超高效液相色谱-串联质谱同时检测水稻中3种农药及2种代谢物残留

张斌斌; 董丰收; 潘兴鲁; 吴小虎; 徐军; 郑永权

doi:10.16801/j.issn.1008-7303.2022.0047

超高效液相色谱-串联质谱同时检测水稻中3种农药及2种代谢物残留

doi: 10.16801/j.issn.1008-7303.2022.0047

1.
中国农业科学院植物保护研究所，植物病虫害生物学国家重点实验室，北京 100193

基金项目: 国家自然科学基金(31901922)

详细信息

作者简介:
张斌斌，zbb35387@163.com

通讯作者:
董丰收，dongfengshou@caas.cn

中图分类号: TQ450.2；O657.63
计量
- 文章访问数: 224
- HTML全文浏览量: 56
- PDF下载量: 42
- 被引次数: 0
出版历程
- 收稿日期: 2022-01-16
- 录用日期: 2022-04-19
- 网络出版日期: 2022-08-23
- 刊出日期: 2022-12-10

Determination of the residues of three pesticides and two metabolites in rice by ultra-high performance liquid chromatography-tandem mass spectrometry

1.
Institute of Plant Protection, Chinese Academy of Agricultural Sciences, State Key Laboratory for Biology of Plant Diseases and Insect Pests, Beijing 100193, China

Funds: National Nature Science Foundation of China (31901922)

摘要

摘要: 建立并优化了基于超高效液相色谱-串联质谱 (UPLC-MS/MS) 同时检测水稻中甲氨基阿维菌素苯甲酸盐、氯虫苯甲酰胺、氟唑菌酰胺及其代谢物M700F008、M700F048的分析方法。稻壳和糙米经0.2%甲酸-乙腈提取后用C₁₈净化，稻秆经1%甲酸-乙腈提取后用N-丙基乙二胺 (PSA) 和石墨化碳黑 (GCB) 净化，流动相为甲醇和0.2%甲酸水溶液，梯度洗脱，采用T3色谱柱进行分离。电喷雾正离子模式 (ESI⁺) 采集，多反应监测 (MRM)，外标法定量。结果表明：5种供试化合物的质量浓度与峰面积间线性关系良好，决定系数均大于0.9955；5种化合物在水稻稻壳、糙米及稻秆3种基质中的平均回收率均在71%~119%之间，相对标准偏差 (RSD) 在1%~11%之间，满足残留分析的要求。在3地供试糙米样品中，甲氨基阿维菌素苯甲酸盐、氯虫苯甲酰胺和氟唑菌酰胺的残留量均低于中国规定的最大残留限量标准。
- 超高效液相色谱-串联质谱 /
- 水稻 /
- 甲氨基阿维菌素苯甲酸盐 /
- 氯虫苯甲酰胺 /
- 氟唑菌酰胺 /
- 代谢物 /
- 残留
Abstract: An analytical method based on ultra-high performance liquid chromatography-tandem mass spectrometry was established and optimized for simultaneous determination of emamectin benzoate, chlorantraniliprole, fluxapyroxad and its metabolites M700F008 and M700F048 in rice. Rice husk and brown rice were extracted with 0.2% formic acid-acetonitrile and cleaned up with C₁₈, while rice straw was extracted with 1% formic acid-acetonitrile and cleaned up with N-propyl ethylenediamine (PSA) and graphitized carbon black (GCB). T3 chromatographic column was used for separation and the mobile phase was methanol and 0.2% formic acid water. Electrospray ionization in positive ion mode was selected, the detection mode was multi-reaction monitoring, and external standard method was used for quantification. The results showed that there was a good linear relationship between the mass concentration and peak area of the five tested compounds, and the determination coefficient was greater than 0.9955. The average recoveries of the five compounds in rice husk, brown rice and rice straw were in range of 71%-119% with the relative standard deviations (RSD) of 1%-11%, which met the requirements of residue analysis. The residues of emamectin benzoate, chlorantraniliprole and fluxapyroxad in brown rice samples collected from three locations were all lower than MRLs in China.
- ultra-high performance liquid chromatography-tandem mass spectrometry /
- emamectin benzoate /
- chlorantraniliprole /
- fluxapyroxad /
- metabolite /
- rice /
- residue

