RESUMO
A stable nanoscaled single-excitation ratiometric luminescent pH sensor (MPDB-PCN) over a broad pH range from 2.5 to 8.6 is fabricated through post-synthetic modification of PCN-224 with naphthalimide-derived molecules. Due to the rapid, sensitive and linear response to pH, MPDB-PCN is capable of detecting 3-nitropropionic acid (3-NPA), an acid neurotoxin in food safety, with a low detection limit of 15 µM in sugarcane juice.
Assuntos
Substâncias Luminescentes/química , Estruturas Metalorgânicas/química , Micotoxinas/análise , Nanopartículas/química , Contaminação de Alimentos/análise , Concentração de Íons de Hidrogênio , Limite de Detecção , Neurotoxinas/análise , Nitrocompostos/análise , Extratos Vegetais/análise , Propionatos/análise , Saccharum/química , Espectrometria de Fluorescência/métodos , Zircônio/químicaRESUMO
The neurotoxins of venomous scorpion act on ion channels. Whether these neurotoxins are retained in processed Buthus martensii Karsch scorpions used in traditional Chinese medicine materials is unknown. Comprehensive mass spectrometry-based proteomic characterization of functionally active toxins in the processed medicinal scorpion material revealed 22 full-length and 44 truncated thermostable potassium channel-modulatory toxins that preserved six conserved cysteine residues capable of forming the three disulfide bonds necessary for toxicity. Additionally, a broad spectrum of degraded toxin fragments was found, indicating their relative thermal instability which enabled toxicity reduction. Furthermore, the suppression of interleukin-2 (IL-2) production in Jurkat cells and the reduced delayed-type hypersensitivity (DTH) response demonstrated that the extracts have immunoregulatory activity both in vitro and in vivo. Our work describes the first "map" of functionally active scorpion toxins in processed scorpion medicinal material, which is helpful to unveil the pharmaceutical basis of the processed scorpion medicinal material in traditional Chinese medicine. BIOLOGICAL SIGNIFICANCE: Scorpions have been used as medicinal materials in China for more than one thousand years. This is an example of the well-known "Combat poison with poison" strategy common to traditional Chinese medicine. In the past 30â¯years, extensive investigations of Chinese scorpions have indicated that the neurotoxins in the scorpion venom are the main toxic components and they target various ion channels in cell membranes. However, whether these neurotoxins are retained in processed Buthus martensii Karsch scorpions used for traditional Chinese medicine remains unknown. Our study described the thermal stability and instability of potassium channel-modulatory neurotoxins in processed scorpions and helps to understand the pharmaceutical basis underling the strategy of "combat poison with poison to cure diseases".
Assuntos
Medicina Tradicional Chinesa , Neurotoxinas/análise , Bloqueadores dos Canais de Potássio/análise , Proteoma/análise , Venenos de Escorpião/análise , Animais , Estabilidade de Medicamentos , Feminino , Células HEK293 , Humanos , Células Jurkat , Neurotoxinas/metabolismo , Peptídeos/análise , Peptídeos/metabolismo , Preparações Farmacêuticas/análise , Preparações Farmacêuticas/química , Preparações Farmacêuticas/metabolismo , Bloqueadores dos Canais de Potássio/isolamento & purificação , Bloqueadores dos Canais de Potássio/metabolismo , Canais de Potássio/metabolismo , Estabilidade Proteica , Proteoma/metabolismo , Proteômica/métodos , Ratos , Ratos Endogâmicos Lew , Venenos de Escorpião/química , Venenos de Escorpião/metabolismo , Escorpiões/química , Escorpiões/metabolismo , TemperaturaRESUMO
Cyanobacteria are photosynthetic microorganisms distributed globally in aquatic and terrestrial environments. They are also industrially cultivated to be used as dietary supplements, as they have a high nutritional value; however, they are also known to produce a wide range of toxic secondary metabolites, called cyanotoxins. BMAA (ß-methylamino-l-alanine) and its most common structural isomers, DAB (2,4-diaminobutyric acid) and AEG (N-2-aminoethylglycine) produced by cyanobacteria, are non-proteinogenic amino acids that have been associated with neurodegenerative diseases. A possible route of exposure to those amino acids is through consumption of food supplements based on cyanobacteria. The review critically discusses existing reports regarding the occurrence of BMAA, DAB and AEG in cyanobacteria and cyanobacteria-based food supplements. It is shown that inconsistencies in reported results could be attributed to performance of different methods of extraction and analysis applied and in ambiguities regarding determination of soluble and bound fractions of the compounds. The critical aspect of this review aims to grow awareness of human intake of neurotoxic amino acids, while results presented in literature concerning dietary supplements aim to promote further research, quality control as well as development of guidelines for cyanotoxins in food products.
