CN105911170B - 一种六堡茶鲜叶中拟除虫菊酯类农药残留的检测方法 - Google Patents
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Abstract
本发明公开了一种六堡茶鲜叶中拟除虫菊酯类农药残留的检测方法，旨在提供一种检测灵敏度高、对于茶叶质量具有重要的意义；其技术方法：1)标准样品的制备；2)样品制备与提取，称取捣碎的样品10.0mg，加入50mL体积比1:1的***和乙酸乙酯于捣碎机中匀浆提取，真空抽滤并用10mL混合液洗涤滤渣，采用过饱和硫酸钠加入分液漏斗中萃取，分出有机相后浓缩至2mL上液相色谱仪检测；属于化学检测技术领域。
Description
技术领域
本发明涉及一种拟除虫菊酯类农药残留的检测方法，具体地说，是一种六堡茶鲜叶中拟除虫菊酯类农药残留的检测方法，属于化学检测技术领域。
背景技术
六堡茶属于黑茶类，主产于广西梧州六堡镇，该品种茶叶具有红、浓、陈、醇的特点，具有消暑祛湿、明目清心、帮助消化的功效，因为六堡茶所含脂肪分解酵素高于其他茶类，因此具有健胃养神、减肥健身的功效，是一种广受欢迎的茶类。影响六堡茶质量的主要是农药残留，国际上一些茶叶进口国制定了严格的农药残留限量标准，对于农药超标的茶叶进行就地封存、销毁或退回原产地过的政策，对于茶叶贸易具有很大的影响。拟除虫菊酯农药用量少，杀虫效果好，是当前广泛应用的茶叶农药(茶叶除虫剂)，为了从源头上控制(茶叶产品)农药残留，需要直接对鲜叶进行检测，以实现农药源头控制，对于茶园用药进行有效管理，以此减少农药违规造成的环境污染，提升饮茶安全性。因为鲜叶中含有较多的干扰物，因此采用有机溶剂提取，层析净化后进行液相色谱检验，能够依此建立有效的六堡茶鲜叶的农药检测方法，为提升饮茶安全性提供保障。
拟除虫菊酯的杀虫机理
拟除虫菊酯是一种仿生药物，该药物具有悠久的使用历史。拟除虫菊酯通过接触昆虫后迅速渗入虫体，与昆虫神经***作用从而改变神经突触膜对离子的通透性，引发昆虫持续兴奋，最后麻痹而死。相比于常用的杀虫剂，拟除虫菊酯的杀虫力能够高出1-2个数量级，击倒力强，对于农林、园艺、畜牧等地方的开冲均由良好的防治效果，而且该药物的毒性远低于有机磷的危害，在自然界易分解，在动物体内转化迅速，对于环境影响较小。因为该药物杀虫作用快、广谱性、易降解，因此广泛的应用于茶园、果园、农田等害虫的防治，是广泛应用的农药，但是因为其抗性和环境安全性，因此被列为多种农产品农药残留的重点检测对象。
拟除虫菊酯的危害与残留
拟除虫菊酯是一种神经毒剂，对于神经***的毒害作用是多环节的，可能会引发神经损伤，导致淋巴和脾脏受损，甚至诱发癌变。当前该药物已经占全世界化学农药总产量的25％左右。因为贸易壁垒的原因，各国对于拟除虫菊酯的残留量规定不一致，因此对茶叶贸易造成影响。
拟除虫菊酯检测技术
因为拟除虫菊酯残留量微少，需要高灵敏性的检测技术进行残留量的检测，当前常用的方法包括气相色谱法、气相色谱质谱法、高效液相色谱法、免疫分析法等多种方法。为了确保检测的准确性，所选择的方法具有较强的重复性与准确度，确保检测方式满足大批量样品多组分快速测定要求。
发明内容
针对上述问题，本发明提供的目的是提供一种检测灵敏度高、对于茶叶质量具有重要的意义的六堡茶鲜叶中拟除虫菊酯类农药残留的检测方法。
为解决上述技术问题，本发明提供的技术方案是这样的：
一种六堡茶鲜叶中拟除虫菊酯类农药残留的检测方法，包括下述步骤：
1)标准样品的制备
准确称取联苯菊酯、甲氰菊酯、三氟氯氰菊酯、氯氰菊酯、氰戊菊酯、S-氰戊菊酯，采用氯仿溶解配制成为500μg/mL的储备液待用；
3)样品制备与提取
从茶园取未使用任何农药的鲜叶样品100g，置于组织捣碎机中捣碎，称取捣碎的样品10.0mg，加入50mL体积比1:1的***和乙酸乙酯于捣碎机中匀浆提取，真空抽滤并用10mL混合液洗涤滤渣，采用过饱和硫酸钠加入分液漏斗中萃取，分出有机相后浓缩至2mL上高效液相色谱；
色谱柱：Thermo AccliaimTM Mixed-Mode WCX-1柱；流动相：A：甲醇；B:水；梯度洗脱程序：0～6min，90％A，10％B、6～11min，50％A、50％B；11～15min，40％A、60％B；15～19min，30％A、70％B,19～25min，20％A、80％B、流速：0.8mL/min；柱温：40℃；进样量：2μl；检测波长：230nm为主要检测波长；254nm、280nm辅助定性。
与现有技术相比，本发明提供的技术方案，检测结果线性良好、操作简便、抗基质干扰、结果可靠，为食品的质量监管提供了有力的技术支撑。
附图说明
图1是本发明提供的液相色谱曲线图。
具体实施方式
下面结合具体实施方式，对本发明的权利要求做进一步的详细说明，但不构成对本发明的任何限制，任何在本发明权利要求保护范围内所做的有限次的修改，仍在本发明的权利要求保护范围之内。