HTML全文

1 甲维盐、氯虫苯甲酰胺、氟唑菌酰胺及其代谢物M700F008及M700F048结构式

1. The structural formula of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048

下载: 全尺寸图片幻灯片

图 1 甲维盐、氯虫苯甲酰胺、氟唑菌酰胺、M700F008和M700F048的质谱图及其在不同基质中的空白样品添加色谱图

A-甲维盐 emamectin benzoate；B-氯虫苯甲酰胺 chlorantraniliprole；C-氟唑菌酰胺 fluxapyroxad；D-M700F008；E-M700F048。0-标准溶液 standard solution；1-糙米添加 brown rice (1 mg/kg)；2-稻壳添加 rice husk (1 mg/kg)；3-稻秆添加 rice straw (1 mg/kg)。

Figure 1. Mass spectrogram of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048 and chromatogram of different blank matrix samples spiked with the five compounds

下载: 全尺寸图片幻灯片

图 2 甲维盐、氯虫苯甲酰胺、氟唑菌酰胺、M700F008及M700F048在不同基质和提取溶剂中的平均回收率

Figure 2. Average recovery of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048 in different matrices and extraction solvents

下载: 全尺寸图片幻灯片

图 3 不同净化剂对糙米、稻壳和稻秆中甲维盐、氯虫苯甲酰胺、氟唑菌酰胺、M700F008及M700F048回收率的影响

Figure 3. Effects of different cleanup sorbents on recoveries of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048 in brown rice, rice husk and rice straw

下载: 全尺寸图片幻灯片

表 1 甲维盐、氯虫苯甲酰胺、氟唑菌酰胺、M700F008和M700F048主要质谱检测参数

Table 1. Mass spectrometric parameters for the detection of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048

化合物 Compound	保留时间 Retention time/min	母离子 Precursor ion	子离子 Product ion	去簇电压 Declustering potential/V	碰撞能 Collision energy/eV
甲维盐 emamectin benzoate	2.44	886.5	158.1^*	110.00	40.00
甲维盐 emamectin benzoate	2.44	886.5	126.1	110.00	75.00
氯虫苯甲酰胺 chlorantraniliprole	2.42	483.9	452.9^*	80.00	28.00
氯虫苯甲酰胺 chlorantraniliprole	2.42	483.9	285.9	80.00	28.00
氟唑菌酰胺 fluxapyroxad	2.46	382.2	342.0	125.03	33.85
氟唑菌酰胺 fluxapyroxad	2.46	382.2	314.0^*	104.89	36.30
M700F008	2.47	368.2	348.1^*	93.44	23.22
M700F008	2.47	368.2	328.1	109.01	34.35
M700F048	2.40	530.1	368.1^*	82.29	21.79
M700F048	2.40	530.1	348.0	82.29	39.00
注：^* 为定量离子。Note: ^* represents quantitative ion.

下载: 导出CSV

表 2 甲维盐、氯虫苯甲酰胺、氟唑菌酰胺、M700F008和M700F048的线性回归方程、决定系数和基质效应

Table 2. Linear equation, determination coefficients, and matrix effects of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048

