超高效液相色谱-串联质谱法检测绵羊组织中地昔尼尔残留的方法学研究

doi:10.16431/j.cnki.1671-7236.2023.08.038

摘要/Abstract

摘要： 【目的】建立一种绵羊组织中地昔尼尔及其代谢产物2,4,6-三氨基-5-氰基嘧啶质量浓度的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。【方法】对采集的绵羊肌肉、脂肪、肝脏、肾脏组织进行前处理,以乙腈和七氟丁酸水溶液为流动相,利用Acquity UPLC Beh C18色谱柱进行梯度洗脱;流速为0.15 mL/min,柱温为30 ℃,进样量为5 μL。在正离子扫描模式下通过多离子反应监测采集数据,采用外标法测定地昔尼尔及其代谢产物浓度,并考察该检测方法的线性范围、检测限、定量限、回收率、批内与批间精密度、准确度和稳定性。【结果】建立的检测方法灵敏度高,检测限为10 μg/kg,定量限为20 μg/kg,地昔尼尔和2,4,6-三氨基-5-氰基嘧啶的添加浓度为20~1 000 μg/kg,标准曲线的线性关系良好,相关系数均≥0.99。将地昔尼尔和2,4,6-三氨基-5-氰基嘧啶以高、中、低、定量限4个水平分别添加到各空白组织中,测得地昔尼尔和2,4,6-三氨基-5-氰基嘧啶的回收率为70%~110%,批内和批间变异系数均<15%。在考察的条件下,地昔尼尔和2,4,6-三氨基-5-氰基嘧在各组织中的稳定性良好。【结论】建立的UPLC-MS/MS检测方法专属性强、重复性好、灵敏度高,可适用于地昔尼尔乳剂在绵羊组织内的药物代谢动力学和残留研究。

关键词: 地昔尼尔; 2,4,6-三氨基-5-氰基嘧啶; 超高效液相色谱-串联质谱法; 绵羊

Abstract: 【Objective】 This study was constructed to establish a method for the determination of the mass concentration of dicyclanil and its metabolites 2,4,6-triamino-5-pyrimidinecarbonitrile in sheep tissue by ultra-high liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).【Method】 Sheep tissues (muscle,fat,liver and kidney) were pretreated.Acetonitrile and heptafluorobutyric acid aqueous solution were used as mobile phases,Acquity UPLC Beh C18 column was used for gradient elution.The flow rate was 0.15 mL/min,the column temperature was 30 ℃ and the injection volume was 5 μL.The data were collected by multi-reaction monitoring in positive ion scanning mode,and the concentrations of dicyclanil and its metabolites were determined by external standard method.The linear range,detection limit,quantitative limit,recovery rate,intra-assay and inter-assay precision,accuracy and stability of the method were investigated.【Result】 The established detection method had high sensitivity,the detection limit was 10 μg/kg,and the limit of quantitation was 20 μg/kg.The addition concentrations of dicyclanil and 2,4,6-triamino-5-pyrimidinecarbonitrile were in the range of 20 to 1 000 μg/kg,and the linear relationship between the standard curves was good,with correlation coefficients ≥0.99.The recoveries of dicyclanil and 2,4,6-triamino-5-pyrimidinecarbonitrile were 70% to 110% when they were added to the blank tissues at high,medium,low and limited quantification levels,and the coefficient of variation within and between batches was less than 15%.The stability of dicyclanil and 2,4,6-triamino-5-pyrimidinecarbonitrile in different tissues was good under the conditions of investigation.【Conclusion】 The established UPLC-MS/MS assay was specific,reproducible and sensitive,and could be used for pharmacokinetic and residual studies of dicyclanil emulsion in sheep tissues.

Key words: dicyclanil; 2,4,6-triamino-5-pyrimidinecarbonitrile; UPLC-MS/MS; sheep

中图分类号:

S859.84

仇亚伟, 季辉, 彭麟, 江善祥, 高修歌. 超高效液相色谱-串联质谱法检测绵羊组织中地昔尼尔残留的方法学研究[J]. 中国畜牧兽医, 2023, 50(8): 3403-3412.

QIU Yawei, JI Hui, PENG Lin, JIANG Shanxiang, GAO Xiuge. Methodology Study on Determination of Dicyclanil Residual in Sheep Tissue by Ultra-performance Liquid Chromatography-tandem Mass Spectrometry[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3403-3412.

