近年来,随着水产养殖规模日益扩大,养殖过程中过度依赖药物、滥用化学药品及抗生素的问题越来越严重,导致水产品质量和食用安全问题日渐突出,严重威胁水产养殖业的可持续健康发展.因此必须建立和健全养殖过程安全控制技术,从养殖源头上消除水产品食用安全问题.本研究总结了水产品中药物残留检测技术的研究进展.

本研究制备了一种用于稻谷中镉检测的时间分辨荧光免疫层析定量检测卡.采用铕纳米微球作为荧光探针,标记抗Cd-EDTA的单克隆抗体,基于竞争抑制原理建立免疫层析定量方法并对其进行方法学研究.结果 表明:定量检测卡的灵敏度为0.088 ng/mL,标准曲线线性范围为0.1～5.0 ng/mL,检出限和定量限分别为0.026 mg/kg和0.064 mg/kg,对有证标准物质的正确度在81.0％ ～91.7％,对实际样本的检测与ICP-MS检测结果一致性较高.该法重复性较好,变异系数在10％以内.结果 显示,该法灵敏度高、操作简便、成本低廉,因此,非常适合基层单位大量样本的快速初步筛查.

本实验采用抗DON杂交瘤细胞株SW-3 F4、SW-3 G12,分别制备出两种不同特性的单抗,然后与蛋白A-琼脂糖凝胶定向偶联,制备出了能同时分离净化DON、3-Ac-DON和15-Ac-DON的脱氧雪腐镰刀菌烯醇(DON)及其乙酰化衍生物的三合一免疫亲和柱(IAC),并对其性能进行了评价.结果表明:抗体1和抗体2均为IgG1亚型,亲和常数分别为3.0×108 L/mol和2.0×109L/mol,与DON、3-Ac-DON和15-Ac-DON的交叉反应率分别为100％、0.1％、453％和100％、101％、6％.制备的IAC柱,单标过柱的平均柱容量DON、3-Ac-DON和15-Ac-DON依次为3 243 ng/0.25 mL、1 285 ng/0.25 mL和1 527 ng/0.25 mL,非特异性吸附率小于2.3％,3种毒素在小麦基质中平均加标回收率均大于89％.本实验制备的IAC柱容量大、回收率高,适于DON及其乙酰化衍生物的纯化分离.

将初步纯化的抗黄曲霉毒素B1单克隆抗体,定向偶联于蛋白A-琼脂糖凝胶,制备成免疫亲和层析柱.结果显示,辛酸-饱和硫酸铵纯化后的单抗质量浓度为17.5 mg/mL,纯度为45.1％,抗体亚型为IgG1.取纯化后抗体48μL与1mL蛋白A凝胶偶联,偶联率达98.1％,高效液相色谱测定其平均相对柱容量为105 ng/0.125 mL凝胶、柱空白为0,不同样品(玉米粉、面粉、花生油、酱油、醋等)平均加标回收率在80％以上.该亲和柱在2～8℃保存12个月后,相对柱容量仍可达72 ng/0.125 mL凝胶.该法制备的亲和柱相对柱容量大,稳定性好,可用于样品中黄曲霉毒素B1分离净化的要求.

Mycotoxins are fungal secondary metabolites,which can cause food and feed contamination.Its analyze methods are maturing.This review introduced preparation methods and principle of immunoaffinity column clean-up techniques,which had been widely used in analysis of mycotoxins.Moreover,applications of immunoaffinity columns in analysis of normal mycotoxins had been summarized.

Methods of detection and determination of ergot alkaloids in grain and its products were reviewed.They included traditional detection methods:such as colorimetry and thin layer chromatography,instrumental procedures:such as liquid chromatography with fluorescence,mass spectrometric(MS) or MS/MS detection,capillary zone electrophoresis,immunological detection methods:such as radioimmunoassay,enzyme-linked immunosorbent assay(ELISA). The research progress of ergot alkaloids analysis was summarized.

The hapten ergonovine was synthesized by Succinic anhydride method.And the hapten was conjugated with carrier proteins with carbodiimide(EDC) method.The coupling ratio of the artificial antigen was determined by Ultraviolet spectrogram(UV) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).The coupling ratio was 12∶1 and 17∶1,respectively.In addition,infrared spectroscopy(IR) identification results further show that the success of artificial antigen synthesis.The antibody with high titer was produced after immuning to New Zealand rabbits.The results showed that the artificial antigen of ergonovine had been synthesized successfully.

The paper aimed to improve the extraction method of AFB1 in maize by enzyme linked immunosorbent assay(ELISA) so as to minimize the error caused by the affecting factors of the samples.The results showed that the sample detection result was accurate,and the recovery rate was above 83% when the standard work solution for ELISA quantitative detection was equivalent to the components of a sample extraction solution.