该研究建立了一种基于双频超声辅助酶提取水产品中重金属的前处理方法,该法利用蛋白酶的水解作用以及超声探头的空化效应和超声水浴的控温功能,在6 min内即可实现对水产品及其制品中重金属Cd(Ⅱ)的快速提取。通过单因素实验考察了双频超声辅助酶提取方法中超声模式,缓冲溶液种类,酶用量,体系pH值以及料液比对重金属Cd(Ⅱ)提取效率的影响,得到最佳提取条件,获得水产品及其制品中重金属Cd(Ⅱ)的最优前处理方法。结果表明,牡蛎样品中镉的最佳提取条件为:在超声水浴与超声探头相结合的超声模式下,以Tris作为缓冲溶液,0.7 g风味蛋白酶与0.3 g胰蛋白酶为复配酶配方,pH值为7,料液比为1∶5(g∶mL),在此条件下,牡蛎中重金属Cd(Ⅱ)的提取率可达104.76%。在上述最佳提取条件的基础上,只需改变超声探头的功率,也可用此方法实现对蟹肉、带鱼、鱿鱼丝、皮皮虾和虾滑样品中重金属Cd(Ⅱ)的提取,其提取率可达75%～110%,表明该前处理方法可实现对水产品及其制品中重金属Cd(Ⅱ)的快速、高效提取。该前处理方法所用缓冲溶液为中性体系,环境友好,且与重金属离子速测方法具有良好的兼容性,在多种水产品及其制品的重金属Cd(Ⅱ)前处理及快速检测中具有广阔的应用前景。
A pre-treatment method based on dual-frequency ultrasonic-assisted enzyme extraction of heavy metal from aquatic products was establish in this study, using protease hydrolysis, cavitation effect of ultrasonic probe and temperature control of ultrasonic water bath. The rapid extraction of heavy metal Cd(Ⅱ) from aquatic products and its products could be achieved within 6 min. The effects of ultrasonic mode, buffer solution type, enzyme dosage, system pH value and solid-liquid ratio on the extraction efficiency were investigated by single factor experiment to obtain the optimal extraction conditions and pre-treatment method. The optimal extraction conditions of cadmium from oyster samples were as follows: ultrasonic mode combining ultrasonic water bath (UB) and ultrasonic probe (UP), Tris as buffer solution, 0.7 g flavoring protease and 0.3 g trypsin as complex enzyme formula, pH 7, solid-liquid ratio 1∶5(g∶mL). Under these conditions, the extraction rate of heavy metal Cd(Ⅱ) from oyster reached 104.76%. Based on the optimal extraction conditions above, the method could be applied in other matrix such as crab meat, hairtail, squid, shrimp and shrimp slide only by changing the power of ultrasonic probe, and the extraction rate of Cd(Ⅱ) reached 75%-110% in those aquatic samples. The pretreatment method could realize rapid and efficient extraction of heavy metal Cd(Ⅱ) from aquatic products and their products. Compared with the strong acid digestion method specified in the national standard, the buffer solution used in this pretreatment was neutral, environmentally friendly, and had good compatibility with the rapid determination of heavy metal ions. It has a broad application prospect in the pretreatment and rapid detection of heavy metal Cd(Ⅱ) in various aquatic products and their products.
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