Lead (Pb) is the number one heavy metal pollutant in the environment. The high cost and environmental concerns of conventional remediation technologies has led to an emerging alternative technology for heavy metal remediation: phytoremediation. This study was set out to advance Pb phytoremediation by investigating plant-associated factors (e.g. polyphenol levels, Pb-tannin chelation, and superoxide dismutase activity) and chemical-based factors (e.g. concentration of Pb, and the type and dosage of chelating agents in treatments) that may affect Pb accumulation. Using a hydroponic system, sand-grown Symphytum officinale L. plants were exposed to nutrient solutions with or without lead nitrate (Pb(NO₃)₂) and ethylenediamine tetraacetic acid (EDTA). Using flame atomic absorption spectroscopy (to measure Pb content) and bovine serum albumin-protein precipitation (to measure polyphenol and tannin levels), a significant in vivo correlation between tannin level and Pb accumulation level was observed in roots of plants exposed to all Pb treatments. Higher tannin containing-lateral roots accumulated significantly more Pb than lowertannin main roots. Transmission electron micrographs of unchelated Pb-treated plants supported these findings, whilst dialysis-based in vitro Pb-chelation studies with crude S. officinale root polyphenol extracts did not. The dialysis method was likely to be subject to fructan interference. A new, more accurate and simple method based on tannin immobilisation was consequently developed. Results using this method supported the hydroponic trends. This new method was also verified with purified tannic acid (from Sigma). Together, these findings demonstrate that S. officinale root tannins have the ability to chelate Pb. This may be a mechanism to cope with Pb stress (adaptive tolerance). Despite the typical signs of Pb stress at root level (e.g. root growth inhibition, and degraded cytoplasms), shoots showed no signs of stress under any Pb treatments. Most importantly, since this chelation-based tolerance mechanism also influences the accumulation levels, the phytochemical composition of plants should also be considered when screening plants for phytoremediation. The level of Pb accumulated in the shoots depended on the concentration of Pb(NO₃)₂ and presence of chelating agents (EDTA or N-[2 acetamido] iminodiacetic acid (ADA)) in the nutrient solution. The highest level of Pb in shoots was between 0.05-0.06% (d.w. on average) using EDTA or ADA, well short of the 1% (d.w.) shoot accumulation target for Pb phytoextraction. The highest level of Pb in the roots (and of all measurements) was with unchelated 500 µM Pb(NO₃)₂; on average 2% (d.w.) accumulated in root. Overall, since S. officinale accumulated Pb predominately in the roots, it is most suited for rhizofiltration and phytostabilisation. Whilst chelating agents enhanced Pb accumulation in shoots, root levels were unexpectedly reduced compared to unchelated Pb treatments. The level of Pb translocated did not completely account for this loss. Minor factors relating to EDTA desorption of roots, EDTA specificity, and charge repulsion of the PbEDTA complex may account for some of the loss, but the main cause remains unclear. In vitro S. officinale cultures were developed and somaclonal variation (involving Pb pre-treatment of petioles) was used as a tool to further investigate, and attempt to improve its Pb phytoremediation potential. The shoots and roots of plants produced from petioles pre-treated with Pb(NO₃)₂ appeared more stressed than those without Pb pre-treatment. After re-treatment with Pb (Pb(NO₃)₂ or PbADA), plants developed from most Pb pre-treated petioles appeared to have reduced Pb accumulation and polyphenol levels, and increased superoxide dismutase activity in roots (although no statistically significant trends were found). Overall, plants produced from Pb pre-treated petioles in this study may have less phytoremediation potential.

Habilitationsschrift--Technische Hochschule, Karlsruhe. / Sonderabdruck aus "Flora oder Allgem. bot. Zeitung," Ergänzungsband 1902.

