分散固相萃取-超高效液相色谱-串联质谱法测定海龙中11种磺胺类抗菌药物残留

王小乔; 李坚; 许晓辉; 李剑勇; 吴福祥; 张虹艳; 潘秀丽; 李赟

doi:10.16495/j.1006-3757.2023.01.003

分散固相萃取-超高效液相色谱-串联质谱法测定海龙中11种磺胺类抗菌药物残留

doi: 10.16495/j.1006-3757.2023.01.003

1.
兰州市食品药品检验检测研究院/食品中农药兽药残留监控国家市场监管重点实验室，甘肃兰州　730050
2.
中国农业科学院兰州畜牧与兽药研究所，甘肃兰州　730050

基金项目: 甘肃省市场监督管理局科技计划资助项目（SSCJG-SP-A202204）

详细信息

作者简介:
王小乔（1973−），女，本科，研究方向：食品药品检验检测与研究，E-mail：378510575@qq.com

通讯作者:
李坚（1969−），男，本科，主管药师，研究方向：食品药品检验检测与研究，E-mail：237561297@qq.com

中图分类号: O657. 63
计量
- 文章访问数: 134
- HTML全文浏览量: 66
- PDF下载量: 88
- 被引次数: 0
出版历程
- 收稿日期: 2022-10-19
- 录用日期: 2023-01-08
- 修回日期: 2023-01-08
- 刊出日期: 2023-03-31

Determination of 11 Sulfonamide Antibacterial Drug Residues in Syngnathus by Ultra-Performance Liquid Chromatography-Mass Spectrometry Combined with Dispersed Solid Phase Extraction

1.
Lanzhou Institute for Food and Drug Control/Key Laboratory of Pesticides and Veterinary Drugs Monitoring for State Market Regulation, Lanzhou 730050, China
2.
Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS, Lanzhou 730050, China

摘要

摘要: 采用超高效液相色谱串联三重四极杆质谱（UPLC-MS/MS）技术，建立了测定海龙中11种磺胺类抗菌药物残留量的分析方法. 样品经2%甲酸-乙腈超声提取，增强型脂质去除分散固相萃取净化管（EMR-Lipid dSPE）净化，采用Waters ACQUITY BEH C₁₈色谱柱（1.7 μm，2.1 mm×100 mm）分离，电喷雾离子源电离，正离子多反应监测模式（MRM）检测，以含2 mmol/L乙酸铵-0.1%甲酸的水溶液和含0.1%甲酸的乙腈溶液为流动相进行梯度洗脱，外标法定量. 11种磺胺类抗菌药物在0～30 ng/mL的质量浓度范围内线性关系良好，相关系数（R²）均不低于0.995 9，检出限（LOD）与定量限（LOQ）分别在0.11～1.30 µg/kg和0.37～4.40 µg/kg之间，11种磺胺类抗菌药物在质量浓度为20.0、50.0、100.0 µg/kg时的三个加标水平的平均回收率为62.5%～118.1%，相对标准偏差为2.1%～9.2%. 方法前处理简便、准确性好，可用于海龙中11种磺胺类抗菌药物残留的快速筛查.
- 海龙 /
- 超高效液相色谱-三重四极杆质谱联用法 /
- 磺胺类药物 /
- 分散固相萃取 /
- 残留量
Abstract: An analytical method for the determination of 11 sulfonamide antibacterial drug residues in Syngnathus by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) has been established. The samples were extracted by ultrasonication with 2% formic acid-acetonitrile, purified by an enhanced matrix removal lipid-dispersed solid phase extraction (EMR-Lipid dSPE), and separated on a Waters ACQUITY BEH C₁₈ column (1.7 μm, 2.1 mm×100 mm). The analytes were detected by mass spectrometry using the positive electrospray ionization mode under multiple reaction monitoring mode (MRM), with 2 mmol/L ammonium acetate-0.1% formic acid in aqueous solution and 0.1% formic acid in acetonitrile as mobile phases by a gradient elution program, and quantified by the external standard method. The results showed that the 11 antibacterial sulfonamides had good linearity in the range of 0~30 ng/mL, the correlation coefficient (R²) were all greater than 0.9959, the limit of detection (LOD) and quantitation (LOQ) were in the range of 0.11~1.30 µg/kg and 0.37~4.40 µg/kg, respectively. At three spiked levels of 20.0, 50.0 and 100.0 µg/kg, the spiked average recoveries of the analytes were in the range of 62.5%~118.1%, and the relative standard deviation was 2.1%~9.2%. The method is accurate, straightforward, and is suitable for the rapid screening of 11 sulfonamide antibacterial drug residues in Syngnathus.
- Syngnathus /
- UPLC-MS/MS /
- sulfonamides /
- solid phase extraction /
- residues

