Analysis of variance (ANOVA)-simultaneous component analysis (ASCA) is a method of choice for factorial design studies of environmental impacts on plant metabolomes and can be used to quantitatively carry out this comparative analysis. The impacts of seven mixture design extractor systems made up of ethanol, dichloromethane, and hexane and their 1:1 binary and 1:1:1 ternary solvents for several replicate experiments (n = 3, 4, 5) were assessed using ASCA models determined from Fourier-transform infrared (FTIR), near-infrared (NIR), and UV-Vis spectroscopic measurements on yerba mate leaf extracts. This analysis considered two-factor effects: secondary sexual dimorphism (male and female plants) and cultivation systems (monoculture and agroforestry), as well as their interaction effect. The three binary solvents were found to be more efficient extractor systems for all four detectors as they found 83 main and interaction effects significant at or above the 95% confidence level compared with only 47 for the pure solvent extracts. Binary solvent extracts resulted in averages between 44.04 and 86.61% for the ASCA total effect variances compared with 40.62 to 71.07% for pure extractors. Of the 60 significant effects found for experiments with 5 repetitions 53 or 88% were obtained with only triplicate determinations. The choice of spectroscopic technique and solvent system have large impacts on metabolomic analysis results. ANOVAsimultaneous ANOVA simultaneous (ASCA ethanol dichloromethane 11 1 1: 111 1:1: n 3 4 Fouriertransform Fourier transform FTIR, FTIR , (FTIR) nearinfrared near NIR, NIR (NIR) UVVis UV Vis twofactor two factor male plants monoculture agroforestry, agroforestry agroforestry) 8 95 4404 44 04 44.0 8661 86 61 86.61 4062 40 62 40.6 7107 71 07 71.07 extractors 6 88 determinations results (FTIR (NIR 9 440 0 44. 866 86.6 406 40. 710 7 71.0 86. 71.

The effects of water-deficit stress on irrigated and unirrigated field plants of Coffea arabica L. genotype IAPAR 59 were investigated. Plant extracts were obtained following an ethanol-dichloromethane-hexane statistical mixture design. Proton nuclear magnetic resonance (1H NMR) fingerprints of the extracts were discriminated using factor analysis (FA) and hierarchical clustering techniques. Extracts from the 1:1:1 ternary mixture presented the largest discriminations compared with those from the pure solvents or their 1:1 binary mixtures. Metabolites resulting from fermentation processes and nutritional deficiencies as well as senescence and abscission precursors such as lactate, arginine and methionine were prevalent in unirrigated plants that can provoke expressive decreases in bean productivity as well as premature plant aging. Amino acids that control regulatory, physiological processes and soil salinization have higher concentrations in the irrigated plants. The NMR assignments of eighteen substances observed here were confirmed by electrospray ionization mass spectrometry.

Rollinia mucosa fruit has generated great interest due to the presence of bioactive compounds, which exhibit promising biological activities. Fingerprinting analytical techniques, chromatographic-diode array detection (DAD), infrared, mass spectrometric, 1H and 13C nuclear magnetic resonance (NMR) spectra, associated with chemometric analysis and mixture design, were used to determine chemical discriminations of sun-exposed and self-shaded leaves of Rollinia mucosa in each of the four seasons. Sunlit leaves in winter presented higher metabolite signals related to the acetogenins, whereas shaded leaves have higher abundances of carbohydrates and terpenes. Sunlit leaves harvested in the summer had the smallest metabolite abundances. Fingerprints confirmed the presence of two acetogenins, annonacin A and annonastatin. Ternary and quaternary mixtures of the statistical mixture design were most effective for revealing important discriminations.

