Correlation Analysis Between Color and Content Changes of Five Components of Wine-processed Polygonatum kingianum Rhizoma During Processing / 中国实验方剂学杂志

ObjectiveTo study the correlation between the appearance color of the sample powder and the contents of five non-sugar components of wine-processed Polygonatum kingianum rhizoma during processing， and determine the feasibility of color quantitative value for judging the processing end point of the wine-processed products， and to screen steroidal saponins and flavonoids as markers for the control of the wine-processed products during processing. MethodThe changes of apparent color of the sample powder at different time points of the wine-processed products were measured by colorimeter， and the total color value （E*ab），the total color difference value （ΔE*ab） were calculated. The contents of protodioscin， pseudoprotodioscin， dioscin， diosgenin and narcissoside in the wine-processed products （No. S0-S10） after processing for 0， 5， 10， 14， 16， 18， 20， 22， 24， 26， 28 h were determined simultaneously by ultra-high performance liquid chromatography-tandem mass spectrometry （UHPLC-MS/MS）. Cluster analysis （HCA）， principal component analysis （PCA）， partial least squares discriminant analysis （PLS-DA） and Pearson correlation analysis were used to analyze the chromaticity value of the sample powder and the content of the five components. ResultDuring processing of wine-processed P. kingianum rhizoma， E*ab of the sample powder showed a decreasing trend and the apparent color changed from light yellow to lacquer black. The contents of the five components showed an obvious dynamic change trend with time， and showed different laws. HCA results showed that the processing process of the wine-processed products could be divided into three stages， namely， the early stage （samples S0-S1）， the middle stage （samples S2-S4） and the late stage （samples S5-S10）. PCA results showed that there were significant differences in color and contents of five components between the initial sample and the processing samples， and the difference between samples S8 and S9 was the smallest. PLS-DA results showed that the variable importance in the projection （VIP） values of b*， the contents of pseudoprotodioscin， narcissoside， diosgenin and protodioscin were >1. Pearson correlation analysis showed that the contents of protodioscin， diosgenin and narcissoside had a significant positive correlation with E*ab （P<0.01）， the content of diosgenin had a significant negative correlation with E*ab （P<0.01）， while the content of pseudoprotodioscin had no linear correlation with E*ab. ConclusionIn the process of wine-processed P. kingianum rhizoma， there is a certain linear correlation between color quantitative value and chemical composition， and the processing end point can be determined objectively. It can be considered that protodioscin can be used as a marker for the control of the wine-processed products.

Protodioscin levels in Brachiaria spp. in a sheep production system and a brief review of the literature of Brachiaria spp. poisoning in ruminants / Níveis de protodioscina em Brachiaria spp. em um sistema de produção de ovinos e uma breve revisão da literatura da intoxicação por Brachiaria spp. em ruminantes

Plants of the genus Brachiaria, used in several countries as forage, are poisonous to some livestock species. Their toxic principle is protodioscin, and the main form of clinical presentation of the toxicosis is hepatogenous photosensitization. Here we compare protodioscin levels in B. decumbens and B. brizantha and review the literature on the concentrations and methodologies of collection and analysis of the toxic principle in Brachiaria spp. and the risk of contamination of pastures by more toxic species that may facilitate poisoning by plants of this genus in sheep. The experiment was conducted in pastures originally formed by B. brizantha, with many B. decumbens invasion points. The occurrence of cases of poisoning by Brachiaria spp. was the criterion for confirming pasture toxicity. The forage samples were collected at ten random points every 28 days through manual grazing simulation. The samples were analyzed for protodioscin by high-performance liquid chromatography (HPLC) with light scattering by evaporation (ELSD) after being dried and crushed. In the flock of 69 sheep, five poisoning cases occurred, three sheep died, and two recovered. The protodioscin levels found in the evaluated pastures ranged from 0.70 to 0.45%; higher levels appeared in B. decumbens (7.09%) compared to 1.04% in B. brizantha. We suggest that Brachiaria spp. should be avoided in pastures where sheep are grazing.(AU)

