Tacinga inamoena vegetal drug characterization using phytochemistry, pharmacopoeial methods and thermoanalytical techniques

Tacinga inamoena (K. Schum.) N.P. Taylor & Stuppy, also known as quipá, is a native cactus of the Caatinga used in traditional medicine to treat urethral infections and inflammation. This study aimed to determine the physicochemical characteristics of vegetal drug obtained from the roots of T. inamoena. Analytical techniques and phytochemical tests were used, such as thermal analysis, qualitative and semiquantitative determination of secondary metabolites and spectroscopy at the infrared region. The powder of the vegetal drug met the parameters established by the Brazilian Pharmacopoeia, except for compressibility, which was low. On the thermogravimetric curve, three events related to the mass loss were verified, which correlate with the vegetal drug quality control and play a part in their standardization. The qualitative screening suggested the presence of alkaloids, flavonoids and terpenes. The infrared spectrum reinforced the presence of hydroxyl, carbonyl, and ether groups. In the semiquantitative screening, a concentration for total polyphenols of 65 mg equivalent to gallic acid g-1 to the crude ethanol extract (CEE) was obtained. On the correlation of flavonoid content to seasonality, a concentration was obtained of 3.3 mg equivalent to quercetin g-1 to the CEE obtained during the drought period and of 10.6 mg equivalent to quercetin g-1 to the CCE obtained during the rainy season. In T. inamoena, the presence of important classes of secondary metabolites, which are associated with the pharmacognostic characterization, aids the authentication and quality control of vegetal drugs of importance in traditional Brazilian medicine.

Desenvolvimento e aplicação de método analí­tico para detecção de estimulantes em suplementos nutricionais adulterados / The development and application of analytical method for the detection of stimulants in adulterated nutritional supplements

A adição fraudulenta de ativos farmacêuticos em suplementos nutricionais é um problema mundial. É comum encontrar mensagens sobre perda de peso, aumento da capacidade intelectual e/ou física, e estímulo sexual na embalagem de suplementos adulterados com fármacos sintéticos ocultos em formulações aparentemente inofensivas para os usuários. No Brasil, a disponibilidade de dados sobre a adulteração de suplementos nutricionais é escassa. No presente trabalho, foi desenvolvido e aplicado um método analítico empregando cromatografia gasosa com detector de nitrogênio fósforo (GC-NPD) para a detecção, identificação e quantificação de estimulantes/anorexígenos não declarados nos rótulos de suplementos alimentares, tais como: cafeína, femproporex, anfepramona, fenfluramina, sibutramina, fentermina, efedrina, fenilpropanolamina, pseudoefedrina e 4- metilhexan-2-amina. A técnica de extração/solubilização com metanol foi utilizada, ressaltando a utilização de baixa quantidade de amostra, solvente e padrões de estimulantes. Após o desenvolvimento e validação do método, as análises foram aplicadas em amostras de suplementos nutricionais obtidos em lojas especializadas em suplementos, de diversas partes do estado de São Paulo (n=125). Das 125 amostras de suplemento nutricional analisadas, 38 delas (30%) apresentaram resultado positivo para alguma das substâncias de interesse, dentre elas, sibutramina, cafeína e efedrina mediante a metodologia escolhida. As amostras positivas foram posteriormente analisadas qualitativamente por LC-MS/MS, no propósito de confirmar o resultado positivo obtido. A técnica analítica empregada proporciona seletividade, linearidade, precisão, exatidão, recuperação e limites em conformidade ao objetivo que foram destinadas. Os métodos de preparo de amostra desenvolvidos e validados demonstraram ser simples, práticos, eficientes e diferenciados pelo baixo uso de amostra e solvente

The fraudulent addition of active pharmaceutical compounds in nutritional supplements is, indeed, a worldwide problem. Often, it can be found several advertisements on various supplement packaging assuring weight loss, increased intellectual and/or physical capacity and sexual stimulation. These products may have been 'spiked' with synthetic drugs containing formulations which are apparently harmless to users. In Brazil, the availability of data about adulteration of nutritional supplements is scarce. In the present work, an analytical method using gas chromatography coupled with a nitrogen-phosphorus detector (GC-NPD) was developed and applied for the detection, identification and quantification of undeclared stimulants and/or anorectic agents in food supplement labels, such as: caffeine, fenproporex, amfepramone, fenfluramine, sibutramine, phentermine, ephedrine, phenylpropanolamine, pseudoephedrine e 4- metilhexan -2- amine. The extraction/solubilization with methanol presented satisfactory results, emphasizing the use of low amount of sample, solvent and standards of analytes. After the development and validation, the method was applied in samples of nutritional supplements obtained from specialty stores in various parts of the state of São Paulo (n = 125). From the 125 nutritional supplement samples analyzed, 38 of them (30%) presented positive results for some of the substances of interest, among them sibutramine, caffeine and ephedrine according to the chosen methodology. The positive samples were subsequently analyzed qualitatively by LCMS / MS, in order to confirm the positive result obtained. The analytical technique employed provides selectivity, linearity, precision, accuracy, recovery and limits in accordance with the intended purpose. The sample preparation methods developed and validated to be simple, practical, efficient and differentiated by the low sample and solvent usage

Validação de metodologia analítica para determinação de taninos pelo método de difusão radial / Validation of the analytical methodology for the determination of tannins by radial diffusion method

A metodologia para doseamento de taninos através de difusão radial desenvolvido por Hagerman (1987) vem sendo utilizada em laboratórios de fitoterápicos devido, principalmente, à sua simplicidade de execução, rapidez e baixo custo; contudo, não há relato na literatura sobre a submissão desta metodologia a um estudo de validação. Baseando-se neste fato, o presente estudo visou validar a metodologia de Difusão Radial para o doseamento de taninos. Todos os parâmetros obrigatórios exigidos pela ANVISA foram avaliados. O método foi considerado linear e com alta sensibilidade de quantificação (27,72 µg/poço). Mostrou-se também robusto e com recuperação aceitável (85,96%). Os resultados obtidos para repetibilidade (intra-corrida) e precisão intermediária (inter-corridas), certificaram a precisão do método, obtendo-se valores entre 1,89 e 7,03%. Para a exatidão, valores entre 100,47 e 105,26% foram obtidos, os quais se encontram dentro dos limites preconizados pela ANVISA. O método foi considerado preciso, exato e reprodutível, além de ser de fácil execução e de baixo custo.

The method to determine tannins by radial diffusion developed by Hagerman (1987) has been explicitly used in herbal laboratories mainly because of its simplicity to implement, speed and low cost; yet, there are no reports in the literature on the submission of this method to a validation study. Based on this fact, this study sought to validate the method of Radial Diffusion in the tannin determination. All mandatory parameters required by ANVISA were evaluated. The method was considered linear and with high sensitivity quantitation (27.72 µg / well). It also showed robust and acceptable recovery (85.96%). The results obtained for repeatability (within-run) and intermediate precision (inter-run) certified the accuracy of the method, obtaining values between 1.89 and 7.03%. For accuracy, values between 100.47 and 105.26% were obtained, which is within the limits recommended by ANVISA. Thus, the method was considered as precise, accurate and reproducible, and is easy to perform and inexpensive.