RESUMEN
RATIONALE: Polysorbate 80 (PS80) is an amphipathic, nonionic surfactant that is commonly used to stabilize proteins in biopharmaceutical formulations. PS80 undergoes oxidative and/or enzymatic degradation. However, because PS80 is a complex mixture consisting of many constituents, comprehensive evaluations of its oxidative degradation products are difficult and insufficient. METHODS: Our previously reported comprehensive liquid chromatography-tandem mass spectrometry (LC/MS/MS)-based method for PS80 effectively provides an overall profile of PS80 components under simple LC conditions. In this study, we attempted to shorten the analysis time. Furthermore, PS80 was oxidatively degraded in a solution containing histidine and iron, and the oxidative degradation products were evaluated using a modified LC/MS/MS method. In addition, enzymatically hydrolyzed PS80 samples were analyzed. RESULTS: We succeeded in shortening the analysis time from 70 to 20 min while maintaining the resolution of the PS80 components of the same selected reaction monitoring transition. Both the previously reported oxidative degradation products and the newly discovered products were successfully detected, and their composition ratios and changes over time were observed. Changes in the hydrolysates over time are shown in the analysis of the hydrolyzed PS80 samples. CONCLUSIONS: This study clearly showed the presence of changes in PS80 oxidative and/or enzymatic degradation products, including those previously unreported. These results demonstrate that a detailed profiling of PS80 degradation products can be performed using LC/MS/MS, which is less expensive and more generally adopted than high-resolution MS.
Asunto(s)
Polisorbatos , Espectrometría de Masas en Tándem , Polisorbatos/análisis , Polisorbatos/química , Polisorbatos/metabolismo , Espectrometría de Masas en Tándem/métodos , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Liquida , Estrés OxidativoRESUMEN
Abstract Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300 g L-1), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142 g L-1 after 5 days, which corresponded to 0.47 g g-1 yield and productivity of 1.1 g L-1 h-1. Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol.
Asunto(s)
Tensoactivos/metabolismo , Medios de Cultivo/metabolismo , Yarrowia/metabolismo , Eritritol/biosíntesis , Manitol/metabolismo , Polisorbatos/análisis , Polisorbatos/metabolismo , Tensoactivos/análisis , Octoxinol/análisis , Octoxinol/metabolismo , Medios de Cultivo/química , Eritritol/análisis , Manitol/análisisRESUMEN
Asunto(s)
Antibacterianos/metabolismo , Bacteriocinas/metabolismo , Medios de Cultivo/metabolismo , Lactobacillus/crecimiento & desarrollo , Lactobacillus/metabolismo , Brasil , Glucosa/metabolismo , Lactobacillus/clasificación , Pruebas de Sensibilidad Microbiana , Polisorbatos/metabolismo , Carne Roja/microbiología , Propiedades de Superficie , TemperaturaRESUMEN
El proceso de liberación de hidrocortisona de los hidrogeles con la adición del 1 por ciento y del 3 por ciento del polisorbato 20 o polisorbato 80, en la presencia de propilenglicol - 1,2 o PEG 200, tiene dos fases. Durante la primera fase las velocidades de liberación son más altas, comparando con la segunda fase. La segunda fase de liberación corresponde a la cinética de primer orden. Los periodos de semiliberación en el transcurso de esta fase oscilan entre 15,67 y 23,50 (AU)