Uso de tecnologias limpas para a obtenção de biosurfactantes e carboidratos prebióticos a partir do Ginseng brasileiro (Pfaffia glomerata)

Orientadores: Maria Angela de Almeida Meireles Petenate, Diego Tresinari dos SantosTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: O ginseng brasileiro (Pfaffia glomerata) é uma planta nativa do Brasil que apresenta uma rica composição de compostos bioativos, incluindo a beta-ecdisona que apresenta propriedades estimulantes, as saponinas com atividade surfactante e açúcares prebióticos que possuem atividade prebiótica. Diante destes aspectos, diferentes processos de extração foram estudados a fim de maximizar a recuperação dos compostos bioativos de interesse, bem como separá-los em diferentes frações, utilizando apenas solventes não tóxicos. Inicialmente, realizou-se um estudo técnico e econômico da extração de beta-ecdisona e açúcares prebióticos das raízes e partes aéreas do ginseng brasileiro empregando extração com água subcrítica. Os resultados demonstraram que é possível obter extratos das raízes com concentração de beta-ecdisona de 0,7% (base seca, b.s.), enquanto que os extratos das partes aéreas apresentaram apenas 0,3% (b.s.) de beta-ecdisona. Em relação à extração de açúcares prebióticos, os extratos obtidos das raízes apresentaram um teor de frutooligossacarídeos de até 8,8% (b.s.), o que faz desta matéria-prima uma importante fonte desses compostos. A avaliação econômica do processo de extração com água subcrítica considerou principalmente o teor de beta-ecdisona dos extratos obtidos e demonstrou que o processamento das raízes é técnica e economicamente mais favorável do que o processamento das partes aéreas. Considerando os resultados obtidos neste estudo e dados da literatura sobre extração de compostos bioativos do ginseng brasileiro, um processo de extração intensificado realizado em duas etapas foi proposto com o intuito de aumentar a recuperação de beta-ecdisona, saponinas e açúcares prebióticos das raízes de ginseng brasileiro. Realizou-se a primeira etapa com etanol e a segunda com água, ambas a 333 K e avaliou-se o efeito da pressão neste processo. Observou-se que para a obtenção de compostos bioativos das raízes de ginseng brasileiro, processos realizados a pressão ambiente apresentam maior recuperação dos compostos do que quando realizados a alta pressão. Nesse processo obteve-se um extrato etanólico com 5,6% (b.s.) de beta-ecdisona e 47% (b.s.) de saponinas com concentração micelar crítica (CMC) = 6 mg?mL-1. O extrato etanólico não apresentou açúcares prebióticos. Já extrato aquoso apresentou um teor de frutooligossacarídeos de 9% (b.s.), além de 0,5% (b.s.) de beta-ecdisona e 24% (b.s.) de saponinas com CMC = 18 mg?mL-1. Desta forma, o processo de extração intensificado permitiu a extração e fracionamento dos compostos bioativos das raízes de ginseng brasileiro, uma vez que foi possível obter um extrato etanólico rico em beta-ecdisona e saponinas e um extrato aquoso rico em açúcares prebióticos. A partir destes resultados, um estudo econômico foi realizado a fim de comparar diferentes cenários de produção para avaliar a viabilidade econômica do processo de extração intensificado. Esse estudo confirmou o processo intensificado como sendo a rota de produção de extratos das raízes de ginseng brasileiro economicamente mais viável. Ao final do desenvolvimento deste trabalho demonstrou-se que é possível substituir processos de extração convencionais por processos inovadores que não utilizam solventes tóxicos, minimizam a geração de resíduos através do