RESUMO
Pharmaceutical and food products are fortified with pantothenic acid (PA) to address potential deficiency. Therefore, its fast, reliable, and accurate detection is of great importance to the quality control. Here, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and a gold nanoparticle-based lateral flow immunoassay (LFIA) were established for the determination of PA based on an anti-PA monoclonal antibody (mAb). The ic-ELISA displayed a limit of detection (LOD) of 32.22 ng/mL, and the linear range was 64.44-628.84 ng/mL. Average recoveries of PA in fortified samples were 88.60-110.11% when using the ic-ELISA and a good correlation between the ic-ELISA and LC-MS/MS was obtained when analyzing samples. Furthermore, the developed LFIA strip showed a calculated LOD of 71.99, 115.80, and 240.12 ng/mL in B-complex Vitamin tablets, energy drink and infant milk powder samples, respectively. All the results demonstrated that both of these immunoassays are suitable for determining PA in pharmaceutical and food products.
Assuntos
Imunoensaio/métodos , Ácido Pantotênico/análise , Preparações Farmacêuticas/química , Animais , Anticorpos Monoclonais/imunologia , Bebidas Energéticas/análise , Análise de Alimentos , Ouro/química , Limite de Detecção , Nanopartículas Metálicas/química , Ácido Pantotênico/imunologia , Comprimidos/química , Vitaminas/químicaRESUMO
INTRODUCTION: The available immunohistochemical techniques have documented restricted distribution of vitamins in the mammalian brain. The aim of the study was to develop a highly specific antiserum directed against pantothenic acid to explore the presence of this vitamin in the mammalian brain. MATERIAL AND METHODS: According to ELISA tests, the anti-pantothenic acid antiserum used showed a good affinity (10-8 M) and specificity. The antiserum was raised in rabbits. Using an indirect immunoperoxidase technique, the mapping of pantothenic acid-immunoreactive structures was carried out in the rat brain. RESULTS: Pantothenic acid-immunoreactive perikarya were exclusively found in the intermediate part of the lateral septal nucleus. These cells were generally small, round, fusiform or pyramidal and showed 2-3 long (50-100 µm) immunoreactive dendrites. Any immunoreactive axons containing pantothenic acid were detected. CONCLUSIONS: The very restricted anatomical distribution of the pantothenic acid suggests that this vitamin could be involved in some specific neurophysiological mechanisms.
Assuntos
Anticorpos/imunologia , Neurônios/imunologia , Ácido Pantotênico/imunologia , Núcleos Septais/imunologia , Animais , Anticorpos/sangue , Formação de Anticorpos , Especificidade de Anticorpos , Axônios/imunologia , Encéfalo/imunologia , Ensaio de Imunoadsorção Enzimática , Soros Imunes/imunologia , Técnicas Imunoenzimáticas , Imuno-Histoquímica , Masculino , Neurônios/citologia , Coelhos , Ratos , Ratos Wistar , Núcleos Septais/citologiaRESUMO
Immunological assays appear to be the only alternative to the microbiological method for analysis of pantothenic acid in foods and blood. In order to evaluate the influence of the linker on the immunogenicity of the hapten, we have tried to raise antisera against pantothenic acid in rabbits using different conjugates. The hapten was coupled to a carrier protein (BSA or thyroglobulin) using adipoyl dichloride (adipoyl conjugate) or bromoacetyl bromide (acetyl conjugate). Only the acetyl conjugate has induced the production of a specific antibody. With this antibody, an assay on microplate using the ELISA inhibition technique was developed to measure pantothenic acid. The use of pantothenic acid coupled to thyroglobulin with adipoyl dichloride as the capture antigen has improved the sensitivity of the ELISA. This assay was applied to food products and blood.
Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Análise de Alimentos , Ácido Pantotênico/análise , Animais , Anticorpos/isolamento & purificação , Cromatografia , Haptenos , Soros Imunes , Modelos Lineares , Ácido Pantotênico/sangue , Ácido Pantotênico/imunologia , Coelhos , Sensibilidade e Especificidade , Soroalbumina Bovina , TireoglobulinaRESUMO
In a patient with contact dermatitis, dexpanthenol was found to be the causative allergen. There was a positive reaction to dexpanthenol on patch testing. Controls did not show any positive reactions to dexpanthenol on patch testing. Additionally, an LTT was performed. After preincubation with dexpanthenol-modified microsomes, we observed an increase in lymphocyte proliferation to dexpanthenol, in comparison to dexpanthenol without microsomes, suggesting that microsomal metabolism plays a rôle in the pathogenesis of dexpanthenol sensitization, because microsomes are known to possess drug metabolizing enzymes such as cytochrome P450.
