高压脉冲电场处理室内多物理场对微生物灭活作用的研究
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摘要
液态食品的安全问题历来就受到人们的广泛关注,无论新鲜果蔬汁还是奶制品,甚至是酒精饮品,其产品的品质和安全很大程度上取决于杀菌工序是否可靠。传统的热力杀菌方法在导致微生物失活的过程中,食品受热会发生物理和化学性质的变化,使其色、香、味、组织结构发生改变,营养价值下降,产生各种反应,甚至检测出有毒物质,不仅降低了产品的新鲜度,还严重影响了食品的质量。高压脉冲电场(Pulsed electric field,PEF)技术作为最近几年研究最为热门的非热加工技术之一,不但具有良好的杀菌、钝酶效果,还能最大限度地保持食品的新鲜度,处理过程时间短、温度低,能量消耗少。因而,该技术以其优良的处理效果和低廉的运行费用在液态食品加工领域展示了诱人的应用前景。
本文综合利用数值模拟技术、计算流体动力学、微生物计量学、化学计量学和荧光光纤传感技术等诸多领域的知识,研究高压脉冲电场处理室结构参数和处理过程工作参数对于高压脉冲电场技术用于食品中微生物灭活作用的影响。通过建立不同结构的高压脉冲电场杀菌处理室的多物理场耦合模型,分析处理室结构参数对于内部物理场分布规律的影响；研究处理介质的流量和初始温度对于准方波脉冲波形的影响,探索微生物形态特性对于微生物跨膜电势的影响,通过建立杀菌动力学模型分析处理过程工作参数对高压脉冲电场技术处理效率的影响,并通过多物理场耦合模型和杀菌动力学模型的融合,构建基于处理室结构参数和处理过程工作参数的高压脉冲电场技术对微生物作用的定量预测模型,为优化处理室结构参数和优选处理工作参数提供理论依据,为研发具有自主知识产权的工业化大规模高压脉冲电场处理系统提供方法依据,为高压脉冲电场处理技术的推广应用提供一种新的思路,对于提高处理效率和降低生产成本具有深远的现实意义。
主要研究内容、结果和结论如下：
(1)分析了不同处理室结构对高压脉冲电场处理室内部物理场分布规律的影响。采用数值模拟方法构建了高压脉冲电场处理室的多物理场耦合仿真模型,结果表明：1)平行板型静态处理室中电场强度处处相等,温度随处理时间上升很快。2)同轴型连续式处理室中,电场分布呈现不均匀性,电极间距越小,其电场分布均匀性越好。温度随着轴向距离的增加呈现线性升高,在径向方向上,温度的波动较小。湍流动能随着液体流动方向的轴向距离的增加而减弱,随着电极间距的增加而减弱。3)共场型连续式处理室中,相对电场强度和相对能量输入变化趋势一致,与电极半径和电极间距的比值(r/d)之间存在线性关系,随着r/d值的增大而增大；共场型处理室内部的温度随着物料的流动方向逐渐升高,处理腔二的温度比处理腔一的温度高。在每个处理腔的出口位置,电极和绝缘材料的交界处的物料温度显著上升,出现温度峰值。结果表明：通过建立多物理场耦合模型,可以对高压脉冲电场处理室内部的物理量进行分析,得到电场强度、能量输入和温度升高与处理室结构参数的关系。
(2)分析了层流和湍流状态下优化的绝缘环形状对共场型连续式处理室内部各物理场分布规律的影响。对三种不同绝缘环形状(矩形内嵌式、矩形内嵌倒圆角式和圆弧形内嵌式)的共场型处理室进行仿真,结果表明：1)在层流状态下,绝缘环的内嵌式结构并不能有效地改善处理室内部的处理均匀性,反而会导致电场峰值和温度峰值的产生。2)在湍流状态下,改进后的处理室中电场分布较之原结构得到改善,峰值电场强度减弱,处理室径向的流速相比于层流状态更为均匀。湍流状态对于处理室内温度控制的作用较为明显,与原结构相比有显著提高。3)在两种状态下,相对电场强度和相对能量输入的变化趋势一致,随着内嵌深度和电极半径的比值(i/r)的增大而减小,随着电极半径和电极间距的比值(r/d)的增大而增大。4)在两种状态下,相对温升与电极间距(d)、内嵌深度和电极半径的比值(i/r)及电极半径和电极间距的比值(r/d)都有关。湍流状态下的相对温升拟合度(0.921 (3)分析了处理介质的流量和初始温度对于准方波脉冲波形的影响。结果表明：1)通过数值模拟方法得到处理介质的电导率和处理室截面电流值变化趋势一致,随着初始温度的升高,二者逐渐上升,而随着处理介质的流量增大,二者逐渐下降；而脉冲的下降时间随着初始温度的升高而减小,随着处理介质流量的增大而增大。2)建立了处理室等效电阻关于流量和初始温度的函数关系,无论是对单个处理腔的等效电阻还是对两个处理腔的等效并联电阻进行拟合,其决定系数R2均达到0.980以上,RMSE值均小于0.003。3)通过示波器和NI Labview SignalExpress软件采集真实波形与仿真所得结果进行比较。仿真所得的脉冲波形的下降时间与实验所得变化趋势一致,但是,脉冲波形的真实下降时间大于仿真结果。结果表明：通过数值模拟方法,可以建立准方波脉冲波形的下降时间与处理介质流量和初始温度之间的相关性。