Assuntos
Diamino Aminoácidos/análise , Cianobactérias/química , Suplementos Nutricionais/análise , Neurotoxinas/análise , Diamino Aminoácidos/química , Diamino Aminoácidos/toxicidade , Animais , Toxinas de Cianobactérias , Monitoramento Ambiental , Humanos , Isomerismo , Neurotoxinas/química , Neurotoxinas/toxicidadeRESUMO
There is a paucity of information concerning the developmental neurotoxicity (DNT) hazard posed by industrial and environmental chemicals. New testing approaches will most likely be based on batteries of alternative and complementary (non-animal) tests. As DNT is assumed to result from the modulation of fundamental neurodevelopmental processes (such as neuronal differentiation, precursor cell migration or neuronal network formation) by chemicals, the first generation of alternative DNT tests target these processes. The advantage of such types of assays is that they capture toxicants with multiple targets and modes-of-action. Moreover, the processes modelled by the assays can be linked to toxicity endophenotypes, i.e., alterations in neural connectivity that form the basis for neurofunctional deficits in man. The authors of this review convened in a workshop to define criteria for the selection of positive/negative controls, to prepare recommendations on their use, and to initiate the setup of a directory of reference chemicals. For initial technical optimization of tests, a set of > 50 endpoint-specific control compounds was identified. For further test development, an additional "test" set of 33 chemicals considered to act directly as bona fide DNT toxicants is proposed, and each chemical is annotated to the extent it fulfills these criteria. A tabular compilation of the original literature used to select the test set chemicals provides information on statistical procedures, and toxic/non-toxic doses (both for pups and dams). Suggestions are provided on how to use the > 100 compounds (including negative controls) compiled here to address specificity, adversity and use of alternative test systems.
Assuntos
Alternativas aos Testes com Animais , Avaliação Pré-Clínica de Medicamentos/métodos , Neurônios/efeitos dos fármacos , Neurotoxinas/análise , Testes de Toxicidade/métodos , Animais , Congressos como Assunto , Feminino , Substâncias Perigosas , Humanos , Masculino , Neurônios/citologia , Síndromes Neurotóxicas/etiologia , Gravidez , Efeitos Tardios da Exposição Pré-NatalRESUMO
Recent reports of the widespread occurrence of the neurotoxin ß-N-methylamino-L-alanine (BMAA) in cyanobacteria and particularly seafood have raised concerns for public health. LC-MS/MS is currently the analytical method of choice for BMAA determinations but incomplete separation of isomeric and isobaric compounds, matrix suppression and conjugated forms are plausible limitations. In this study, capillary electrophoresis (CE) coupled with MS/MS has been developed as an alternative method for the quantitative determination of free BMAA. Using a bare fused silica capillary, a phosphate buffer (250 mM, pH 3.0) and UV detection, it was possible to separate BMAA from four isomers, but the limit of detection (LOD) of 0.25 µg mL-1 proved insufficient for analysis of typical samples. Coupling the CE to a triple quadrupole MS was accomplished using a custom sheath-flow interface. The best separation was achieved with a 5 M formic acid in water/acetonitrile (9:1) background electrolyte. Strong acid hydrolysis of lyophilized samples was used to release BMAA from conjugated forms. Field-amplified stacking after injection was achieved by lowering sample ionic strength with a cation-exchange cleanup procedure. Quantitation was accomplished using isotope dilution with deuterium-labelled BMAA as internal standard. An LOD for BMAA in solution of 0.8 ng mL-1 was attained, which was equivalent to 16 ng g-1 dry mass in samples using the specified extraction procedure. This was comparable with LC-MS/MS methods. The method displayed excellent resolution of amino acid isomers and had no interference from matrix components. The presence of BMAA in cycad, mussel and lobster samples was confirmed by CE-MS/MS, but not in an in-house cyanobacterial reference material, with quantitative results agreeing with those from LC-MS/MS. Graphical Abstract CE-MS separation and detection of BMAA, its isomers and the internal standard BMAA-d3.