Plantas do gênero Brachiaria, utilizadas em vários países como forragem, são tóxicas para várias espécies pecuárias. Seu princípio tóxico é a protodioscina, e a principal forma de apresentação clínica da toxicose é a fotossensibilização hepatógena. Este estudo teve como objetivo comparar os níveis de protodioscina em B. decumbens e B. brizantha e revisar a literatura sobre as concentrações e metodologias de coleta e análise do princípio tóxico em Brachiaria spp. e o risco de contaminação das pastagens por espécies mais tóxicas que podem facilitar a intoxicação por plantas desse gênero em ovinos. O experimento foi conduzido em pastagens originalmente formadas por B. brizantha, com diversos pontos de invasão por B. decumbens. Ocorrência de casos de intoxicação por Brachiaria spp. foi o critério para confirmação da toxicidade da pastagem. As amostras de forragem foram coletadas a cada 28 dias em dez pontos aleatórios por meio de simulação de pastejo manual. As amostras foram analisadas para protodioscina por cromatografia líquida de alta eficiência (HPLC) com dispersão de luz por evaporação (ELSD) após serem secadas e trituradas. No rebanho de 69 ovelhas, cinco desenvolveram a intoxicação, três morreram e duas se recuperaram. Os níveis de protodioscina encontrados nas pastagens avaliadas variaram de 0,70 a 0,45%; níveis mais elevados apareceram em B. decumbens (7,09%) em comparação com 1,04% em B. brizantha. Sugerimos que Brachiaria spp. deve ser evitada no pasto de ovelhas em pastejo.(AU)

Protodioscin levels in Brachiaria spp. in a sheep production system and a brief review of the literature of Brachiaria spp. poisoning in ruminants

ABSTRACT: Plants of the genus Brachiaria, used in several countries as forage, are poisonous to some livestock species. Their toxic principle is protodioscin, and the main form of clinical presentation of the toxicosis is hepatogenous photosensitization. Here we compare protodioscin levels in B. decumbens and B. brizantha and review the literature on the concentrations and methodologies of collection and analysis of the toxic principle in Brachiaria spp. and the risk of contamination of pastures by more toxic species that may facilitate poisoning by plants of this genus in sheep. The experiment was conducted in pastures originally formed by B. brizantha, with many B. decumbens invasion points. The occurrence of cases of poisoning by Brachiaria spp. was the criterion for confirming pasture toxicity. The forage samples were collected at ten random points every 28 days through manual grazing simulation. The samples were analyzed for protodioscin by high-performance liquid chromatography (HPLC) with light scattering by evaporation (ELSD) after being dried and crushed. In the flock of 69 sheep, five poisoning cases occurred, three sheep died, and two recovered. The protodioscin levels found in the evaluated pastures ranged from 0.70 to 0.45%; higher levels appeared in B. decumbens (7.09%) compared to 1.04% in B. brizantha. We suggest that Brachiaria spp. should be avoided in pastures where sheep are grazing.

RESUMO: Plantas do gênero Brachiaria, utilizadas em vários países como forragem, são tóxicas para várias espécies pecuárias. Seu princípio tóxico é a protodioscina, e a principal forma de apresentação clínica da toxicose é a fotossensibilização hepatógena. Este estudo teve como objetivo comparar os níveis de protodioscina em B. decumbens e B. brizantha e revisar a literatura sobre as concentrações e metodologias de coleta e análise do princípio tóxico em Brachiaria spp. e o risco de contaminação das pastagens por espécies mais tóxicas que podem facilitar a intoxicação por plantas desse gênero em ovinos. O experimento foi conduzido em pastagens originalmente formadas por B. brizantha, com diversos pontos de invasão por B. decumbens. Ocorrência de casos de intoxicação por Brachiaria spp. foi o critério para confirmação da toxicidade da pastagem. As amostras de forragem foram coletadas a cada 28 dias em dez pontos aleatórios por meio de simulação de pastejo manual. As amostras foram analisadas para protodioscina por cromatografia líquida de alta eficiência (HPLC) com dispersão de luz por evaporação (ELSD) após serem secadas e trituradas. No rebanho de 69 ovelhas, cinco desenvolveram a intoxicação, três morreram e duas se recuperaram. Os níveis de protodioscina encontrados nas pastagens avaliadas variaram de 0,70 a 0,45%; níveis mais elevados apareceram em B. decumbens (7,09%) em comparação com 1,04% em B. brizantha. Sugerimos que Brachiaria spp. deve ser evitada no pasto de ovelhas em pastejo.

Analysis on Quality Evaluation Methods of Asparagi Radix Decoction Pieces and Its Standard Decoction / 中国实验方剂学杂志