化合物 Compound	基质 Matrix	线性回归方程 Linear regression equation	决定系数 R²	斜率比值 k	线性范围 Linear range/(mg/kg)
甲维盐 emamectin benzoate	乙腈 Acetonitrile	y = 35248600x + 427415	0.9978	—	0.01~1
	糙米 Brown rice	y = 46968500x + 420911	0.9982	1.33
	稻壳 Rice husk	y = 21927591x + 221375	0.9996	0.62
	稻秆 Rice straw	y = 30013600x + 320622	0.9990	0.85
氯虫苯甲酰胺 chlorantraniliprole	乙腈 Acetonitrile	y = 13607100x-37150	0.9985	—	0.02~1
	糙米 Brown rice	y = 18372800x + 23839	0.9985	1.35
	稻壳 Rice husk	y = 7352155x + 31114	0.9997	0.54
	稻秆 Rice straw	y = 8747939x + 224273	0.9992	0.64
氟唑菌酰胺 fluxapyroxad	乙腈 Acetonitrile	y = 22437160x + 28345	0.9955	—	0.02~1
	糙米 Brown rice	y = 25435816x + 20798	0.9981	1.13
	稻壳 Rice husk	y = 5488118x + 12804	0.9998	0.24
	稻秆 Rice straw	y = 11379354x + 25084	0.9996	0.51
M700F008	乙腈 Acetonitrile	y = 31321500x-24314	0.9998	—	0.02~1
	糙米 Brown rice	y = 28296164x + 15514	0.9994	0.90
	稻壳 Rice husk	y = 3299129x + 19778	0.9998	0.11
	稻秆 Rice straw	y = 12350021x + 15281	0.9994	0.39	0.05~1
M700F048	乙腈 Acetonitrile	y = 20029051x-84841	0.9978	—	0.02~1
	糙米 Brown rice	y = 22392939x + 14154	0.9993	1.12
	稻壳 Rice husk	y = 3825229x + 19778	0.9991	0.19
	稻秆 Rice straw	y = 7487584x + 12745	0.9996	0.37	0.05~1

下载: 导出CSV

表 3 甲维盐、氯虫苯甲酰胺、氟唑菌酰胺、M700F008及M700F048在糙米、稻壳和稻秆中的添加回收率及RSD

Table 3. Recovery and RSDs of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048 in three matrices at 3 spiked levels

化合物 Compound	糙米 Brown rice			稻壳 Rice husk			稻秆 Rice straw
化合物 Compound	添加水平 Spiked level/ (mg/kg)	回收率 Recovery/ %	相对标准偏差 RSD/%	添加水平 Spiked level/ (mg/kg)	回收率 Recovery/ %	相对标准偏差 RSD/%	添加水平 Spiked level/ (mg/kg)	回收率 Recovery/ %	相对标准偏差 RSD/%
甲维盐 emamectin benzoate	0.01	85	3.0	0.01	98	3.7	0.01	107	2.5
	0.02	92	1.4	0.02	94	5.0	0.02	97	1.5
	1	85	2.8	1	97	4.2	1	102	2.4
氯虫苯甲酰胺 chlorantraniliprole	0.02	105	3.9	0.02	106	4.0	0.02	93	4.1
	0.5	92	1.9	0.5	106	1.9	0.5	104	6.2
	1	95	2.7	1	107	2.8	1	97	8.8
氟唑菌酰胺 fluxapyroxad	0.02	103	5.0	0.02	119	3.7	0.02	99	5.0
	1	97	6.1	1	104	4.7	1	102	7.0
	5	97	3.0	5	107	1.7	5	108	4.4
M700F008	0.02	104	4.4	0.02	112	5.7	0.05	79	4.7
	1	92	2.3	1	98	5.3	1	83	4.2
	5	96	1.4	5	107	1.1	5	87	4.3
M700F048	0.02	88	2.4	0.02	104	10.7	0.05	73	2.0
	1	83	2.3	1	78	9.5	1	71	1.1
	5	87	1.7	5	81	5.9	5	74	4.3

下载: 导出CSV

表 4 甲维盐、氯虫苯甲酰胺、氟唑菌酰胺、M700F008及M700F048在糙米、稻壳和稻秆中的最终残留量(mg/kg)

Table 4. The terminal residues of emamectin benzoate, chlorantraniliprole, fluxapyroxad, M700F008 and M700F048 in rice samples (mg/kg)

