Chemometric optimization of trypsin digestion method applying infrared, microwave and ultrasound energies for determinat
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ORIGINAL ARTICLE
Chemometric optimization of trypsin digestion method applying infrared, microwave and ultrasound energies for determination of caseins and ovalbumin in wines Jessy Pavo´n-Pe´rez1 • Karem Henriquez-Aedo2,3 • Ricardo Salazar4 Miguel Herrero5 • Mario Aranda6
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Revised: 20 August 2020 / Accepted: 12 September 2020 Ó Association of Food Scientists & Technologists (India) 2020
Abstract Caseins and ovalbumin are frequently used as wine fining agents to remove undesirable compounds like polymeric phenols. Their presence in wines is a subject of concern because may cause adverse effects on susceptible consumers, especially when their presence is not labeled. A key step for its determination is trypsin digestion, which is considered the bottleneck of bottom-up approach workflow because usually requires several hours. To reduce this time, the objective of this work was to carry out a chemometric optimization of trypsin digestion method applying infrared,
& Mario Aranda [email protected]; [email protected]
microwave and ultrasound energies to determine caseins and ovalbumin in wines. The conditions of each accelerated digestion method were optimized using a Response Surface Methodology based on central composite design. The parameters optimized were digestion time and trypsin: protein ratio. The response variable evaluated was digestion yield, which was determined through the peak area of each protein transition determined by liquid chromatography-mass spectrometry. The most effective technique was microwave followed by ultrasound and infrared. Since optimal values of microwave and ultrasound-assisted digestion were the same, the later was chosen considering sample preparation and cost. Applying the proposed approach, a reduction of ca. 140 and 240-fold on digestion time was achieved compared with optimized and non-optimized conventional methods, respectively. With this workflow, both proteins were digested in a single 3 min process allowing its detection by liquid chromatographymass spectrometry at lg L-1 level, which is ca. 60 times lower than the current limit of 0.25 mg L-1.
1
Departamento de Ciencia Y Tecnologı´a de Los Alimentos, Facultad de Farmacia, Universidad de Concepcio´n. Barrio Universitario S/N, Concepcio´n, Chile
2
Laboratorio de Biotecnologı´a Y Gene´tica de Alimentos, Departamento de Ciencia Y Tecnologı´a de Los Alimentos, Facultad de Farmacia, Universidad de Concepcio´n. Barrio Universitario S/N, Concepcio´n, Chile
3
Laboratorio de Alimentos Funcionales, Centro de Biotecnologı´a, Universidad de Concepcio´n. Barrio Universitario S/N, Concepcio´n, Chile
Keywords Fining agents Mass spectrometry Central composite design Response surface methodology
4
Laboratorio de Electroquı´mica del Medio Ambiente, LEQMA, Departamento de Quı´mica de Los Materiales, Universidad de Santiago de Chile, USACH, Casilla 40, C.P. 33, Av. Libertador Bernardo O’Higgins, 3363, Estacio´n Central, Santiago, Chile
Introduction
5
Instituto de Investigacio´n en Ciencias de La Alimentacio´n CIAL (CSIC-UA
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