应用固化酶检测黄瓜中氨基甲酸酯类农药

通过向固定液中加入适量明胶构成BSA-GA-gelatin系统,从而对传统交联固化酶法进行了改进,不仅提高了固化乙酰胆碱酯酶的活性而且经固定化的乙酰胆碱酯酶具有较好的稳定性,在常温下保存45d后仍未见酶活性有明显下降.在研究过程中分别对BSA、GA、明胶的最佳浓度进行了考察,最佳使用浓度分别为5%、0.5%、和0.1%.把乙酰胆碱酯酶固定到96孔酶标板上并与酶标仪联用减少了因目视颜色变化带来的误差,提高了检测结果的准确性.针对以往农药残留速测法普遍采用的表面冲洗法进行农药残留提取的方法进行了改进,使提取效率大大提高,对大多数残留农药的提取效率可达到80%以上而且方法快速、简便.该方法用来检测氨基甲酸酯类农药具有简便、快速、准确、灵敏度较高的特点,对黄瓜中5种氨基甲酸酯类农药的检测灵敏度在0.1～0.2 mg/kg范围内.

Detection Carbamate Pesticides Residue in Cucumber by Immobilized Acetylcholinesterase Enzyme

A novel method was described for detection carbamate pesticides residue in cucumber by immobilizing acetylcholinesterase enzyme in a micro-screen plate with 96 wells based on GA-BSA-gelatin gels. The concentrations of BSA, GA and gelatin in enzyme immobilization solution were optimized and the best concentrations were 5%, 0.5% and 0.1%, respectively. To analyze the pesticides residue 5g cucumber sample was homogenized with 10 mL acetone then an aliquot of 5mL extract was collected in a 10 mL test tube with a cap, into which 2g sodium chloride and 2mL dichloromethane were added in sequence. After shaking, 1 mL of the supernatant aliquot was evaporated by a blower in a small beaker and dissolved by 20% methanol-water solution then 50 microL was piped to a sample well. After incubation 10 minutes the absorbance was detected. The proposed method offered a rapid, simple and inexpensive means to in screen of batch samples. The minimum detection limit of this method was in a range of 0.1-0.2 mg/kg for cucumber samples.