Lead (Pb) is the number one heavy metal pollutant in the environment. The high cost and environmental concerns of conventional remediation technologies has led to an emerging alternative technology for heavy metal remediation: phytoremediation. This study was set out to advance Pb phytoremediation by investigating plant-associated factors (e.g. polyphenol levels, Pb-tannin chelation, and superoxide dismutase activity) and chemical-based factors (e.g. concentration of Pb, and the type and dosage of chelating agents in treatments) that may affect Pb accumulation. Using a hydroponic system, sand-grown Symphytum officinale L. plants were exposed to nutrient solutions with or without lead nitrate (Pb(NO₃)₂) and ethylenediamine tetraacetic acid (EDTA). Using flame atomic absorption spectroscopy (to measure Pb content) and bovine serum albumin-protein precipitation (to measure polyphenol and tannin levels), a significant in vivo correlation between tannin level and Pb accumulation level was observed in roots of plants exposed to all Pb treatments. Higher tannin containing-lateral roots accumulated significantly more Pb than lowertannin main roots. Transmission electron micrographs of unchelated Pb-treated plants supported these findings, whilst dialysis-based in vitro Pb-chelation studies with crude S. officinale root polyphenol extracts did not. The dialysis method was likely to be subject to fructan interference. A new, more accurate and simple method based on tannin immobilisation was consequently developed. Results using this method supported the hydroponic trends. This new method was also verified with purified tannic acid (from Sigma). Together, these findings demonstrate that S. officinale root tannins have the ability to chelate Pb. This may be a mechanism to cope with Pb stress (adaptive tolerance). Despite the typical signs of Pb stress at root level (e.g. root growth inhibition, and degraded cytoplasms), shoots showed no signs of stress under any Pb treatments. Most importantly, since this chelation-based tolerance mechanism also influences the accumulation levels, the phytochemical composition of plants should also be considered when screening plants for phytoremediation. The level of Pb accumulated in the shoots depended on the concentration of Pb(NO₃)₂ and presence of chelating agents (EDTA or N-[2 acetamido] iminodiacetic acid (ADA)) in the nutrient solution. The highest level of Pb in shoots was between 0.05-0.06% (d.w. on average) using EDTA or ADA, well short of the 1% (d.w.) shoot accumulation target for Pb phytoextraction. The highest level of Pb in the roots (and of all measurements) was with unchelated 500 µM Pb(NO₃)₂; on average 2% (d.w.) accumulated in root. Overall, since S. officinale accumulated Pb predominately in the roots, it is most suited for rhizofiltration and phytostabilisation. Whilst chelating agents enhanced Pb accumulation in shoots, root levels were unexpectedly reduced compared to unchelated Pb treatments. The level of Pb translocated did not completely account for this loss. Minor factors relating to EDTA desorption of roots, EDTA specificity, and charge repulsion of the PbEDTA complex may account for some of the loss, but the main cause remains unclear. In vitro S. officinale cultures were developed and somaclonal variation (involving Pb pre-treatment of petioles) was used as a tool to further investigate, and attempt to improve its Pb phytoremediation potential. The shoots and roots of plants produced from petioles pre-treated with Pb(NO₃)₂ appeared more stressed than those without Pb pre-treatment. After re-treatment with Pb (Pb(NO₃)₂ or PbADA), plants developed from most Pb pre-treated petioles appeared to have reduced Pb accumulation and polyphenol levels, and increased superoxide dismutase activity in roots (although no statistically significant trends were found). Overall, plants produced from Pb pre-treated petioles in this study may have less phytoremediation potential.

Lead (Pb) is the number one heavy metal pollutant in the environment. The high cost and environmental concerns of conventional remediation technologies has led to an emerging alternative technology for heavy metal remediation: phytoremediation. This study was set out to advance Pb phytoremediation by investigating plant-associated factors (e.g. polyphenol levels, Pb-tannin chelation, and superoxide dismutase activity) and chemical-based factors (e.g. concentration of Pb, and the type and dosage of chelating agents in treatments) that may affect Pb accumulation. Using a hydroponic system, sand-grown Symphytum officinale L. plants were exposed to nutrient solutions with or without lead nitrate (Pb(NO₃)₂) and ethylenediamine tetraacetic acid (EDTA). Using flame atomic absorption spectroscopy (to measure Pb content) and bovine serum albumin-protein precipitation (to measure polyphenol and tannin levels), a significant in vivo correlation between tannin level and Pb accumulation level was observed in roots of plants exposed to all Pb treatments. Higher tannin containing-lateral