化合物 Compound	样品采收地 Sample collection site	糙米 Brown rice		稻壳 Rice husk		稻秆 Rice straw
化合物 Compound	样品采收地 Sample collection site	21 d	28 d	21 d	28 d	21 d	28 d
甲维盐 emamectin benzoate	湖北 Hubei	< LOQ	< LOQ	< LOQ	< LOQ	< LOQ	< LOQ
	广西 Guangxi	< LOQ	<LOQ	< LOQ	< LOQ	< LOQ	< LOQ
	广东 Guangdong	< LOQ	< LOQ	< LOQ	< LOQ	< LOQ	< LOQ
氯虫苯甲酰胺 chlorantraniliprole	湖北 Hubei	< LOQ	0.02	3.12	1.29	1.15	0.29
	广西 Guangxi	< LOQ	< LOQ	2.40	2.56	0.17	0.23
	广东 Guangdong	0.03	< LOQ	3.11	1.29	0.36	0.16
氟唑菌酰胺^a fluxapyroxad^a	湖北 Hubei	0.08	0.07	2.50	1.36	3.64	1.48
氟唑菌酰胺^a fluxapyroxad^a	广西 Guangxi	0.07	0.06	4.09	3.60	0.56	0.24
注：^a为总残留量 (氟唑菌酰胺总残留量 = 氟唑菌酰胺残留量 + M700F008残留量 × 1.038 + M700F048残留量 × 0.72^[15]，残留量小于定量限以定量限计)。Note: ^a represents the total residues (total residue of fluxapyroxad = fluxapyroxad residue + M700F008 residue × 1.038 + M700F048 residue × 0.72, the limit of quantification was used for the calculation when the residue was lower than the limit of quantification).

下载: 导出CSV

参考文献(15)