Assuntos
Diamino Aminoácidos/análise , Eletroforese Capilar/métodos , Contaminação de Alimentos/análise , Neurotoxinas/análise , Frutos do Mar/análise , Espectrometria de Massas em Tandem/métodos , Animais , Bivalves/química , Cianobactérias/química , Toxinas de Cianobactérias , Eletroforese Capilar/instrumentação , Desenho de Equipamento , Limite de Detecção , Nephropidae/química , Espectrometria de Massas em Tandem/instrumentaçãoRESUMO
In this study, based on web database, 324 neurotoxic compounds and 234 non-neurotoxic compounds were selected as a data set for neurotoxicity discriminative model. 6 122 molecular descriptors, including charge distribution, physicochemical and geometrical descriptors,were calculated to characterize the molecular structure of neurotoxic compounds. The combination of Cfs Subset Evaluation and Best First-D1-N5 searching was used to select molecular descriptors. A discrimination model with high accuracy was built based on the support vector machine (SVM) approach. Meanwhile, the model accuracy, sensitivity and specificity were all above 80%. Besides, 30 traditional Chinese medicine compositions with neurotoxicity were set as external validation to further verify the model accuracy,with anaccuracy of 73.333%. Using the model, 13 potential neurotoxic compounds were screened from Sophorae subprostrate Radix,4 of them were verified by literatures. The results demonstrated that the discrimination model can be applied to screen neurotoxic compounds from Chinese medicinal materials.
Assuntos
Medicina Tradicional Chinesa/métodos , Neurotoxinas/análise , Máquina de Vetores de Suporte , Simulação por Computador , Avaliação Pré-Clínica de Medicamentos/métodos , Modelos Teóricos , Neurotoxinas/química , Reprodutibilidade dos TestesRESUMO
Introducción: Las cícadas son plantas que en algunas partes del mundo son empleadas como alimento fresco o materia prima para la elaboración de harina con alto valor nutricional. Sin embargo, contienen principios activos como metilazoximetanol, β-metilamino-L-alanina, β-oxalilamino-L-alanina y cicasina, entre otros, que pueden producir efectos neurotóxicos. El consumo de cícadas y sus derivados se ha asociado con enfermedades neurodegenerativas, como el complejo demencia-parkinsonismo-esclerosis-lateral amiotrófica y otras enfermedades caracterizadas por alteraciones en la motricidad. Por lo tanto, no debemos perder de vista que todo producto, aunque sea de origen natural, puede ser benéfico o perjudicial para la salud, lo cual dependerá de sus componentes químicos y de la vulnerabilidad de quienes los consumen. Desarrollo: Se realizó un análisis de la literatura sobre las propiedades neurotóxicas de las cícadas y su asociación con enfermedades neurológicas, con el fin de proporcionar información estructurada a la población para contribuir a la prevención de problemas de salud en quienes interactúan con estas plantas. Conclusión: Las cícadas contienen neurotóxicos que contribuyen al desarrollo de enfermedades neurológicas cuando son ingeridas inadecuadamente, por lo que debemos considerar que si bien algunos vegetales pueden tener un alto valor nutricional y subsanar el déficit alimentario en las poblaciones vulnerables, también pueden ser tóxicos e impactar negativamente sobre la salud
Introduction: Cycads are ornamental plants that in some parts of the world are used as fresh food or raw material for producing flour with a high nutritional value. However, they also contain active compounds, including methylazoxymethanol, β-methylamino-L-alanine, β-alanine-L-oxalylamino and cycasin, which may produce neurotoxic effects. Some studies have associated consuming cycads and their derivatives with neurodegenerative diseases such as amyotrophic lateral sclerosis/Parkinsonism dementia complex, and other diseases characterised by motor impairment. Therefore, we must not forget that any product, no matter how natural, may present health risks or benefits depending on the chemical compounds it contains and the susceptibility of those who consume it. Development: We completed a literature analysis to evaluate the neurotoxic properties of cycads and their association with neurological diseases in order to provide structured scientific information that may contribute to preventing health problems in people who use these plants. Conclusion: Cycads contain neurotoxic compounds that may contribute to the development of neurological diseases when ingested improperly. We must be mindful of the fact that while some plants have a high nutritional value and may fill the food gap for vulnerable populations, they can also be toxic and have a negative impact on health
