基于酶生物传感器法检测果蔬中毒死蜱残留的提取工艺研究
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摘要
使用自主研制的酶生物传感器型农残检测仪进行农药残留检测,通过筛选可测果蔬种类、调整优化样品处理大小、样品加标后静置时间、样品与提取液比例和振荡提取速度,提高检测用酶对农药的抑制敏感性,从而达到降低农药检出限、提高回收率的目的。主要设置的参数如下:样品处理方式分为打碎、切碎(切成1 cm×1 cm和1. 5 cm×3 cm大小);加标后静置时间为5 min、15min、30 min、60 min和90 min;料液比为1∶1、1∶2. 5、1∶5、1∶7. 5和1∶10;振荡提取速度为0 r/min、110 r/min、190 r/min和225 r/min;主要测定的农药为毒死蜱。结果发现,不同果蔬品种对固定化酶的抑制率影响小,样品大小为1 cm×1 cm、加标后静置时间为5 min、料液比为1∶1、加入提取剂后的振荡速度为190 r/min为最佳前处理方式组合。酶生物传感器农残检测仪能够满足快速检测果蔬中有机磷农药残留的需要。
The pesticide residue detection was carried out by using a self-developed enzyme biosensor type detector. The inhibition sensitivity of enzyme to pesticide was improved by screening measurable fruit and vegetable species,adjusting and optimizing sample treatment size,static time after sample addition,ratio of sample to extract solution and oscillating extraction speed,so as to achieve the purpose of reducing the detection limit of pesticides and improving the recovery rate.The main parameters set were as follows: the sample treatment was divided into breaking and shredding( cut into 1 cm ×1 cm and 1. 5 cm × 3 cm); the static time after adding standard was 5 min,15 min,30 min,60 min and 90 min; the ratio of material to liquid was 1∶ 1,1∶ 2. 5,1∶ 5,1∶ 7. 5; the oscillating extraction speed was 0 r/min,110 r/min,190 r/min and 225 r/min; and the main pesticide was chlorpyrifos. The results showed that different fruit and vegetable varieties had little effect on the inhibition rate of immobilized enzyme. The optimum combination of pretreatment methods were as follows: the sample size was 1 cm × 1 cm; the static time was 5 min after adding the standard; the ratio of material to liquid was 1 ∶ 1; the oscillating speed was 190 r/min. The self-developed enzyme biosensor for detecting agricultural residues could meet the needs of rapid detecting organophosphorus pesticide residues in fruits and vegetables.
The pesticide residue detection was carried out by using a self-developed enzyme biosensor type detector. The inhibition sensitivity of enzyme to pesticide was improved by screening measurable fruit and vegetable species,adjusting and optimizing sample treatment size,static time after sample addition,ratio of sample to extract solution and oscillating extraction speed,so as to achieve the purpose of reducing the detection limit of pesticides and improving the recovery rate.The main parameters set were as follows: the sample treatment was divided into breaking and shredding( cut into 1 cm ×1 cm and 1. 5 cm × 3 cm); the static time after adding standard was 5 min,15 min,30 min,60 min and 90 min; the ratio of material to liquid was 1∶ 1,1∶ 2. 5,1∶ 5,1∶ 7. 5; the oscillating extraction speed was 0 r/min,110 r/min,190 r/min and 225 r/min; and the main pesticide was chlorpyrifos. The results showed that different fruit and vegetable varieties had little effect on the inhibition rate of immobilized enzyme. The optimum combination of pretreatment methods were as follows: the sample size was 1 cm × 1 cm; the static time was 5 min after adding the standard; the ratio of material to liquid was 1 ∶ 1; the oscillating speed was 190 r/min. The self-developed enzyme biosensor for detecting agricultural residues could meet the needs of rapid detecting organophosphorus pesticide residues in fruits and vegetables.
引文
[1]曾家源,黄俊硕.有机磷农药检测方法的研究进展[J].广东化工,2015,42(5):70-71.
[2]李流川,李德华,王卓,等.2015-2016年达州市108份果蔬和茶叶中农药残留监测结果分析[J].现代预防医学,2018,45(9):1568-1573.
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[4]张亚佳,李忠海.果蔬中有机磷农药残留检测方法研究进展[J].食品与机械,2016,32(2):173-182.
[5]孙元欣,陈炳太,易森,等.有机磷农药残留的快速定量检测方法研究[J].光谱学与光谱分析,2014,34(5):1338-1342.
[6]李萌,于劲松,徐斐,等.酶生物传感器法和国标方法检测农药残留的抗干扰效果对比研究[J].食品与发酵科技,2016,52(1):101-105.
[7]戴莹,王纪华,韩平,等.食品中毒死蜱残留量检测技术进展[J].食品安全质量检测学报.2015,6(7):2696-2701.
[8]王青,黄铮.食品中拟除虫菊酯类农药残留检测前处理技术研究进展[J].食品研究与开发,2018,39(11):186-191.
