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呋喃唑酮等硝基呋喃类药物残留是海参重要的安全危害之一,研究其胶体金免疫层析快速筛选检测技术具有重要的现实意义,但是关于海参基质对检测的干扰及消除方法目前尚未见报道。本研究发现海参基质中蛋白、多糖等组分的存在,会使呋喃唑酮代谢物(AOZ)在胶体金免疫层析过程中C线和T线的显色程度、均匀性和稳定性均明显降低,甚至出现色带断裂的情况,导致检测结果无法准确判断。针对这一问题具体研究了不同的样本前处理方法,发现将海参样本在衍生化前经80℃加热5 min,并结合0.10~0.15 mol/L盐酸处理后,可将样本中的蛋白质浓度降低到0.06 mg/m L以下,海参岩藻糖几乎消除,检测卡的主要性能基本恢复正常,对阴性海参添加0.5、1μg/kg的AOZ标准液的样本能够实现有效检测;同时省略了有机溶剂萃取、氮吹、净化等步骤,前处理操作步骤较目前AOZ检测的同类技术更为简便、快速。本研究可以为下一步研究建立海参中硝基呋喃等药残的胶体金免疫层析现场快速检测技术提供基础和依据。
Nitrofurans and other furazolidone residues are one of the important safety hazards of sea cucumber. It is of great practical significance to study the rapid screening and detection of colloidal gold immunochromatography. However, no reports have been reported on the interference and elimination of sea cucumber matrix. The present study found that the presence of proteins and polysaccharides in sea cucumber matrix will significantly reduce the color, uniformity and stability of furazolidone metabolite (AOZ) during colloidal gold immunochromatography. Even ribbon breakage, resulting in the test results can not be accurately judged. In order to solve this problem, we studied different sample pretreatment methods and found that when the sea cucumber samples were heated at 80 ℃ for 5 min and 0.10 ~ 0.15 mol / L hydrochloric acid before derivatization, the protein concentration in the samples could be reduced to 0.06 mg / m L or less, the sea cucumber fucose almost eliminated, the main performance of the test card returned to normal, the negative sea cucumber added 0.5,1μg / kg AOZ standard samples can be effectively detected; the same time omit the organic solvent extraction, Nitrogen blowing, purification and other steps, pre-treatment steps AOZ detection than the current similar technology is more simple and fast. This study can provide the basis and basis for the further research on the establishment of rapid detection technology of colloidal gold immunochromatographic drug residues such as nitrofurans in sea cucumber.