HTML全文

图 1 不同提取溶剂的提取效率

Figure 1. Extraction efficiencies of different extraction solvents

下载: 全尺寸图片幻灯片

图 2 11种磺胺类药物对照品提取离子流图

Figure 2. Extract ion chromatogram of 11 standards of sulfonamides

下载: 全尺寸图片幻灯片

图 3 11种磺胺类药物的基质效应

Figure 3. Matrix effects of 11 sulfonamides

下载: 全尺寸图片幻灯片

图 4 阴性样品总离子流图

Figure 4. Total ion chromatogram of negative sample

下载: 全尺寸图片幻灯片

表 1 11种磺胺类药物的质谱参数及保留时间

Table 1. Mass spectrum parameters and retention times of 11 sulfonamides

序号	名称	保留时间/min	母离子/（m/z）	子离子/（m/z）	碎裂电压/V	碰撞能量/eV
1	磺胺甲噁唑	4.665	254.1	156.0^*/108.0	100	10/25
2	磺胺氯哒嗪	4.449	285.0	156.0^*/108.0	100	10/25
3	磺胺间甲氧嘧啶	4.006	281.1	156.0^*/108.0	105	15/25
4	磺胺甲基嘧啶	3.607	265.1	172.0/156.0^*	110	12/15
5	磺胺邻二甲氧嘧啶	4.585	311.1	156.0^*/92.0	120	15/30
6	磺胺间二甲氧嘧啶	5.067	311.0	156.0^*/108.0	130	20/26
7	磺胺二甲嘧啶	3.950	279.1	186.1^*/156.1	120	15/16
8	磺胺嘧啶	2.902	251.1	156.0^*/108.0	100	10/22
9	磺胺甲二唑	3.969	271.0	156.0^*/108.0	90	10/22
10	磺胺噻唑	3.208	256.0	156.0^*/108.0	100	10/21
11	磺胺甲氧哒嗪	4.287	281.1	156.0^*/108.0	105	15/25
注：“^*”为定量离子

下载: 导出CSV

表 2 11种磺胺类药物的线性关系、检出限、定量限

Table 2. Linear relationship, LOD, LOQ of 11 sulfonamides

序号	名称	线性方程	R²	线性范围/（ng/mL）	LOD/（µg/kg）	LOQ/（µg/kg）
1	磺胺甲噁唑	y=139.010x+28.158 7	0.995 9	0～30	0.48	1.60
2	磺胺氯哒嗪	y=107.763x+15.615 7	0.997 9	0～30	0.84	2.80
3	磺胺间甲氧嘧啶	y=171.804x+35.380 5	0.998 5	0～30	0.73	2.40
4	磺胺甲基嘧啶	y=141.980x+7.901 7	0.998 6	0～30	1.30	4.40
5	磺胺邻二甲氧嘧啶	y=1 038.220x+227.398 0	0.998 1	0～30	0.11	0.37
6	磺胺间二甲氧嘧啶	y=596.926x+90.787 3	0.998 6	0～30	0.38	1.30
7	磺胺二甲嘧啶	y=325.171x+42.142 3	0.998 8	0～30	0.17	0.54
8	磺胺嘧啶	y=146.238x+62.685 0	0.999 2	0～30	0.22	0.70
9	磺胺甲二唑	y=120.498x-3.966 5	0.999 2	0～30	0.95	3.10
10	磺胺噻唑	y=117.540x+14.036 4	0.999 5	0～30	0.32	0.97
11	磺胺甲氧哒嗪	y=165.395x+35.380 5	0.998 5	0～30	0.76	2.50