Objective:To establish the quality evaluation methods of Asparagi Radix decoction pieces and its standard decoction. Method:Ten batches of Asparagi Radix standard decoction were prepared. High performance liquid chromatography-evaporative light scattering detection method (HPLC-ELSD) was established for the determination of protodioscin and protoneodioscin in Asparagi Radix decoction pieces and its standard decoction, and the fingerprint detection of Asparagi Radix decoction pieces with acetonitrile-water as mobile phase for gradient elution. UHPLC-LTQ-Orbitrap-MS/MS was used to identify ten main common peaks in the fingerprint with acetonitrile-0.1% formic acid solution as mobile phase for gradient elution, electrospray ionization (ESI) and positive and negative ion mode scanning were employed, the detection range was m/z 100-1 400. Result:The total content of protodioscin and protoneodioscin in Asparagi Radix decoction pieces was 0.41%-0.72%, and their total content in Asparagi Radix standard decoction was 0.33%-0.59%, the transfer rate of these two components was 73.6%-98.3%. The dry extract yield of the standard decoction was 59.0%-73.0%, and its pH was 4.9-5.6. There were 10 common peaks in the fingerprint, and all of them were saponins, including protoneodioscin, protodioscin, aspacochioside A and its isomer, methyl protodioscin, asparagoside F, (25R)-26-O-β-D-glucopyranosyl-furostan-5, 20-diene-3β, 26-diol-3-O-[α-L-rhamnopyranosyl (1→2)]-[β-D-glucopyranosyl (1→4)-α-L-rhamnopyranosyl (1→4)]-β-D-glucopyranoside, 26-O-β-D-glucopyranosyl-furostan-20 (22)-ene-3β, 26-diol-3-O-[α-L-rhamnopyranosyl (1→2)]-[α-L-rhamnopyranosyl (1→4)]-β-D-glucopyranoside, pseudodiosgenin, aspacochioside C. Conclusion:In this paper, the quality evaluation methods of Asparagi Radix decoction pieces and its standard decoction are established, and these methods are stable and feasible, which can provide reference for the quality control of pharmaceutical preparations containing Asparagi Radix.

Comparison of two collection methods to determine protodioscin concentrations in Brachiaria spp. and to monitor Brachiaria spp. poisoning / Comparação entre dois métodos de coleta para determinar as concentrações de protodioscina em Brachiaria spp. e monitorar a intoxicação por Brachiaria spp

The objective of this study was to evaluate the methods of collecting Brachiaria spp. (Simulated Grazing vs. Total Collection) for the determination of protodioscin concentrations and to monitor the intoxication in sheep susceptible and resistant to poisoning by Brachiaria spp. The collection of Brachiaria spp. was done through two different methods. The first one consisted in throwing a metallic square at 10 random picket points. The second was performed simulating, with the hands, the grazing behavior of sheep. The leaf blades obtained by the two methods were conditioned in a forced ventilation oven. The extraction and quantification of protodioscin from the dry milled plant material was done by reverse-phase high-performance liquid chromatography (HPLC). The collections were carried out at three stages of the life of experimental lamb (suckling, rearing, and finishing). The lambs were divided into resistant and susceptible to intoxication by Brachiaria spp. The criterion for a lamb to be considered sensitive was to present some clinical signs at some stage of its life. No significant differences were observed between the general averages of the two collections when the evaluation was done in the whole period. However, a significant difference was found at the onset of the clinical signs. The concentrations of protodioscin in this period were higher in Brachiaria spp. collected by the simulating grazing method. Both methodologies were efficient to determine the level of protodioscin in the pastures of Brachiaria spp. However, during the periods when the outbreaks occurred, the simulated grazing method was more reliable.(AU)

Objetivou-se avaliar dois métodos de coleta de Brachiaria spp. (Pastejo Simulado vs Coleta Total) para determinação das concentrações de protodioscina e efetuar o monitoramento da intoxicação em rebanhos sensíveis e resistentes à intoxicação por Brachiaria spp. Foram realizadas coletas de Brachiaria spp. através de duas metodologias. A primeira consistiu em fazer o lançamento de um quadrado metálico em 10 pontos aleatórios do piquete. A segunda foi realizada simulando com as mãos o comportamento de pastejo dos ovinos. As lâminas foliares obtidas pelas duas metodologias foram acondicionadas em estufa de ventilação forçada. O material seco e moído foi encaminhado para extração e quantificação da protodioscina por cromatografia líquida de alto desempenho (HPLC) de fase reversa. As coletas foram realizadas em três fases de vida dos cordeiros (cria, recria e terminação). Os cordeiros foram divididos em resistentes e sensíveis a intoxicação por Brachiaria spp. O critério para o ovino ser considerado sensível foi apresentar algum sinal clínico da intoxicação em alguma fase de sua vida. Não foram observadas diferenças significativas entre a média geral das duas coletas quando a avaliação foi feita no período total. Porém, foi observada diferença significativa no período de aparecimento dos casos. As concentrações de protodioscina neste período foram superiores pelo método do pastejo simulado. Ambas as metodologias mostraram-se eficientes para determinação da concentração da protodioscina nas pastagens de Brachiaria spp. No entanto, nos períodos em que ocorreram os surtos, o método do pastejo simulado foi mais fidedigno.(AU)

Ensayos de masculinización de trucha arcoíris Oncorhynchus mykiss suministrando Tribulus terrestris en el alimento / Masculinization assays of rainbow trout Oncorhynchus mykiss feeding with Tribulus terrestris / Ensaios de masculinização da truta arco-íris Oncorhynchus mykiss alimentação com Tribulus terrestres

