分散固相萃取-液相色谱-串联质谱法测定蜂蜜中新型烟碱类杀虫剂及其代谢物的残留量
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摘要
称取蜂蜜样品2.00g至具塞离心管中,加入100μL同位素内标溶液(200μg·L~(-1))和10mL水,涡旋混匀,加入甲醇至20mL,涡旋混匀,以8 500转·min~(-1)离心5min,移取上清液0.50mL,用甲醇定容至10.0mL,涡旋混匀后,以8 500转·min~(-1)离心5min,取1.0mL上清液转移至具塞离心管中(内含混合均匀的吸附剂:30mg N-丙基乙二胺、15mg C_(18)粉末和50mg无水MgSO_4),涡旋混匀进行吸附净化,以8 500转·min~(-1)离心5min,转移全部上清液,于40℃下用氮气吹至近干,用甲醇-0.15%(体积分数,下同)甲酸溶液(1+9)混合液溶解残渣并定容至1.0mL,过0.22μm滤膜后供液相色谱-串联质谱分析。采用Agilent Eclipse XDB-C_(18)色谱柱进行分离,以不同比例的含5mmol·L~(-1)乙酸铵的0.15%甲酸溶液和甲醇为流动相进行梯度洗脱。质谱中采用电喷雾离子源正离子扫描方式和多反应监测模式,采用同位素内标法和外标法进行定量。吡蚜酮、呋虫胺、烯啶虫胺、噻虫嗪、氟啶虫酰胺、吡虫啉、噻虫胺、氯噻啉、啶虫脒、噻虫啉、4-(三氟甲基)烟酰胺和N-去甲基啶虫脒的质量浓度在一定范围内呈线性,测定下限(10S/N)在2.5～12.5μg·kg~(-1)之间。对空白蜂蜜样品进行加标回收试验,回收率在79.9%～108%之间,测定值的相对标准偏差(n=6)在1.7%～15%之间。
The honey sample(2.00 g) was added into in a centrifuge tube with a cover,and 100μL of isotope internal standard solution(200μg·L~(-1)) and 10 mL water were added.After mixing well by vortex,the mixture was made up to 20 mL with methanol,and mixed well,centrifuged at 8 500 r·min~(-1) for 5 min.0.50 mL of the supernatant was taken and diluted to 10.0 mL with methanol,and the mixture was mixed well by vortex,then centrifuged at 8 500 r·min~(-1) for 5 min.1.0 mL of the supernatant was transferred into a centrifuge tube containing a homogeneously mixed adsorbent(30 mg of N-propyl ethylenediamine,15 mg of C_(18) powder and 50 mg of anhydrous MgSO_4).After vortexing and mixing for adsorption purification,the tube was centrifuged at 8 500 r·min~(-1) for 5 min.All the supernatant was blowed to near dryness using nitrogen at 40 ℃,and the residue was disolved and made up to 1.0 mL with a mixture of methanol and 0.15%(volume fraction)formic acid solution(1+9).The solution was passed through a 0.22μm filter,and the filtrate was analyzed by liquid chromatographytandem mass spectrometry.Chromatographic separation was carried out using an Agilent Eclipse XDB-C_(18) column,with a mixture in varying proportions of 0.15% formic acid solution containing 5 mmol·L~(-1) ammonium acetate and methanol as mobile phase for gradient elution.Electrospray ion source,positive ion scanning mode and multireaction monitoring mode were used in the mass spectrometry,and the isotope dilution internal standard method and external standard method were used for quantification.The mass concentration of pyridoxine,dinotefuran,nitenpyram,thiame-thoxam,flonicamid,imidacloprid,clothianidin,chlorothiazide,acetamiprid,thiacloprid,4-(trifluoromethyl) nicotinamide and N-desmethylpyrazole,was linear within definite ranges,and the lower limits of determination(10 S/N) were between 2.5-12.5μg·kg~(-1).Tests for recovery was made by standard addition method to blank honey samples,giving results in the range of 79.9%-108%,and RSDs(n=6) were between1.7% and 15%.