roots accumulated significantly more Pb than lowertannin main roots. Transmission electron micrographs of unchelated Pb-treated plants supported these findings, whilst dialysis-based in vitro Pb-chelation studies with crude S. officinale root polyphenol extracts did not. The dialysis method was likely to be subject to fructan interference. A new, more accurate and simple method based on tannin immobilisation was consequently developed. Results using this method supported the hydroponic trends. This new method was also verified with purified tannic acid (from Sigma). Together, these findings demonstrate that S. officinale root tannins have the ability to chelate Pb. This may be a mechanism to cope with Pb stress (adaptive tolerance). Despite the typical signs of Pb stress at root level (e.g. root growth inhibition, and degraded cytoplasms), shoots showed no signs of stress under any Pb treatments. Most importantly, since this chelation-based tolerance mechanism also influences the accumulation levels, the phytochemical composition of plants should also be considered when screening plants for phytoremediation. The level of Pb accumulated in the shoots depended on the concentration of Pb(NO₃)₂ and presence of chelating agents (EDTA or N-[2 acetamido] iminodiacetic acid (ADA)) in the nutrient solution. The highest level of Pb in shoots was between 0.05-0.06% (d.w. on average) using EDTA or ADA, well short of the 1% (d.w.) shoot accumulation target for Pb phytoextraction. The highest level of Pb in the roots (and of all measurements) was with unchelated 500 µM Pb(NO₃)₂; on average 2% (d.w.) accumulated in root. Overall, since S. officinale accumulated Pb predominately in the roots, it is most suited for rhizofiltration and phytostabilisation. Whilst chelating agents enhanced Pb accumulation in shoots, root levels were unexpectedly reduced compared to unchelated Pb treatments. The level of Pb translocated did not completely account for this loss. Minor factors relating to EDTA desorption of roots, EDTA specificity, and charge repulsion of the PbEDTA complex may account for some of the loss, but the main cause remains unclear. In vitro S. officinale cultures were developed and somaclonal variation (involving Pb pre-treatment of petioles) was used as a tool to further investigate, and attempt to improve its Pb phytoremediation potential. The shoots and roots of plants produced from petioles pre-treated with Pb(NO₃)₂ appeared more stressed than those without Pb pre-treatment. After re-treatment with Pb (Pb(NO₃)₂ or PbADA), plants developed from most Pb pre-treated petioles appeared to have reduced Pb accumulation and polyphenol levels, and increased superoxide dismutase activity in roots (although no statistically significant trends were found). Overall, plants produced from Pb pre-treated petioles in this study may have less phytoremediation potential.

Habilitationsschrift--Technische Hochschule, Karlsruhe. / Sonderabdruck aus "Flora oder Allgem. bot. Zeitung," Ergänzungsband 1902.

Programa de Pós-Graduação em Ciências Farmacêuticas. CIPHARMA, Escola de Farmácia, Universidade Federal de Ouro Preto. / Submitted by giuliana silveira (giulianagphoto@gmail.com) on 2016-01-28T17:32:41Z No. of bitstreams: 1 DISSERTAÇÃO_AvaliaçãoAtividadeAntiinflamatória.pdf: 1345173 bytes, checksum: 36238855dd90974856f2b7447113f6ec (MD5) / Approved for entry into archive by Oliveira Flávia (flavia@sisbin.ufop.br) on 2016-02-01T15:49:22Z (GMT) No. of bitstreams: 1 DISSERTAÇÃO_AvaliaçãoAtividadeAntiinflamatória.pdf: 1345173 bytes, checksum: 36238855dd90974856f2b7447113f6ec (MD5) / Made available in DSpace on 2016-02-02T12:57:14Z (GMT). No. of bitstreams: 1 DISSERTAÇÃO_AvaliaçãoAtividadeAntiinflamatória.pdf: 1345173 bytes, checksum: 36238855dd90974856f2b7447113f6ec (MD5) Previous issue date: 2015 / O processo inflamatório é uma resposta dos tecidos a estímulos nocivos através de alterações celulares e vasculares. Em geral são processos dolorosos e incômodos para os pacientes e o tratamento convencional é realizado com a utilização dos Antiinflamatórios Não Esteroidais (AINEs), apesar das reações adversas que provocam. No Brasil e em todo o mundo plantas medicinais e medicamentos fitoterápicos são utilizados pela população para tratamento e prevenção de doenças, incluindo as inflamatórias. É o que ocorre com as espécies brasileiras, Lychnophora pinaster Mart. e Symphytum officinale L., utilizadas para tratamento de inflamação, dor e reumatismo, e que já tiveram a ação antiinflamatória demonstrada. Com base no exposto, este trabalho teve por objetivo a avaliação da atividade antiinflamatória de formulações de uso tópico, como gel, emulgel e nanoemulsão, contendo extratos de Lychnophora pinaster ou Symphytum officinale. A avaliação da atividade das formulações foi realizada em modelo de edema de pata induzido pela carragenina. Todas as formulações obtidas apresentaram aspecto homogêneo e odor característicos. A medida da viscosidade para as formulações base obtidas para o gel de hidroxietilcelulose 7 %, gel