melhor aproveitamento das matérias-primas e são mais eficientes do ponto de vista energético além de serem altamente promissores do ponto de vista econômicoAbstract: Brazilian ginseng (Pfaffia glomerata) is a native plant from Brazil that contains a rich composition of bioactive compounds, including beta-ecdysone with stimulating effects, saponins with surfactant activity and prebiotic sugars with prebiotic effects. In this context, different extraction process were studied to maximize the bioactive compounds recovery, as well as to fractionate them, using only non-toxic solvents. Firstly, a techno-economic evaluation of the extraction of beta-ecdysone from roots and aerial parts of Brazilian ginseng using subcritical water was performed. The results showed that is possible to obtain extracts from Brazilian ginseng roots with 0.7% (dry basis, d.b.) of beta-ecdysone, while the Brazilian ginseng aerial parts extracts yielded 0.3% (d.b.) of beta-ecdysone. In terms of prebiotic sugars, the extracts from Brazilian ginseng roots showed a fructooligosaccharides content of 8.8% (d.b.), which makes this raw material an important source of such compounds. Since to date the beta-ecdysone is the main compound with commercial value obtained from Brazilian ginseng, the economic evaluation of the subcritical water extraction process accounted only the beta-ecdysone content in the extracts. The economic evaluation showed that the manufacturing of roots was a great opportunity of business, while the manufacturing of the aerial parts should not be undertaken. Considering the results obtained in this study and data from literature about extraction of bioactive compounds from Brazilian ginseng, an intensified process was proposed to increase the beta-ecdysone, saponins and prebiotic sugars recovery from Brazilian ginseng roots. The intensified process was performed in two steps: the first step used ethanol as solvent and the second one used water as solvent, both at 333 K. The effect of pressure on this process was evaluated. It was observed that to obtain bioactive compounds from Brazilian ginseng roots, the use of ambient pressure yielded greater results than those obtained at high pressure. In this process, an ethanolic extract containing 5.6% (d.b.) of beta-ecdysone, 47% (d.b.) of saponins with critical micellar concentration (CMC) = 6 mg?mL-1 was obtained. No prebiotic sugar was detected in the ethanolic extract, otherwise, the aqueous extract showed a fructooligosaccharides content of 9% (d.b.) besides 0.5% (d.b.) of beta-ecdysone and 24% (d.b.) of saponins with CMC = 18 mg?mL-1. In this way, the intensified process allowed the extraction and fractionating of the bioactive compounds from Brazilian ginseng roots, since it was possible to obtain an ethanolic extract rich in beta-ecdysone and saponins and an aqueous extract rich in prebiotic sugars. Based on the experimental results, an economic study was developed aiming to compare different production scenarios to evaluate the economic viability of the intensified process. The study confirmed that the intensified process is the best way to produce Brazilian ginseng extracts. At the end of the development of this work, it was showed that it is possible replace conventional extraction processes by innovative processes, which use non-toxic solvents, reduces the residues generation and are more efficient from the energetic point of view. Furthermore, the proposed processes showed high economic feasibilityDoutoradoEngenharia de AlimentosDoutora em Engenharia de Alimentos2013/17260-5FAPESPCNP