Resumen Tribulus terrestris (TT) es una planta herbácea que induce la producción de testosterona una vez es ingerida. Con el fin de determinar el porcentaje de neomachos de trucha arcoíris, obtenidos a partir de la utilización de Tribulus terrestris en el alimento; se trabajó con 1.200 larvas 100% hembras adquiridas comercialmente, que se repartieron equitativamente en 12 tanques de concreto. Se suministró alimento balanceado comercial (50% de PB) desde el día 15 poseclosión y durante 96 días con los siguientes tratamientos: 3.000 mg de TT/kg de alimento (tratamiento A), 5.000 mg TT/kg de alimento (tratamiento B), control positivo 3 mg de MT/kg de alimento (tratamiento C) y control negativo con etanol (tratamiento D). Se determinaron parámetros productivos y diferenciación gonadal mediante la técnica de squash. Se midieron parámetros fisicoquímicos del agua. Con cada tratamiento A y B se obtuvo un porcentaje de machos de 2,67% (P>0,05) y hubo presencia de intersexos pero sin significancia estadística. Las ganancias diarias de peso fueron 0,128, 0,132, 0,109 y 0,088 g/d para los tratamientos A, B, C y D respectivamente. Las ganancias de talla se encontraron entre 0,063 cm (D) y 0,077 cm (B); ambas variables tuvieron diferencia estadística significativa (P<0,05). La sobrevivencia, el factor de conversión alimenticia y la incidencia de costos no estuvieron afectados significativamente por los tratamientos. Se concluyó que Tribulus terrestris mezclado con alimento balanceado en dosis de 3.000 mg/kg y de 5.000 mg/kg no induce la masculinización de la trucha arcoíris; sin embargo, no incide negativamente en la sobrevivencia de la misma, ejerce un efecto anabólico significativo, reflejado en ganancias de peso y talla y no resulta costoso utilizarlo comparado con el método convencional de la hormona 17α-metil testerona. No se recomienda su uso para este fin por la ineficiencia. Este estudio es el primer reporte en Colombia al respecto de este tema.

Abstract Tribulus terrestris (TT) is herbaceous plant that induces testosterone production once ingested. In order to determine the percentage of rainbow trout neomales, obtained from feeding with Tribulus terrestris; 1,200 larvae commercially acquired 100% females, was distributed in 12 concrete tanks and fed commercial balanced food (50% CP) from day 15 poseclosión for 96 days with the following treatments: 3.000 mg TT / kg feed (treatment A), 5,000 mg TT / kg feed (treatment B) control MT positive 3mg / kg feed (treatment C) and negative control with ethanol (treatment D). Productive parameters and gonadal differentiation were determined by squash technique. In water, physico-chemical parameters were measured. With each treatment A and B a percentage of males 2.67% (P <0.05) was obtained and there was the presence of intersex but without statistical significance. The daily weight gains were 0.128, 0.132, 0.109 and 0.088 g/d for treatments A, B, C and D respectively. Size gains found from 0.063 cm (D) until 0.077 cm (B); both variables had statistically significant difference (P <0.05). Survival, feed conversion factor and the incidence of costs were not significantly affected by the treatments. In conclusion, Tribulus terrestris mixed with meal doses of 3,000 mg / kg and 5,000 mg / kg does not induce masculinization of rainbow trout; however, no negative impact on the survival of it, has a significant anabolic effect, reflected in gains weight and height and is not expensive to use compared to the conventional method of hormone 17α-methyl testerone. Its use for this purpose is not recommended for inefficiency. It is the first study reported in Colombia in this regard.

Resumo Tribulus terrestris (TT) é uma planta herbácea que induz a produção de testosterona, uma vez ingerida. Com a fim de determinar a percentagem de neomales de truta arco-íris, obtidos a partir de a utilização de Tribulus terrestris no alimento; realizado com 1.200 larvas comercialmente adquidas 100% do sexo feminino, que foram uniformemente distribuídos em 12 tanques de concreto. Uma dieta comercial foi fornecida (50% PB) a partir do dia 15 poseclosión e durante 96 dias com os seguintes tratamentos: 3.000 mg TT / kg de ração (tratamento A), 5,000 mg TT / kg de ração (B tratamento) controle MT positiva 3mg / kg de ração (tratamento C) e controle negativo com etanol (tratamento D). Os parâmetros de produção e diferenciação gonadal foram determinados pela técnica de polpa. Parâmetros físico-químicos da água foram medidos. Com cada tratamento A e B, se chegou o um percentagem de machos 2,67% (P <0,05) e houvo a presença de intersexo mas sem significância estatística. Os pesos alacanzados diário foram 0,128, 0,132, 0,109 e 0,088 g/d para os tratamentos A, B, C e D, respectivamente. Os tamanhos obtidos encontrados a partir de 0,063 cm (D) e 0,077 cm (b); ambas variáveis apresentaram diferença estatisticamente significativa (P <0,05). A sobrevivência, o fator de conversão do alimentação e a incidência de custos não foram significativamente afetados pelos tratamentos. Em conclusão a Tribulus terrestris misturado com doses concentradas de 3,000 mg / kg e 5000 mg / kg não induzem masculinização de truta arco-íris; mas, não tem nenhum impacto negativo sobre a sobrevivência da mesma, tem um efeito anabólico significativo, mostrandose em peso e lucro de altura e não é costoso para utilizar em comparação com o método convencional de hormônio 17α-metil testerona. A sua utilização para esse fim não é recomendado pela ineficiência. É o primeiro estudo relatado na Colômbia no este respeito.

