腈菌唑在三七和土壤中的消解动态与残留分析
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摘要
腈菌唑(myclobutanil)被认为是防治三七黑斑病和圆斑病等病害应用前景较好的杀菌剂,但其在三七中残留检测和残留消解动态研究尚未见报道。为建立三七中腈菌唑残留检测和残留消解动态的检测方法,利用高效液相色谱-质谱联用技术(HPLC-MS)研究40%腈菌唑可湿性粉剂在三七(Panax notoginseng)根、茎、叶和土壤中的消解动态及喷施前后的皂苷含量变化,并对其可能产生的膳食安全风险进行评估。三七样品经丙酮提取,SPE固相萃取柱净化后,再由BEH-C18色谱柱(2.1 mm×50 mm,1.7μm)分离,以体积比为70∶30的乙腈水溶液为流动相进行洗脱,利用正离子扫描,多反应监测进行定量分析,外标法定量;采用高效液相色谱法测定三七块根中主要皂苷成分的含量。结果表明,建立的三七中腈菌唑残留量检测方法在0.1～5.0 mg/kg水平的平均添加回收率为80.9%～91.9%,相对标准偏差为5.05%～9.3%,腈菌唑检测极限为0.1μg/kg。在大棚内喷施135 g/hm2的腈菌唑时,在三七及土壤中的降解过程均符合一级动力学方程,腈菌唑在三七根、茎、叶和土壤中的降解半衰期分别为6.77 d、6.70 d、6.63 d和6.03 d,其风险商值为9.5×10-5,远小于1,风险较低,处于安全水平;随着生育期的增加,三七根中人参皂苷Rg1、人参皂苷Rb1的含量不断增加。因此,建议我国腈菌唑在三七中的最大残留量值可暂定为0.2 mg/kg,安全间隔期为35 d。研究结果表明腈菌唑对三七使用安全,为腈菌唑在三七上的安全合理使用提供科学依据。
Myclobutanil is regarded as a fungicide against Panax notoginseng round spot and black spot disease with good application prospect. The method for the determination and residue dynamics of myclobutanil in P. notoginseng has not been reported. In order to establish the method for the determination and residue dynamics of myclobutanil in P. notoginseng,the residue dynamic of myclobutanile in root,stem,leaf of P. notoginseng and its soil were studied by HPLC-MS,moreover the dietary safety and risk assessment were conducted based on the field trial data. The samples were separated by acetone,purified by SPE solid phase extraction,separated on a BEH-C18 column(2.1 mm×50 mm,1.7 μm),eluted with 70 ∶ 30 acetonitrile aqueous solution as the mobile phase. Quantitative analysis was performed using positive ion scanning and multi-reaction monitoring. The content of saponins in P. notoginseng root was determined by high performance liquid chromatography(HPLC). The fortified recovery rate at level of 0.1 ～ 5.0 mg/kg ranged from 80.9% ～ 91.9%,with relative standard deviation of 5.05% ～ 9.3%,and the limit of quantification for myclobutanil in P. notoginseng is 0.1 μg/kg. The results showed that the decline of myclobutanil in P. notoginseng and soil followed the first-kinetic equation. The half-life of myclobutanil in P. notoginseng root,stem,leaf and soil was 6.77 d,6.70 d,6.63 d,and 6.03 d,respectively,when the spaying dosage was 135 g/hm2. The risk quotient was 9.5×10-5,which was much less than 1,suggested the risk was very low with a safe level. With the development of P. notoginseng,the contents of notoginsenoside Rg1 and notoginsenoside Rb1 were increasing. It was suggested that the maximum residual value of myclobutanil in P. notoginseng could be set at level of 0. 2 mg/kg and the safety interval were 35 d. The results showed that myclobutanil was safe for use in controlling diseases in P. notoginseng.