Intensification Of Bioactive Compounds Extraction From Medicinal Plants Using Ultrasonic Irradiation.

Extraction processes are largely used in many chemical, biotechnological and pharmaceutical industries for recovery of bioactive compounds from medicinal plants. To replace the conventional extraction techniques, new techniques as high-pressure extraction processes that use environment friendly solvents have been developed. However, these techniques, sometimes, are associated with low extraction rate. The ultrasound can be effectively used to improve the extraction rate by the increasing the mass transfer and possible rupture of cell wall due the formation of microcavities leading to higher product yields with reduced processing time and solvent consumption. This review presents a brief survey about the mechanism and aspects that affecting the ultrasound assisted extraction focusing on the use of ultrasound irradiation for high-pressure extraction processes intensification.888-9

Extraction of valuable compounds from granadilla (Passiflora ligularis Juss) peel using pressurized fluids technologies

Valuable products, including carotenoids, phenolic compounds, and pectin were obtained from granadilla peel using green technologies based on pressurized fluids. The processes of pressurized liquid extraction (PLE) and CO2-expanded ethanol (GXE) were evaluated to recover the carotenoids and phenolic compounds, while pressurized hot water extraction (PHWE) was evaluated to extract pectin from the granadilla peel. The results demonstrated that although PLE and GXE had no significant difference in the carotenoids yield and antioxidant activity of the extracts obtained, PLE was faster than GXE to achieve the highest extraction yield. The pectin yield obtained by PHWE was affected by the process temperature with the highest value obtained at 160 °C. Further studies are needed to characterize the chemical composition of the pectin obtained, but the present results enable establishing a biorefinery for the complete use of granadilla wastes (seeds and peel).The authors thank the financial support of the Ministry of Education (MECESUP) through the Program Strengthening Featuring MECESUP-ANT 1999 (Attraction of Advanced Human Capital from Abroad short-term visits) and the FONDEQUIP EQM160073 Project of the National Agency for Research and Development (ANID) ex-National Commission for Scientific and Technological Research(CONICYT) of Chile. Renata Vardanegathanks Horizon 2020 MSCA grant agreement No 101062938 for the postdoctoral fellowship.info:eu-repo/semantics/publishedVersio

Obtaining saponins from Brazilian ginseng roots (Pfaffia glomerata) by dynamic low-pressure solvent extraction assisted by ultrasound