Assuntos
Humanos , Cycas/efeitos adversos , Extratos Vegetais/efeitos adversos , Doenças Neurodegenerativas/induzido quimicamente , Neurotoxinas/análise , Acetato de Metilazoximetanol/análiseRESUMO
Shark cartilage products are marketed as dietary supplements with claimed health benefits for animal and human use. Shark fin and cartilage products sold as extracts, dry powders and in capsules are marketed based on traditional Chinese medicine claims that it nourishes the blood, enhances appetite, and energizes multiple internal organs. Shark cartilage contains a mixture of chondroitin and glucosamine, a popular nutritional supplement ingested to improve cartilage function. Sharks are long-lived apex predators, that bioaccumulate environmental marine toxins and methylmercury from dietary exposures. We recently reported detection of the cyanobacterial toxin ß-N-methylamino-l-alanine (BMAA) in the fins of seven different species of sharks from South Florida coastal waters. Since BMAA has been linked to degenerative brain diseases, the consumption of shark products may pose a human risk for BMAA exposures. In this report, we tested sixteen commercial shark cartilage supplements for BMAA by high performance liquid chromatography (HPLC-FD) with fluorescence detection and ultra performance liquid chromatography/mass spectrometry/mass spectrometry (UPLC-MS/MS). Total mercury (Hg) levels were measured in the same shark cartilage products by cold vapor atomic fluorescence spectrometry (CVAFS). We report here that BMAA was detected in fifteen out of sixteen products with concentrations ranging from 86 to 265µg/g (dry weight). All of the shark fin products contained low concentrations of Hg. While Hg contamination is a known risk, the results of the present study demonstrate that shark cartilage products also may contain the neurotoxin BMAA. Although the neurotoxic potential of dietary exposure to BMAA is currently unknown, the results demonstrate that shark cartilage products may contain two environmental neurotoxins that have synergistic toxicities.
Assuntos
Diamino Aminoácidos/toxicidade , Suplementos Nutricionais , Contaminação de Medicamentos , Mercúrio/toxicidade , Neurotoxinas/toxicidade , Extratos de Tecidos/toxicidade , Diamino Aminoácidos/análise , Toxinas Bacterianas/toxicidade , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Cianobactérias/química , Toxinas de Cianobactérias , Meio Ambiente , Toxinas Marinhas/toxicidade , Mercúrio/análise , Microcistinas/toxicidade , Neurotoxinas/análise , Espectrometria de Massas em Tandem , Extratos de Tecidos/químicaRESUMO
The aim of this work was to develop an analytical method capable of determining the presence of anisatin in star anise. This neurotoxin may induce severe side effects such as epileptic convulsions. It is therefore of prime importance to have rapid and accurate analytical methods able to detect and quantify anisatin in samples that are purportedly edible star anise. The sample preparation combined an automated accelerated solvent extraction with a solid-supported liquid-liquid purification step on EXtrelut®. Samples were analysed on a porous graphitic carbon HPLC column and quantified by tandem mass spectrometry operating in the negative ionisation mode. The quantification range of anisatin was between 0.2 and 8 mg kg⻹. The applicability of this validated method was demonstrated by the analysis of several Illicium species and star anise samples purchased on the Swiss market. High levels of anisatin were measured in Illicium lanceolatum, I. majus and I. anisatum, which may cause health concerns if they are misidentified or mixed with edible Illicium verum.
