herbarium and the essential oil was obtained from the leaves by hydrodistillation for 2 h, using a Clevenger-type apparatus. The oil was analyzed by GC-FID and GC-MS on Agilent 7890A and 5975C systems, both with HP-5MS fused silica capillary columns (30 m x 0.25 mm x 0.25 μm). Oil components were identified by comparison of both mass spectra and linear retention indices with spectral libraries and literature. Oil yield was 0.2%. Major compounds present were α-phellandrene (34.2%), p-cymene (10.6%), germacrene D (8.5%), (E)-β-caryophyllene (6.3%) and δ-elemene (4.6%). The oil composition was quite different from other Calea, like C. clematidea, rich in clematerol, a terpenic epoxide [3]. Although subject to systematic phytochemical studies since the 1980's, with the identification of sesquiterpene lactones, chromones, benzopyrans, flavonoids, chalcones and even acetylenes, very few studies have been published regarding the essential oil of Calea species [2]. To the best of our knowledge, this is the first analysis on the essential oil from Calea hymenolepis

Green and roasted arabica coffees differentiated by ripeness, process and cup quality via electrospray ionization mass spectrometry fingerprinting

Direct infusion electrospray ionization mass spectrometry in both the negative ESI(-)-MS and positive ESI(+)-MS ion modes are investigated to differentiate green and roasted Arabica coffees with different stages of ripeness (green, ripe and overripe), post-harvesting process (dry, wet and semi-wet) and coffees with different cup qualities. In the ESI(-)-MS of green coffees, ions from deprotonated fatty acids and chlorogenic acids are the most important for ripeness discrimination. In the ESI(+)-MS, maturity is differentiated by ions from protonated caffeine, chlorogenic acids and K+ adducts of fatty acids. To differentiate between post-harvesting process in both ionization modes, ions from fatty acids, chlorogenic acids, sugars and carboxylic acids generated in the fermentation process are the most representative. Roasted Arabica coffees are also well discriminated: in the ESI(-)-MS, ions from chlorogenic acids and short-chain organic acids derived from sugars are important. In the ESI(+)-MS, discrimination are mainly performed by low m/z ions such as protonated pyridine and alkylpiridines formed via trigonelline degradation. Both ESI(+)-MS and ESI(-)-MS are able to differentiate cup quality for Arabica roasted coffees and the ions used to perform discrimination are the same ones described in ripeness and post-harvesting processes.A habilidade da técnica de espectrometria de massas com infusão direta e ionização por eletronebulização (IES-EM), nos modos de íons positivos e negativos, foi avaliada na diferenciação de cafés Arábica verdes e torrados e com diferentes estágios de amadurecimento (verde, maduro e passado), processo pós-colheita (seco, úmido e semi-úmido) e cafés classificados por prova de xícara. No modo negativo, a análise dos cafés verdes mostrou que os íons correspondentes aos ácidos graxos e ácidos clorogênicos desprotonados são os mais importantes para a discriminação da maturidade. No modo positivo, a maturidade é diferenciada através de íons correspondentes a cafeína, ácidos clorogênicos protonados e adutos de K+ de ácidos graxos. Na diferenciação da pós-colheita, em ambos os modos de ionização, são mais importantes os íons correspondentes aos ácidos graxos, ácidos clorogênicos, açúcares e ácidos carboxílicos formados da fermentação. Cafés Arábica torrados também são discriminados com eficiência. No modo negativo, são importantes os íons correspondentes aos ácidos clorogênicos e ácidos orgânicos de cadeia curta, derivados de açúcares. No modo positivo, a discriminação é realizada por íons de baixa m/z tais como piridina e alquil piridinas protonadas, formadas através da degradação da trigonelina. Ambos os IES(+)-EM e IES(-)-EM são capazes de discriminar diferentes cafés Arábica torrados classificados por prova de xícara e os íons que permitem esta diferenciação são os mesmos descritos para a maturidade e processos pós-colheita.313321Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Óleos essenciais no Brasil: aspectos gerais, desenvolvimento e perspectivas

Essential oils are extracted by steam distillation of plants or cold pressing of citrus fruit pericarp. They are used in food, cosmetic, personal care and pharmaceutical industries. In Brazil, oils from orange and related products contribute to near 97% to the positive commercial performance of the sector. Predatory exploitation and the availability of new sources of raw materials, with more attracting prices, changed the paradigm. Prospective studies, sustainable use of Brazilian biodiversity, domestication of exotic species with commercial relevance, the use of breeding techniques and the development of new applications for essential oils are thematic lines, usually multidisciplinary, which have been prompting the expansion of the research on essential oils. This paper presents an analysis on essential oils balance trade from 2005 to 2008 and some historical data on research and production of essential oils in Brazil