下载: 导出CSV

表 3 11种磺胺类药物的回收率及RSD（n=6）

Table 3. Recoveries and RSD of 11 sulfonamides

序号	名称	20.0 µg/kg				50.0 µg/kg				100.0 µg/kg
序号	名称	加入质量/ng	测得质量/ng	平均回收率/%	RSD/ %	加入质量/ng	测得质量/ng	平均回收率/%	RSD/ %	加入质量/ng	测得质量/ng	平均回收率/%	RSD/ %
1	磺胺甲噁唑	20.68	14.08	68.1	9.2	51.70	38.88	75.2	4.3	103.40	82.41	79.7	3.9
2	磺胺氯哒嗪	21.76	18.54	85.2	7.7	54.40	46.51	85.5	3.7	108.80	88.45	81.3	2.6
3	磺胺间甲氧嘧啶	21.68	16.19	74.7	4.8	54.20	52.46	96.8	3.2	108.40	83.36	76.9	2.4
4	磺胺甲基嘧啶	22.04	16.18	73.4	3.8	55.10	42.87	77.8	6.9	110.20	85.07	77.2	6.5
5	磺胺邻二甲氧嘧啶	21.70	18.44	85.0	2.7	54.25	47.68	87.9	3.0	108.50	98.08	90.4	2.6
6	磺胺间二甲氧嘧啶	21.82	19.72	90.4	4.5	54.55	55.80	102.3	2.5	109.10	118.05	108.2	2.1
7	磺胺二甲嘧啶	21.76	21.37	98.2	3.0	54.40	53.91	99.1	3.4	108.80	108.04	99.3	2.8
8	磺胺嘧啶	22.76	17.41	76.5	4.6	56.90	45.63	80.2	5.8	113.80	84.89	74.6	4.3
9	磺胺甲二唑	21.00	24.42	116.3	6.9	52.50	62.00	118.1	3.1	105.00	116.02	110.5	4.7
10	磺胺噻唑	20.80	13.00	62.5	5.0	52.00	32.92	63.3	4.9	104.00	65.21	62.7	6.0
11	磺胺甲氧哒嗪	22.52	15.67	69.6	4.3	56.30	37.10	65.9	2.9	112.60	74.65	66.3	4.3

下载: 导出CSV

参考文献(14)