(4)分析了不同微生物细胞的形态特性对于高压脉冲电场技术杀菌效果的影响。1)通过数值模拟方法分析了三种微生物细胞(金黄色葡萄球菌、大肠杆菌DH5α和酿酒酵母菌)在高压脉冲电场作用下的跨膜电势,比较其对高强度电场的抵抗能力。结果表明,在同一电场强度下,酿酒酵母菌比大肠杆菌DH5a拥有更大的跨膜电势,金黄色葡萄球菌的跨膜电势最小。2)通过高压脉冲电场技术对三种微生物杀菌实验表明,酿酒酵母菌对高压脉冲电场处理过程的抵抗力最弱,大肠杆菌DH5α次之,金黄色葡萄球菌最强,与跨膜电势的仿真结果一致。3)较大的细胞尺寸可以增加跨膜电势和细胞膜的电场强度,使细胞对于高压脉冲电场处理更为敏感。4)不同的细胞形态对于跨膜电势也有较大影响。杆形细胞的跨膜电势和电场强度均大于拥有相同长径和短径的椭球形细胞。5)随着细胞膜厚度增加,跨膜电势增大非常微小,但是细胞膜的电场强度下降十分明显。结果表明：通过数值模拟方法,可以对不同微生物的跨膜电势进行分析,比较不同微生物对高压脉冲电场处理过程的抵抗力。
(5)分析了高压脉冲电场技术的不同处理工作参数对微生物的致死规律。1)在一定范围的处理强度(电场强度为12-21kV/cm,处理时间为30-180μs)和初始温度(25-35℃)下,研究高压脉冲电场技术对黄酒中酿酒酵母菌的致死作用,得到最大的杀菌效果为5.5个对数级。2)从AFM成像的结果可以看出,经过处理后微生物细胞的数量减少,图像呈现模糊的边界,细胞边缘塌陷,表明高压脉冲电场技术引起了酵母菌细胞膜的破坏。3)采用Log-linear模型、Weibull模型和多元回归模型分别对实验数据进行拟合,通过另一组独立实验分别对根据模型预测得到的杀菌数量和5-D值进行验证,对于杀菌数量的预测,Weibull模型的Af和Bf值分别为1.082和1.019,多元回归模型的Af和Bf值分别为1.152和0.961。对于5-D值的预测,Weibull模型的Af和Bf值分别为1.169和0.996,Log-linear模型的Af和Bf值分别为1.773和1.773。结果表明,Weibull模型和多元回归模型都可用于预测高压脉冲电场技术对黄酒中酿酒酵母的杀菌效果。
(7)探索了将多物理场耦合模型和杀菌动力学模型相结合,构建基于处理室结构参数和处理过程工作参数的高压脉冲电场技术的杀菌效果预测模型的可行性。结果表明：将杀菌物理场和前文所述的多物理场相结合,得到杀菌效果的预测模型,所得到的结果与真实测量所得到的酵母菌的存活数量基本一致。通过所构建的杀菌效果预测模型,提出针对处理室结构参数和处理过程工作参数的优化方法。结果表明：构建基于处理室结构参数和处理过程工作参数的高压脉冲电场技术的杀菌效果预测模型是可行的。
The safety issue of liquid foods is always attracting widespread interest. The quality and safety of products, including not only fruit and vegetable juice, but also milk products and alcoholic liquor, largely depend on that if the pasteurization producer is reliable. Although the traditional thermal pasteurization method is capable to inactivating the microorganisms, the physical and chemical properties of treated food products would change. It causes the variation of color, flavor, taste and nutrients, occurring of chemical reactions, and degrading of freshness, which seriously affects the quality of the food. As an innovative nonthermal processing technology mainly for liquid and pumpable foods, pulsed electric field (PEF) technology is predominantly used for the inactivation of microorganisms and enzymes at ambient or mild temperature, thereby preserving the fresh flavor, functional properties, and integrity of heat-sensitive compounds. It presents an attractive prospect in the field of liquid food processing with excellent treatment efficient and low operating costs.