de hidroxietilcelulose 14 % e emulgel foi respectivamente de 1299782,00, 555632,00 e 373067,20 cP. Os emulgéis obtidos (base e contendo os diferentes extratos vegetais) apresentaram pH igual a 6,5, para os géis preparados com a utilização de hidroxietilcelulose a 7 % e 14 % (base e contendo os extratos ou nanoemulsão) a medida deste parâmetro foi 6,0 e 6,5 respectivamente. As formulações emulgel e géis base foram avaliadas durante 180 dias de estocagem, e durante este período não apresentaram alterações nas características observadas. Todas estas formulações foram de fácil aplicação. As nanoemulsões foram obtidas como suspensões coloidais com tamanho da partícula variando entre 233,0 ± 9,39 e 245,1 ± 8,17 nm e apresentaram distribuição de tamanho unimodal, com índice de polidispersão menor que 0,184 ± 0,041. A administração tópica das formulações emulgel contendo L. pinaster nas duas concentrações avaliadas (2,0 % e 5,0 %) e gel contendo nanoemulsão de L. pinaster a 2,0 % apresentou maior atividade que o Cataflam Emulgel (cada 1,16 g equivale a 1,0 g de diclofenaco potássico), as nanoemulsões contendo L. pinaster a 1,25 % e 2,0 % mostraram atividade antiinflamatória similar ao medicamento Cataflam Emulgel, enquanto que a nanoemulsão contendo L. pinaster a 0,625% e o gel contendo L. pinaster nas duas concentrações avaliadas (2,0 % e 5,0 %) não demonstraram atividade farmacológica. A administração tópica das formulações gel e emulgel contendo S. officinale a 10,0 % e 16,0 % mostraram atividade antiinflamatória similar ao Cataflam em emulgel, as formulações emulgel contendo S. officinale a 2,0 % e 5,0 % não apresentaram atividade. A comparação entre as formulações mostra que o emulgel e a nanoemulsão contendo Lychnophora pinaster na concentração de 2,0 % e 1,25 %, respectivamente foram as formulações mais interessantes e satisfatórias dentre as analisadas para esta espécie vegetal, visto que apresentaram atividade antiinflamatória significativa com baixas concentrações de extrato, além da facilidade de produção e administração. Em relação ao Symphytum officinale, as formulações emulgel e gel contendo o extrato bruto glicólico da espécie nas concentrações 10,0 e 16,0 % também foram consideradas satisfatórias, pois além da atividade antiinflamatória semelhante ao cataflam em emulgel, elas também se apresentaram de fácil produção e administração. ____________________________________________________________________________________ / ABSTRACT: The inflammation process is a tissue response to noxious stimuli through cellular and vascular changes. Usually, these processes are painful and uncomfortable for patients and the conventional treatment is accomplished with the use of Non Steroidal Anti-inflammatory Drugs (NSAIDs), despite the adverse reactions that they cause. In Brazil and throughout the world medicinal plants and phytoterapic drugs are used to treat and prevent diseases, including inflammatory ones. The Brazilian species, Lychnophora pinaster Mart. and Symphytum officinale L. are used to treat inflammatory conditions, pain and rheumatism, and their anti-inflammatory activity were already demonstrated. The aim of the present work was to evaluate the anti-inflammatory activity of topical formulations such as gels, emulgel and nanoemulsion containing Lychnophora pinaster or Symphytum officinale extracts. The formulations activity evaluation was performed using the acute paw oedema induced by carrageenan. All obtained formulations presented homogeneous appearance and characteristic smell. The viscosity measurement for the obtained pharmaceutical base forms gel hydroxyethylcellulose 7 %, gel hydroxyethylcellulose 14 % and emulgel was respectively 1299782,00, 555632,00 e 373067,20 cP. The obtained emulgel (base and containing plant extracts) showed pH of 6.5, for the prepared gels with the use of hydroxyethylcellulose 7 % and 14 % (base and containing extracts or nanoemulsion) the mensuration of this parameter was 6.0 and 6.5 respectively. The base formulations gels and emulgel were evaluated during 180 storage days, and showed no changes of the evaluated characteristics. All these formulations were easy for topical administration. The nanoemulsions were obtained as a colloidal suspension with particle size ranging from 233.0 ± 9.39 to 245.1 ± 8.17 nm and showed unimodal size distribution with size distribution less than 0.184 ± 0.041. The topical administration of the formulations emulgel containing L. pinaster in both concentrations (2.0 % and 5.0 %) showed greater activity than Cataflam , the nanoemulsion containing L. pinaster 1.25 % and 2.0 % and gel containing nanoemulsion of L. pinaster 2.0 % showed anti-inflammatory activity similar to Cataflam  while the nanoemulsion containing L. pinaster 0.625 % and the gels containing L. pinaster in both concentrations (2.0 % and 5.0 %) showed no pharmacological activity. The topical formulations emulgel and gel containing S. officinale 10.0 % and 16.0 % showed anti-inflammatory activity similar to Cataflam, the formulations emulgel containing S. officinale 2.0 % and 5.0 % showed no activity. Comparing the formulations, emulgel and nanoemulsion containing Lychnophora pinaster at a concentration of 2.0 % and 1.25 % respectively were the most interesting and satisfactory one among those analyzed for this plant species, since they showed significant anti-inflammatory activity with lower concentrations of extract, and easy production and administration. Regarding Symphytum officinale formulations, emulgel and gel containing the crude glycolic extract at the concentrations 10.0 and 16.0% were considered satisfactory, because besides the anti-inflammatory activity similar to Cataflam, they also presented easy production and administration.