Coffee bean chemical compositions has been extensively studied. However, there is a small amount of research on other parts of the coffee plant, including leaves. Fourier transform infrared (FTIR) spectral profiles of Coffea arabica L. cv. IAPAR 59 leaf extracts from a simplex-centroid design were studied by principal component analysis (PCA) to evaluate the effect of solvent extractor on its metabolites. PCA indicated that the extractor solvents containing ethanol were the most suitable for this study. FTIR spectra in conjunction with orthogonal signal correction and partial least squares-discrimination analysis (OSC-PLS-DA) were used to classify and discriminate the leaves of irrigated and non-irrigated plants by bands related to carbohydrates, amino acids and lipids. Leaves receiving different intensities of solar radiation were also discriminated by bands corresponding to caffeine, carbohydrates and lipids. FTIR spectral profile analyzed with chemometric tools showed to be a useful, powerful and simple procedure to discriminate coffee leaves collected from different microclimate conditions.

Ultraviolet characteristic profiles and infrared spectroscopic (FTIR) fingerprints of green bean extracts of Coffea arabica L., cv. IAPAR 59, cultivated in two planting patterns, rectangular and square, and at two different densities, 10,000 and 6,000 plants ha-1, identified as R10, R6, S10, and S6 were analyzed with principal component and hierarchical cluster analyses. A simplex centroid design for four solvents (ethanol, acetone, dichloromethane, hexane) was used for sample extraction. The largest chlorogenic acid (CGA) contents were found at the lower planting density. The dichloromethane extracts of the S10 treatment showed the highest levels of unsaponifiable lipids (cafestol and kahweol). The R6 treatment showed a slightly higher content of cafestol and kahweol. Cluster analysis of FTIR fingerprints confirmed that the CGA and caffeine levels differentiate the spatial arrangements. The FTIR fingerprints suggest that green beans from S6 and R10 were richer in lipids and the other two treatments had more sugars and proteins.

Vernonia condensata Baker leaves have different uses in Brazilian folk medicine, including as analgesic and anti-inflammatory agents. The purpose of this study was to evaluate the seasonal effects on their high performance liquid chromatography with a diode array detector (HPLC-DAD-UV) and ultra-performance liquid chromatography coupled to a mass spectrometer with an electrospray interface (UPLC-ESI-MS) fingerprints, as well as their analgesic activities in mice. There were significant seasonal effects on the relative abundances of the metabolites of the V. condensate leaves as well as on their activities. Analgesic activities in the writhing test were observed with the polar fraction of the leaf extracts collected in autumn, winter and summer (400 mg kg-1); and with the intermediate fraction of leaves collected in autumn (25 and 400 mg kg-1) and in the summer (100 mg kg-1). In conclusion, the results confirm peripherally-mediated anti-inflammatory and analgesic activities for V. condensata leaves and suggest that these are influenced by the harvesting season. N-oxides alkaloids as well as vernonioside play important roles in determining this activity.

A statistical mixture-design technique was used to study the effects of different solvents and their mixtures on the yield, total polyphenol content, and antioxidant capacity of the crude extracts from the bark of Schinus terebinthifolius Raddi (Anacardiaceae). The experimental results and their response-surface models showed that ternary mixtures with equal portions of all the three solvents (water, ethanol and acetone) were better than the binary mixtures in generating crude extracts with the highest yield (22.04 ± 0.48%), total polyphenol content (29.39 ± 0.39%), and antioxidant capacity (6.38 ± 0.21). An analytical method was developed and validated for the determination of total polyphenols in the extracts. Optimal conditions for the various parameters in this analytical method, namely, the time for the chromophoric reaction to stabilize, wavelength of the absorption maxima to be monitored, the reference standard and the concentration of sodium carbonate were determined to be 5 min, 780 nm, pyrogallol, and 14.06% w v-1, respectively. UV-Vis spectrophotometric monitoring of the reaction under these conditions proved the method to be linear, specific, precise, accurate, reproducible, robust, and easy to perform.