Utilizing the knowledge of different fields, such as numerical simulation, computational fluid dynamics, microbial metrology, chemometrics and fluorescent optical fiber sensing technology, this research focuses on the impacts of configuration parameters of treatment system and operating parameters of PEF processing on the inactivation of pathogenic and spoilage microorganisms. The multiphysics coupled models with different geometries were developed for analyzing the influence of chamber configuration on the physical environment in the treatment chamber. The impact of flow rate and initial temperature of treated medium was studied to calculate the fall time of square pulses. The relationship between microbial morphological characteristics and PEF treatment efficiency was also analyzed. The kinetic models of microbial inactivation were established base on PEF experiments to study the effects of operating parameters on the microorganisms. In order to describe the microbial reduction based on the distribution discipline of various physical fields and processing operation parameters, the quantitative prediction model is constructed through a combination of both two models mentioned above:the multiphysics coupled model and the kinetic model for microbial inactivation. This study is to prove the feasibility of optimizing the configuration of treatment chamber and preferring operating parameters of PEF processing based on numerical simulation method. It may provide new ideas and theoretical basis for the wide application of PEF technology in the liquid food processing. It has profound effects on improving processing efficiency and reducing production costs.
The main results and conclusions were listed as follows:
(1) The influence of different configurations on the distribution of physical field in the treatment chamber was analyzed. Based on the numerical simulation method, multiphysics coupled models for each kind of chamber were developed. The results indicated that:1) In the parallel plate static treatment chambers, the electric field distribution was uniform, however, the temperature of treated medium obviously raised as the increasing of treatment time.2) In the coaxial continuous treatment chambers, the distribution of electric field was not homogeneous. The uniformity of electric field depended on the radial difference between inner and outer electrodes. With the axial distance increasing, the temperature in the coaxial chamber increased linearly. However, the temperature fluctuation was weak in the radial direction. Besides, the turbulent kinetic energy in the coaxial chamber became lower with the increasing of electrode distance.3) In the co-field continuous chambers, the axial distribution of electric field strength along the centerline was more uniform than that at0.1mm distance from the wall of the insulator. The high field intensity peaks were observed in the vicinity of edge interfaces of the electrodes and insulator. Both the relative electric field strength and the relative specific energy input linearly increase with increasing radius to gap ratio(r/d). The temperature increased with the flow direction of treated medium, which in the first treatment zone was higher than the second treatment zone. At each outlet of treatment zone, the temperature increased significantly in the vicinity of edge interfaces of the electrodes and insulator, where the peak temperature was observed. The results indicate that based on the multiphysics coupled models, the physical variables in the PEF treatment chambers could be analyzed, achieving the relationship between geometric parameters of electrodes and electric field, energy input, as well as temperature.
