Mass Spectrometry Determination of Fining-Related Allergen Proteins in Chilean Wines
Casein (milk powder) and ovalbumin (egg white powder) are often used as wine fining agents with the objective to promote interactions with undesirable compounds, e.g., polymeric phenols. This kind of proteins may trigger allergic reactions in susceptible individuals; therefore, their occurrence in wines could become a human health risk, moreover when their presence is not reported. The objective of this work was to establish a mass spectrometry method to determine casein and ovalbumin in Chilean wines. To the best of our knowledge, the present work reports for the first time the presence of these proteins in Chilean wines. Proteins were extracted combining the use of ultrafiltration membranes and protein precipitation with organic solvents. Thereafter, proteins were digested with trypsin during 7 h with a 1:10 enzyme protein ratio. Chromatographic separation was carried out on a Kinetex XB C18 (100 × 4.6 mm, 2.6 μm) column set at 35 °C using an acidified (0.1% v/v formic acid) mobile phase composed of ultrapure water (A) and acetonitrile (B). For a highly selective evaluation, quantification was carried out by mass spectrometry applying multiple reaction monitoring (MRM). Validation was established according to the International Conference Harmonization guidelines. Calibration data (n = 6) fitted a linear regression model with determination coefficients (R2) higher than 0.99. Repeatability (n = 6) and intermediate precision (n = 3) showed RSD values ≤ 1.36% and ≤ 1.53%, respectively. Recovery (n = 3) at three levels ranged from 86.40 to 106.98%. Detection and quantification limits ranged from 4.70 to 8.50 μg L−1 and 10 to 20 μg L−1, respectively. Twenty Chilean wine samples were analyzed; 18 samples showed quantifiable levels, from which three samples presented a total casein and ovalbumin concentration (0.24–0.26 mg L−1) closer or higher to the European limit for mandatory labeling (0.25 mg L−1).
KeywordsCasein Ovalbumin Mass spectrometry Wines, fining agents
This work is part of Jessy Pavón Pérez thesis to obtain the degree of Doctor in Science and Analytical Technology from the University of Concepcion, Chile. Authors want to thank to the National Commission of Scientific and Technological Research (CONICYT) of the Chilean Government for the doctoral scholarship granted.
This study was funded by the National Fund for Scientific and Technological Development (FONDECYT) project no. 1171857 and by the Fund for Scientific and Technological Equipment (FONDEQUIP) project no. 130209 and University of Concepcion.
Compliance with Ethical Standards
Conflict of Interest
Jessy Pavon declares that she has no conflict of interest. Karem Henríquez-Aedo declares that she has no conflict of interest. Mario Aranda declares that he has no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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