水环境中36种农药残留的同时分析方法及其应用
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摘要
农业生产中广泛应用的农药通过沉降、地表径流等途径进入水环境后,引起水资源污染、水生生态系统破坏、食品安全等一系列问题。随着高毒的有机氯、有机磷农药的禁用和限用,拟除虫菊酯、酰胺、苯胺、唑类杂环等“新一代”农药的用量不断加大,但是这些农药在水环境中的存在、行为和污染效应研究尚未受到足够关注。本论文以固相萃取(SPE)为富集和净化手段,应用气相色谱检测技术建立了水环境(表层水和沉积物)中36种常用农药准确、快速、高灵敏的分析方法,并将其成功用于九龙江口和厦门西海域水环境中农药污染状况的调查。主要研究内容和结果如下:
(1)建立了河水和海水中36种目标农药的固相萃取-气相色谱分析方法
实验以气相色谱-电子捕获检测器(GC-ECD)作为初步定性及定量手段,气相色谱-质谱联用(GC-MS)作为确证方法,通过优化色谱条件,建立了36种目标农药的高灵敏、准确可靠的仪器分析方法。优化后,仪器的测定限为0.09～2.66μg/L,线性范围为0.9～2600.0μg/L。
实验在比较5种固相萃取柱萃取效果的基础上,选择Oasis~(?)HLB作为水样中36种农药的固相萃取柱,考察了洗脱溶剂、上样速度、穿透体积、pH值和离子强度等因素对HLB固相萃取柱萃取效果的影响。方法优化后确定:分析实际水样时不调整水样pH值和氯化钠浓度,分析水样体积为1.0 L,上样速度为4mL/min,通过在富集水样后的HLB上串联除水用无水硫酸钠SPE柱和净化用LC-NH_2柱,以2 mL/min的流速,用3 mL乙腈、10 mLl:1(vol.)丙酮-正己烷及10 mL乙酸乙酯进行洗脱和净化。以四氯间二甲苯、环氟菌胺和氟丙菊酯为替代物,PCB103为内标物,分析九龙江河口表层水样,得到基底加标回收率和相对标准偏差(RSD,n=3)分别为62.2%～124.1%,0.2%～9.6%(淡水)和64.5%～131.3%,0.2%～10.3%(海水);方法检出限为0.06～0.98 ng/L。
(2)建立了表层沉积物中目标农药的分析方法
以乙腈为萃取溶剂、均质和超声波辅助萃取、Carb-NH_2串联固相萃取柱净化,建立了表层沉积物样品中35种农药残留的分析方法。在优化的条件下,15 mL乙腈均质萃取3 min再超声萃取3 min(两次)能很好地萃取表层沉积物中的35种目标农药,串联的Carb-NH_2柱用4次每次3 mL的1:1(vol.)丙酮-正己烷洗脱,能够很好地净化萃取液,除丙环唑外35种农药的回收率在63.7%～104.2%之间,且相对标准偏差(RSD,n=3)在0.1%～10.3%之间,具有较好的重现性;方法检测限为0.01～0.56μg/kg(干重)。
(3)运用所建立的方法调查九龙江口及厦门西海域水环境中目标农药的污染状况
运用建立的水和沉积物中多种农药残留的同时分析方法,于2007年4月对九龙江口及厦门西海域表层水和沉积物中6类36种农药的污染状况进行了调查。发现有2种有机氯、7种有机磷、2种苯胺、3种酰胺、4种唑类杂环和8种菊酯类农药在表层水中被检出,检出的农药总量在24.8 ng/L～609.5 ng/L之间;有3种有机氯、5种有机磷、3种酰胺、3种唑类杂环和7种菊酯类农药在沉积物中被检出,检出的农药总量在14.1～122.6μg/kg(干重)之间。通过与已有研究的结果比较,发现部分酰胺、苯胺、唑类杂环和菊酯类等“新一代”农药的检出浓度和范围远高于传统的有机氯、有机磷农药,其对水环境的污染状况已经不可忽视。随着有机氯、有机磷农药的削减和禁用,“新一代”农药必将得到进一步推广和应用,对水环境的污染也将随之增大,十分必要对其环境行为和效应开展深入研究。
A series of environmental problems, such as water resource pollution, aquatic ecosystem destroy as well as food safety, are being caused by pesticides widely used in agriculture after entering aquatic environment through dust deposition, rainfall etc. With the prohibition and limitation of organophosphorous pesticides (OPPs) and organochlorine pesticides (OCPs), "new generation" pesticides labeled "low poison" such as pyrethroids, amides, nitrodiphenylamines and heterozoles, are increasingly used as the alternatives. However, researches about their existence, behavior and pollution effects are of seldom concern. In this dissertation, an accurate and sensitive method for the determination of 36 pesticides in aquatic environment (surface water and sediment), using solid phase extraction (SPE) for extraction, enrichment and cleanup and gas chromatography (GC) for detection, has been established and successfully applied to survey the pesticide pollution status in Jiulong River Estuary and Western Xiamen Bay. The detailed contents and results are as the followings:
(1) Development of determination method for 36 pesticides in river water and seawater samples
Instrumental analysis method was established using GC with electron capture detection (GC-ECD) for fast screening and quantitative and gas chromatography-mass spectrometry (GOMS) for identification. Under the optimized conditions, instrumental detection limits were in the range of 0. 09～2. 66μg/L, linearity ranges were between 0. 9～2600μg/L.