Myclobutanil is regarded as a fungicide against Panax notoginseng round spot and black spot disease with good application prospect. The method for the determination and residue dynamics of myclobutanil in P. notoginseng has not been reported. In order to establish the method for the determination and residue dynamics of myclobutanil in P. notoginseng,the residue dynamic of myclobutanile in root,stem,leaf of P. notoginseng and its soil were studied by HPLC-MS,moreover the dietary safety and risk assessment were conducted based on the field trial data. The samples were separated by acetone,purified by SPE solid phase extraction,separated on a BEH-C18 column(2.1 mm×50 mm,1.7 μm),eluted with 70 ∶ 30 acetonitrile aqueous solution as the mobile phase. Quantitative analysis was performed using positive ion scanning and multi-reaction monitoring. The content of saponins in P. notoginseng root was determined by high performance liquid chromatography(HPLC). The fortified recovery rate at level of 0.1 ～ 5.0 mg/kg ranged from 80.9% ～ 91.9%,with relative standard deviation of 5.05% ～ 9.3%,and the limit of quantification for myclobutanil in P. notoginseng is 0.1 μg/kg. The results showed that the decline of myclobutanil in P. notoginseng and soil followed the first-kinetic equation. The half-life of myclobutanil in P. notoginseng root,stem,leaf and soil was 6.77 d,6.70 d,6.63 d,and 6.03 d,respectively,when the spaying dosage was 135 g/hm2. The risk quotient was 9.5×10-5,which was much less than 1,suggested the risk was very low with a safe level. With the development of P. notoginseng,the contents of notoginsenoside Rg1 and notoginsenoside Rb1 were increasing. It was suggested that the maximum residual value of myclobutanil in P. notoginseng could be set at level of 0. 2 mg/kg and the safety interval were 35 d. The results showed that myclobutanil was safe for use in controlling diseases in P. notoginseng.
引文
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[4]王勇,谢健杨,李保同.25%腈菌唑乳油对水稻纹枯病和稻曲病的防治效果[J].农药,2016,55(2):141-142.Wang Y,Xie J Y,Li B T. Efficacy of myclobutanil 25%EC against rice sheath blight and rice false smut in paddy field[J]. Agrochemicals,2016,55(2):141-142.
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[6]龚耀刚,王兴文,程莉,等.水提三七有效部位重金属及农药残留的研究[J].云南中医学院学报,2004,27(4):11-13.Gong Y G,Wang X W,Cheng L,et al.. Determination of oleanlic acid in Micromeria biflora from diferent districts of Yunnan Province[J]. J. Yunnan Coll. Trad. Chin. Med.,2004,27(4):11-13.
[7]姜涛,陈林明,林岱滨.超高效液相色谱法测定三七中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的方法研究[J].中国现代药物应用,2014,20:15-16.Jiang T,Chen L M,Lin D B. Method study of ultra high performance liquid chromatography in determination of notoginsenoside R1 and ginsenoside Rg1 and ginsenoside Rb1in Panax notoginseng[J]. Chin. J. Modern Drug Appl.,2014,20:15-16.
[8]王春伟,王燕,许允成,等. SPE-HPLC-MS/MS法测定人参及土壤中氟硅唑的残留及风险评估[J].华南农业大学学报,2014,6:58-62.Wang C W,Wang Y,Xu Y C,et al.. Residue determination by SPE-HPLC-MS/MS and risk assessment of flusilazole in the ginseng and soil[J]. J. South China Agric. Univ.,2014,6:58-62.
[9]王燕,王春伟,高洁,等.烯酰吗啉在人参和土壤中的残留动态及膳食风险评估[J].食品科学,2014,14:170-174.Wang Y,Wang C W,Gao J,et al.. Degradation dynamics and dietary risk assessment of dimethomorph in ginseng and soil[J]. Food Sci.,2014,14:170-174.
