Carotenoid analysis in citrus is still a challenge due to their complex carotenoid profile and matrix acidity, which can promote isomerization and epoxide-furanoid rearrangements of these compounds. A new method for carotenoid extraction from freeze-dried citrus was developed. Extraction was carried out by magnetic stirring using ethyl acetate and methanol as solvents and Na2CO3 to neutralize organic acids. Liquid-liquid partition was achieved by centrifugation. Carotenoid quantification was accomplished by both UV-Vis and high-performance liquid chromatography-diode array detection (HPLC-DAD). Method repeatability (2.9 to 8.5%) and recovery (82 to 88%) were similar to reported values for procedures including all the carotenoid analysis steps. In 'Valencia' orange pulp, xanthophylls accounted for 98% of the total carotenoid content and the predominant carotenoids were (9Z)-violaxanthin, (all-E)-lutein and (9Z)-antheraxanthin. This method was further applied for carotenoid analysis in freeze-dried 'Tommy Atkins' mango pulp. The developed method is precise, accurate, fast and able to maintain epoxy-carotenoids stability during analysis.

Caveol e cafestol, diterpenos da fração lipídica do café, têm efeitos conhecidos na saúde humana, como atividade anticarcinogênica e hipercolesterolêmica. Existem divergências quanto às concentrações desses compostos reportadas para café torrado, provavelmente devido aos processos de extração empregados. Assim, quatro métodos de preparo de amostra foram estudados: saponificação direta a quente (SDQ), saponificação direta a frio (SDF); e extração por Bligh e Dyer (BD) ou soxhlet (SO) seguida de saponificação. Teores dos diterpenos e seus dehidroderivados obtidos por cromatografia líquida de alta eficiência acoplada a detector de arranjo de diodos e espectrometria de massa (HPLC-DAD-MS/MS) e perfis cromatográficos de café torrado, obtidos pelos quatro métodos, foram comparados. SDQ foi mais eficiente quanto à extração, mostrando melhor separação dos picos cromatográficos e teores de 930,2 (± 36,8), 113,2 (± 4,7), 568,6 (± 16,6) e 87,1 (± 3,7) mg 100 g-1 para caveol, dehidrocaveol, cafestol e dehidrocafestol, respectivamente. O extrato SDQ apresentou teores de diterpenos (caveol e cafestol) 15% superiores àqueles obtidos por SDF e até 88% maiores que pelos métodos SO e BD.

Kahweol and cafestol, diterpenes from the unsaponifiable fraction of coffee, present known effects on human health such as anticarcinogenic and hipercholesterolemic activities. There are discrepancies regarding the levels reported for these compounds in roasted coffee, probably due to the extraction processes. Therefore, four sample preparation methods were studied: direct hot saponification (DHS), direct cold saponification (DCS); and Bligh and Dyer (BD) or Soxhlet (SO) extraction followed by saponification. The levels of diterpenes and their dehydro derivatives obtained by high performance liquid chromatography with diode array and mass spectrometry detectors (HPLC-DAD-MS/MS) and the chromatographic profiles of roasted coffee, obtained by these four methods, were compared. DHS was more efficient for extraction, showing better separation of chromatographic peaks and levels of 930.2 (± 36.8), 113.2 (± 4.7), 568.6 (± 16.6) and 87.1 (± 3.7) mg 100 g-1 for kahweol, dehydrokahweol, cafestol and dehydrocafestol, respectively. The DHS extract presented a diterpene content (kahweol and cafestol) 15% superior to that of DCS and up to 88% superior than using SO and BD methods.

Although soybean isoflavones naturally accumulate in their conjugated forms, the beneficial effects on human health of soybean-containing foods have been credited to their aglycone forms. In the present study we analyzed the effects of a chemical agent, sodium nitroprusside (SNP), in eliciting the exudation of non-conjugated isoflavones from intact soybean seeds, embrionary axes and cotyledons. The isoflavones in the exudates were determined by high performance liquid chromatography and mass spectrometry. The effect of the exudates on the emission of nitric oxide (NO) and on the proliferation of breast carcinoma cells (MCF-7) was also evaluated. SNP elicitation increased the production of the aglycone forms dose- and time-dependently. Exudates of embrionary axes and cotyledons stimulated NO emission and showed biphasic effects on viability of MCF-7 cells. At lower concentrations both extracts presented proliferative effects (10-25%), and at higher concentrations inhibited (15%) cell proliferation. The biphasic effect might be due to the action of isoflavone aglycones in activating estrogen receptors which in turn stimulate the production of NO. Overall, the results suggest that soybean extracts enriched in isoflavone aglycones by elicitation with SNP could be exploited as a functional ingredient in the food industry.