参考文献

[1] SCHMID H R,TULDER G V,JUNQUERA P.Field efficacy of the insect growth regulator dicyclanil for flystrike prevention on lambs[J].Veterinary Parasitology,1999,86(2):147-151.
[2] 詹希美.人体寄生虫学[M].北京:人民卫生出版社,2001. ZHAN X M.Human Parasitology[M].Beijing:People's Medical Publishing House,2001.(in Chinese)
[3] SALES N,SUANN M,KOEFORD K.Dicyclanil resistance in the Australian sheep blowfly,Lucilia cuprina,substantially reduces flystrike protection by dicyclanil and cyromazine based products[J].International Journal for Parasitology:Drugs and Drug Resistance,2020,14:118-125.
[4] GIFFORD L J,DUNSIRE J P.Absorption,distribution,and excretion of[2-14C] Pyrimidyl CGA 183893 after topical administration to sheep[R].Report No.10067,Inveresk Research International,Trenent,Scotland,1995.
[5] MACLEAN C L,DUNSIRE J P.Absorption,distribution,and excretion of[2-14C] Pyrimidyl CGA 183893 after pour-on administration to sheep[R].Report No.1337,Inveresk Research International,Trenent,Scotland,1998.
[6] ANDERSON A R,SPEIRS G C.The absorption,distribution,and excretion of[Pyrimidyl-2-14C] CGA 183893 after pour-on administration to sheep[R].Report No.14072,Inveresk Research International,Trenent,Scotland,1998.
[7] 吴海霞,冯培生,张素霞,等.高效液相色谱法测定动物性食品中地昔尼尔残留量[J].中国畜牧兽医,2010,37(11):224-226. WU H X,FENG P S,ZHANG S X,et al.Determination of residual dicyclanil in animal food by HPLC[J].China Animal Husbandry&Veterinary Medicine,2010,37(11):224-226.(in Chinese)
[8] 中华人民共和国农业农村部.中华人民共和国农业农村部公告第326号.兽药残留消除试验指导原则[EB/OL].(2020-08-26)[2023-03-01] .http://www.moa.gov.cn/nybgb/2020/202009/202011/t20201124_6356888.htm. MINISTRY OF AGRICULTURE AND RURAL AFFAIRS OF THE PEOPLE'S REPUBLIC OF CHINA.Announcement No.326 of the Ministry of Agriculture and Rural Affairs of the People's Republic of China.Guidelines for veterinary drug residue elimination testing[EB/OL].(2020-08-26)[2023-03-01] .http://www.moa.gov.cn/nybgb/2020/202009/202011/t20201124_6356888.htm.(in Chinese)
[9] 中国国家标准化管理委员会.中华人民共和国出入境检验检疫行业标准:SN/T 2153-2008[S].北京:中华人民共和国国家质量监督检验检疫总局,2008:7. STANDARDIZATION ADMINISTRATION OF THE PEOPLE'S REPUBLIC OF CHINA.The People's Republic of China entry-exit inspection and quarantine industry standard:SN/T 2153-2008[S].Beijing:General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China,2008:7.(in Chinese)
[10] 方忠意,刘素梅.超高效液相色谱法测定饲料中地昔尼尔[J].中国饲料,2010,6:38-40. FANG Z Y,LIU S M.The determination of dicyclanil in feed by ultra performance liquid chromatographic[J].China Feed,2010,6:38-40.(in Chinese)
[11] 方忠意,刘素梅,班付国,等.超高效液相色谱-电喷雾串联质谱法测定饲料中地昔尼尔[J].饲料工业,2009,30(21):38-40. FANG Z Y,LIU S M,BAN F G,et al.Determination of dicyclanil in feeds using ultra-performance liquid chromatography-electrospray tandem mass spectrometry[J].Feed Industry,2009,30(21):38-40.(in Chinese)
[12] 樊苑牧,俞雪钧,顾晓俊,等.高效液相色谱法测定含脂羊毛中灭蝇胺和环虫腈[J].分析化学,2010,38(1):113-116. FAN Y M,YU X J,GU X J,et al.Determination of cyromazine and dicyclanil residues on greasy wool by high performance liquid chromatography[J].Analytical Chemistry,2010,38(1):113-116.(in Chinese)