[10]韦荣昌,唐春风,冯世鑫,等.广西不同年限三七中皂苷含量的HPLC分析[J].河南农业科学,2014,3:154-157.Wei R C,Tang C F,Feng S X,et al.. HPLC analysis of saponins content from different growth years of Panax notoginseng in Guangxi[J]. 2014,3:154-157.
[11]钱永忠,李耘.农产品质量安全风险评估:原理、方法和应用[M].北京:中国标准出版社,2007,232-233.
[12]段丽芳,朴秀英,简秋,等.食品中85种农药最大残留限量国家标准介绍[J].农药科学与管理,2012,7:21-24.
[13]杨大进,王竹天,宋书锋,等.中药材类保健食品中添加剂限量标准研制[J].中国公共卫生,2008,24(3):358-359.
[14]岳永德.农药残留分析[M].北京:中国农业出版社,2014.
[15]中国农业科学院农业质量标准与检测技术研究所.农产品质量安全风险评估:原理、方法和应用[M].北京:中国标准出版社,2007.
[16] Sun M J,Liu D H, Zhou G X, et al.. Enantioselective degradation and chiral stability of malathion in environmental samples[J]. J. Agric. Food Chem.,2012,60(1):372-379.
[17]崔淑华,王开运,杨娟,等.腈菌唑在大葱中的残留分析与消解动态[J].农药,2011,50(4):283-285.Cui S H,Wang K Y,Yang J,et al.. Residue dynamics of myclobutanil in onion[J]. Agrochemicals,2011,50(4):283-285.
[18]王思威,孙海滨,曾繁娟,等.腈菌唑在荔枝及土壤中的残留动态研究[J].农学学报,2011,6:28-31.Wang S W,Sun H B,Zeng F J,et al.. Residual dynamics of myclobutanil fungicides in litchi and soil[J]. J. Agric.,2011,6:28-31.
[19]张志恒,李红叶,吴珉,等.百菌清、腈菌唑和吡唑醚菌酯在草莓中的残留及其风险评估[J].农药学学报,2009,11(4):449-455.Zhang Z H, Li H Y, Wu M, et al.. Residue and risk assessment of chlorothalonil,myclobutanil and pyraclostrobin in greenhouse strawberry[J]. Chin. J. Pesticide Sci.,2009,11(4):449-455.
[20]杨淑娟,郭建辉,蔡恩兴,等.菜用大豆和豌豆中腈菌唑残留动态研究[J].安全与环境学报,2008,8(6):1-4.Yang S J,Guo J H,Cai E X,et al.. Residue dynamics of myclobutanil in vegetable soybean and pea[J]. J. Safety Environ.,2008,8(6):1-4.
[21]楼正云,汤富彬,罗逢建,等.腈菌唑在黄瓜和土壤中的残留动态[J].农药,2008,47(3):205-207.Lou Z Y,Tang F B,Luo F J,et al.. Residue dynamics of myclobutanil in cucumber and soil[J]. Agrochemicals,2008,47(3):205-207.
[22] Chung H W,Ha Y G,Im M H,et al.. Establishment of 22pesticide mrls in agricultural products based on risk assessment[J]. Korean J. Environ. Agric.,2011,30(2):166-172.
[23]韩丙军,汤建彪,彭黎旭,等.腈菌唑在香蕉上的降解残留研究[J].农业环境科学学报,2007(26):197-200.Han B J, Tang J B, Peng L X, et al.. Degradation of myclobutanil residue in banana[J]. J. Agro-Environ. Sci.,2007(26):197-200.
[24] Authority E F S. Review of the existing maximum residue levels(MRLs)for diuron according to article 12 of regulation(EC)No 396/2005[J]. Efsa J.,2011,9(7):2324.
[25] Nagayama T. Positive list system for agricultural chemical residues in foods[J]. J. Pesticide Sci.,2005,30(4):418-425.
[26]中华人民共和国卫生部.中华人民共和国农业部GB 2763-2016食品安全国家标准食品中农药最大残留限量[S].北京:中国标准出版社,2016.