Resumo: O Symphytum officinale (confrei) é uma planta usada desde a antiguidade para o tratamento da consolidação de fraturas. Existem relatos de seu uso in natura , como fitoterápico ou como medicamento homeopático. Recentemente foram relatados casos de intoxicação em humanos e em animais que consumiram a planta e desenvolveram lesões hepáticas graves que foram associadas à presença de alcalóides pirrolizidínicos. Tais fatos levaram à proibição do uso interno da planta. No entanto, não há relato de efeitos adversos associados à sua formulação homeopática. O objetivo deste trabalho foi comparar os efeitos do Symphytum officinale, preparado como medicamento fitoterápico e homeopático, avaliando sua toxicidade hepática e renal. Foram utilizados 42 ratos adultos saudáveis, divididos em três grupos de acordo com os diferentes tratamentos: Symphytum officinale 500mg/kg (F), Symphytum officinale 6CH 2 glóbulos (H) e água como placebo no grupo controle negativo (C), que foram administrados diariamente a cada animal por via oral por gavagem. Os sacrifícios foram realizados 30 e 60 dias após o início do tratamento. A avaliação toxicológica foi feita através de análise bioquímica da atividade enzimática da fosfatase alcalina (FA), aspartato aminotransferase (AST), alanina aminotransferase (ALT), gama glutamil transferase (GGT) e da concentração plasmática de uréia e creatinina. Também foi realizada análise histológica e histomorfométrica de fígado e rim. Os dados foram submetidos aos testes estatísticos de t (Student) e ANOVA 1 fator (5%). Tanto as análises bioquímicas quanto histológicas indicaram discretas alterações hepáticas e evidenciaram ausência de alterações renais. As modificações no perfil de atividade enzimática, na uréia e creatinina e no peso dos animais sugeriram alterações no metabolismo hepático interferindo na síntese... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Symphytum officinale (comfrey) is a plant used over thousands of years to treat bone fractures consolidation. There are reports of its use in natura, as phytotherapic and as homeopathic medication. Recent scientific reports described cases of human and animal intoxication, with development of severe hepatic lesions after consumption of this plant, associated with the presence of pyrrolizidine alkaloids. Because of these facts its internal use was prohibited. However, there are no reports of adverse effects of Symphytum officinale 's homeopathic formulation. The aim of this work was to compare the effects of Symphytum officinale used as phytotherapic and as homeopathic formulations, evaluating its hepatic and renal toxicity. Adult healthy rats (n=42) were divided in 3 groups according to treatment with Symphytum officinale 500mg/kg (F), Symphytum officinale 6CH 2 globules (H), and water as placebo for negative control (C), given daily orally by gavage to each animal. They were sacrificed 30 and 60 days after beginning of treatments. The toxicological evaluation was realized by biochemical analysis of the enzymatic activity of alkaline phosphatase (AF), aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT) and of creatinine and urea plasmatic concentration. Histological and histomorphometric analysis of liver and kidney was also conducted. Data were submitted to statistical t (Student) and ANOVA 1 factor (5%) tests. Both biochemical and histological analysis showed mild hepatic alterations and no renal alterations. The modifications of the enzymatic activity, of urea and creatinine, and of rats' weight suggested hepatic metabolic alterations interfering on protein synthesis. Histological analysis was compatible with chronic mild hepatitis with development of important fibrosis only in the F group. The phytotherapic formulation used was... (Complete abstract click electronic access below) / Orientador: Adriana Aigotti Haberbeck Bandão / Coorientador: Maria Nadir Gasparoto Mancini / Banca: Rosilene Fernandes da Rocha / Banca: João Ernesto de Carvalho / Mestre