[1]	ANE N U, HUSSAIN M. Diversity of insect pests in major rice growing areas of the world[J]. J Entomol Zool Stud, 2015, 4(1): 36-41.
[2]	MEW T W, LEUNG H, SAVARY S, et al. Looking ahead in rice disease research and management[J]. Crit Rev Plant Sci, 2004, 23(2): 103-127. doi: 10.1080/07352680490433231
[3]	蒲小明, 王爱臣, 林壁润, 等. 5% 甲维盐水分散粒剂防治鳞翅目害虫田间效果[J]. 广东农业科学, 2021, 48(4): 106-110. PU X M, WANG A C, LIN B R, et al. Control effect of 5% emamectin benzoate water dispersible granule against Lepidoptera pests in field[J]. Guangdong Agric Sci, 2021, 48(4): 106-110.
[4]	林炳松. 甲氨基阿维菌素苯甲酸盐和氯虫苯甲酰胺环保剂型研制[D]. 保定: 河北农业大学, 2020. LIN B S. Development of environmental friendly formulations of emamectin benzoate and chlorantraniliprole[D]. Baoding: Hebei Agricultural University, 2020.
[5]	伏荣桃, 王剑, 陈诚, 等. 氯虫苯甲酰胺防控水稻二化螟的应用效果评价[J]. 中国植保导刊, 2020, 40(1): 78-81. doi: 10.3969/j.issn.1672-6820.2020.01.015 FU R T, WANG J, CHEN C, et al. Effect evaluation of chlorantraniliprole applied in rice Chilo suppressalis Walker control[J]. China Plant Prot, 2020, 40(1): 78-81. doi: 10.3969/j.issn.1672-6820.2020.01.015
[6]	路彩红, 杨林, 申流柱, 等. 超高效液相色谱-串联质谱法检测水稻和土壤中氟唑菌酰胺的残留及消解动态[J]. 植物保护, 2016, 42(4): 132-136. doi: 10.3969/j.issn.0529-1542.2016.04.021 LU C H, YANG L, SHEN L Z, et al. Residues and degradation determination of fluxapyroxad in rice and soil by UPLC-MS/MS[J]. Plant Prot, 2016, 42(4): 132-136. doi: 10.3969/j.issn.0529-1542.2016.04.021
[7]	王刚, 吕亮, 刘吉永, 等. 氟唑菌酰胺的合成与生物活性[J]. 现代农药, 2017, 16(4): 12-14. doi: 10.3969/j.issn.1671-5284.2017.04.003 WANG G, LV L, LIU J Y, et al. Synthesis and bioactivity of fluxapyroxad[J]. Mod Agrochem, 2017, 16(4): 12-14. doi: 10.3969/j.issn.1671-5284.2017.04.003
[8]	国家食品安全标准食品中农药最大残留限量: GB 2763—2021. [S]. 北京: 中国农业出版社, 2021. National Food Safety Standard−Maximum residue limits for pesticides in food: GB 2763—2021. [S]. Beijing: China Agriculture Press, 2021.
[9]	DONG F S, CHEN X, LIU X G, et al. Simultaneous determination of five pyrazole fungicides in cereals, vegetables and fruits using liquid chromatography/tandem mass spectrometry[J]. J Chromatogr A, 2012, 1262: 98-106. doi: 10.1016/j.chroma.2012.08.100
[10]	CHEN X X, DONG F S, XU J, et al. Effective monitoring of fluxapyroxad and its three biologically active metabolites in vegetables, fruits, and cereals by optimized QuEChERS treatment based on UPLC-MS/MS[J]. J Agric Food Chem, 2016, 64(46): 8935-8943. doi: 10.1021/acs.jafc.6b03253
[11]	农作物中农药残留试验准则: NY/T 788—2018[S]. 北京: 中国农业出版社, 2018. Guideline for the testing of pesticide residues in crops: NY/T 788—2018[S]. Beijing: China Agriculture Press, 2018.
[12]	齐晓雪, 董崭, 冯义志, 等. 超高效液相色谱串联质谱法检测水稻中五氟磺草胺、吡嘧磺隆和丙草胺的残留量[J]. 农药, 2021, 60(4): 290-293. doi: 10.16820/j.cnki.1006-0413.2021.04.015 QI X X, DONG Z, FENG Y Z, et al. Determination of the residues of penoxsulam, pyrazosulfuron-ethyl and pretilachlor in rice by ultra high performance liquid chromatography-tandem mass spectrometry[J]. Agrochemicals, 2021, 60(4): 290-293. doi: 10.16820/j.cnki.1006-0413.2021.04.015
[13]	刘亚伟, 董一威, 孙宝利, 等. QuEChERS 在食品中农药多残留检测的应用研究进展[J]. 食品科学, 2009, 30(9): 285-289. doi: 10.3321/j.issn:1002-6630.2009.09.067 LIU Y W, DONG Y W, SUN B L, et al. Summary of application of QuEChERS method in multi-residue determination of pesticides in food[J]. Food Sci, 2009, 30(9): 285-289. doi: 10.3321/j.issn:1002-6630.2009.09.067
[14]	LI S S, LIU X G, ZHU Y L, et al. A statistical approach to determine fluxapyroxad and its three metabolites in soils, sediment and sludge based on a combination of chemometric tools and a modified quick, easy, cheap, effective, rugged and safe method[J]. J Chromatogr A, 2014, 1358: 46-51. doi: 10.1016/j.chroma.2014.06.088
[15]	郭亚歌, 徐军, 董丰收, 等. 关于 JMPR 植物源产品残留定义中含有代谢产物的农药在进行膳食风险评估时残留数据计算的介绍[J]. 植物保护, 2021, 47(2): 169-178. doi: 10.16688/j.zwbh.2020401 GUO Y G, XU J, DONG F S, et al. Conversion of residue data for pesticide residue definition containing metabolites in dietary risk assessment for plant commodities of JMPR[J]. Plant Prot, 2021, 47(2): 169-178. doi: 10.16688/j.zwbh.2020401