Orientadores: Maria Angela de Almeida Meireles, Diego Tresinari dos SantosDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: Espécies do gênero Pfaffia (Amaranthaceae) têm sido comercializadas como substitutas para Panax (ginseng, Araliaceae) e em função da morfologia similar de suas raízes são popularmente conhecidas como ginseng brasileiro. Dentre as espécies de Pfaffia conhecidas, a Pfaffia glomerata é a mais importante, uma vez que esta é a única que possui o composto ?-ecdisona, o qual representa a saponina de maior interesse desta planta, devido a seus efeitos terapêuticos. Atualmente, cápsulas contendo extratos de P. glomerata são comercializadas sob indicação de auxiliar da memória e tônico. Além destas indicações, as raízes desta planta apresentam teores expressivos de saponinas (10-17%), o que faz com que o extrato das raízes de P. glomerata apresente um grande potencial para aplicação como emulsificante/surfactante. Comercialmente, extratos de P. glomerata são obtidos por percolação em leito fixo, no entanto, com baixos rendimentos. O objetivo deste estudo foi obter um extrato de ginseng brasileiro (P. glomerata) rico em saponinas empregando o processo de extração por percolação a baixa pressão, o qual é um processo de extração já estabelecido comercialmente, sob a assistência do ultrassom, que vem se destacando como uma técnica emergente na melhoria de processos de extração. Deste modo, as variáveis resposta estudadas foram o rendimento global de extração (X0) e as propriedades emulsificantes/surfactantes dos extratos obtidos: Índice de emulsificação (E24) e redução da tensão superficial da água (RTSA); os parâmetros avaliados foram: método de extração (percolação, PE; percolação assistida por ultrassom durante toda extração, PEU; percolação assistida por ultrassom durante o início da extração, PEUI e percolação assistida por ultrassom com pulsos, PEUP), temperatura (40 e 60°C) e solventes (água; etanol:água [35:65, v/v]; etanol:água [70:30, v/v], isopropanol:água [35:65, v/v] e isopropanol:água [70:30, v/v]). As condições de extração que proporcionaram maiores valores de X0 (62,8%), E24 (54,4%) e RTSA (36,3%) foram água, 60°C, PEUP; isopropanol:água (70:30, v/v), 60°C, PEUP e isopropanol:água (70:30, v/v), 60°C, PEUI, respectivamente. Estas condições foram selecionadas para estudo cinético e obtenção de parâmetros cinéticos ajustados que mostraram que, quando água foi empregada como solvente de extração sob ultrassom, obteve-se a maior recuperação de extrato com menor tCER (período de taxa de extração constante). Portanto, a condição de extração com água, 60°C, PE, representa a condição ótima em termos de rendimento. Ainda, realizou-se a quantificação do teor de ?- ison nos xtr tos o ti os n s in ti s on o t v -s -14% (base seca) de ?-ecdisona nos extratos das raízes de P. glomerata. A condição de extração que proporcionou os extratos com maior teor de ?-ecdisona foi a isopropanol:água (70:30), 60°C, PEUI, comprovando a maior seletividade de extração da mistura de solventes isopropanol:água (70:30, v/v), já que este foi também o solvente que apresentou extratos com maiores propriedades surfactantes. Portanto, pode-se concluir que ao final deste estudo, ao empregar o método de percolação em leito fixo assistida por ultrassom no início da extração utilizando isopropanol:água (70:30) como solvente de extração a 60°C, obteve-se uma condição de extração otimizada que fornece extratos das raízes de P. glomerata com alta recuperação de compostos bioativosAbstract: Species of the genus Pfaffia (Amaranthaceae) have been commercialized as substitute to Panax (ginseng, Araliaceae). Due to the similar morphology of its roots to those of ginseng, they are popularly known as Brazilian ginseng. Among the species of Pfaffia, the P. glomerata is the most important because it is the only one that contains ?- ecdysone; which is the most important saponin from this plant due their therapeutic effects. Nowadays, capsules containing P. glomerata roots extracts are commercialized under indication to of improving the memory and as a tonic. Besides these indications, the P. glomerata roots exhibit expressive saponins content (10-17%); therefore, its extract can potentially be used as an emulsifier/ surfactant. Commercially, P. glomerata roots extracts are obtained by low-pressure solvent extraction in fixed bed extractors, however, with low yields. The aim of this study was to obtain an extract from Brazilian ginseng roots (P. glomerata) rich in saponins applying the process of low-pressure solvent extraction, which is an extraction process commercially established, assisted by ultrasound ; ultrasound assisted extraction is an emerging technique used to improve extraction process. The effects of the following parameters were evaluated on the overall extraction yield (X0) and surfactant properties (Emulsification index, E24 and reduction of water surface tension, RTSA) of P. glomerata roots extracts: extraction method (percolation, PE; ultrasound assisted percolation during the entire process, PEU; ultrasound assisted percolation at the beginning of the process, PEUI and ultrasound assisted percolation with pulses, PEUP), temperature (40 and 60°C) and extracting solvents (water, ethanol:water [35:65], ethanol:water [70:30], isopropanol:water [35:65] and isopropanol:water [70:30]). The extraction conditions that provided higher values to X0 (water, 60°, PEUP), E24 (isopropanol:water [70:30], 60°C, PEUP) and RTSA (isopropanol:water [70:30], 60°C, PEUI) were selected for the kinetic study; the kinetic parameters were estimated fitting the experimental data to a linear spline with 3 straight lines. The kinetic parameters showed that when water was used as extracting solvent assisted by ultrasound, the higher extract recovery was obtained with the shortest tCER (constant extraction rate period). Therefore, the extraction condition with water, 60°C, PEUP is the optimum condition with respect to the X0. The content of ?-ecdysone was evaluated on the extracts obtained from kinetic study. The content of ?-ecdysone in the P. glomerata roots xtr ts w r -14% (dry base, d.b.). The extraction condition that provided the higher extraction of ?-ecdysone was that using isopropanol:water [70:30], 60°C, PEUI; the extracts with the best surfactant properties were obtained in this extracting condition . Therefore, it was concluded that applying ultrasound at the beginning of process using IsoC3OH:water (70:30) as extracting solvent at 60°C, was the optimized extraction condition to obtain extracts from P. glomerata rootsMestradoEngenharia de AlimentosMestra em Engenharia de Alimento

Proposal For Fractionating Brazilian Ginseng Extracts: Process Intensification Approach