A fractional factorial design 2(5-1) was used to evaluate the effect of temperature, pH, and concentrations of yeast extract, tryptone and Nitsch's trace elements on the biomass, total carotenoids and protection against singlet oxygen by carotenoid extracts of the bacterium Thermus filiformis. In addition, the carotenoid composition was determined by high-performance liquid chromatography connected to a diode array and mass spectrometer detectors (HPLC-DAD-MS/MS). The production of biomass ranged from 0.113 to 0.658 g/L, the total carotenoid from 137.6 to 1,517.4 mg/g and the protection against singlet oxygen from 4.3 to 85.1 %. Results of the fractional factorial design showed that temperature had a negative effect on biomass production and a positive effect on carotenoid content and protection against singlet oxygen, besides, high levels of pH value, concentrations of yeast extract and tryptone had a positive effect on biomass production only at lower temperatures. The main carotenoids of T. filiformis were thermozeaxanthins. In the tested conditions, changes in the levels of the variables influenced the biomass, carotenoid production, and protection against singlet oxygen, although they did not influence the carotenoid profile. The results of this study provide a better understanding on the interactions among certain nutritional and cultivation conditions of a thermophile bacterium, Thermus filiformis, on biomass and carotenoid amounts, as well as on the antioxidant capacity.

O alto consumo de frutas tem sido associado à baixa incidência de doenças crônico-degenerativas, provavelmente devido à presença de compostos bioativos, como os antioxidantes, nestes alimentos. Os teores de compostos bioativos -ácido ascórbico, fenólicos totais, flavonóides e carotenóides totais - foram determinados em polpas obtidas a partir de 18 frutas tropicais adquiridas no Brasil. A atividade anti-radical livre foi avaliada pelo método ABTS. As frutas que apresentaram maior teor de compostos bioativos foram buriti, cajá-manga, canistel, murici, physalis, piquia e tucumã. Considerando a composição dos compostos bioativos analisados e as atividades anti-radical livre, as frutas foram divididas em 5 grupos, segundo Análise Hierárquica de Agrupamentos. Aplicando a Análise de Componentes Principais, a atividade anti-radical livre mostrou alta correlação com compostos fenólicos totais (r = 0,99) e flavonóides (r = 0,86); porém, a correlação encontrada foi muito pequena para ácido ascórbico (r = 0,02) e carotenóides totais (r = 0,08).

High ingestion of fruits has been associated with low incidence of chronic-degenerative diseases, probably due to the presence of bioactive compounds in these foods, such as antioxidants. The levels of bioactive compounds - ascorbic acid, total phenolics, flavonoids and carotenoids - were determined in 18 pulps obtained from tropical fruits acquired in Brazil. The free radical scavenger activity was evaluated by the ABTS assay. The fruits that showed higher levels of bioactive compounds were buriti, otaheite apple, egg-fruit, golden spoon, physalis, piquia and star nut palm. Considering the composition of bioactive compounds and free radical scavenger capacities the fruits were divided into five groups, according to Hierarchical Cluster Analysis. Applying Principal Component Analysis, free radical scavenger showed high correlation with total phenolic compounds (r = 0.99) and flavonoids (r = 0.86); however, the correlation was found to be very poor with ascorbic acid (r = 0.02) and with total carotenoids levels (r = 0.08).

As amoras-pretas apresentam coloração atrativa devido à presença de antocianinas, porém a utilização destes pigmentos como corante natural é difícil, pois alguns fatores, especialmente pH>3, provocam sua descoloração. Por outro lado, a microencapsulação tem sido utilizada na indústria para proteger ingredientes ativos. Os objetivos deste estudo foram estabelecer as condições de encapsulação de antocianinas de amora-preta, avaliar as características das cápsulas, bem como a liberação de antocianinas. Três polissacarídeos (curdlana, pectina e alginato de sódio) foram avaliados como material de parede para obter cápsulas por gelificação. A curdlana foi escolhida, pois foi a única goma capaz de formar um gel firme sob baixo pH. As cápsulas apresentaram forma esférica e multinucleada, e a eficiência de encapsulação variou entre 80,3 e 96,7%. As curvas de liberação apresentaram comportamento cinético de primeira ordem, com forte efeito "burst" com 80 a 100% das antocianinas liberadas em solução pH 1 após 20 min.