[27] Manivannan P, Jaleel C A, Somasundaram R, et al..Osmoregulation and antioxidant metabolism in drought-stressed Helianthus annuus under triadimefon drenching[J]. CR Biol.,2008,331(6):418-425.
[2]陆燕,潘旭.三七总皂苷中代表性皂苷含量测定方法学的建立及稳定性研究[J].中国药物应用与监测,2014,2:88-91.Lu Y, Pan X. Study on the methodology establishment of content determination and stability of representative saponins in Panax notoginseng saponins[J]. Chin. J. Drug Appl.Monitor.,2014,2:88-91.
[3]王燕,王春伟,高洁,等.液相色谱-串联质谱法研究腈菌唑在人参根、茎、叶和土壤中的残留动态[J].中国中药杂志,2014,39(13):2464-2468.Wang Y, Wang C W, Gao J, et al.. Determination of myclobutanil 25%WG degradation dynamics in ginseng root,stem,leaf and soil by HPLC-MS/MS[J]. China J. Chin.Mater. Med.,2014,39(13):2464-2468.
[4]王勇,谢健杨,李保同.25%腈菌唑乳油对水稻纹枯病和稻曲病的防治效果[J].农药,2016,55(2):141-142.Wang Y,Xie J Y,Li B T. Efficacy of myclobutanil 25%EC against rice sheath blight and rice false smut in paddy field[J]. Agrochemicals,2016,55(2):141-142.
[5]王勇,陈昱君,杨建忠,等.几种杀菌剂对三七黑斑病防效及与三七质量关系研究[J].文山学院学报,2013,26(6):4-7.Wang Y,Chen Y J,Yang J Z,et al.. Prevention effect of several fungicides on black spot disease and its influence on the quafity of Panax notoginseng[J]. J. Wen Shan Univ.,2013,26(6):4-7.
[6]龚耀刚,王兴文,程莉,等.水提三七有效部位重金属及农药残留的研究[J].云南中医学院学报,2004,27(4):11-13.Gong Y G,Wang X W,Cheng L,et al.. Determination of oleanlic acid in Micromeria biflora from diferent districts of Yunnan Province[J]. J. Yunnan Coll. Trad. Chin. Med.,2004,27(4):11-13.
[7]姜涛,陈林明,林岱滨.超高效液相色谱法测定三七中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的方法研究[J].中国现代药物应用,2014,20:15-16.Jiang T,Chen L M,Lin D B. Method study of ultra high performance liquid chromatography in determination of notoginsenoside R1 and ginsenoside Rg1 and ginsenoside Rb1in Panax notoginseng[J]. Chin. J. Modern Drug Appl.,2014,20:15-16.
[8]王春伟,王燕,许允成,等. SPE-HPLC-MS/MS法测定人参及土壤中氟硅唑的残留及风险评估[J].华南农业大学学报,2014,6:58-62.Wang C W,Wang Y,Xu Y C,et al.. Residue determination by SPE-HPLC-MS/MS and risk assessment of flusilazole in the ginseng and soil[J]. J. South China Agric. Univ.,2014,6:58-62.
[9]王燕,王春伟,高洁,等.烯酰吗啉在人参和土壤中的残留动态及膳食风险评估[J].食品科学,2014,14:170-174.Wang Y,Wang C W,Gao J,et al.. Degradation dynamics and dietary risk assessment of dimethomorph in ginseng and soil[J]. Food Sci.,2014,14:170-174.
[10]韦荣昌,唐春风,冯世鑫,等.广西不同年限三七中皂苷含量的HPLC分析[J].河南农业科学,2014,3:154-157.Wei R C,Tang C F,Feng S X,et al.. HPLC analysis of saponins content from different growth years of Panax notoginseng in Guangxi[J]. 2014,3:154-157.