In vitro cytotoxic activity of five commercial samples of Tribulus terrestris Linn in Espírito Santo (Brazil)

ABSTRACT The objective was to investigate the total saponin and protodioscin concentrations and the cytotoxicity in vitro, of five samples of the plant Tribulus terrestris, commercially available in the metropolitan region of Vitória - Espirito Santo, Brazil, and to compare them with the aqueous extract of the plant. The chromatographic profile and quantification of protodioscin in commercial samples and plant extract were evaluated by LC-MS/MS. The percentage of total saponins were determined by the colorimetric method. Extracts and protodioscin cytotoxicity were analyzed by the MTT assay in three cell lineages: fibroblasts (L929), ovarian cancer (Ovcar3) and murine hepatoma (Hepa1c1c7). All extracts displayed high levels of total saponins (207.2 to 780.3 mg g-1 of dry extract). The chromatographic profile revealed a wide diversity of compounds, and the saponin protodioscin was detected in only two extracts. One extract displayed high cytotoxicity, with IC50 values of 157.0, 38.2 and 7.4 µg mL-1 for the Ovcar3, Hepa1c1c7 and L929 cell lines, respectively. The other extracts displayed cytotoxic effects only at concentrations equal to or greater than 125.0 µg mL-1. Surprisingly, the most cytotoxic extract displayed the highest protodioscin concentration. Therefore, it is suggested that these products be marketed with caution, and followed-up by a certified healthcare professional.

Study of the Biological Activities Preparation with Tribulus Terrestris l. on Free Serum Testosterone Changes in Male Rats / Монголын эм зүй, эм судлал

Wistar nonlinear rats weighing 170-220g. Rats were divided 5 groups, including control group, group-1, group-2, group -3 and reference group. Dried thistle extract and raw bovine testicle were contained by 1:1, 1:2 and 2:1 ratio. Each 0.1g ratio was dissolved in 20 ml distilled water and administered 2 times per day. Blood sampling was done for each rat after 3, 7, 14, 21, 28 and 35 days. Their testosterone level was measured by ELISA Kit. Results: The results indicated that free serum testosterone level in male rats increase and decrease in 7 days frequency. All tested groups showed gradual increase in the level of free serum testosterone when compared to that of corresponding control (p<0.05). Statistical comparison of all groups revealed that the maximum level was found in group 1. Moreover, group 3 was showed gradually increase in level of free serum testosterone, irrelative with period of decrease testosterone level. Conclusion: According to our results and previous study, it is suggested that preparation with Tribulus terrestris L. extract could be used in the androgen deficiency and erectile dysfunctions. Keywords: Tribulus Terrestris L, Free testosterone, Dihydrotestosterone, Protodioscin

Determination of six kinds of index components in Dieda Zhitong San by HPLC-DVD variable wavelength method / 中草药

Objective: To develop an HPLC-DVD wavelength switching combined with gradient elution method for the determination of the contents of hydroxysafflor yellow A, protodioscin, dioscin, methylprotodioscin, pseudoprotodioscin, and gracillin in Dieda Zhitong San (DZS) simultaneously. Methods: The chromatographic separation was achieved on a Diamonsil C18 (250 mm × 4.6 mm, 5 μm) with acetonitrile (A)-0.4% formic acid solution (B) as mobile phase at the flow rate of 0.9 mL/min for gradient elution; Hydroxysafflor yellow A was detected at 403 nm; Protodioscin, dioscin, methylprotodioscin, pseudoprotodioscin, and gracillin were detected at 203 nm; Sample quantity was 20 μL. Results: The six active components were well separated and showed good linearity, such as hydroxysafflor yellow A 3.46-69.20 μg/mL (r = 0.999 4), protodioscin 9.52-190.40 μg/mL (r = 0.999 7), dioscin 8.74-174.80 μg/mL (r = 0.999 6), methylprotodioscin 4.45-89.00 μg/mL (r = 0.999 9), pseudoprotodioscin 2.64-52.80 μg/mL (r = 0.999 5), and gracillin 3.28-65.60 μg/mL (r = 0.999 8). The precision and repeatability were good, and RSD values were less than 2.0%. The stability was good in 12 h. The average recoveries and the corresponding RSD values were 98.57% (1.59%), 97.64% (1.28%), 99.43% (1.07%), 97.98% (1.64%), 98.57% (1.16%), and 97.17% (1.37%), respectively. Conclusion: An HPLC wavelength switching combined with gradient elution method has been successfully established for simultaneous determination of six components in DZS. The method is simple, quick, accurate, and helpful for the quality control of DZS.