Blackberry shows an attractive color due to the presence of anthocyanins; however the use of these pigments as natural colorant is difficult since some external agents, specially pH>3, cause their discoloration. On the other hand, encapsulation techniques have been widely used in the industry to protect active ingredients. The aims of this study were to establish the encapsulation conditions of anthocyanins from blackberry, to evaluate the capsules characteristics, as well as the anthocyanins release. Three polysaccharides (curdlan, pectin and sodium alginate) were evaluated as wall material to obtain capsules by gelation. Curdlan was chosen since it was the only gum capable to form hard gel under low pH value. The capsules showed spherical form and multinucleated appearance, and encapsulation efficiency ranged from 80.3 to 96.7%. The release curves followed first order kinetics, with a strong burst effect, 80 to 100% of the anthocyanins released in solution at pH 1 after 20 min.

As pesquisas com licopeno têm sido intensificadas uma vez que o consumo de alimentos ricos neste caroteno, como tomate e seus produtos, tem sido associado com o menor risco de desenvolvimento de câncer de próstata. Os objetivos deste trabalho foram (1) desenvolver e otimizar um método para extração de licopeno a partir de descarte de tomate, (2) utilizar este método na obtenção de licopeno a partir de diferentes coletas de tomate, (3) obter cristais de licopeno puro e (4) acompanhar o processo de purificação de licopeno por cromatografia líquida de alta eficiência em coluna C30. O método desenvolvido e otimizado através de planejamento experimental fatorial para a obtenção de licopeno consistiu de uma etapa preliminar para retirada de água do tomate, com 4 extrações de 30 minutos, cada uma com 30mL de etanol, seguido por 4 extrações de 120 minutos com acetato de etila e relação massa/volume de 1:0,7. O teor de carotenóides médio (calculado como licopeno) obtido de 6 lotes de tomate foi de 59,2 ± 21,8µg/g. Através de duas cristalizações sucessivas obteve-se licopeno com 98% de pureza. A análise por CLAE mostrou a presença de traços dos isômeros 5-cis, 5-cis, 5'-cis- e 13-cis-licopeno no cristal de licopeno.

Over the past few years, research regarding lycopene has intensified since the consumption of lycopene rich foods such as tomato and tomato products has been associated with reduced risks of developing prostrate cancer. Thus the objectives of this research were (1) develop and optimise methodology for the extraction of lycopene from tomato waste, (2) use the methodology to obtain lycopene from different tomato batches, (3) produce high purity lycopene crystals and (4) follow the purification process of lycopene using high performance liquid chromatography on a C30 column. The method developed to obtain lycopene from tomato residue, and optimised using a factorial experimental design, consisted of a preliminary water removal step with four 30-minute extractions, each with 30mL ethanol, followed by four 120-minute extractions with ethyl acetate, each extraction with a mass/volume ratio of 1:0.7. The mean carotenoid content (calculated as lycopene), obtained from 6 tomato pickings, was 59.2 ± 21.8µg/g. Lycopene with 98% purity was obtained from two successive crystallisation processes. HPLC analysis showed the presence of traces of the 5-cis-, 5-cis,5'-cis- and 13-cis-lycopene isomers in the lycopene crystal.

Um método por cromatografia líquida de alta eficiência (CLAE) foi otimizado para a análise da composição de carotenóides de cinco linhagens de Rhodotorula.A extração com ruptura mecânica da parede celular da levedura com areia tratada mostrou ser mais eficiente que a ruptura química com dimetilsulfóxido. Os carotenóides foram separados e quantificados por CLAE em coluna de C18 utilizando como fase móvel acetonitrila/metanol (0,1% trietilamina)/acetato de etila (75:15:10) e 100% metanol (0,1% trietilamina) entre as injeções, com vazão de 1 mL/min. Em todas as linhagens, os carotenóides majoritários encontrados foram torularrodina, toruleno, ³-caroteno e ²-caroteno. Os teores totais de carotenóides, em µg/g, foram de 251,7 em R. glutinis,123,5 em R. rubra,113,2 em R. araucariae,105,8 em R. lactosa ede 103,7 em R. minuta.

A method for extraction and HPLC separation of carotenoids from fiveRhodotorula strains was optimized. The extraction by mechanical disruption of the yeast cell wall with fine treated sand was shown to be more efficient than chemical disruption with dimethylsulfoxide. The carotenoids were separated and quantified by HPLC on a C18 column using as mobile phase acetonitrile/methanol (0.1% triethylamina)/ethyl acetate (75:15:10) with 100% methanol (0.1% triethylamine) between the injections, at a flow rate of 1.0 mL/min. In all strains, the major carotenoids found were torularhodin, torulene, ³-carotene and ²-carotene. The total carotenoid contents, in µg/g, obtained were 251.7 for R. glutinis,123.5 for R. rubra,113.2 for R. araucariae,105.8 for R. lactosaand 103.7 for R. minuta.