[9]LI R,WEI W,HE L,et al.Chlorpyrifos residual behaviors in field crops and transfers during duck pellet feed processing[J].JAgric Food Chem,2014,62(42):10215-10221.
[10]UJAN A A,KHANZAD M A,SHAHZAD S.Efficiency of Metarhizium spp.(Sorokin)strains and insecticides against cotton mealybug Phenaeoccus solenopsis(Tinsley)[J].Pak J Zool,2014,46(6):100-105.
[11]OTIENO P,OKINDA OWUOR P,LALAH J O,et al.Monitoring the occurrence and distribution of selected organophosphates and carbamate pesticide residues in the ecosystem of Lake Naivasha,Kenya[J].Toxico Environ Chem,2015,97:51-61.
[12]徐斐,许学勤,张慧君,等.鸡肝酯酶在有机磷和氨基甲酸酯类农药检测中的应用研究[J].食品科学,2007,28(5):237-240.
[13]中华人民共和国卫生部,中国国家标准化管理委员会.GB-T5009.199-2003蔬菜中有机磷和氨基甲酸酯类农药残留量的快速检测[S].北京:中国标准出版社,2004.
[14]张丽青,闫艳春,姜红霞,等.高酯酶活性工程菌的构建及其对有机磷农药的降解[J].工业微生物,2008,38(3):47-50.
[15]扶继军.酶抑制率法农残速测技术及常见问题探究[J].农业与技术,2014,34(2):29-32.
[16]顾亚萍,乔方,林慧纯,等.4种蔬菜中20种农药残留的基质效应的研究[J].食品研究与开发,2017,38(22):148-152.
[17]CHEN D,WANG Y J,XU Y,et al.Highly sensitive detection of organophosphorus pesticides by acetylcholinesterase-coated thin film bulk acoustic resonatormass-loading sensor[J].Biosens Bioelectron,2013,41(15):163-167.
[2]李流川,李德华,王卓,等.2015-2016年达州市108份果蔬和茶叶中农药残留监测结果分析[J].现代预防医学,2018,45(9):1568-1573.
[3]郭巍.食品中常见有机磷农药残留的检测方法[J].现代食品,2018,37(1):117-123.
[4]张亚佳,李忠海.果蔬中有机磷农药残留检测方法研究进展[J].食品与机械,2016,32(2):173-182.
[5]孙元欣,陈炳太,易森,等.有机磷农药残留的快速定量检测方法研究[J].光谱学与光谱分析,2014,34(5):1338-1342.
[6]李萌,于劲松,徐斐,等.酶生物传感器法和国标方法检测农药残留的抗干扰效果对比研究[J].食品与发酵科技,2016,52(1):101-105.
[7]戴莹,王纪华,韩平,等.食品中毒死蜱残留量检测技术进展[J].食品安全质量检测学报.2015,6(7):2696-2701.
[8]王青,黄铮.食品中拟除虫菊酯类农药残留检测前处理技术研究进展[J].食品研究与开发,2018,39(11):186-191.
[9]LI R,WEI W,HE L,et al.Chlorpyrifos residual behaviors in field crops and transfers during duck pellet feed processing[J].JAgric Food Chem,2014,62(42):10215-10221.
[10]UJAN A A,KHANZAD M A,SHAHZAD S.Efficiency of Metarhizium spp.(Sorokin)strains and insecticides against cotton mealybug Phenaeoccus solenopsis(Tinsley)[J].Pak J Zool,2014,46(6):100-105.
[11]OTIENO P,OKINDA OWUOR P,LALAH J O,et al.Monitoring the occurrence and distribution of selected organophosphates and carbamate pesticide residues in the ecosystem of Lake Naivasha,Kenya[J].Toxico Environ Chem,2015,97:51-61.
[12]徐斐,许学勤,张慧君,等.鸡肝酯酶在有机磷和氨基甲酸酯类农药检测中的应用研究[J].食品科学,2007,28(5):237-240.
[13]中华人民共和国卫生部,中国国家标准化管理委员会.GB-T5009.199-2003蔬菜中有机磷和氨基甲酸酯类农药残留量的快速检测[S].北京:中国标准出版社,2004.
[14]张丽青,闫艳春,姜红霞,等.高酯酶活性工程菌的构建及其对有机磷农药的降解[J].工业微生物,2008,38(3):47-50.
[15]扶继军.酶抑制率法农残速测技术及常见问题探究[J].农业与技术,2014,34(2):29-32.
[16]顾亚萍,乔方,林慧纯,等.4种蔬菜中20种农药残留的基质效应的研究[J].食品研究与开发,2017,38(22):148-152.
[17]CHEN D,WANG Y J,XU Y,et al.Highly sensitive detection of organophosphorus pesticides by acetylcholinesterase-coated thin film bulk acoustic resonatormass-loading sensor[J].Biosens Bioelectron,2013,41(15):163-167.