Content Determination of Protodioscin and Diosgenin in Dioscoreae Hypoglaucae Rhizoma from Different Habitats by HPLC / 中国中医药信息杂志

Objective To establish an HPLC method for the content determination of protodioscin and diosgenin in Dioscoreae Hypoglaucae Rhizoma from different habitats.Methods The chromatographic separation was performed on an Agilent C18 column (4.6 mm×150 mm, 5μm) with mobile phase of acetonitrile-water solution with gradient elution at the flow rate of 0.8 mL/min; the detection wavelength was 208 nm; the column temperature was 30℃; the injection volume was 20μL.Results Protodioscin showed a good linear relationship among the range of 1.73–8.64 μg (r=0.999 6), with the average recovery of 101.98% (RSD=1.53%); Diosgenin showed a good linear relationship among the range of 1.03–8.20μg (r=0.999 1), with the average recovery of 101.60% (RSD=2.41%). The contents of protodioscin and diosgenin in Dioscoreae Hypoglaucae Rhizoma from 10 different habitats were in the range of 0.89%–2.24% and 0.75%–3.22%, respectively.Conclusion The method is simple, accurate and with repeatability, which can be used as quality control method of Dioscoreae Hypoglaucae Rhizoma.

The effect of ensiling and haymaking on the concentrations of steroidal saponin in two Brachiaria grass species / O efeito da ensilagem e fenação sobre as concentrações de saponina esteroide em duas espécies de gramínea Brachiaria

Brachiaria (signalgrass) is now the most widely used tropical grass genus in Central and South America. However, Brachiaria spp. can cause hepatogenous photosensitization in livestock. Steroidal saponins, specifically protodioscin, present in Brachiaria spp. may be responsible for liver injury and subsequent photosensitization. The objective of this study was to determine the effect of ensiling Brachiaria decumbens and Brachiaria brizantha or making hay from Brachiaria decumbens on the concentrations of steroidal saponin in these grasses. Brachiaria grass had no detectable levels of the saponin protodioscin after 24 days of ensiling. In addition, in Brachiaria decumbens, the concentration of the protodioscin decreased 48% over the first three days after haymaking and then remained constant. These results suggest that livestock consuming Brachiaria either as silage or hay may have reduced risk of intoxication by protodioscin. .

Brachiaria atualmente é o gênero de gramínea tropical mais amplamente utilizado na América Central e do Sul. Entretanto, as espécies de Brachiaria podem causar fotossensibilização hepatógena em animais de produção. Saponinas esteroides, especificamente protodioscina, presentes em Brachiaria, podem ser responsáveis pelo dano hepático e consequentemente fotossensibilização. O objetivo deste estudo foi determinar o efeito da ensilagem de Brachiaria decumbens e Brachiaria brizantha e fenação de Brachiaria decumbens sobre a concentração de saponina esteroide. As amostras de Brachiaria ensiladas não tiveram concentrações detectáveis de saponina protodioscina depois de 24 dias de ensilagem. Assim como em Brachiaria decumbens, a concentração de protodioscina reduziu 48% nos três primeiros dias após a fenação e posteriormente manteve-se constante. Estes resultados sugerem que animais de produção, ao consumirem Brachiaria tanto como silagem ou feno, podem ter o risco de intoxicação por protodioscina reduzido.