[13] 刘新辉,王情情,张瑗,等.Oasis PRiME HLB净化法测定动物源性食品中的兽药多残留[J].农产品质量与安全,2019,101(5):15-20. LIU X H,WANG Q Q,ZHANG Y,et al.Oasis PRiME HLB purification method for the determination of veterinary drug residues in foods of animal origin[J].Quality and Safety of Agro-Products,2019,101(5):15-20.(in Chinese)
[14] 王丹,谷翠梅,李琳,等.EMR-Lipid QuEChERS UPLC-MS/MS法测定动物源性食品中残留氯霉素及4种硝基呋喃代谢物[J].中国卫生工程学,2023,22(1):17-19. WANG D,GU C M,LI L,et al.Determination of chloramphenicol residues and four nitrofuran metabolites in foods of animal origin using EMR-Lipid QuEChERS UPLC-MS/MS[J].Chinese Journal of Public Health Engineering,2023,22(1):17-19.(in Chinese)
[15] 李宁,肖琎,王雪松,等.高效液相色谱-串联质谱法同时测定动物源性食品中15种禁限兽药残留[J].中国饲料,2023,2:97-100. LI N,XIAO J,WANG X S,et al.Simultaneous determination of 15 prohibited and restricted veterinary drug residues in animal derived food by high performance liquid chromatographytandem mass spectrometry[J].China Feed,2023,2:97-100.(in Chinese)
[16] 梁晶晶,沈丹,徐潇颖,等.超高效液相色谱-串联质谱法测定动物源性食品中12种青霉素类药物残留量[J].食品与发酵工业,2018,44(5):265-269. LIANG J J,SHEN D,XU X Y,et al.Determination of 12 penicillins antibiotics residues in foods from animal origin by UPLC-MS/MS[J].Food and Fermentation Industries,2018,44(5):265-269.(in Chinese)
[17] 杨昊.固相萃取-超高效液相色谱-串联质谱同时检测动物源性食品兽药残留[J].当代化工,2017,46(11):2386-2388. YANG H.Simultaneous detection of animal origin veterinary drug residues by solid-phase extraction-ultra-performance liquid chromatography-tandem mass spectrometry[J].Contemporary Chemical Industry,2017,46(11):2386-2388.(in Chinese)
[18] 高洁,朱莉萍,倪永付,等.超高效液相色谱-串联质谱法测定多脂肪类动物源性食品中硝基呋喃代谢物[J].食品安全质量检测学报,2018,9(6):1362-1368. GAO J,ZHU L P,NI Y F,et al.Determination of nitrofuran metabolites in foods of multi-fat animal origin by ultra-high performance liquid chromatography-tandem mass spectrometry[J].Journal of Food Safety&Quality,2018,9(6):1362-1368.(in Chinese)
[19] 翟莉芳,刘战花.UPLC-MS/MS快速检测动物源性食品中多种兽药残留[J].惠州学院学报,2018,38(3):37-42. ZHAI L F,LIU Z H.Rapid detection of multiple veterinary drug residues in animal derived foods by UPLC-MS/MS[J].Journal of Huizhou University,2018,38(3):37-42.(in Chinese)
[20] 张勇.地昔尼尔的药理学研究进展[J].四川畜牧兽医,2013,40(6):31-32. ZHANG Y.Progress in pharmacological research of decicinil[J].Sichuan Animal&Veterinary Sciences,2013,40(6):31-32.(in Chinese)
[21] 王雅娟,张杨杨,郭亚文,等.动物源性食品中抗球虫药物残留的色谱和质谱检测技术研究进展[J].中国兽医学报,2018,38(12):8. WANG Y J,ZHANG Y Y,GUO Y W,et al.An overview on chromatographic and mass spectrometry analytical methods for the multiresidue detection of anticoccidiosis drug in tissues of animal origin[J].Chinese Journal of Veterinary Science,2018,38(12):8.(in Chinese)
[22] 杨爽.高效液相色谱-串联质谱法检测动物血清、尿液中硝基呋喃代谢物残留[J].中国畜牧杂志,2018,54(7):5. YANG S.High performance liquid chromatography-tandem mass spectrometry for the detection of nitrofuran metabolite residues in animal serum and urine[J].Chinese Journal of Animal Science,2018,54(7):5.(in Chinese)
[23] 王根林,孟颖.高效液相色谱法测定吡虫啉杀虫剂中4种关键杂质残留量[J].理化检验:化学分册,2021,57(6):4. WANG G L,MENG Y.HPLC determination of 4 key impurities residues in lmidacloprid insecticide[J].Physical Testing and Chemical Analysis (Part B:Chemical Analysis),2021,57(6):4.(in Chinese)