A composição de carotenóides de maracujá-amarelo in natura (Passiflora edulis) comercializado em Campinas, Brasil, foi avaliada utilizando-se cromatografia líquida de alta eficiência. Em todas as amostras foram encontrados os seguintes carotenóides: beta-criptoxantina, prolicopeno, cis-zeta-caroteno, zeta-caroteno, beta-caroteno e 13-cis-beta-caroteno, além de neurosporeno e gama-caroteno em alguns lotes. Em geral, os teores de carotenóides nos diversos lotes foram significativamente diferentes. Em dois dos cinco lotes analisados, o z-caroteno foi o carotenóide principal, contribuindo com 41 e 62% do total de carotenóides, enquanto que em outros dois lotes, o beta-caroteno foi o majoritário, com 55 e 74% do total e em um dos lotes o cis-zeta-caroteno esteve presente em maior concentração, com 29%. Este tipo de inversão entre os carotenóides majoritários não é comumente encontrado em frutas e vegetais.

The carotenoid composition of fresh yellow passion fruit (Passiflora edulis) commercialized in Campinas, Brazil, was evaluated by high performance liquid chromatography. In all the samples, the following carotenoids were found: beta-cryptoxanthin, prolycopene, cis-zeta-carotene, zeta-carotene, beta-carotene, 13-cis-beta-carotene. Besides that neurosporene and gamma-carotene were also found in some lots. In general, the carotenoid contents were significantly different amongst the lots. In two of the five lots, zeta-carotene was the major carotenoid, contributing with 41 and 62% of the total carotenoid content, while in other two lots beta-carotene was the main carotenoid, with 55 and 74% of the total carotenoid, and in one of the lots cis-zeta-carotene was present as the major carotenoid, with 29%. The inversion occurring between the main carotenoids is not frequently found in fruits and vegetables.

Numerosos trabalhos comprovaram que os carotenóides principais de folhas verdes são invariavelmente luteína, beta-caroteno, violaxantina e neoxantina. No entanto, há discordância em torno dos carotenóides minoritários. Portanto, a espectrometria de massas por impacto de elétrons e cromatografia líquida de alta eficiência com detector de arranjo de diodos foram utilizados para confirmar a identidade de carotenóides minoritários com atividade provitamínica A em verduras folhosas brasileiras. Os carotenóides pró-vitamínicos A, incluindo os isômeros cis e trans de beta-caroteno, foram separados em coluna de C18 polimérica, Vydac 201TP54, com metanol/água (98:2) como fase móvel. Os espectros UV-visível e de massas confirmaram o carotenóide monoidroxilado como sendo alfa-criptoxantina e não beta-criptoxantina como aponta a literatura internacional. Todas as onze folhas analisadas (agrião, alface crespa, alface lisa, almeirão, caruru, chicória, couve, espinafre, rúcula, salsinha e taioba) apresentaram alfa-criptoxantina, 13-cis-beta-caroteno e 9-cis-beta-caroteno, enquanto que alfa-caroteno foi encontrado em apenas quatro folhas (caruru, couve, salsinha e taioba).

The main carotenoids from green leafy vegetables have been consistently found to be lutein, beta-carotene, violaxanthin and neoxanthin. However, there is a controversy about the identity of minor carotenoids. Therefore, electron impact mass spectrometry and high performance liquid chromatography with a diode array detector were used in order to confirm the identity of the minor provitamin A carotenoids in Brazilian green leaves. The provitamin A carotenoids, including the cis and trans isomers of beta-carotene, were separated on a polymeric C18 column, Vydac 201TP54, with MeOH/H2O (98:2) as mobile phase. The UV-visible and mass spectra confirmed that the monohydroxy carotenoid present in Brazilian green leafy vegetable to be alpha-cryptoxanthin, and not beta-cryptoxanthin as reported in the international literature. All eleven green leaves analyzed (water-cress, unheaded lettuce, lettuce, wild chicory, "caruru", common chicory, kale, spinach, endive, roquette, parsley, and "taioba") had alpha-cryptoxanthin, 9-cis and 13-cis- beta-carotene, whereas alpha-carotene was found in only four of these leaves ("caruru", kale, salsinha and "taioba").