Chemical analysis of animal and plants origin raw materials to improve body potential and strength / Монголын Анагаах Ухаан

IntroductionOur country imported drugs that are contain androgen and testosterone with high selling cost. Therefore,we have to made new body potential and strength biologically activity product which have natural, lowcost and high effective.GoalThe main purpose of study was to determine chemical composition of dried testicle powder and maincompounds of Tribulus terrestris, Astragalus mongolicus.Material and MethodsThe bovine testicle used in this research was purchased from “Makh Market” Co.Ltd in 2013. T.terrestriswas collected from Gurvansaikhan, Dundgobi province July 20, 2014 and A. mongolicus was collectedfrom Botanical garden of Medicinal Plant of Drug Research Institute in September, 2014. Testicles wereremoved from skin and other parts than cut in a mechanical cutting machine. It was freeze dried at -500Cby Labconco freezone12 freeze drier. 500 g of the finely powdered T. terrestris was extracted three timeswith 5000 ml 70% ethanol for 72 hours. All extracts were combined and evaporated by vacuum rotary till2500 ml. 50 g of the powdered A. mongolicus was extracted three times with 500 ml of distilled water for72 hours. Extract was heated until 800C for 24 hours. Extract were collected and evaporated by vacuumrotary till 200 ml. Protodioscin was determined by high performance liquid chromatography (HPLC) wasachieved by using reversed-phase (RP-18) column, ultraviolet detector (UV) and water, acetonitrilegradient as mobile phase, polysaccharide was determined spectrophotometric method, protein wasanalyzed by Kjeldahl method, moisture was measured by Moisture balance 6KD-50K instrument, totalfat was analyzed by Soxhlet apparatus.ResultThe analyses of testicle powder showed 69.8% protein, 8.0% ashes, 5.42% moisture, 15.6% total fatcontent and protodioscin content 1.12% in T.terrestris extract. In A.mongolicus water extract the 7.26%polysaccharide content was found. We were determined to chemical composition of bovine testiclepowder and results were agreed with MNS 5775:2007. More over, high content of polysaccharide andprotodioscin were found T.terrestris and A.mongolicus. Therefore, those raw materials can use forpotential and strength biological activity product.

Steroidal saponin concentrations in Brachiaria decumbens and B. brizantha at different developmental stages / Concentrações de saponina esteroidal em diferentes fases do desenvolvimento de Brachiaria decumbens e B. brizantha

Brachiaria species contain steroidal saponins and are involved in outbreaks of hepatogenous photosensitization. This research presents the levels of a steroidal saponin, protodioscin, in the seeds and aerial parts of B. brizantha and B. decumbens during different developmental stages (growth, bloom, fructification and seed fall). The butanolic fraction of the ethanolic extract of each stage was submitted to thin layer chromatography (TLC) and spectrophotometric analysis through the Ehrlich reagent in 515nm. The chromatograms in TLC of the butanolic fraction of B. brizantha and B. decumbens showed similar spots as the protodioscin standard. The estimated level of protodioscin isomers in B. brizantha and B. decumbens ranged from 0.5 percent to 2.1 percent, having the highest level at the end of their developmental stages during seed falling comparison with the previous one. Protodioscin was not detected in the seeds. Outbreaks of Brachiaria spp. poisoning in central Western Brazil are frequently observed in pastures that had been more than 30 days without animals grazing, and also during the growing or blooming stage of the pastures. Other saponin determinations in toxic and non toxic pastures are necessary to determine the saponin concentrations that cause intoxication.

As Brachiaria spp contêm saponinas esteroidais envolvidas no desenvolvimento de fotossensibilizacão hepatógena. No presente trabalho foram determinados os teores da saponina esteroidal protodioscina nas partes aéreas de B. brizantha e B. decumbens, durante as diferentes fases do desenvolvimento (crescimento, floração, frutificação e queda das sementes) e nas sementes. A fração butanólica do extrato etanólico de cada fase foi submetida à cromatografia de camada delgada (CCD) e à análise espectrofotométrica por meio do reagente de Ehrlich em 515nm. Os cromatogramas em CCD da fração butanólica de B. brizantha e B. decumbens, de cada fase do desenvolvimento, apresentaram manchas similares ao padrão de protodioscina. Por meio da análise de protodioscina por espectrofotometria, os teores de protodioscina em B. brizantha e B. decumbens variaram entre 0.5 por cento e 2.1 por cento, sendo mais altos na fase de queda das sementes. Nas sementes não foi encontrada protodioscina. Surtos de intoxicação por Brachiaria spp. no Centro-Oeste brasileiro são freqüentemente observados em pastagens diferidas por mais de 30 dias e também durante as fases de crescimento e florescimento. São necessárias outras dosagens de saponinas em pastagens tóxicas e não-tóxicas para determinar as concentrações capazes de causar intoxicação.

Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil: [revision] / Intoxicação por Panicum dicotomiflorum em ovinos na região nordeste: [revisão]

Different species of Panicum, including P. dichotomiflorum,have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to ÕH and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

Diferentes espécies de Panicum, incluindo P. dichotomiflorum,causam fotossensibilização em ovinos, eqüinos, bovinos e caprinos. Um surto de fotossensibilização hepatógena ocorreu em 3 rebanhos de ovinos no município de Casa Nova, Bahia. Oitenta e um de 365 ovinos adoeceram e 39 morreram. Cordeiros lactentes e ovinos de menos de um ano foram mais afetados. Jumentos, cabras e bovinos que permaneciam nas mesmas pastagens não foram afetados. Clinicamente observou-se edema da face, seguido por dermatite, principalmente da face, orelhas e garupa, corrimento ocular, opacidade da córnea com cegueira, e avermelhamento do rodete coronário e casco. Na necropsia de um cordeiro afetado o fígado estava amarelado. Na histologia observou-se necrose aleatória de hepatócitos no fígado e áreas focais de necrose de miócitos no miocárdio. Amostras de P. dicotomiflorum foram analisadas por cromatografia em camada delgada e aquelas contendo saponinas foram isoladas por HPLC utilizando uma coluna RP-C18 com mistura de metanol e água. Os compostos isolados foram submetidos a espectroscopia de ressonância magnética nuclear de hidrogênio (RMN ÕH) e de carbono (RMN 13C). As reações foram positivas para saponinas furostanólicas com o mesmo Rf que os padrões de protodioscina (0,21) e metilprotodioscina (0,32). Este resultado e os dados espectroscópicos sugerem a presença, em P. dichotomiflorum, de metilprotodioscina e uma mistura de isômeros (25R)- e (25S)- de protodioscina em uma proporção de 3:1.

Effect of MPD on Acute Myocardial Ischemia in Dogs / 中药新药与临床药理

Objective To study the protecitve effect of methy Protodioscin(MPD)on myocardial infarction in dogs. Methods Dog models of myocardial infarction were induced by ligation of coronary artery. The degree of myocardial ischemia was calculated by measuring the epicardial electrogram, and the myocardial infarction area was detected with N-BT histochemistry staining method. The changes of ET, 6-Keto-PGF1a, TXB2 in blood plasma were observed with radioimmunoassay. Meanwhile, the coronary blood flow was measured. Results MPD can obviously relieve the pathological changes of the acute myocardial infarction, decrease the infarction size, reduce the myocardium ischemic degree, dilate the coronary artery , and increase the myocardial blood supply. Meanwhile, MPD can improve the vascular endothelial cell function. Conclusion MPD had obvious effect for relieving acute myocardial infarction in dogs.

Effects of MPD on Heart Hemodynamics in Normal Anesthetic Dogs / 中药新药与临床药理

Objective To investigate the pharmacological effects of MPD on heart hemodynamics and myocardial oxygen consumption in dogs, and to study its mechanism. Methods The changes of parameters such as blood pressure (BP), coronary blood flow, myocardial oxygen consumption and heart rates were observed in normal anesthetic dogs. Results Compared to those in the normal group, MPD can obviously lower the BP and peripheral vascular resistance, dilate the coronary and peripheral artery, increase the coronary blood flow and improve the left ventricle function and myocardium oxygen consumption rate. Conclusion MPD can improve the heart hemodynamics and regulate myocardium oxygen consumption.

Therapeutic Effects of Methyl Protodioscin for Myocardial Infarction in Rats / 中药新药与临床药理

Objective To study the therapeutic effects of methyl protodioscin (MPD ) on myocardial infarction in rats. Methods Rat models of myocardial infarction were induced by ligation of coranary artery. Then myocardium infarction area and the vasoactive substance were measured to evaluate the therapeutic effect of MPD on acute myocardial infarction in rats. Results Compared with the control group,MPD lessened the myocardial infarction size dramatically,inhibited the increase of CK and LDH ,lowered the increased MDA content level and improved the activity of SOD and NO. Conclusion MPD reduces the level of myocardium enzyme and the myocardial infarction size,and increases the capability of clearing oxygen free radical and function of the vascular endothelial cell. MPD by intravenous injection has a better effect than that by oral use.

Effects of Methyl Protodioscin on In-Vivo and In-Vitro Thrombosis and Blood Viscosity in Rats / 中药新药与临床药理

Objective To investigate the effects of methyl protodioscin (MPD ) on in-vitro and in-vivo thrombosis and blood viscosity in rats. Methods The vitro thrombus was induced by Chandler method,and the length,wet and dry weight of the thrombus were measured. Thrombosis instrument was to observe the in-vivo occlusion time (OT). At the same time,determined the high-,middle-,low-shear blood viscosity as well as the plasma viscosity in rats was determined .Results Compared to normal group,middle-dose MPD group can delay the OT,and the high-dose group can decrease the length,wet and dry weight of in-vitro thrombus. The blood viscosity is reduced in all groups. Conclusion MPD can inhibit the in-vitro thrombosis,decrease the dry and wet weight of thrombus and delay the OT. Moreover,MPD has the effects of lowering the whole blood viscosity and plasma viscosity.