Assuntos
Qualidade de Produtos para o Consumidor , Inspeção de Alimentos/métodos , Frutas/química , Illicium/química , Lactonas/análise , Neurotoxinas/análise , Sesquiterpenos/análise , Compostos de Espiro/análise , Automação Laboratorial , Bebidas/efeitos adversos , Bebidas/análise , Cromatografia Líquida de Alta Pressão , Rotulagem de Alimentos , Doenças Transmitidas por Alimentos/etiologia , Doenças Transmitidas por Alimentos/prevenção & controle , Frutas/efeitos adversos , Frutas/classificação , Frutas/economia , Humanos , Illicium/efeitos adversos , Illicium/classificação , Lactonas/química , Lactonas/toxicidade , Medicina Tradicional , Neurotoxinas/química , Neurotoxinas/toxicidade , Plantas Comestíveis/efeitos adversos , Plantas Comestíveis/química , Plantas Tóxicas/efeitos adversos , Plantas Tóxicas/química , Reprodutibilidade dos Testes , Sesquiterpenos/química , Sesquiterpenos/toxicidade , Especificidade da Espécie , Espectrometria de Massas por Ionização por Electrospray , Compostos de Espiro/química , Compostos de Espiro/toxicidade , Suíça , Espectrometria de Massas em TandemRESUMO
The simultaneous determination of harman and norharman using second derivative synchronous fluorescence method has been developed based on their natural fluorescence. Due to their similar molecular structures, it is difficult to determine them simultaneously in the mixture using conventional fluorimetry. Overlapping of fluorescence spectra was resolved by using a constant second derivative synchronous fluorimetry. The derivative synchronous spectrum, maintaining a constant difference of Δλ=150 nm between emission and excitation for both the compounds, has been selected for the analysis. The range of application is between 0.182 and 18.2 µg/mL (correlation coefficient, R=0.9982) for harman and between 0.504 and 16.8 µg/mL (correlation coefficient, R=0.9962) for norharman. The recovery ranges of 98.5-101.1% for harman and 97.5-99.1% for norharman from their synthetic mixture was reported. The detection limits are 0.016 µg/mL and 0.038 µg/mL for harman and norharman, respectively. Similarly, the quantitation limit of the two compounds was found to be 0.049 and 0.109 µg/mL, respectively. The method was applied to the simultaneous determination of both compounds in coffee samples with satisfactory results.
