响应面法优化斑马鱼中微塑料分离、提取的工艺

Process optimization on extraction and isolation of microplastics in zebrafish using response surface methodology

中文关键词: 斑马鱼  微塑料  消化液  响应面分析  工艺优化

英文关键词: Zebrafish  Microplastic  Digestive compounds  Response surface analysis  Process optimization

中文摘要:

      微塑料广泛存在于海洋中，是全球十大新兴环境问题之一。微塑料可以被鱼类、贝类和底栖动物等海洋生物摄入，危害海洋生命。为了更好地从海洋生物中分离和检测微塑料，本研究优化了一种新方案。以斑马鱼(Danio rerio)为原料，对其中的微塑料进行分离提取。首先，通过消化率、回收率、拉曼光谱分析和扫描电镜等方法对KOH、H2O2、HNO3和胰蛋白酶4种消化液进行选择，选定KOH溶液为最适消化液。然后通过单因素实验和Box-Behnken响应面实验设计优化KOH溶液为消化液的最佳消解条件。研究表明，消解最适条件为浓度4%、温度47℃、时间20 h，在此条件下的消化率为97.38%。KOH消化液显著提高了消解效率，为微塑料的进一步检测分析提供了技术支持。

英文摘要:

      Recently, with the increase of global plastic production, microplastic accumulation in aquatic environments is increasing. This seriously endangers the living environment and aquatic organism health. Furthermore, it endangers human health through the food chain. Microplastic detection in aquatic products is fundamental to understanding microplastics. Using digestive compounds to extract and separate microplastic from aquatic organisms is an effective means. It is particularly important to choose the correct digestive compounds and conditions. In the present experiment, zebrafish was used as an aquatic model, and polystyrene was used as the raw material. Zebrafish and microplastics were mixed in proportion. The microplastics were separated and extracted using different digestive compounds. Initially, 10% KOH, 30% H2O2, 69% HNO3, and trypsin were selected based on digestibility, recovery rate, Raman spectroscopic analysis, and scanning electron microscopy. The results showed that 30% H2O2 has the lowest digestibility and can change its chemical structure. Trypsin has the lowest recovery rate. Scanning electron microscopy and stereoscopic microscopy showed that 69% HNO3 changed the microplastics color and corroded the surface. Therefore, 10% KOH solution was selected as the optimal digestive compound. Then, the optimal digestion conditions of KOH solution were determined using single factor and Box-Behnken design tests. The results showed that the optimum digestion conditions were: concentration – 4%, temperature – 47℃, time – 20 h, with 97.38% digestibility. The digestive compounds and conditions screened in this study could significantly improve zebrafish digestion efficiency. The results provided a new method for the digestion of aquatic products and further detection and analysis of microplastics in aquatic products.