(2) The influence of insulator shape on the on the distribution of physical fields in the co-field treatment chamber was analyzed under laminar and turbulent flow conditions. Three geometries of the insulator configuration of the optimized co-field treatment chambers were modeled:rectangular, chamfer edge, and circular arc. The results indicated that:1) Under laminar flow condition, the intensity distribution of physical fields was not improved in the optimized co-field chambers, on the contrary, both electric field and temperature peak values were higher than those in the original configuration.2) Under turbulent flow condition, the distribution of electric field strength was improved in all three optimized treatment chambers. The electric field peaks still existed at the same location, however, the strength was significantly weakened. Compared with laminar flow, the flow rate in the chamber was more homogeneous, the difference of which along the centerline and at0.1mm distance from the wall of the insulator was reduced significantly. The effect of turbulent kinetic energy on the temperature controlling was obvious, and the temperature in the treatment chambers (B) and (C) was much lower than original configuration.3) Under both laminar and turbulent flow conditions, both the relative electric field strengthand the relative specific energy input depended on the ratio of inset depth to electrode radius (i/r), as well as the ratio of electrode radius to gap distance(r/d).4) Under both laminar and turbulent flow conditions, the relative temperature increasing was a function of gap distance (d), the ratio of inset depth to electrode radius (i/r), and the ratio of electrode radius to gap distance (r/d). The goodness of fitting of relative temperature increasing under turbulent condition (0.921 (3) The influence of flow rate and initial temperature of treated medium on the waveform of square pulse was analyzed. The results indicated that:1) Based on the multiphysics model for co-field treatment chamber, the impacts of flow rate and initial temperature on the electrical conductivity of treated medium, the current through the cross-section of efficient treatment area, and the equivalent parallel resistance of treatment chamber were studied. The simulated results indicated that the variation tendency of electrical conductivity of treated medium and current through the cross-section were similar, both of which increased gradually with the increasing of initial temperature, but decreased as the fluid flow rate increased. On the contrary, the fall time of pulse was directly proportional to flow rate but inversely proportional to initial temperature.2) The relationship between equivalent resistance and two tested operating parameters was established. The results indicated that the fitting goodness of the model was available whether for single treatment zone or for parallel-connected chambers, and the determination coefficients were higher than0.980, as well as the RMSE values were lower than0.003.3) The real waveform was acquisited using oscilloscope and NI SignalExpress software, and the fall time of square pulse was calculated and compared to the simulated results. The variation trend of experimental fall time was consistent with simulated results, however, the experimental values were slightly larger than the calculated values.4) The practical significance of shorten fall time of pulse was proposed, which was benefit to controlling the temperature increasing and reducing the energy consumption. The results indicate the fall time of square pulse is related to the flow rate and initial temperature of treated medium.
(4) The relationship between microbial morphological characteristics and PEF treatment efficiency was also analyzed.1) The influence of cell size and shape as well as the membrane thickness on the transmembrane potential of different microorganisms such as Staphyloccocus aureus, Escherichia coli DH5a and Saccharomyces cerevisiae were studied by means of numerical simulation method. The results indicated that with the same intensity of electric field, S. cerevisiae presented a largest transmembrane potential, E. coli DH5a the second, S. aureus the third.2) The PEF resistance of three strains of microorganisms in grape juice was investigated by applying treatments ranging from12to24kV/cm and from30to180μs at an initial treatment temperature of30℃. In agreement with the simulation results of transmembrane potential, S. cerevisiae exhibited the least resistance to PEF treatments, and S.aureus presented least sensitivity to PEF.3) The larger cell size could increase the transmembrane potential and induced electric field strength in the cell, hence led to more sensitivity to PEF treatment.4) With the same major-axis and minor-axis diameters, the rod cells had higher transmembrane potential and induced field strength than elliptical cells.5) As the membrane thickness increased, the transmembrane potential increased slightly, however, the induced electric field strength decreased obviously. The concentrated electric field in the cell membrane enhanced the sensitivity to PEF treatment, which caused more reduction of microorganisms. The results indicate that numerical simulation could be applied to calculate the transmembrane potential in different microorganisms and predict the resistance to PEF treatment.