The honey sample(2.00 g) was added into in a centrifuge tube with a cover,and 100μL of isotope internal standard solution(200μg·L~(-1)) and 10 mL water were added.After mixing well by vortex,the mixture was made up to 20 mL with methanol,and mixed well,centrifuged at 8 500 r·min~(-1) for 5 min.0.50 mL of the supernatant was taken and diluted to 10.0 mL with methanol,and the mixture was mixed well by vortex,then centrifuged at 8 500 r·min~(-1) for 5 min.1.0 mL of the supernatant was transferred into a centrifuge tube containing a homogeneously mixed adsorbent(30 mg of N-propyl ethylenediamine,15 mg of C_(18) powder and 50 mg of anhydrous MgSO_4).After vortexing and mixing for adsorption purification,the tube was centrifuged at 8 500 r·min~(-1) for 5 min.All the supernatant was blowed to near dryness using nitrogen at 40 ℃,and the residue was disolved and made up to 1.0 mL with a mixture of methanol and 0.15%(volume fraction)formic acid solution(1+9).The solution was passed through a 0.22μm filter,and the filtrate was analyzed by liquid chromatographytandem mass spectrometry.Chromatographic separation was carried out using an Agilent Eclipse XDB-C_(18) column,with a mixture in varying proportions of 0.15% formic acid solution containing 5 mmol·L~(-1) ammonium acetate and methanol as mobile phase for gradient elution.Electrospray ion source,positive ion scanning mode and multireaction monitoring mode were used in the mass spectrometry,and the isotope dilution internal standard method and external standard method were used for quantification.The mass concentration of pyridoxine,dinotefuran,nitenpyram,thiame-thoxam,flonicamid,imidacloprid,clothianidin,chlorothiazide,acetamiprid,thiacloprid,4-(trifluoromethyl) nicotinamide and N-desmethylpyrazole,was linear within definite ranges,and the lower limits of determination(10 S/N) were between 2.5-12.5μg·kg~(-1).Tests for recovery was made by standard addition method to blank honey samples,giving results in the range of 79.9%-108%,and RSDs(n=6) were between1.7% and 15%.
引文
[1] PASTOR-BELDA M,GARRIDO I,CAMPILLO N,et al.Determination of spirocyclic tetronic/tetramic acid derivatives and neonicotinoid insecticides in fruits and vegetables by liquid chromatography and mass spectrometry after dispersive liquid-liquid microextraction[J].Food Chemitry,2016,202:389-395.
[2] CHAGNON M,KREUTZWEISER D,MITCHELL E A,et al.Risks of large scale use of systemic insecticides to ecosystem functioning and services[J].Environmental Science and Pollution Research,2012,22:119-134.
[3] LU C,WARCHOL K M,CALLAHAN R A.In situ replication of honey bee colony collapse disorder[J].Bulletin of Insectology,2012,65:99-106.
[4] LAURINO D,MANINO A,PATETTA A,et al.Toxicity of neonicotinoid insecticides on different honey bee genotypes[J].Bulletin of Insectology,2013,66(1):119-126.
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[6] MITCHELL E A D,MULHAUSER B,MULOT M,et al.A world wide survey of neonicotinoids in honey[J].Science,2017,358:109-111.
[7] The European Union.EU pesticides data-base[EB/OL].[2018-05-01]http://ec.europa.eu/food/plant/pesticides.
[8] Japan.Positive list system for agricultural chemical residues in foods[EB/OL].[2018-05-01]http://www.mhlw. go. jp/english/topics/foodsafety/positivelist060228/2018/05/01.
[9] 许秀莹,施海燕,王鸣华.气相色谱-质谱联用测定大米中6种烟碱类农药残留[J].质谱学报,2012,33(2):99-103.
[10] 侯如燕,蔡荟梅,张正竹,等.液相色谱法检测水果蔬菜中的烟碱类农药残留[J].分析试验室,2010,29(2):59-63.
[11] 侯如燕,卞红正,赵秀霞,等.固相萃取-液相色谱测定复杂基质蔬菜中9种烟碱类残留[J].分析测试学报,2011,30(1):58-63.
[12] KO A Y,RAHMAN M M,El-ATY A M A,et al.Development of a simple extraction and oxidation procedure for the residue analysis of imidacloprid and its metabolites in lettuce using gas chromatography[J].Food Chemistry,2014,148:402-409.
[13] GB/T 19648-2006 水果和蔬菜中500种农药及相关化学品残留量的测定气相色谱-质谱法[S].
