Bioscience Methods 2025, Vol.16, No.2, 70-82 http://bioscipublisher.com/index.php/bm 74 3.3 Limitations of traditional technologies Traditional separation technologies, especially wet separation, face issues such as high energy consumption and the need for large amounts of water. Additionally, research by Assatory et al. (2019) indicates that such processes typically require long drying times and the use of organic solvents, which may damage the natural structure and functionality of the separated starch and gluten. In contrast, dry techniques such as flow classification and electrostatic separation reduce energy and water consumption, but still have shortcomings in achieving high-purity separation (Assatory et al., 2019; Silventoinen et al., 2020). A major limitation of traditional techniques is the trade-off between achieving high purity in the separation and maintaining the functional properties of the components. Although dry methods can preserve the natural structure and functionality of the separated components, they often struggle to produce high-purity products (greater than 90%) due to issues like powder agglomeration and low yield. Additionally, mechanical forces involved in processes like milling may alter the structural properties of gluten, affecting its functional quality (Liu et al., 2021). 4 Innovative Separation Technology for Wheat Starch and Gluten Powder 4.1 Ultrasound assisted separation Ultrasonic assisted separation is an innovative technology that utilizes high-frequency sound waves to disrupt the composite interface between starch and gluten powder. Its principle is to generate cavitation bubbles in the medium by applying ultrasonic wave. When the foam collapses violently, micro jet flow and shock wave will be generated, which can physically destroy the matrix of starch and gluten, thus promoting the separation of starch and gluten. The separation efficiency of breaking the bond between starch and gluten powder using the mechanical action of ultrasound is higher than traditional methods (Van Der Borght et al., 2005; Peigabardoust et al., 2008). The efficiency of ultrasound assisted separation is related to the intensity of ultrasound. Higher ultrasound intensity increases the energy input of the system, enhances cavitation, and thus improves separation efficiency. However, when the ultrasound intensity is within the optimal range, excessively high intensity may lead to degradation of gluten powder quality or excessive fragmentation of starch particles, thereby negatively affecting the purity and yield of the isolate (Zalm et al., 2009; Liu et al., 2021). 4.2 Enzymatic separation Enzymatic separation utilizes selective enzymatic hydrolysis to disrupt the protein starch network. Specific enzymes, such as proteases, target peptide bonds in gluten proteins, weaken the gluten network, and promote the release of starch granules. The advantage of this method is that it can finely regulate specific components of the gluten network without affecting the starch, thereby maintaining the integrity and functionality of the separated starch (Van Der Borght et al., 2005). Optimizing the combination of enzymes can improve separation efficiency by selecting appropriate types and concentrations of enzymes, customizing the enzymatic hydrolysis process, and achieving maximum separation efficiency in the shortest possible time. At the same time, factors such as enzyme activity, temperature, pH value, and reaction time must be taken into account to ensure that the enzyme can effectively decompose the gluten network while maintaining starch quality (Van Der Borght et al., 2005; Al-Hakkak and Al-Hakkak, 2007). 4.3 Microfluidic separation technology Microfluidic separation technology is an efficient and promising method for separating starch and gluten particles. It utilizes microfluidic channels to achieve nanoscale separation, which has advantages in maintaining the functional characteristics of separation components. This technology is based on passive sorting by particle size, which helps to avoid the common clogging problem in traditional separation methods. By changing the boundary interfaces in microfluidic arrays, uniform flow patterns can be achieved, thereby improving the separation efficiency of the entire array (Inglis, 2009). This precise control at the nanoscale ensures that the functional properties of starch and gluten powder are preserved, making it an efficient technology that requires high-purity and high-quality separation components for application.
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