5) The lethal effects of various PEF processing parameters on the microorganism were analyzed.1) PEF was applied at the initial temperature of25-35℃, the electric field strengths of12-21kV/cm, with respective treatment times of30-180μs to evaluate the efficiency of this technology on the inactivation of spoilage yeast S. cerevisiae commonly associated with rice wine. The results indicated that the highest inactivation value was approximately5.5-log cycles.2) The results from atomic force microscope imaging of yeast cells indicated that PEF treatment induced the destruction of cell membrane structures, which supported by the decrease of yeast number, the blurring of images, and the flattened yeast border.3) Based on fitting with log-linear model, Weibull model and polynomial regression model, the count of survival yeast was fitted. The model validation results showed that the predicted count of survival yeast calculated from Weibull model (Af=1.082and Bf=1.019) was slightly accurater than polynomial regression model (Af=1.152and Bf=0.961), and the5-D values calculated from the Weibull model (Af=1.169and Bf=0.996) provided more accuracy than those from the log-linear model (Af=1.773and Bf=1.773). The results suggest that both Weibull model and polynomial regression were more suitable for calculating the survival ratio of PEF-treated spoilage microorganisms in Chinese rice wine.
(7) The feasibility of estibalishing the quantitative prediction model through a combination of both multiphysics coupled model and kinetic model was demonstrated. In order to describe the microbial reduction based on the distribution discipline of various physical fields and processing operation parameters, the quantitative prediction model is constructed through a combination of both two models mentioned above:the multi-physics coupled model and the kinetic model for microbial inactivation. Selecting S. cerevisiae in Chinese rice wine as research subject, the kinetic model suitable for model conbining was developed based on polynomial regression method, and was transferred to the partial differential equation about the time variation. Comparison of the simulated and measured count of survival yeast showed that implementation was successful in predicting the treatment efficiency. It could be concluded that coupling the inactivation field to the three physical fields (electric field, thermal field, and fluid flow field) using a finite element method was feasible.
本文综合利用数值模拟技术、计算流体动力学、微生物计量学、化学计量学和荧光光纤传感技术等诸多领域的知识,研究高压脉冲电场处理室结构参数和处理过程工作参数对于高压脉冲电场技术用于食品中微生物灭活作用的影响。通过建立不同结构的高压脉冲电场杀菌处理室的多物理场耦合模型,分析处理室结构参数对于内部物理场分布规律的影响；研究处理介质的流量和初始温度对于准方波脉冲波形的影响,探索微生物形态特性对于微生物跨膜电势的影响,通过建立杀菌动力学模型分析处理过程工作参数对高压脉冲电场技术处理效率的影响,并通过多物理场耦合模型和杀菌动力学模型的融合,构建基于处理室结构参数和处理过程工作参数的高压脉冲电场技术对微生物作用的定量预测模型,为优化处理室结构参数和优选处理工作参数提供理论依据,为研发具有自主知识产权的工业化大规模高压脉冲电场处理系统提供方法依据,为高压脉冲电场处理技术的推广应用提供一种新的思路,对于提高处理效率和降低生产成本具有深远的现实意义。
主要研究内容、结果和结论如下：
