RESUMEN

The combination of QuEChERS-LC-QTOFMS and matrix-matched calibration (MMC) to simultaneously determinate legislated and emerging mycotoxins in malt and beer was evaluated for the first time. The method performance was satisfactory displaying suitable linearity (R2 >0.99) and recovery (71-102%). The lowest values (in µg kg-1) of LOD (0.01) and LOQ (0.05) were found for enniatins, while the highest LOD (15) and LOQ (50) were reported for fumonisin B1. Precision and sensitivity (RSD <10%) were in accordance with the different guidelines of method validation. MMC was important to avoid inaccurate quantification of all mycotoxins due to signal enhancement or suppression. Another advantage was the enhanced throughput, requiring 1.2 min of analysis per analyte. The detection of legislated (aflatoxins, ochratoxin A, deoxynivalenol, fumonisin B1, zearalanone, T-2 and HT-2 toxin) and emerging mycotoxins (enniatins, beauvericin, moniliformin and sterigmatocystin) allowed verifying compliance with legislation and generating data to support the establishment of limits for emerging mycotoxins.

Asunto(s)

Cerveza/análisis , Análisis de los Alimentos/métodos , Contaminación de Alimentos/análisis , Hordeum/química , Micotoxinas/análisis , Espectrometría de Masas en Tándem/métodos , Calibración , Cromatografía Líquida de Alta Presión/métodos

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Acmella oleracea (L.) R. K. Jansen (Asteraceae), known as jambú in Brazil, is used in traditional medicine as analgesic and for inflammatory conditions, characterized by the presence of N-alkylamides, mainly spilanthol. This bioactive compound is responsible for the above-described pharmacological properties, including sialagogue and anesthetic. AIM OF THE STUDY: This study aimed to characterize the anti-inflammatory effects of A. oleracea leaves (AOEE-L) and flowers (AOEE-F) extracts, including an isolated alkylamide (spilanthol), using in vitro and in vivo models. The mechanism underlying this effect was also investigated. MATERIALS AND METHODS: Extracts were analyzed by HPLC-ESI-MS/MS in order to characterize the N-alkylamides content. AOEE-L, AOEE-F (25-100 µg/mL) and spilanthol (50-200 µM) were tested in vitro on VSMC after stimulation with hyperglycemic medium (25 mM glucose). Their effects over nitric oxide (NO) generation, chymase inhibition and expression, catalase (CAT), superoxide anion (SOD) radical activity were evaluated. After an acute administration of extracts (10-100 mg/mL) and spilanthol (6.2 mg/mL), the anti-inflammatory effects were evaluated by applying the formalin test in rats. Blood was collected to measure serum aminotransferases activities, NO activity, creatinine and urea. RESULTS: A number of distinct N-alkylamides were detected and quantified in AOEE-L and AOEE-F. Spilanthol was identified in both extracts and selected for experimental tests. Hyperglycemic stimulation in VSMC promoted the expression of inflammatory parameters, including chymase, NO, CAT and SOD activity and chymase expression, all of them attenuated by the presence of the extracts and spilanthol. The administration of extracts or spilanthol significantly inhibited edema formation, NO production and cell tissue infiltration in the formalin test, without causing kidney and liver toxicity. CONCLUSION: Taken together, these results provide evidence for the anti-inflammatory activity of leaves and flowers extracts of jambú associated distinctly with their chemical profile. The effects appear to be associated with the inhibition of chymase activity, suppression of the proinflammatory cytokine NO and antioxidant activities.

Asunto(s)

Antiinflamatorios/farmacología , Antioxidantes/farmacología , Asteraceae/química , Quimasas/antagonistas & inhibidores , Extractos Vegetales/farmacología , Alcamidas Poliinsaturadas/farmacología , Animales , Antiinflamatorios/uso terapéutico , Antioxidantes/química , Antioxidantes/uso terapéutico , Brasil , Línea Celular , Quimasas/metabolismo , Edema/inducido químicamente , Edema/tratamiento farmacológico , Edema/patología , Etanol/química , Flores/química , Formaldehído/toxicidad , Inflamación/inducido químicamente , Inflamación/tratamiento farmacológico , Inflamación/patología , Masculino , Medicina Tradicional , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/metabolismo , Óxido Nítrico/antagonistas & inhibidores , Óxido Nítrico/metabolismo , Extractos Vegetales/química , Extractos Vegetales/uso terapéutico , Hojas de la Planta/química , Alcamidas Poliinsaturadas/uso terapéutico , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo

RESUMEN

Primarily formed by the microbial decarboxylation of the amino acid histidine, histamine is the leading global cause of food poisoning from fish consumption worldwide. In the present work, the quality of 12 fresh and 12 frozen marketed sardines (Sardinella brasiliensis) were evaluated for histamine concentration using High-performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD), while the detection and quantification of histamine-producing bacteria were performed via quantitative Polymerase Chain Reaction (qPCR), and the microbiota composition of sardines was assessed through amplification of the 16S rRNA gene using high-throughput sequencing (HTS). According to the results obtained by HPLC-DAD, histamine concentration ranged from 226.14 to 583.87 mg kg-1. The histidine decarboxylase (hdc) genes from gram-negative bacteria (Morganella morganii, and Enterobacter aerogenes) were identified. The most abundant microorganisms present in fresh sardines belong to the genera Macrococcus spp., Acinetobacter spp., and Pseudomonas spp., while the genera Phyllobacterium spp., Pseudomonas spp., and Acinetobacter spp. were most abundant in frozen sardines.