Made available in DSpace on 2014-06-11T19:27:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-07-21Bitstream added on 2014-06-13T19:15:38Z : No. of bitstreams: 1 lima_ap_me_sjc.pdf: 840973 bytes, checksum: 2f4db3725d3a6941b05476631d72d7de (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O Symphytum officinale (confrei) é uma planta usada desde a antiguidade para o tratamento da consolidação de fraturas. Existem relatos de seu uso in natura , como fitoterápico ou como medicamento homeopático. Recentemente foram relatados casos de intoxicação em humanos e em animais que consumiram a planta e desenvolveram lesões hepáticas graves que foram associadas à presença de alcalóides pirrolizidínicos. Tais fatos levaram à proibição do uso interno da planta. No entanto, não há relato de efeitos adversos associados à sua formulação homeopática. O objetivo deste trabalho foi comparar os efeitos do Symphytum officinale, preparado como medicamento fitoterápico e homeopático, avaliando sua toxicidade hepática e renal. Foram utilizados 42 ratos adultos saudáveis, divididos em três grupos de acordo com os diferentes tratamentos: Symphytum officinale 500mg/kg (F), Symphytum officinale 6CH 2 glóbulos (H) e água como placebo no grupo controle negativo (C), que foram administrados diariamente a cada animal por via oral por gavagem. Os sacrifícios foram realizados 30 e 60 dias após o início do tratamento. A avaliação toxicológica foi feita através de análise bioquímica da atividade enzimática da fosfatase alcalina (FA), aspartato aminotransferase (AST), alanina aminotransferase (ALT), gama glutamil transferase (GGT) e da concentração plasmática de uréia e creatinina. Também foi realizada análise histológica e histomorfométrica de fígado e rim. Os dados foram submetidos aos testes estatísticos de t (Student) e ANOVA 1 fator (5%). Tanto as análises bioquímicas quanto histológicas indicaram discretas alterações hepáticas e evidenciaram ausência de alterações renais. As modificações no perfil de atividade enzimática, na uréia e creatinina e no peso dos animais sugeriram alterações no metabolismo hepático interferindo na síntese... / Symphytum officinale (comfrey) is a plant used over thousands of years to treat bone fractures consolidation. There are reports of its use in natura, as phytotherapic and as homeopathic medication. Recent scientific reports described cases of human and animal intoxication, with development of severe hepatic lesions after consumption of this plant, associated with the presence of pyrrolizidine alkaloids. Because of these facts its internal use was prohibited. However, there are no reports of adverse effects of Symphytum officinale ´s homeopathic formulation. The aim of this work was to compare the effects of Symphytum officinale used as phytotherapic and as homeopathic formulations, evaluating its hepatic and renal toxicity. Adult healthy rats (n=42) were divided in 3 groups according to treatment with Symphytum officinale 500mg/kg (F), Symphytum officinale 6CH 2 globules (H), and water as placebo for negative control (C), given daily orally by gavage to each animal. They were sacrificed 30 and 60 days after beginning of treatments. The toxicological evaluation was realized by biochemical analysis of the enzymatic activity of alkaline phosphatase (AF), aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT) and of creatinine and urea plasmatic concentration. Histological and histomorphometric analysis of liver and kidney was also conducted. Data were submitted to statistical t (Student) and ANOVA 1 factor (5%) tests. Both biochemical and histological analysis showed mild hepatic alterations and no renal alterations. The modifications of the enzymatic activity, of urea and creatinine, and of rats’ weight suggested hepatic metabolic alterations interfering on protein synthesis. Histological analysis was compatible with chronic mild hepatitis with development of important fibrosis only in the F group. The phytotherapic formulation used was... (Complete abstract click electronic access below)

This diploma thesis deals with macerates of comfrey (Symphytum officinale) in selected fats (food lard, cosmetic lard and almond oil). Selected fats were first characterized by dry matter content, saponification, acid, iodine number, peroxide value. At the same time, total and free fatty acids were determined using GC-FID. Macerates were prepared by extraction of comfrey roots with selected fats. In prepared macerates the change in acid and peroxide value was monitored, as well as the content of selected bioactive compounds. The content of total phytosterol and total carotenoid content was determined by UV-VIS spectrometry, phytosterols and carotenoids were also analysed using HPLC-DAD. Total phenolic content was measured using Folin-Ciocalteu reagent and antioxidant activity by ABTS assay. The measured properties were then compared with industrially produced comfrey ointment. In macerated fats the increase in acid and peroxide value was determined. Furthermore, an increase in the content of total phytosterols and total carotenoids was observed. Using HPLC-DAD the content of -sitosterol and stigmasterol was determined, but carotenoids were not detected. Of the tocopherols, only DL--tocopherol acetate was detected. During maceration, the content of total phenolic compound in fat increased, which caused a change in antioxidant activity.