(1)分析了不同处理室结构对高压脉冲电场处理室内部物理场分布规律的影响。采用数值模拟方法构建了高压脉冲电场处理室的多物理场耦合仿真模型,结果表明：1)平行板型静态处理室中电场强度处处相等,温度随处理时间上升很快。2)同轴型连续式处理室中,电场分布呈现不均匀性,电极间距越小,其电场分布均匀性越好。温度随着轴向距离的增加呈现线性升高,在径向方向上,温度的波动较小。湍流动能随着液体流动方向的轴向距离的增加而减弱,随着电极间距的增加而减弱。3)共场型连续式处理室中,相对电场强度和相对能量输入变化趋势一致,与电极半径和电极间距的比值(r/d)之间存在线性关系,随着r/d值的增大而增大；共场型处理室内部的温度随着物料的流动方向逐渐升高,处理腔二的温度比处理腔一的温度高。在每个处理腔的出口位置,电极和绝缘材料的交界处的物料温度显著上升,出现温度峰值。结果表明：通过建立多物理场耦合模型,可以对高压脉冲电场处理室内部的物理量进行分析,得到电场强度、能量输入和温度升高与处理室结构参数的关系。
(2)分析了层流和湍流状态下优化的绝缘环形状对共场型连续式处理室内部各物理场分布规律的影响。对三种不同绝缘环形状(矩形内嵌式、矩形内嵌倒圆角式和圆弧形内嵌式)的共场型处理室进行仿真,结果表明：1)在层流状态下,绝缘环的内嵌式结构并不能有效地改善处理室内部的处理均匀性,反而会导致电场峰值和温度峰值的产生。2)在湍流状态下,改进后的处理室中电场分布较之原结构得到改善,峰值电场强度减弱,处理室径向的流速相比于层流状态更为均匀。湍流状态对于处理室内温度控制的作用较为明显,与原结构相比有显著提高。3)在两种状态下,相对电场强度和相对能量输入的变化趋势一致,随着内嵌深度和电极半径的比值(i/r)的增大而减小,随着电极半径和电极间距的比值(r/d)的增大而增大。4)在两种状态下,相对温升与电极间距(d)、内嵌深度和电极半径的比值(i/r)及电极半径和电极间距的比值(r/d)都有关。湍流状态下的相对温升拟合度(0.921
(4)分析了不同微生物细胞的形态特性对于高压脉冲电场技术杀菌效果的影响。1)通过数值模拟方法分析了三种微生物细胞(金黄色葡萄球菌、大肠杆菌DH5α和酿酒酵母菌)在高压脉冲电场作用下的跨膜电势,比较其对高强度电场的抵抗能力。结果表明,在同一电场强度下,酿酒酵母菌比大肠杆菌DH5a拥有更大的跨膜电势,金黄色葡萄球菌的跨膜电势最小。2)通过高压脉冲电场技术对三种微生物杀菌实验表明,酿酒酵母菌对高压脉冲电场处理过程的抵抗力最弱,大肠杆菌DH5α次之,金黄色葡萄球菌最强,与跨膜电势的仿真结果一致。3)较大的细胞尺寸可以增加跨膜电势和细胞膜的电场强度,使细胞对于高压脉冲电场处理更为敏感。4)不同的细胞形态对于跨膜电势也有较大影响。杆形细胞的跨膜电势和电场强度均大于拥有相同长径和短径的椭球形细胞。5)随着细胞膜厚度增加,跨膜电势增大非常微小,但是细胞膜的电场强度下降十分明显。结果表明：通过数值模拟方法,可以对不同微生物的跨膜电势进行分析,比较不同微生物对高压脉冲电场处理过程的抵抗力。
(5)分析了高压脉冲电场技术的不同处理工作参数对微生物的致死规律。1)在一定范围的处理强度(电场强度为12-21kV/cm,处理时间为30-180μs)和初始温度(25-35℃)下,研究高压脉冲电场技术对黄酒中酿酒酵母菌的致死作用,得到最大的杀菌效果为5.5个对数级。2)从AFM成像的结果可以看出,经过处理后微生物细胞的数量减少,图像呈现模糊的边界,细胞边缘塌陷,表明高压脉冲电场技术引起了酵母菌细胞膜的破坏。3)采用Log-linear模型、Weibull模型和多元回归模型分别对实验数据进行拟合,通过另一组独立实验分别对根据模型预测得到的杀菌数量和5-D值进行验证,对于杀菌数量的预测,Weibull模型的Af和Bf值分别为1.082和1.019,多元回归模型的Af和Bf值分别为1.152和0.961。对于5-D值的预测,Weibull模型的Af和Bf值分别为1.169和0.996,Log-linear模型的Af和Bf值分别为1.773和1.773。结果表明,Weibull模型和多元回归模型都可用于预测高压脉冲电场技术对黄酒中酿酒酵母的杀菌效果。
(7)探索了将多物理场耦合模型和杀菌动力学模型相结合,构建基于处理室结构参数和处理过程工作参数的高压脉冲电场技术的杀菌效果预测模型的可行性。结果表明：将杀菌物理场和前文所述的多物理场相结合,得到杀菌效果的预测模型,所得到的结果与真实测量所得到的酵母菌的存活数量基本一致。通过所构建的杀菌效果预测模型,提出针对处理室结构参数和处理过程工作参数的优化方法。结果表明：构建基于处理室结构参数和处理过程工作参数的高压脉冲电场技术的杀菌效果预测模型是可行的。
The safety issue of liquid foods is always attracting widespread interest. The quality and safety of products, including not only fruit and vegetable juice, but also milk products and alcoholic liquor, largely depend on that if the pasteurization producer is reliable. Although the traditional thermal pasteurization method is capable to inactivating the microorganisms, the physical and chemical properties of treated food products would change. It causes the variation of color, flavor, taste and nutrients, occurring of chemical reactions, and degrading of freshness, which seriously affects the quality of the food. As an innovative nonthermal processing technology mainly for liquid and pumpable foods, pulsed electric field (PEF) technology is predominantly used for the inactivation of microorganisms and enzymes at ambient or mild temperature, thereby preserving the fresh flavor, functional properties, and integrity of heat-sensitive compounds. It presents an attractive prospect in the field of liquid food processing with excellent treatment efficient and low operating costs.