Um método computacional interativo foi desenvolvido para a separação eletroforética de 13 compostos fenólicos de azeite de oliva extravirgem, usando valores individuais de resposta para cada pico. Um planejamento composto central foi executado para a otimização da concentração de tetraborato de sódio, pH e voltagem aplicada. Foram determinados modelos estatísticos para oito respostas de resolução e treze de mobilidades efetivas. Seis modelos de resolução apresentaram significativa falta de ajuste após ANOVA, o que limitou sua acurácia para uso nas funções de desejabilidade de Derringer-Suich na busca pelas condições ótimas de separação. Nenhum dos 13 modelos de mobilidade efetiva apresentou falta de ajuste significativa. Visto que não foi possível definir valores alvos para as funções de desejabilidade, um programa de computador interativo, desenvolvido em nossos laboratórios, foi aplicado aos modelos individuais de cada pico. Movimentos do mouse ou do cursor foram executados para definir as condições experimentais nas simulações dos eletroferogramas. Essas simulações resultaram em uma melhor separação dos picos, especialmente para os picos de apigenina e luteolina, em 35 min, comparado aos obtidos para cerca de 50 min com os modelos de resolução. Experimentos de verificação executados 2 e 3 anos depois confirmaram a robustez dos modelos.

An interactive computer method is proposed for the electrophoretic separation of 13 phenolic compounds from extra-virgin olive oil using single peak response values. A central composite design was executed for optimization of the sodium tetraborate concentration, pH and applied voltage. Statistical models were determined for eight resolution responses and thirteen effective mobilities. Six of the resolution models had highly significant ANOVA lack of fit values, limiting their accuracies for use in Derringer´s desirability function search for optimal separation conditions. None of the 13 effective mobility models suffered from significant lack of fit. Since it is not possible to define effective mobility target values for the desirability function, an interactive computer program developed in our laboratories was applied to the single peak models. Mouse or cursor movements were executed to define experimental conditions in model simulations of the electropherogram. These simulations resulted in superior peak separations, especially for the apigenin and luteolin peaks, in 35 min, compared with those obtained in close to 50 min with the resolution models. Verification experiments performed 2 and 3 years later confirmed the robustness of the models.

O desenvolvimento de fármacos para terapia foto-dinâmica (TFD) é uma importante área de pesquisa devido ao seu crescente uso em aplicações médicas. Portanto, é importante desenvolver novos métodos de bioensaios para TFD fotossensibilizadores que sejam de baixo custo, de fácil execução e altamente sensíveis às condições do meio. Corantes xantênicos (fluoresceína, rosa de bengala B, eritrosina B e eosina Y) iluminados com luz LED foram investigados usando Artêmia salina como bioindicador da atividade fotodinâmica. Neste estudo, três fatores foram investigados (i) concentração do fotossensibilizador; (ii) o tempo de irradiação LED e (iii) o tempo de espera entre a adição do fotossensibilizador e o início da irradiação. Para analisar a foto-mortandade da A. salina foi aplicado o planejamento fatorial 23 completo. A morte da A. salinafoi relacionada à estrutura do corante e à interação entre o tempo de irradiação e a concentração do fotossensibilizador. Cerca de 60% da morte do crustáceo foi obtida usando rosa de bengala B, que apresenta o maior rendimento quântico de oxigênio singlete devido aos átomos de iodo substituintes no anel xantênico. O bioensaio proposto usando A. salina, corantes xantenos e irradiação LED foi apropriado para a avaliação quantitativa de fármacos para TFD.

The development of drugs for photodynamic therapy (PDT) is an important area of research due to their growing use in medical applications. Therefore, it is important to develop new bioassay methods for PDT photosensitizers that are inexpensive, easy to handle and highly sensitive to environmental conditions. Xanthene dyes (fluorescein, rose bengal B, erythrosine B and eosin Y) with LED light sources were investigated using Artemia salina as a bioindicator of photodynamic activity. In this study, three factors were investigated: (i) photosensitizers concentration, (ii) the LED irradiation time and (iii) the waiting time between the addition of the photosensitizers and the beginning of the irradiation. To analyze the photo-killing of A. salina, it was employed a 23 full factorial design. The death of A. salina was related to dye structure and the interaction between the irradiation time and the photosensitizers concentration. About 60% of crustaceans death was obtained using rose bengal B, which presentes the highest quantum yield of singlet oxygen due to the number of iodide substituents in the xanthenes ring. The proposed bioassay using A. salina, xanthene dyes and LED irradiation was found suitable for quantitative PDT drug evaluation.