<p>U okviru ove doktorske disertacije ispitane su i<br />poređene različite ekstrakcione tehnike korena<br />gaveza, lista crnog duda i peteljki tre&scaron;nje. Poređene<br />su konvencionalne i savremene ekstrakcione<br />tehnike i izvedena je optimizacija ekstrakcionih<br />postupaka s ciljem dobijanja ekstrakata ispitivanih<br />biljnih vrsta sa najvećom bioaktivno&scaron;ću. Za svaku<br />ekstrakcionu tehniku ispitan je uticaj najznačajnijih<br />parametara: maceracija (rastvarač i vreme<br />ekstrakcije), ubrzana ekstrakcija (temperatura i<br />rastvarač), supekritična ekstrakcija (pritisak i<br />koncentracija ko-rastvarača) i ekstrakcija<br />subkritičnom vodom (temperatura, pritisak, vreme<br />ekstrakcije i brzina me&scaron;anja). Ekstrakcija polifenola<br />korena gaveza, lista crnog duda i peteljki tre&scaron;nje je<br />bila najefikasnija ekstrakcijom subkritičnom<br />vodom. Primenjene su tehnika visokopritisne tečne<br />hromatografije spregnute sa masenom<br />spektrometrijom uz primenu različitih masenih<br />analizatora, masenog analizatora na bazi vremena<br />preleta jona i tandema kvadrupolnog i analizatora<br />na bazi vremena preleta jona. Za karakterizaciju<br />dobijenih ekstrakata instrumentalnom analizom su<br />određene kvalitativne i kvantitativne karakteristike dobijenih ekstrakata korena gaveza, lista crnog duda i peteljki tre&scaron;nje. HPLC-ESI-QTOF-MS/MS analizom ekstrakata identifikovano je vi&scaron;e od 27 novih jedinjenja različitih hemijskih klasa u svakom od ispitivanih biljnih matriksa.</p> / <p>Within the scope of this doctoral dissertation,<br />different extraction techniques of comfrey root,<br />black mulberry leaves and sweet cherry stems were<br />applied and compared. Conventional and novel<br />extraction techniques were optimized and<br />compared in order to obtain plant extracts with the<br />highest content of bioactive compounds. The<br />influence of the most important parameters was<br />investigated for each extraction technique:<br />maceration (solvent and extraction time),<br />accelerated solvent extraction (temperature and<br />solvent), supercritical fluid extraction (co-solvent<br />percentage and pressure) and subcritical water<br />extraction (temperature, pressure, extraction time<br />and agitation rate). Extraction of polyphenolic<br />compounds from comfrey root, black mulberry<br />leaves and sweet cherry stems was the most<br />effective using subcritical water. High-pressure<br />liquid chromatography coupled to mass<br />spectrometry techniques using time-of-flight and<br />quadrupole-time-of-flight mass analyzers have<br />been used to characterize bioactive compounds.<br />Qualitative and quantitative characteristics of the<br />obtained comfrey root, black mulberry leaves and sweet cherry stem extracts were determined. HPLC-ESI-QTOF-MS/MS revealed the presence of more than 27 compounds of different chemical classes that have been identified for the first time in&nbsp; the plant matrix.</p>