Utilizing the knowledge of different fields, such as numerical simulation, computational fluid dynamics, microbial metrology, chemometrics and fluorescent optical fiber sensing technology, this research focuses on the impacts of configuration parameters of treatment system and operating parameters of PEF processing on the inactivation of pathogenic and spoilage microorganisms. The multiphysics coupled models with different geometries were developed for analyzing the influence of chamber configuration on the physical environment in the treatment chamber. The impact of flow rate and initial temperature of treated medium was studied to calculate the fall time of square pulses. The relationship between microbial morphological characteristics and PEF treatment efficiency was also analyzed. The kinetic models of microbial inactivation were established base on PEF experiments to study the effects of operating parameters on the microorganisms. In order to describe the microbial reduction based on the distribution discipline of various physical fields and processing operation parameters, the quantitative prediction model is constructed through a combination of both two models mentioned above:the multiphysics coupled model and the kinetic model for microbial inactivation. This study is to prove the feasibility of optimizing the configuration of treatment chamber and preferring operating parameters of PEF processing based on numerical simulation method. It may provide new ideas and theoretical basis for the wide application of PEF technology in the liquid food processing. It has profound effects on improving processing efficiency and reducing production costs.
The main results and conclusions were listed as follows:
(1) The influence of different configurations on the distribution of physical field in the treatment chamber was analyzed. Based on the numerical simulation method, multiphysics coupled models for each kind of chamber were developed. The results indicated that:1) In the parallel plate static treatment chambers, the electric field distribution was uniform, however, the temperature of treated medium obviously raised as the increasing of treatment time.2) In the coaxial continuous treatment chambers, the distribution of electric field was not homogeneous. The uniformity of electric field depended on the radial difference between inner and outer electrodes. With the axial distance increasing, the temperature in the coaxial chamber increased linearly. However, the temperature fluctuation was weak in the radial direction. Besides, the turbulent kinetic energy in the coaxial chamber became lower with the increasing of electrode distance.3) In the co-field continuous chambers, the axial distribution of electric field strength along the centerline was more uniform than that at0.1mm distance from the wall of the insulator. The high field intensity peaks were observed in the vicinity of edge interfaces of the electrodes and insulator. Both the relative electric field strength and the relative specific energy input linearly increase with increasing radius to gap ratio(r/d). The temperature increased with the flow direction of treated medium, which in the first treatment zone was higher than the second treatment zone. At each outlet of treatment zone, the temperature increased significantly in the vicinity of edge interfaces of the electrodes and insulator, where the peak temperature was observed. The results indicate that based on the multiphysics coupled models, the physical variables in the PEF treatment chambers could be analyzed, achieving the relationship between geometric parameters of electrodes and electric field, energy input, as well as temperature.