Assuntos
Café/química , Harmina/análogos & derivados , Neurotoxinas/análise , Espectrometria de Fluorescência/métodos , Carbolinas , Harmina/análise , Limite de DetecçãoRESUMO
After ingestion, products containing Chinese star anise (Illicium verum) contaminated or adulterated with Japanese star anise (Illicium anisatum) or other Illicium species, can cause epilepsy, hallucinations, and nausea due to the rare neurotoxic sesquiterpene dilactone anisatin that is present in Japanese star anise. Thus a rapid, simple and unambiguous method for distinguishing between the morphologically similar Chinese star anise and toxic Japanese star anise is important for food safety issues. Direct Analysis in Real Time (DART) ambient ionisation coupled with orbitrap high resolution mass spectrometry allowed the recording of mass spectra of anisatin in solid star anise fruits in seconds without any prior sample pretreatment. Spectra could be obtained in both positive ([M+NH(4)](+) at m/z 346.1496, C(15)H(24)NO(8)) and negative mode ([M-H](-) at m/z 327.1074, C(15)H(19)O(8)) and gave the same outcome provided a mass resolution of at least 27,000 is available. The anisatin signal was typically >1000 times larger in Japanese star anise than in Chinese star anise thus allowing an unequivocal qualitative determination. Herbal teas containing star anise fragments too small to be visually recognised, could be analysed by preparing a tea in 6 min and subsequently sampling â¼2 µL of tea on a glass rod. None of the 8 investigated retail teas contained significant quantities of anisatin. Spiking a complex herbal tea containing Chinese star anise with an equally concentrated tea prepared from Japanese star anise provided a linear calibration curve (R(2) ≥ 0.995) after normalising on a native constituent of Chinese star anise (standard addition method). This showed that adulteration down to 1% (w/w) is still measurable. Compared with existing PCR, TLC, GC-MS and HPLC-ESI-MS/MS procedures, the proposed DART-HRMS procedure is faster and simpler and moreover measures the actual biotoxin.
Assuntos
Frutas/química , Illicium/química , Lactonas/análise , Espectrometria de Massas/métodos , Neurotoxinas/análise , Sesquiterpenos/análise , Compostos de Espiro/análise , Chá/química , Cromatografia Líquida de Alta Pressão , Limite de Detecção , Modelos LinearesRESUMO
Guadeloupean Parkinsonism has been linked epidemiologically to the consumption of Annonaceae fruits. These were proposed to be etiological agents for sporadic atypical Parkinsonism worldwide, because of their content of neurotoxins such as isoquinolinic alkaloids and Annonaceous acetogenins. The pulp of Annona cherimolia Mill. from Spain was screened for these toxic molecules using Matrix-Assisted Laser Desorption Ionisation - Time of Flight mass spectrometry (MALDI-TOF MS) and it was found not to be a source of exposure. However, kaurenoic acid, a diterpene considered to be cytotoxic, was detected in high amounts (66 mg/fresh fruit). Treatment of rat embryonic striatal primary cultures, up to a high concentration (50 µM), did not cause neuronal death nor astrogliosis, suggesting that this molecule is not at risk of implication in human neurodegenerative diseases.
Assuntos
Annona/efeitos adversos , Diterpenos/análise , Transtornos Parkinsonianos/induzido quimicamente , Animais , Annona/química , Astrócitos/efeitos dos fármacos , Células Cultivadas , Diterpenos/isolamento & purificação , Frutas/química , Neurotoxinas/análise , Neurotoxinas/isolamento & purificação , Ratos , Ratos Wistar , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
The aim of this research was to study the antioxidant and nutritional (selected objects) properties of protein isolates obtained from grass pea seedlings as compared with soaked and raw seeds. Two percent extract of isolate from 5-day-old seedlings showed the highest total antioxidant activity (25%) and the ability to chelate Fe²+ (2.35 mg/g d.m.) as compared with other isolates. Protein isolates from grass pea seeds had on average 89% total protein, 87% in vitro protein bioavailability, about 5574 TIU/g (d.m.) (trypsin inhibitors activity) and did not contain ODAP. Germination of seeds for 5 days considerably improved the in vitro bioavailability of isolates, by 12%, and profile of sulfur amino acids by 42%, in comparison with isolates obtained from the raw seeds. Isolates from 5-day-old grass pea seedlings had the best antioxidant properties and improved nutritional parameters (as compared with raw seeds), which makes them worthy of being considered as a potential food additive.
