Leaves of Eugenia florida DC. have been used in folk medicine as hypotensive, hypoglycemic, antipyretic and for gastrointestinal disorders, among others. In this study dried leaves were extracted with ethanol and the extract was partitioned with hexane, CHCl3, EtOAc, BuOH and water to afford triterpenoid- and polyphenol-rich fractions which were analyzed by GC-MS and UFLC-DAD-ESI-Ion Trap-MSn. All fractions were tested against the proliferation of leukemic K562 and human melanoma SK-Mel-28 cell lines and for their ability to scavenge DPPH radical. Twenty-nine compounds were fully identified (among triterpenic and phenolic acids, flavonol- and dihydroflavonol-monoglyclosides) based on mass fragmentation patterns and comparison to standards and/or literature data. The ability to scavenge DPPH radical resulted in the following order: BuOH ≈ EtOAc > crude extract ≈ Water >> CHCl3 >> hexane; with the two most active reaching half of the TBHQ control potency (EC50 6.59 ± 0.03 µg mL-1). On testing against the proliferation of K562 and SK-Mel-28 cell lines, the most significant inhibitions were achieved on the former by the crude extract (IC50 12 µg mL-1) and the hexane and EtOAc fractions at 15 µg mL-1.

Uncaria tomentosa (Willd.) DC. e Uncaria guianensis (Aubl.) Gmel., conhecidas comercialmente como unha-de-gato, são trepadeiras lenhosas nativas das florestas tropicais amazônica e central-americana. Essas espécies de Rubiaceae enfrentam ultimamente um eminente risco de extinção devido tanto ao extrativismo indiscriminado quanto ao desflorestamento de seus habitats naturais. Esse trabalho descreve métodos de cultivo in vitro para as duas espécies, analisando o perfil e o teor dos alcalóides oxindólicos pentacíclicos marcadores por CLAE no material vegetal obtido. Os resultados mostraram que os métodos desenvolvidos foram capazes de produzir plantas com perfil e teor dos alcalóides alvos similares à plantas nativas e cultivadas in vivo, com a vantagem adicional de produção em menor tempo de mudas próprias para plantio no campo. Os dados evidenciaram que o cultivo in vitro pode ser uma ferramenta viável para a produção das duas espécies com impacto na sua comercialização e na preservação das florestas.

Uncaria tomentosa (Willd.) DC. and Uncaria guianensis (Aubl.) Gmel., commercially known as cat's claw, are large woody vines native of the Amazon and Central America rainforests. These Rubiaceae species face nowadays an imminent risk of extinction due to indiscriminate harvesting in the wild as well as to increasing deforestation of their natural habitat. This work describes in vitro cultivation methods for both species with determination, by HPLC, of the pentacyclic oxindole alkaloid contents and profiles of the plant material. The results have proved that the methods we developed were able to produce plants with alkaloid profiles and contents similar to the wild and in vivo cultivated species, and with the additional advantage of producing suitable young plants in a shorter period of time. The data showed that the in vitro technique can be a feasible tool for the growth of the species, and may thus be important for their commercialization and for their conservation as a forest resource.

O gênero Uncaria (Rubiaceae) é representado na América do Sul e Central por duas espécies: U. tomentosa (Willd.) DC. e U. guianensis (Aubl.) Gmel., conhecidas popularmente como unha-de-gato. Ambas são trepadeiras perenes, sendo empregadas na prevenção e cura de várias doenças. Nessas plantas são encontrados alcalóides oxindólicos e indólicos, triterpenos glicosilados, taninos e flavonóides. Seis alcalóides oxindólicos pentacíclicos, considerados seus marcadores: especiofilina, mitrafilina, uncarina F, isomitrafilina, pteropodina e isopteropodina, são usados na padronização do material vegetal e fitoterápicos derivados. O presente trabalho descreve o desenvolvimento de metodologia analítica qualitativa utilizando cromatografia em camada delgada (CCD) para determinação do perfil dos seis alcalóides oxindólicos pentacíclicos marcadores das espécies. O desenvolvimento do método incluiu a comparação entre o uso do extrato metanólico bruto, e de frações enriquecidas obtidas por partição ácido-base clássica ou pelo uso de resina básica Poliamida 6. Utilizou-se gel de sílica como fase estacionária, e variaram-se alguns parâmetros como: eluentes, concentração da amostra, espaço de eluição e tipos de reveladores. O método desenvolvido em CCD mostrou-se confiável, reprodutível e seletivo para os alcalóides alvos, sendo aplicado na análise de amostras de folhas e caule das duas espécies e também de fitoterápicos comerciais à base de U. tomentosa.

The species Uncaria tomentosa (Willd.) DC. and U. guianensis Gmel. (Rubiaceae), known as cat's claw, are large woody vines occurring in the Amazon rain forest and other tropical areas of South and Central America. It has been used medicinally by indigenous peoples for at least 2,000 years for several diseases. Tetra- and pentacyclic oxindole alkaloids, triterpenoid glycosides, sterols and flavonoids are found in these plants. Among these metabolites, six pentacyclic oxindole alkaloids, speciophylline, mitraphylline, pteropodine, uncarine F, isopteropodine and isomitraphylline, are considered to be the biochemical markers and are used to standardize commercial herbal medicines. The present study describes the development of an analytical methodology to determine the profile of these alkaloid markers through thin layer chromatography (TLC). This development has also included a comparison among the use of the crude methanol extract and fractions obtained through the classical acid-base partition or by using the basic resin Polyamide 6. Silica gel was used as stationary phase with the variation of some parameters such as solvent systems, sample concentration, distance of development and detection method. The TLC method developed was shown to be reliable, reproducible and selective for the target alkaloids. It has been applied to the analysis of leaves and stems from both species as well as phytopharmaceutical derivatives based on U. tomentosa.

Quantitative GC-FID was evaluated for analysis of methylated copaiba oils, using trans-(-)-caryophyllene or methyl copalate as external standards. Analytical curves showed good linearity and reproducibility in terms of correlation coefficients (0.9992 and 0.996, respectively) and relative standard deviation (< 3%). Quantification of sesquiterpenes and diterpenic acids were performed with each standard, separately. When compared with the integrator response normalization, the standardization was statistically similar for the case of methyl copalate, but the response of trans-(-)-caryophyllene was statistically (P < 0.05) different. This method showed to be suitable for classification and quality control of commercial samples of the oils.