[14] GB/T 20769-2008 水果和蔬菜中450种农药及相关化学品残留量的测定液相色谱-串联质谱法[S].
[15] GB/T 23205-2008 茶叶中448种农药及相关化学品残留量的测定液相色谱-串联质谱法[S].
[16] GARCIA M D G,GALERA M M,VALVERDE R S,et al.Column switching liquid chromatography and post-column photochemically fluorescence detection to determine imidacloprid and 6-chloronicotinic acid in honeybees[J].Journal of Chromatography A,2007,1147(1):17-23.
[17] RANCAN M,SABATINI A G,ACHILLI G,et al.Determination of imidacloprid and metabolites by liquid chromatography with an electrochemical detector and post column photochemical reactor[J].Analytica Chimica Acta,2006,555:20-24.
[18] CAMPILLO N,VINAS P,FEREZ-MELGAREJO G,et al.Liquid chromatography with diode array detection and tandem mass spectrometry for the determination of neonicotinoid insecticides in honey samples using dispersive liquid-liquid microextraction[J].Journal of Agricultural and Food Chemistry,2013,61:4799-4805.
[19] 王东,侯传金,赵尔成,等.盐析辅助均相液液萃取/分散固相萃取-超高效液相色谱串联质谱法测定蜂蜜中新烟碱类农药残留[J].分析测试学报,2015,34(6):681-685.
[20] 粟有志,李芳,齐鑫,等.QuEChERS-高效液相色谱/串联质谱法同时测定蜂蜜中9种新烟碱类杀虫剂残留[J].分析科学学报,2015,31(2):203-207.
[21] TANNER G,CZERWENKA C.LC-MS/MS Analysis of neonicotinoid insecticides in honey methodology and residue findings in Austrian honeys[J].Journal of Agricultural and Food Chemistry,2011,59:12271-12277.
[22] JIAO W T,XIAO Y,QIAN X S,et al.Optimized combination of dilution and refined QuEChERS to overcome matrix effects of six types of tea for determination eight neonicotinoid insecticides by ultra performance liquid chromatography-electrospray tandem mass spectrometry[J].Food Chemistry,2016,210:26-34.
[23] GBYLIK-SIKORSKA M,SNIEGOCKI T,POSYNIAK A.Determination of neonicotinoid insecticides and their metabolites in honey bee and honey by liquid chromatography tandem mass spectrometry[J].Journal of Chromatography B,2015,990:132-140.
[24] SUGANTHI A,BHUVANESWARI K,RAMYA M.Determination of neonicotinoid insecticide residues in sugarcane juice using LC-MS/MS[J].Food Chemistry,2018,241:275-280.
[25] STAHNKE H,KITTLAUS S,KEMPE G N,et al.Reduction of matrix effectsin liquid chromatographyelectrospray ionization-mass spectrometry by dilution of the sample extracts:How much dilution is needed?[J].Analytical Chemistry,2012,84(3):1474-1482.
[26] FERRER C,LOZANO A,AGUERA A,et al.Overcoming matrix effects using the dilution approach in multiresidue methods for fruits and vegetables[J].Journal of Chromatography A,2011,1218(42):7634-7639.
[2] CHAGNON M,KREUTZWEISER D,MITCHELL E A,et al.Risks of large scale use of systemic insecticides to ecosystem functioning and services[J].Environmental Science and Pollution Research,2012,22:119-134.
[3] LU C,WARCHOL K M,CALLAHAN R A.In situ replication of honey bee colony collapse disorder[J].Bulletin of Insectology,2012,65:99-106.
[4] LAURINO D,MANINO A,PATETTA A,et al.Toxicity of neonicotinoid insecticides on different honey bee genotypes[J].Bulletin of Insectology,2013,66(1):119-126.
[5] VICHAPONG J,BURAKHAM R,SRIJARANAI S.In-coupled syringe assisted octanol-water partition microextraction coupled with high-performance liquid chromatography for simultaneous determination of neonicotinoid insecticide residues in honey[J].Talanta,2015,139:21-26.
[6] MITCHELL E A D,MULHAUSER B,MULOT M,et al.A world wide survey of neonicotinoids in honey[J].Science,2017,358:109-111.
[7] The European Union.EU pesticides data-base[EB/OL].[2018-05-01]http://ec.europa.eu/food/plant/pesticides.