[1]	国家药典委员会. 中华人民共和国药典(一部2020版)[M]. 北京: 中国医药科技出版社, 2020:306.
[2]	吴伟建, 王燕, 王斌, 等. 基于聚类、主成分和判别分析的海龙红外指纹图谱研究[J]. 中国药学杂志,2013,48(18):1540-1545 WU Weijian, WANG Yan, WANG Bin, et al. Infrared fingerprint analysis of syngnathus' s ethanol extracts coupled with cluster analysis, principal component analysis and discriminant analysis[J]. Chinese Pharmaceutical Journal,2013,48 (18):1540-1545.
[3]	王梦月, 韦静斐, 史海明, 等. 海龙药材及其伪品的HPLC指纹图谱研究[J]. 中国药学杂志,2009,44(24):1847-1851 doi: 10.3321/j.issn:1001-2494.2009.24.002 WANG Mengyue, WEI Jingfei, SHI Haiming, et al. Study on HPLC fingerprint of syngnathus and its adulterants[J]. Chinese Pharmaceutical Journal,2009,44 (24):1847-1851. doi: 10.3321/j.issn:1001-2494.2009.24.002
[4]	蒋超, 袁媛, 李军德, 等. 尖海龙商品调查与《中国药典》中海龙的基原考证[J]. 中国实验方剂学杂志,2019,25(17):104-112 doi: 10.13422/j.cnki.syfjx.20191713 JIANG Chao, YUAN Yuan, LI Junde, et al. Syngnathus acus in herbal markets and zoological origin of syngnathus in China pharmacopoeia[J]. Chinese Journal of Experimental Traditional Medical Formulae,2019,25 (17):104-112. doi: 10.13422/j.cnki.syfjx.20191713
[5]	颜洁. 海龙质量标准的定性研究[D]. 青岛: 中国海洋大学, 2014 YAN Jie. Qualitative studies on quality control of sea dragon[D]. Qingdao: Ocean University of China, 2014.
[6]	丁晴, 仇雅静, 房克慧, 等. 动物来源中药材、饮片质量现状及原因分析[J]. 中国中药杂志,2015,40(21):4309-4312 DING Qing, QIU Yajing, FANG Kehui, et al. Animal drugs quality status and reason analysis[J]. China Journal of Chinese Materia Medica,2015,40 (21):4309-4312.
[7]	许晓辉, 李剑勇, 朱仁愿, 等. 动物源性中药材兽药残留检测技术研究进展[J]. 中国兽药杂志,2021,55(6):45-52 XU Xiaohui, LI Jianyong, ZHU Renyuan, et al. Research progress of detecting technology of veterinary drug residues in animal-derived Chinese medicinal materials[J]. Chinese Journal of Veterinary Drug,2021,55 (6):45-52.
[8]	唐溱. 我国动物相关法律法规对动物类中药材利用与发展的影响[J]. 中草药,2021,52(24):7718-7727 doi: 10.7501/j.issn.0253-2670.2021.24.034 TANG Zhen. Influence of legislations related to animal on utilization and development of animal traditional Chinese medicine in China[J]. Chinese Traditional and Herbal Drugs,2021,52 (24):7718-7727. doi: 10.7501/j.issn.0253-2670.2021.24.034
[9]	缪苗, 黄一心, 沈建, 等. 水产品安全风险危害因素来源的分析研究[J]. 食品安全质量检测学报,2018,9(19):5195-5201 doi: 10.3969/j.issn.2095-0381.2018.19.031 MIAO Miao, HUANG Yixin, SHEN Jian, et al. Analysis and research on hazard sources of aquatic products[J]. Journal of Food Safety & Quality,2018,9 (19):5195-5201. doi: 10.3969/j.issn.2095-0381.2018.19.031
[10]	陈胜军, 李来好, 杨贤庆, 等. 我国水产品安全风险来源与风险评估研究进展[J]. 食品科学,2015,36(17):300-304 doi: 10.7506/spkx1002-6630-201517055 CHEN Shengjun, LI Laihao, YANG Xianqing, et al. Progress in the risk sources and assessment of aquatic products in China[J]. Food Science,2015,36 (17):300-304. doi: 10.7506/spkx1002-6630-201517055
[11]	周瑞铮, 陈锦杭, 张树权, 等. 分散固相萃取结合液相色谱-串联质谱法测定淡水鱼中9种磺胺类和3种喹诺酮类药物残留量[J]. 食品安全质量检测学报,2021,12(17):6946-6952 doi: 10.19812/j.cnki.jfsq11-5956/ts.2021.17.035 ZHOU Ruizheng, CHEN Jinhang, ZHANG Shuquan, et al. Determination of 9 kinds of sulfonamides and 3 kinds of quinolones residues in freshwater fish by dispersive solid phase extraction coupled with liquid chromatography-tandem mass spectrometry[J]. Journal of Food Safety & Quality,2021,12 (17):6946-6952. doi: 10.19812/j.cnki.jfsq11-5956/ts.2021.17.035
[12]	覃玲, 董亚蕾, 王钢力, 等. 分散固相萃取-液相色谱-串联质谱法测定常见动物源性食品中42种兽药残留[J]. 色谱,2018,36(9):880-888 doi: 10.3724/SP.J.1123.2018.04024 QIN Ling, DONG Yalei, WANG Gangli, et al. Determination of 42 veterinary drug residues in common animal derived food by dispersive solid phase extraction coupled with liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography,2018,36 (9):880-888. doi: 10.3724/SP.J.1123.2018.04024
[13]	余丽梅, 张聪, 宋超, 等. 分散固相萃取-超高液相色谱/质谱测定水产品中磺胺类抗生素的残留量[J]. 中国农学通报,2017,33(26):128-135 doi: 10.11924/j.issn.1000-6850.casb16080104 YU Limei, ZHANG Cong, SONG Chao, et al. Dispersive solid-phase extraction-ultra performance liquid chromatography tandem mass spectrometry detecting sulfonamides residues in aquatic products[J]. Chinese Agricultural Science Bulletin,2017,33 (26):128-135. doi: 10.11924/j.issn.1000-6850.casb16080104
[14]	张蓓蓓, 孙慧婧, 吉鑫. 混合型离子交换反相吸附固相萃取柱串联萃取-超高效液相色谱-串联质谱法测定地表水中32种抗生素的含量[J]. 理化检验-化学分册,2022,58(8):893-901 ZHANG Beibei, SUN Huijing, JI Xin. Determination of 32 antibiotics in surface water by ultra-high performance liquid chromatography-tandem mass spectrometry after tandem extraction with mixed ion exchange reversed phase adsorption solid phase extraction columns[J]. Physical Testing and Chemical Analysis (Part B:Chemical Analysis),2022,58 (8):893-901.