A rapid and simple method for the determination of organic acids in proteolytic enzymes by capillary electrophoresis with indirect ultraviolet detection

The use of organic acids (e.g. acetic acid and gluconic acid) as additives during protease production is regarded as one of the simplest alternatives to increase enzyme stability and activity in many industrial processes. However, no methods have been described for the determination of organic acids...

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Detalles Bibliográficos
Autores: Pont Villanueva, Laura, Barbosa Torralbo, José, Benavente Moreno, Fernando J. (Julián)
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2020
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/170728
Acceso en línea:https://hdl.handle.net/2445/170728
Access Level:acceso abierto
Palabra clave:Electroforesi capil·lar
Àcids carboxílics
Capillary electrophoresis
Carboxylic acids
Descripción
Sumario:The use of organic acids (e.g. acetic acid and gluconic acid) as additives during protease production is regarded as one of the simplest alternatives to increase enzyme stability and activity in many industrial processes. However, no methods have been described for the determination of organic acids in proteases and their contents have not been established yet. In this work, a novel, rapid and simple method for the determination of organic acids in proteolytic enzymes by capillary electrophoresis (CE) with indirect ultraviolet (UV) detection has been developed. Under the optimized conditions, the method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ) and intra-day and inter-day repeatability. Later, a sample pretreatment based on a hydroalcoholic microextraction was carefully optimized to obtain good recovery and repeatability and determine acetic and gluconic acids in a commercial protease sample. The complete procedure was validated using the standard-addition calibration method, finding matrix effects on the studied compounds. Finally, acetic acid and gluconic acid were quantified at 80 mg/Kg (0.0080% (m/m)) and 69 mg/Kg (0.0069% (m/m)) in the protease sample, respectively.