Erythrina speciosa Andrews, Fabaceae, é usada na região Sul do Brasil como sedativa e tranquilizante. Neste estudo, uma possível ação central do extrato aquoso (EA) e fração diclorometano (DCM) das folhas da E. speciosa foi avaliada em camundongos machos submetidos a testes comportamentais 1 h após o tratamento (gavage). Também foi avaliada a dose letal 50% (DL50) como indicativa da toxicidade aguda desta planta. O EA diminuiu a locomoção (50, 100 e 400 mg/kg) e o levantar (50 e 400 mg/kg) no teste de campo aberto mas não alterou nenhum dos comportamentos avaliados nos testes de labirinto em cruz elevado, rotarod e sono induzido. A DCM não alterou nenhum dos comportamentos avaliados. A DL50 de ambos os extratos foi estimada como sendo >2000 mg/kg. Os resultados sugerem ausência de efeito ansiolítico e depressor do Sistema Nervoso Central das folhas de E. speciosa. Entretanto, como mecanismos serotonérgicos podem estar envolvidos na ação farmacológica de plantas do gênero Erythrina e o teste de labirinto em cruz elevado não é adequado para avaliar o efeito de drogas serotonérgicas, nossos resultados não invalidam o uso desta planta na medicina popular, mas apontam a necessidade de se investigar o mecanismo de ação envolvido no possível efeito central de plantas do gênero Erythrina.

Erythrina speciosa Andrews, Fabaceae, is used in the South of Brazil as sedative and tranquilizers. In this study, behavioral effects of aqueous (AE) and dichloromethane (DCM) extracts of E. speciosa leaves were evaluated in male mice, as well as their lethal dose 50% (LD50). The extracts were administered by gavage in doses ranging from 50 to 400 mg/kg 1 h before the behavioral tests. AE decreased ambulation (50, 100 and 400 mg/kg) and rearing (50 and 400 mg/kg) in the open-field but did not influence elevated plus maze, rotarod and pentobarbital-induced sleep tests. No behavioral effects were observed after DCM administration. LD50 for both extracts were higher than 2000 mg/kg. The results showed that AE and DCM extracts of E. speciosa leaves do not produce anxiolytic effect in the elevated plus maze nor seems to depress the Central Nervous System. However, since serotonergic mechanisms may be involved in the pharmacological action of Erythrina plants and the elevated plus maze test is not adequate to evaluate serotonergic drugs, our results do not invalidate the use of this plant in folk medicine but suggest that the mechanism involved in a possible central action of Erythrina needs to be clarified.

Chemometric activities in Brazil are described according to three phases: before the existence of microcomputers in the 1970s, through the initial stages of microcomputer use in the 1980s and during the years of extensive microcomputer applications of the ´90s and into this century. Pioneering activities in both the university and industry are emphasized. Active research areas in chemometrics are cited including experimental design, pattern recognition and classification, curve resolution for complex systems and multivariate calibration. New trends in chemometrics, especially higher order methods for treating data, are emphasized.

The equilibria, the spectra and the identities of the species of Cr(VI) that are present in aqueous solution have long been an active subject of discussion in the literature. In this paper, three different chemometric methodologies are applied to sets of UV/Visible spectra of aqueous Cr(VI) solutions, in order to solve a chemical system where there is no available information concerning the composition of the samples nor spectral information about the pure species. Imbrie Q-mode factor analysis, followed by varimax rotation and Imbrie oblique projection, were used to estimate the composition of Cr(VI) equilibrium solutions and, by combining these results with the k-matrix method, to obtain the pure spectra of the species. Evolving factor analysis and self modeling curve resolution were used to confirm the number of the species and the resolution of the system, respectively. Sets of 3.3×10-4 and 3.3×10-5 mol L-1 Cr(VI) solutions, respectively, were analyzed in the pH range from 1 to 12. Two factors were identified, which were related to the chromate ion (CrO4(2-)) and bichromate ion (HCrO4-). The pK of the equilibrium was estimated as 5.8.