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Brazilian ginseng (Pfaffia glomerata) roots contain several bioactive compounds, including beta-ecdysone with several therapeutic effects, saponins with surface activity and fructooligosaccharides (FOS) with prebiotic effects. Regarding this rich composition, a two-step intensified extraction process that uses the same solid-liquid extraction apparatus was developed to obtain and fractionate the bioactive compounds from Brazilian ginseng roots using ethanol and water as the extraction solvents. The intensified process was compared to a conventional extraction process using water as the solvent. The beta-ecdysone and saponins were mainly concentrated in the ethanolic extract obtained in step 1 of the intensified extraction process, while the FOS was isolated in the aqueous extract obtained in step 2. The effect of pressure (0.1, 5 and 10 MPa) on the extraction yield, beta-ecdysone content, saponin content, water surface tension reduction rate and FOS content was evaluated using the analysis of variance (ANOVA) statistical method. The highest beta-ecdysone content (5.6%, d.b.) was obtained at ambient pressure (0.1 MPa), while the highest saponin content (55%, d.b.) was obtained at 5 MPa. However, the extracts that had better surface activity were also obtained at ambient pressure. The total FOS content obtained at ambient pressure was 7.9% (d.b.). The kinetic study showed that suitable process times for the first and second steps were 38 and 110 min, respectively. (C) 2016 Elsevier Ltd. All rights reserved.1967380FAPESP [2013/17260-5, 2010/16485-5, 2012/19304-7, 2012/10685-8, 2013/04304-4]CAPES [7545-15-0]CNPq [301301/2010-7, 470916/2012-5]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Hibiscus sabdariffa L. leaves as an alternative source of bioactive compounds obtained through high pressure technologies

The aim of the present study was to obtain bioactive compounds from hibiscus leaves through supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE). For SFE, temperature, pressure, and co-solvent were evaluated on extraction yield (X0), content of bioactive compounds and antioxidant activity (AA). For PLE, temperature and solvent composition were evaluated on the same responses. The highest X0 was observed at 60 °C, 300bar and 15% of co-solvent for SFE, and at 100 °C and ethanol 75% for PLE. For bioactive compounds, different results were observed. For SFE, the best condition was at 60 ºC, 100bar and 15% of co-solvent, and for PLE at 80 ºC with ethanol 99.5%. Regarding AA, SFE extracts presented highest values at 60 ºC, 100bar and 5% of co-solvent while PLE presented better results at 60 °C and 99.5%. Chlorogenic acid and quercetin were the main compounds identified in the extracts.This research received financial support of São Paulo Research Foundation (FAPESP) (Brazil, Grants 2019/27354-3, and 2021/07639-3). G. Nunes Clímaco thanks Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 for a Ph.D. fellowship. Renata Vardanega thanks European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 101062938 for the postdoctoral fellowship.info:eu-repo/semantics/publishedVersio

Supercritical Carbon Dioxide Technology for Recovering Valuable Phytochemicals from <i>Cannabis sativa</i> L. and Valorization of Its Biomass for Food Applications

Supercritical carbon dioxide (CO2) extraction techniques meet all-new consumer market demands for health-promoting phytochemical compound-rich extracts produced from green and sustainable technology. In this regard, this review is dedicated to discussing is the promise of integrating high-pressure CO2 technologies into the Cannabis sativa L. processing chain to valorize its valuable pharmaceutical properties and food biomass. To do this, the cannabis plant, cannabinoids, and endocannabinoid system were reviewed to understand their therapeutic and side effects. The supercritical fluid extraction (SFE) technique was presented as a smart alternative to producing cannabis bioproducts. The impact of SFE operating conditions on cannabis compound extraction was examined for aerial parts (inflorescences, stems, and leaves), seeds, and byproducts. Furthermore, the opportunities of using non-thermal supercritical CO2 processing on cannabis biomass were addressed for industrial hemp valorization, focusing on its biorefinery to simultaneously produce cannabidiol and new ingredients for food applications as plant-based products

Economical Analysis of a Pressurized Fluid-Based Process Applied for Phytochemicals Recovery in a Sustainable Biorefinery Concept for Brazilian Ginseng Roots