[8] Japan.Positive list system for agricultural chemical residues in foods[EB/OL].[2018-05-01]http://www.mhlw. go. jp/english/topics/foodsafety/positivelist060228/2018/05/01.
[9] 许秀莹,施海燕,王鸣华.气相色谱-质谱联用测定大米中6种烟碱类农药残留[J].质谱学报,2012,33(2):99-103.
[10] 侯如燕,蔡荟梅,张正竹,等.液相色谱法检测水果蔬菜中的烟碱类农药残留[J].分析试验室,2010,29(2):59-63.
[11] 侯如燕,卞红正,赵秀霞,等.固相萃取-液相色谱测定复杂基质蔬菜中9种烟碱类残留[J].分析测试学报,2011,30(1):58-63.
[12] KO A Y,RAHMAN M M,El-ATY A M A,et al.Development of a simple extraction and oxidation procedure for the residue analysis of imidacloprid and its metabolites in lettuce using gas chromatography[J].Food Chemistry,2014,148:402-409.
[13] GB/T 19648-2006 水果和蔬菜中500种农药及相关化学品残留量的测定气相色谱-质谱法[S].
[14] GB/T 20769-2008 水果和蔬菜中450种农药及相关化学品残留量的测定液相色谱-串联质谱法[S].
[15] GB/T 23205-2008 茶叶中448种农药及相关化学品残留量的测定液相色谱-串联质谱法[S].
[16] GARCIA M D G,GALERA M M,VALVERDE R S,et al.Column switching liquid chromatography and post-column photochemically fluorescence detection to determine imidacloprid and 6-chloronicotinic acid in honeybees[J].Journal of Chromatography A,2007,1147(1):17-23.
[17] RANCAN M,SABATINI A G,ACHILLI G,et al.Determination of imidacloprid and metabolites by liquid chromatography with an electrochemical detector and post column photochemical reactor[J].Analytica Chimica Acta,2006,555:20-24.
[18] CAMPILLO N,VINAS P,FEREZ-MELGAREJO G,et al.Liquid chromatography with diode array detection and tandem mass spectrometry for the determination of neonicotinoid insecticides in honey samples using dispersive liquid-liquid microextraction[J].Journal of Agricultural and Food Chemistry,2013,61:4799-4805.
[19] 王东,侯传金,赵尔成,等.盐析辅助均相液液萃取/分散固相萃取-超高效液相色谱串联质谱法测定蜂蜜中新烟碱类农药残留[J].分析测试学报,2015,34(6):681-685.
[20] 粟有志,李芳,齐鑫,等.QuEChERS-高效液相色谱/串联质谱法同时测定蜂蜜中9种新烟碱类杀虫剂残留[J].分析科学学报,2015,31(2):203-207.
[21] TANNER G,CZERWENKA C.LC-MS/MS Analysis of neonicotinoid insecticides in honey methodology and residue findings in Austrian honeys[J].Journal of Agricultural and Food Chemistry,2011,59:12271-12277.
[22] JIAO W T,XIAO Y,QIAN X S,et al.Optimized combination of dilution and refined QuEChERS to overcome matrix effects of six types of tea for determination eight neonicotinoid insecticides by ultra performance liquid chromatography-electrospray tandem mass spectrometry[J].Food Chemistry,2016,210:26-34.
[23] GBYLIK-SIKORSKA M,SNIEGOCKI T,POSYNIAK A.Determination of neonicotinoid insecticides and their metabolites in honey bee and honey by liquid chromatography tandem mass spectrometry[J].Journal of Chromatography B,2015,990:132-140.
[24] SUGANTHI A,BHUVANESWARI K,RAMYA M.Determination of neonicotinoid insecticide residues in sugarcane juice using LC-MS/MS[J].Food Chemistry,2018,241:275-280.
[25] STAHNKE H,KITTLAUS S,KEMPE G N,et al.Reduction of matrix effectsin liquid chromatographyelectrospray ionization-mass spectrometry by dilution of the sample extracts:How much dilution is needed?[J].Analytical Chemistry,2012,84(3):1474-1482.
[26] FERRER C,LOZANO A,AGUERA A,et al.Overcoming matrix effects using the dilution approach in multiresidue methods for fruits and vegetables[J].Journal of Chromatography A,2011,1218(42):7634-7639.