Oasis~(?) HLB was selected to extract and enrich target pesticides from water samples after comparison of 5 different SPE cartridges. Parameters such as elution solvents, loading speed, penetration volume, cleanup cartridges, pH and salinity were investigated. After optimization, the pH and ionic strength of water samples were found had little effect on recoveries, 1.0 L water was load onto HLB SPE cartridge at the speed of 4 mL/min, 5 g Na_2SO_4 and LC-NH_2 SPE cartridges were coupled to HLB for cleanup, 3 mL acetonitrile followed by 10 mL n-hexane/acetone (1:1, vol.) and 10 mL ethyl acetate were used to elute the target pesticides. Using 2,4,5,6-tetrachloro-m-xylene, cyflufenamid, and acrinathrin as surrogates, PCB103 as internal standard, spiked recoveries and relative standard deviation (RSD, n=3) of 36 target pesticides in fresh water and seawater were 62. 2%～124.1%, 0. 2%～9. 6%and 64. 5%～131. 3%, 0.2%～10.3%, respectively. Method detection limits were between 0.06～0.98 ng/L.
(2) Development of determination method for 36 pesticides in surface sediment samples
A method for the determination of target pesticides in surface sediment was proposed with acetonitrile for extraction assisted by homogenization and ultrasound and coupled Carb-NH_2 SPE cartridges for cleanup. Under the optimized conditions, except propiconazol, spiking recoveries of 35 target pesticides were between 63. 7%～104. 2%, RSD (n=3) 0. l%～10. 3%, and method detection limits were between 0. 01～0. 56μg/kg (dry weight).
(3) Pollution status survey of 36 pesticide residues in Jiulong River Estuary and Western Xiamen Bay
The pollution status of 36 target pesticides in Jiulong River Estuary and Western Xiamen Bay was surveyed using the established determination methods. Results showed that 2 OCPs, 8 OPPs, 2 nitrodiphenylamines, 3 amides, 3 heterozoles and 8 pyrethroids pesticides were detected in surface water samples with total concentration between 24.8 ng/L～609. 5 ng/L. 3 OCPs, 5 OPPs, 2 nitrodiphenylamine, 3 amides, 3 heterozoles and 7 pyrethroids pesticides were found in sediment samples with total concentration between 14.1μg/L～122. 6μg/L (dry weight).
Compared with other research results, although the concentrations of OCPs and OPPs in the researching area were lower, the results show that many "new generation" pesticides have been found in aquatic environment at certain concentrations reveal that their existence, environmental behaviors and effects should not be ignored and deserve further study.
(1)建立了河水和海水中36种目标农药的固相萃取-气相色谱分析方法
实验以气相色谱-电子捕获检测器(GC-ECD)作为初步定性及定量手段,气相色谱-质谱联用(GC-MS)作为确证方法,通过优化色谱条件,建立了36种目标农药的高灵敏、准确可靠的仪器分析方法。优化后,仪器的测定限为0.09～2.66μg/L,线性范围为0.9～2600.0μg/L。
实验在比较5种固相萃取柱萃取效果的基础上,选择Oasis~(?)HLB作为水样中36种农药的固相萃取柱,考察了洗脱溶剂、上样速度、穿透体积、pH值和离子强度等因素对HLB固相萃取柱萃取效果的影响。方法优化后确定:分析实际水样时不调整水样pH值和氯化钠浓度,分析水样体积为1.0 L,上样速度为4mL/min,通过在富集水样后的HLB上串联除水用无水硫酸钠SPE柱和净化用LC-NH_2柱,以2 mL/min的流速,用3 mL乙腈、10 mLl:1(vol.)丙酮-正己烷及10 mL乙酸乙酯进行洗脱和净化。以四氯间二甲苯、环氟菌胺和氟丙菊酯为替代物,PCB103为内标物,分析九龙江河口表层水样,得到基底加标回收率和相对标准偏差(RSD,n=3)分别为62.2%～124.1%,0.2%～9.6%(淡水)和64.5%～131.3%,0.2%～10.3%(海水);方法检出限为0.06～0.98 ng/L。
(2)建立了表层沉积物中目标农药的分析方法
以乙腈为萃取溶剂、均质和超声波辅助萃取、Carb-NH_2串联固相萃取柱净化,建立了表层沉积物样品中35种农药残留的分析方法。在优化的条件下,15 mL乙腈均质萃取3 min再超声萃取3 min(两次)能很好地萃取表层沉积物中的35种目标农药,串联的Carb-NH_2柱用4次每次3 mL的1:1(vol.)丙酮-正己烷洗脱,能够很好地净化萃取液,除丙环唑外35种农药的回收率在63.7%～104.2%之间,且相对标准偏差(RSD,n=3)在0.1%～10.3%之间,具有较好的重现性;方法检测限为0.01～0.56μg/kg(干重)。
(3)运用所建立的方法调查九龙江口及厦门西海域水环境中目标农药的污染状况
运用建立的水和沉积物中多种农药残留的同时分析方法,于2007年4月对九龙江口及厦门西海域表层水和沉积物中6类36种农药的污染状况进行了调查。发现有2种有机氯、7种有机磷、2种苯胺、3种酰胺、4种唑类杂环和8种菊酯类农药在表层水中被检出,检出的农药总量在24.8 ng/L～609.5 ng/L之间;有3种有机氯、5种有机磷、3种酰胺、3种唑类杂环和7种菊酯类农药在沉积物中被检出,检出的农药总量在14.1～122.6μg/kg(干重)之间。通过与已有研究的结果比较,发现部分酰胺、苯胺、唑类杂环和菊酯类等“新一代”农药的检出浓度和范围远高于传统的有机氯、有机磷农药,其对水环境的污染状况已经不可忽视。随着有机氯、有机磷农药的削减和禁用,“新一代”农药必将得到进一步推广和应用,对水环境的污染也将随之增大,十分必要对其环境行为和效应开展深入研究。