Assuntos
Antioxidantes/análise , Germinação , Lathyrus/química , Preparações de Plantas/química , Proteínas de Plantas/química , Plântula/química , Sementes/química , Aminoácidos Essenciais/análise , Aminoácidos Sulfúricos/análise , Digestão , Alimentos Fortificados , Quelantes de Ferro/química , Quelantes de Ferro/metabolismo , Quelantes de Ferro/farmacologia , Lathyrus/crescimento & desenvolvimento , Neurotoxinas/análise , Valor Nutritivo , Preparações de Plantas/metabolismo , Preparações de Plantas/farmacologia , Proteínas de Plantas/metabolismo , Proteínas de Plantas/farmacologia , Polônia , Plântula/crescimento & desenvolvimento , Sementes/crescimento & desenvolvimento , Fatores de Tempo , Inibidores da Tripsina/química , Inibidores da Tripsina/metabolismo , Inibidores da Tripsina/farmacologia , beta-Alanina/análogos & derivados , beta-Alanina/análiseRESUMO
Twenty-six Pseudo-nitzschia multistriata cultures were tested for intracellular domoic acid production and fourteen were found to be toxic. Four suboptimal growth conditions were compared with conditions observed to be optimal to explore possible triggers for intracellular domoic acid production. Silica- and phosphate-limitation and low light treatment induced elevated toxin concentrations whereas high temperature appeared to suppress it. Inheritance of the toxin-production ability was investigated by measuring intracellular toxin content in a total of thirty-nine F(1) strains from two different crosses. Results showed radical differences in domoic acid levels among the F(1) offspring from the same parents.
Assuntos
Diatomáceas/química , Agonistas de Aminoácidos Excitatórios/análise , Ácido Caínico/análogos & derivados , Toxinas Marinhas/análise , Neurotoxinas/análise , Cruzamentos Genéticos , Diatomáceas/genética , Diatomáceas/crescimento & desenvolvimento , Diatomáceas/isolamento & purificação , Temperatura Alta , Itália , Ácido Caínico/análise , Luz , Mar Mediterrâneo , Fósforo/deficiência , Intoxicação por Frutos do Mar/etiologia , Silício/deficiência , Especificidade da Espécie , Fatores de TempoRESUMO
Since diverse taxa of cyanobacteria has been linked to biosynthesis of BMAA, a controversy has arisen about the detection of neurotoxic amino acids in cyanobacteria. In this context, a novel LC-MS/MS method was developed for the unambiguous determination of beta-N-methylamino-L-alanine (BMAA) and 2,4-diaminobutyric acid (DAB) in cyanobacteria and selected plant seeds. Both neurotoxic and non-proteinogenic amino acids were analyzed without derivatization considering the total concentration of the free and protein-bound form. The investigation of overall 62 cyanobacterial samples of worldwide origin by application of this method revealed the absence of BMAA, whereas seeds of Cycas revoluta contained 6.96 microg g(-1) of free BMAA. In contrast, the isomer DAB was confirmed in 16 cyanobacterial samples in concentrations of 0.07-0.83 microg g(-1),whereof one sample is distributed as nutritional supplement. In addition, seeds of Lathyrus latifolius contained 4.21 microg g(-1) of free DAB. Limits of detection were for BMAA<1.0 microg g(-1) in the cyanobacterial matrix and<0.14 microg g(-1) in angiosperm seeds. DAB exhibits higher sensitivities of <0.06 microg g(-1) in cyanobacteria and <0.008 microg g(-1) in angiosperm seeds. The highly specific analysis method with increased detection sensitivity eliminates the disadvantages of derivatization-based methods to be discussed.
Assuntos
Diamino Aminoácidos/análise , Aminobutiratos/análise , Cianobactérias/química , Cycas/química , Lathyrus/química , Neurotoxinas/análise , Cromatografia Líquida de Alta Pressão , Toxinas de Cianobactérias , Suplementos Nutricionais/análise , Isomerismo , Sementes/química , Espectrometria de Massas em TandemRESUMO
In the mountains of Peru, globular colonies of Nostoc commune (Nostocales) are collected in the highland lakes by the indigenous people, who call them llullucha. They are consumed locally, traded for maize, or sold, eventually entering the folk markets of Cusco and other neighboring cities. Throughout highland Peru, Nostoc commune is highly salient as a seasonal dietary item, being eaten alone, or in picante -- a local stew -- and is said to be highly nutritious. Nostoc commune has been known to produce unusual amino acids, including those of the mycosporine group, which possibly function to prevent UV damage. We analyzed 21 different Nostoc commune spherical colonies from 7 different market collections in the Cusco area for the presence of beta-N-methylamino-L-alanine (BMAA), a neurotoxic amino acid produced by diverse taxa of cyanobacteria, using four different analytical techniques (HPLC-FD, UPLC-UV, UPLC/MS, LC/MS/MS). We found using all four techniques that BMAA was present in the samples purchased in the Peruvian markets. Since BMAA has been putatively linked to neurodegenerative illness, it would be of interest to know if the occurrence of ALS, Alzheimer's, or Parkinson's Disease is greater among individuals who consume llullucha in Peru.