A growing number of technologies are being developed in order to produce different high added value products, such as phytochemicals, ethanol and other chemicals from biomass, representing an important trend that needs to be evaluated. The use of pressurized-fluid technologies to obtain phytochemicals from certain biomasses has shown encouraging results in terms of selectivity, enabling its use prior bioenergy generation since generally only a small part of the solid material is extracted. The phytochemical compound which is most financially interesting to extract from the Brazilian Ginseng (Pfaffia glomerata) roots is ecdysterone (beta-ecdysone), an ecdysteroid. Ecdysteroids are a class of natural steroids acting as hormones and beta-ecdysone is the most important steroid employed in cosmetic formulations. It is also used in other industries such as the food and pharmaceutical industry. Due to its health properties, large amounts of Pfaffia glomerata roots are being exported from Brazil to countries that use phytochemical therapies widely. This work is part of a project that aims to present a solution to perform phytochemicals recovery step in Brazil envisioning decentralized local-scale production. In this study, first the influence of the extraction temperature (353-413 K) and static extraction time (5-15 min) using pressurized ethyl acetate as extracting solvent on the beta-ecdysone recovery was experimentally evaluated and compared with literature results employing other pressurized fluids. Afterwards, an economical analysis was done for comparison using ASPEN PLUS software and OSMOSE platform. It was observed that the relationship of the beta-ecdysone recovery, extraction temperature and static extraction time was linear. An increase in temperature and static time resulted in enhancement of the beta-ecdysone recovery from the biomass. On the other hand, the increase of the extraction temperature beyond 373 K possibly might enhance the degradation of the beta-ecdysone extracted decreasing its relative amount in the extract. Since for the economical analysis performed the former response variable (beta-ecdysone recovery) is most important than the latter, it was calculated the net present value of this process considering high extraction temperatures. Considering an installation time of two years, a lifetime of 25 years and an interest rate of 15 % the obtained net present value for the different pressurized extracting solvents were in the following order: ethyl acetate > ethanol > CO2-ethanol mixture. The results demonstrate that the inclusion of a pressurized fluid-based process using ethyl acetate or ethanol for beta-ecdysone recovery prior further bioenergy production is very promising

Biorefinery of turmeric (curcuma longa l.) using non-thermal and clean emerging technologies: an update on the curcumin recovery step

In this study, a biorefinery for the processing of turmeric (Curcuma longa L.) based on clean and emerging technologies has been proposed. High-intensity ultrasound (HIUS) technology was evaluated as a promising technique for curcumin recovery aiming to improve its extraction yield and technological properties as a colorant. In addition, we evaluated the effects of process conditions on the turmeric biomass after the extractions. The process variables were the number of stages of extraction with ethanol (1, 3 and 5) and the solvent to feed ratio (S/F) of 3, 5, 7, 9 (w/w). The highest curcumin content (41.6 g/100 g extract) was obtained using 1 wash and a S/F of 5 w/w, while the highest curcumin yield (3.9 g/100 g unflavored turmeric) was obtained using 5 stages and a S/F of 7. The extracts obtained by solid-liquid extraction assisted by HIUS showed a yellow color (157 and 169 of yellowness index) more intense than those obtained by the pressurized liquid extraction technique (101 of yellowness index) and better yield results than low-pressure solid-liquid extraction (using the same processing time). Thus, it was possible to obtain a characteristic yellow colorant with high curcumin yield in a short process time (5 min of extraction) using HIUS technology. Besides that, SEM images and FTIR spectra demonstrated that the turmeric biomasses processed by HIUS technology were not degraded101112121CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP141110/2018-0; 302423/2015-0Sem informação2015/22226-

Identification and quantification of genipin and geniposide from Genipa americana L. by HPLC-DAD using a fused-core column

<div><p>Abstract In this work, it was developed a fast, simple and selective method for quantification of genipin and geniposide from unripe fruits of genipap, which are known as natural colorants, blue and yellow, respectively. The compounds separation was performed in a fused-core C18 column using as mobile phase water (A) and acetonitrile (B) both acidified with 0.1% formic acid, with the following gradient: 0 min, 99% A; 9 min, 75% A; 10 min, 99% A and 13 min, 99% A. The temperature and flow rate that allowed the best chromatographic performance were 35 °C and 1.5 mL/min, respectively, resulting a total run time of 13 min, including column clean-up and re-equilibration. This short analysis time represents an advantage compared to the methods reported in the literature where the running times are 2-5 times greater. The detection wavelength was set at 240 nm. The method validation was performed based on specificity, linearity, detection and quantification limits, precision and accuracy, according to ICH methodology. Finally, the developed method was suitable for monitoring analysis of those compounds content in vegetable samples.</p></div