(2) The influence of insulator shape on the on the distribution of physical fields in the co-field treatment chamber was analyzed under laminar and turbulent flow conditions. Three geometries of the insulator configuration of the optimized co-field treatment chambers were modeled:rectangular, chamfer edge, and circular arc. The results indicated that:1) Under laminar flow condition, the intensity distribution of physical fields was not improved in the optimized co-field chambers, on the contrary, both electric field and temperature peak values were higher than those in the original configuration.2) Under turbulent flow condition, the distribution of electric field strength was improved in all three optimized treatment chambers. The electric field peaks still existed at the same location, however, the strength was significantly weakened. Compared with laminar flow, the flow rate in the chamber was more homogeneous, the difference of which along the centerline and at0.1mm distance from the wall of the insulator was reduced significantly. The effect of turbulent kinetic energy on the temperature controlling was obvious, and the temperature in the treatment chambers (B) and (C) was much lower than original configuration.3) Under both laminar and turbulent flow conditions, both the relative electric field strengthand the relative specific energy input depended on the ratio of inset depth to electrode radius (i/r), as well as the ratio of electrode radius to gap distance(r/d).4) Under both laminar and turbulent flow conditions, the relative temperature increasing was a function of gap distance (d), the ratio of inset depth to electrode radius (i/r), and the ratio of electrode radius to gap distance (r/d). The goodness of fitting of relative temperature increasing under turbulent condition (0.921
(4) The relationship between microbial morphological characteristics and PEF treatment efficiency was also analyzed.1) The influence of cell size and shape as well as the membrane thickness on the transmembrane potential of different microorganisms such as Staphyloccocus aureus, Escherichia coli DH5a and Saccharomyces cerevisiae were studied by means of numerical simulation method. The results indicated that with the same intensity of electric field, S. cerevisiae presented a largest transmembrane potential, E. coli DH5a the second, S. aureus the third.2) The PEF resistance of three strains of microorganisms in grape juice was investigated by applying treatments ranging from12to24kV/cm and from30to180μs at an initial treatment temperature of30℃. In agreement with the simulation results of transmembrane potential, S. cerevisiae exhibited the least resistance to PEF treatments, and S.aureus presented least sensitivity to PEF.3) The larger cell size could increase the transmembrane potential and induced electric field strength in the cell, hence led to more sensitivity to PEF treatment.4) With the same major-axis and minor-axis diameters, the rod cells had higher transmembrane potential and induced field strength than elliptical cells.5) As the membrane thickness increased, the transmembrane potential increased slightly, however, the induced electric field strength decreased obviously. The concentrated electric field in the cell membrane enhanced the sensitivity to PEF treatment, which caused more reduction of microorganisms. The results indicate that numerical simulation could be applied to calculate the transmembrane potential in different microorganisms and predict the resistance to PEF treatment.
5) The lethal effects of various PEF processing parameters on the microorganism were analyzed.1) PEF was applied at the initial temperature of25-35℃, the electric field strengths of12-21kV/cm, with respective treatment times of30-180μs to evaluate the efficiency of this technology on the inactivation of spoilage yeast S. cerevisiae commonly associated with rice wine. The results indicated that the highest inactivation value was approximately5.5-log cycles.2) The results from atomic force microscope imaging of yeast cells indicated that PEF treatment induced the destruction of cell membrane structures, which supported by the decrease of yeast number, the blurring of images, and the flattened yeast border.3) Based on fitting with log-linear model, Weibull model and polynomial regression model, the count of survival yeast was fitted. The model validation results showed that the predicted count of survival yeast calculated from Weibull model (Af=1.082and Bf=1.019) was slightly accurater than polynomial regression model (Af=1.152and Bf=0.961), and the5-D values calculated from the Weibull model (Af=1.169and Bf=0.996) provided more accuracy than those from the log-linear model (Af=1.773and Bf=1.773). The results suggest that both Weibull model and polynomial regression were more suitable for calculating the survival ratio of PEF-treated spoilage microorganisms in Chinese rice wine.
(7) The feasibility of estibalishing the quantitative prediction model through a combination of both multiphysics coupled model and kinetic model was demonstrated. In order to describe the microbial reduction based on the distribution discipline of various physical fields and processing operation parameters, the quantitative prediction model is constructed through a combination of both two models mentioned above:the multi-physics coupled model and the kinetic model for microbial inactivation. Selecting S. cerevisiae in Chinese rice wine as research subject, the kinetic model suitable for model conbining was developed based on polynomial regression method, and was transferred to the partial differential equation about the time variation. Comparison of the simulated and measured count of survival yeast showed that implementation was successful in predicting the treatment efficiency. It could be concluded that coupling the inactivation field to the three physical fields (electric field, thermal field, and fluid flow field) using a finite element method was feasible.
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