Assuntos
Diamino Aminoácidos/metabolismo , Neurotoxinas/metabolismo , Nostoc commune/química , Diamino Aminoácidos/análise , Diamino Aminoácidos/toxicidade , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida/métodos , Toxinas de Cianobactérias , Suplementos Nutricionais , Medicina Tradicional , Neurotoxinas/análise , Neurotoxinas/toxicidade , Peru , Espectrometria de Massas em Tandem/métodosRESUMO
This study was designed to document convulsant and neurotoxic properties of extracts of a tropical tree, Magnistipula butayei subsp. Montana, and to investigate the involvement of the glutamatergic system in these effects. Continuous behavioral observations and electroencephalographic (EEG) records were obtained after per os administration of an aqueous extract of Magnistipula (MBMAE) in rats. MBMAE (800 mg/kg) induced behavioral changes resembling motor limbic seizures: staring and head tremor, automatisms, forelimb clonic movements and violent tonic-clonic seizures leading to death in all animals. Concomitantly, important seizure activity that gradually evolved to epileptiform activity was recorded on the EEG. Moreover, c-Fos immunohistochemistry has revealed an increased c-Fos expression in the dentate gyrus and in piriform, peri- and entorhinal cortices 2 and 4h after treatment. This expression pattern suggested that the mechanism of action for the MBMAE is similar to that observed in glutamate-induced models of epilepsy. The MBMAE increased cell death also in hippocampal cell cultures. Furthermore, the build-up of convulsive activity and epileptic discharges induced by MBMAE in rat were abolished by MK-801, an NMDA receptor antagonist. Our study suggests that MBMAE contains a potent toxin, with a powerful neurotoxic activity in rat, and corresponding to a new natural component(s) that act as an NMDA-mediated convulsant molecule.
Assuntos
Chrysobalanaceae/química , Convulsivantes/toxicidade , Epilepsia Tônico-Clônica/induzido quimicamente , Neurotoxinas/toxicidade , Extratos Vegetais/toxicidade , Análise de Variância , Animais , Convulsivantes/análise , Maleato de Dizocilpina/farmacologia , Eletroencefalografia/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Imuno-Histoquímica , Masculino , Neurotoxinas/análise , Extratos Vegetais/análise , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Wistar , Receptores de N-Metil-D-Aspartato/antagonistas & inibidoresRESUMO
Espresso coffee (EC) brews were analyzed for beta-carboline [norharman (NH) and harman (H)] contents, by RP-HPLC with fluorescence detection. The influence of the coffee species (arabica or robusta), the roast degree, and the brew length was studied. The results show that the content of NH and H in EC is dependent primarily on the coffee species, followed by brew length. The roast degree has only a minor influence on the final content of NH and H in EC. When compared with other coffee brews, EC has an amount of these beta-carbolines (in micrograms per liter) similar to that of mocha coffee, both being more concentrated than filter and press-pot coffees. Therefore, the consumer's preferences will determine the amount of NH and H ingested daily. For the caffeinated 30 mL of EC, the arabica coffees contain about 4.08 microg of NH and 1.54 microg of H. Commercial blends (usually with a maximum of 30% robusta) range from the cited arabica values to 10.37 microg of NH and 4.35 microg of H.