[11]钱永忠,李耘.农产品质量安全风险评估:原理、方法和应用[M].北京:中国标准出版社,2007,232-233.
[12]段丽芳,朴秀英,简秋,等.食品中85种农药最大残留限量国家标准介绍[J].农药科学与管理,2012,7:21-24.
[13]杨大进,王竹天,宋书锋,等.中药材类保健食品中添加剂限量标准研制[J].中国公共卫生,2008,24(3):358-359.
[14]岳永德.农药残留分析[M].北京:中国农业出版社,2014.
[15]中国农业科学院农业质量标准与检测技术研究所.农产品质量安全风险评估:原理、方法和应用[M].北京:中国标准出版社,2007.
[16] Sun M J,Liu D H, Zhou G X, et al.. Enantioselective degradation and chiral stability of malathion in environmental samples[J]. J. Agric. Food Chem.,2012,60(1):372-379.
[17]崔淑华,王开运,杨娟,等.腈菌唑在大葱中的残留分析与消解动态[J].农药,2011,50(4):283-285.Cui S H,Wang K Y,Yang J,et al.. Residue dynamics of myclobutanil in onion[J]. Agrochemicals,2011,50(4):283-285.
[18]王思威,孙海滨,曾繁娟,等.腈菌唑在荔枝及土壤中的残留动态研究[J].农学学报,2011,6:28-31.Wang S W,Sun H B,Zeng F J,et al.. Residual dynamics of myclobutanil fungicides in litchi and soil[J]. J. Agric.,2011,6:28-31.
[19]张志恒,李红叶,吴珉,等.百菌清、腈菌唑和吡唑醚菌酯在草莓中的残留及其风险评估[J].农药学学报,2009,11(4):449-455.Zhang Z H, Li H Y, Wu M, et al.. Residue and risk assessment of chlorothalonil,myclobutanil and pyraclostrobin in greenhouse strawberry[J]. Chin. J. Pesticide Sci.,2009,11(4):449-455.
[20]杨淑娟,郭建辉,蔡恩兴,等.菜用大豆和豌豆中腈菌唑残留动态研究[J].安全与环境学报,2008,8(6):1-4.Yang S J,Guo J H,Cai E X,et al.. Residue dynamics of myclobutanil in vegetable soybean and pea[J]. J. Safety Environ.,2008,8(6):1-4.
[21]楼正云,汤富彬,罗逢建,等.腈菌唑在黄瓜和土壤中的残留动态[J].农药,2008,47(3):205-207.Lou Z Y,Tang F B,Luo F J,et al.. Residue dynamics of myclobutanil in cucumber and soil[J]. Agrochemicals,2008,47(3):205-207.
[22] Chung H W,Ha Y G,Im M H,et al.. Establishment of 22pesticide mrls in agricultural products based on risk assessment[J]. Korean J. Environ. Agric.,2011,30(2):166-172.
[23]韩丙军,汤建彪,彭黎旭,等.腈菌唑在香蕉上的降解残留研究[J].农业环境科学学报,2007(26):197-200.Han B J, Tang J B, Peng L X, et al.. Degradation of myclobutanil residue in banana[J]. J. Agro-Environ. Sci.,2007(26):197-200.
[24] Authority E F S. Review of the existing maximum residue levels(MRLs)for diuron according to article 12 of regulation(EC)No 396/2005[J]. Efsa J.,2011,9(7):2324.
[25] Nagayama T. Positive list system for agricultural chemical residues in foods[J]. J. Pesticide Sci.,2005,30(4):418-425.
[26]中华人民共和国卫生部.中华人民共和国农业部GB 2763-2016食品安全国家标准食品中农药最大残留限量[S].北京:中国标准出版社,2016.
[27] Manivannan P, Jaleel C A, Somasundaram R, et al..Osmoregulation and antioxidant metabolism in drought-stressed Helianthus annuus under triadimefon drenching[J]. CR Biol.,2008,331(6):418-425.