A series of environmental problems, such as water resource pollution, aquatic ecosystem destroy as well as food safety, are being caused by pesticides widely used in agriculture after entering aquatic environment through dust deposition, rainfall etc. With the prohibition and limitation of organophosphorous pesticides (OPPs) and organochlorine pesticides (OCPs), "new generation" pesticides labeled "low poison" such as pyrethroids, amides, nitrodiphenylamines and heterozoles, are increasingly used as the alternatives. However, researches about their existence, behavior and pollution effects are of seldom concern. In this dissertation, an accurate and sensitive method for the determination of 36 pesticides in aquatic environment (surface water and sediment), using solid phase extraction (SPE) for extraction, enrichment and cleanup and gas chromatography (GC) for detection, has been established and successfully applied to survey the pesticide pollution status in Jiulong River Estuary and Western Xiamen Bay. The detailed contents and results are as the followings:
(1) Development of determination method for 36 pesticides in river water and seawater samples
Instrumental analysis method was established using GC with electron capture detection (GC-ECD) for fast screening and quantitative and gas chromatography-mass spectrometry (GOMS) for identification. Under the optimized conditions, instrumental detection limits were in the range of 0. 09～2. 66μg/L, linearity ranges were between 0. 9～2600μg/L.
Oasis~(?) HLB was selected to extract and enrich target pesticides from water samples after comparison of 5 different SPE cartridges. Parameters such as elution solvents, loading speed, penetration volume, cleanup cartridges, pH and salinity were investigated. After optimization, the pH and ionic strength of water samples were found had little effect on recoveries, 1.0 L water was load onto HLB SPE cartridge at the speed of 4 mL/min, 5 g Na_2SO_4 and LC-NH_2 SPE cartridges were coupled to HLB for cleanup, 3 mL acetonitrile followed by 10 mL n-hexane/acetone (1:1, vol.) and 10 mL ethyl acetate were used to elute the target pesticides. Using 2,4,5,6-tetrachloro-m-xylene, cyflufenamid, and acrinathrin as surrogates, PCB103 as internal standard, spiked recoveries and relative standard deviation (RSD, n=3) of 36 target pesticides in fresh water and seawater were 62. 2%～124.1%, 0. 2%～9. 6%and 64. 5%～131. 3%, 0.2%～10.3%, respectively. Method detection limits were between 0.06～0.98 ng/L.
(2) Development of determination method for 36 pesticides in surface sediment samples
A method for the determination of target pesticides in surface sediment was proposed with acetonitrile for extraction assisted by homogenization and ultrasound and coupled Carb-NH_2 SPE cartridges for cleanup. Under the optimized conditions, except propiconazol, spiking recoveries of 35 target pesticides were between 63. 7%～104. 2%, RSD (n=3) 0. l%～10. 3%, and method detection limits were between 0. 01～0. 56μg/kg (dry weight).
(3) Pollution status survey of 36 pesticide residues in Jiulong River Estuary and Western Xiamen Bay
The pollution status of 36 target pesticides in Jiulong River Estuary and Western Xiamen Bay was surveyed using the established determination methods. Results showed that 2 OCPs, 8 OPPs, 2 nitrodiphenylamines, 3 amides, 3 heterozoles and 8 pyrethroids pesticides were detected in surface water samples with total concentration between 24.8 ng/L～609. 5 ng/L. 3 OCPs, 5 OPPs, 2 nitrodiphenylamine, 3 amides, 3 heterozoles and 7 pyrethroids pesticides were found in sediment samples with total concentration between 14.1μg/L～122. 6μg/L (dry weight).
Compared with other research results, although the concentrations of OCPs and OPPs in the researching area were lower, the results show that many "new generation" pesticides have been found in aquatic environment at certain concentrations reveal that their existence, environmental behaviors and effects should not be ignored and deserve further study.
引文
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