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. 2021 May 25;10(6):1192. doi: 10.3390/foods10061192

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Optimization of the chymotryptic release of (left lane, 1) Leg1 and (right lane, 2) Leg2 from chickpea legumin. The following parameters were tested: (a) enzyme/protein ratio, (b) incubation time, (c) incubation temperature, (d) digestion stop (FA, formic acid), (e) digestion additives (DTT, dithiothreitol; IAA, iodoacetamide; DB, digestion buffer). (f) Comparison of the optimized digestion protocol with the standard protocol. The amounts of Leg1 and Leg2 were analyzed by ultrahigh performance-tandem mass spectrometry in scheduled multiple reaction monitoring mode (UHPLC–MS/MS-sMRM) and the results are shown as the index (analyte peak area/standard peak area) mean of triplicates ± standard deviation in percent compared to the standard protocol (Ctl). The levels of significance were ns, non-significant, * p < 0.05, ** p < 0.01.

Optimization of the chymotryptic release of (left lane, 1) Leg1 and (right lane, 2) Leg2 from chickpea legumin. The following parameters were tested: (a) enzyme/protein ratio, (b) incubation time, (c) incubation temperature, (d) digestion stop (FA, formic acid), (e) digestion additives (DTT, dithiothreitol; IAA, iodoacetamide; DB, digestion buffer). (f) Comparison of the optimized digestion protocol with the standard protocol. The amounts of Leg1 and Leg2 were analyzed by ultrahigh performance-tandem mass spectrometry in scheduled multiple reaction monitoring mode (UHPLC–MS/MS-sMRM) and the results are shown as the index (analyte peak area/standard peak area) mean of triplicates ± standard deviation in percent compared to the standard protocol (Ctl). The levels of significance were ns, non-significant, * p < 0.05, ** p < 0.01.