实施例1
本文所采用的六堡茶鲜叶为苍梧县六堡镇某茶业有限公司茶园现摘鲜叶作为试验，采用液相色谱法对六堡茶鲜叶的拟除虫菊酯类农药残留进行检测，该方法具有灵敏度高、假阳性少的特点，是国际上公认的定量确认的方法。
仪器与试剂
采用Agilent液相色谱仪，79型多用高速组织捣碎机，美国Sigma公司硅土，无水硫酸钠、活性炭均经过脱水处理，试剂均为分析纯试剂，所有试剂经过重蒸馏后使用，采用双蒸馏水作为试验用水。
方法
标准样品的制备
联苯菊酯、甲氰菊酯、三氟氯氰菊酯、氯氰菊酯、氰戊菊酯、S-氰戊菊酯均为德国Augsburg产品，准确称取上述标准品，采用氯仿溶解配制成为500μg/mL的储备液待用。
样品制备与提取
从茶园取未使用任何农药的鲜叶样品200g，置于组织捣碎机中捣碎。称取捣碎的样品10.0mg，加入100mL V(***)：V(乙酸乙酯)＝1:1于捣碎机中匀浆提取，真空抽滤并用10mL混合液洗涤滤渣，采用过饱和硫酸钠加入分液漏斗中萃取，分出有机相后浓缩至2mL上净化柱。
色谱柱：Thermo AccliaimTM Mixed-Mode WCX-1柱；流动相：A：甲醇；B:水；梯度洗脱程序：0～6min，90％A，10％B、6～11min，50％A、50％B；11～15min，40％A、60％B；15～19min，30％A、70％B,19～25min，20％A、80％B、流速：0.8mL/min；柱温：40℃；进样量：2μl；检测波长：230nm为主要检测波长；254nm、280nm辅助定性，其检测结果如图1所示。
Claims (1)
1.一种六堡茶鲜叶中拟除虫菊酯类农药残留的检测方法，其特征在于，包括下述步骤：
1)标准样品的制备
准确称取联苯菊酯、甲氰菊酯、三氟氯氰菊酯、氯氰菊酯、氰戊菊酯、S-氰戊菊酯，采用氯仿溶解配制成为500μg/mL的储备液待用；
2)样品制备与提取
从茶园取未使用任何农药的鲜叶样品100g，置于组织捣碎机中捣碎，称取捣碎的样品10.0mg，加入50mL体积比1:1的***和乙酸乙酯于捣碎机中匀浆提取，真空抽滤并用10mL混合液洗涤滤渣，采用过饱和硫酸钠加入分液漏斗中萃取，分出有机相后浓缩至2mL上液相色谱仪；
其中：高效液相色谱法柱色谱条件如下：
色谱柱：ThermoAccliaimTM Mixed-Mode WCX-1柱；流动相：A：甲醇；B:水；梯度洗脱程序：0～6min，90％A，10％B、6～11min，50％A、50％B；11～15min，40％A、60％B；15～19min，30％A、70％B,19～25min，20％A、80％B、流速：0.8mL/min；柱温：40℃；进样量：2μl；检测波长：230nm为主要检测波长；254nm、280nm辅助定性。
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| CN101650343A (zh) * | 2008-12-15 | 2010-02-17 | 河南工程学院 | 水中菊酯类农药的快速检测 |
| WO2012069571A1 (en) * | 2010-11-24 | 2012-05-31 | Givaudan Sa | In vitro method for determining presence of type ii pyrethroids |
| CN104407069A (zh) * | 2014-11-25 | 2015-03-11 | 苏州东辰林达检测技术有限公司 | 一种茯茶中菊酯类农药的检测方法 |
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| 加速溶剂萃取-液相色谱-串级质谱法测定近岸及河口沉积物中的拟除虫菊酯农药;杨琳等;《分析化学》;20100731;第38卷(第7期);第968-972页 * |
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Application publication date: 20160831 Assignee: WUZHOU TIANYU TEA INDUSTRY Co.,Ltd. Assignor: WUZHOU CITY PRODUCT QUALITY INSPECTION Contract record no.: X2023980044738 Denomination of invention: A detection method for pyrethroid pesticide residues in fresh leaves of Liubao tea Granted publication date: 20180831 License type: Common License Record date: 20231030 |