0, 65.0, 70.0, 75.0, 80.0 and 85.0 °C. The kinetic model used to Galunisertib concentration represent the thermal inactivation of indicators POD, ALP and LPO was a first order reaction model of a two-component system (Chen and Wu, 1998, Fujikawa and Itoh, 1996, Murasaki-Aliberti

et al., 2009 and Tribess and Tadini, 2006). According to this model, there are two isoenzymes (with different thermal resistances) that contribute to the enzymic activity. Parameter α represents the fraction of the activity associated with the thermostable isoenzyme; accordingly, (1 − α) represents the contribution of the thermolabile enzyme to the activity (before thermal treatment). The thermal inactivation of the each isoenzyme follows a first order decay kinetic model, which is characterized by the parameters D-value (decimal reduction time) and z-value ABT 737 (temperature change necessary to obtain a tenfold decrease in the D-value). For an isothermal treatment, the residual activity at time θ can be obtained through Eq. (2), where the D-values of the thermostable and thermolabile isoenzymes are obtained from Eq. (3) and Eq. (4), respectively, where Tref is the reference temperature for parameters DS,ref and DL,ref. equation(2) AR=α·alog(−θDS)+(1−α)·alog(−θDL) equation(3) DS=DS,ref·10−(T−Tref)/zSDS=DS,ref·10−(T−Tref)/zS

equation(4) DL=DL,ref·10−(T−Tref)/zLDL=DL,ref·10−(T−Tref)/zL For a non-isothermal treatment, where the time-temperature history T(t) is known, the equivalent processing times for both isoenzymes at the

reference temperature were calculated through Eq. (5a) and Eq. (5b) using the corresponding temperature dependence parameter z-value (zS and zL). equation(5a) θS,ref=∫0∞alog(T(t)−TrefzS)ⅆt equation(5b) θL,ref=∫0∞alog(T(t)−TrefzL)ⅆt Combination of Eq. (2), Eq. (5a) and Eq. (5b) gives the first order reaction model of a two-component system in Eq. (6). The detailed derivation of Eq. (2) and Eq. (6) is presented by Murasaki-Aliberti et al. (2009). This kinetic much model has five parameters, as follows: α (fraction of thermostable component), DS,ref and DL,ref (D-values at reference temperature of thermostable and thermolabile components), and zS and zL (z-values of thermostable and thermolabile components). equation(6) AR=α·alog(−∫0∞alog(T(t)−TrefzS)ⅆtDS,ref)+(1−α)·alog(−∫0∞alog(T(t)−TrefzL)ⅆtDL,ref) The integrals in Eq. (6) were numerically evaluated by the trapezium method using the experimental time-temperature history data. Using an initial guess for the five model parameters, the predicted residual activity could be calculated using software Excel (Microsoft, Redmond USA). For a set of experiments, the sum of squared errors between experimental and predicted residual activities was minimized using Excel Solver to determine the optimal values of the model parameters (Matsui et al., 2008). Before using the Solver, a manual exploration of the parameters was performed to improve the initial guess and to detect large outliners.