通过式固相萃取-超高效液相色谱-串联质谱法测定水产品中地西泮

doi:10.3724/SP.J.1123.2019.11028

摘要/Abstract

摘要：

建立了通过式固相萃取-超高效液相色谱-串联质谱测定水产品中地西泮残留量的分析方法。样品用乙腈直接提取，经Prime HLB通过式固相萃取柱（60 mg/3 mL）净化，以Acquity UPLC BEH C₁₈（100 mm×2.1 mm，1.7 μm）为色谱柱，甲醇-0.1%（v/v）甲酸水溶液为流动相进行梯度洗脱。在多反应监测（MRM）、正离子电离模式下检测，采用基质匹配标准曲线外标法进行定量分析。结果显示：在0.1~10 ng/mL范围内，地西泮线性关系良好（r²>0.99）。在1.5、3.0和15.0 μg/kg 3个加标水平下，地西泮的加标回收率为88.2%~101.1%，日间和日内精密度（RSD）均在10%以下。该法简便快速，灵敏度高，可用于水产品中地西泮的准确测定。

关键词: 超高效液相色谱-串联质谱, 通过式固相萃取, 地西泮, 水产品

Abstract:

A method was developed for the determination of diazepam in aquatic products by pass-through solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The analyte was extracted with acetonitrile directly and purified on a Prime HLB solid phase extraction column (60 mg/3 mL). The separation was performed on an Acquity UPLC BEH C₁₈ column (100 mm×2.1 mm, 1.7 μm)using methanol-0.1% (v/v) formic acid aqueous solution as the mobile phase in gradient elution mode. Qualitative analysis was performed in the multiple reaction monitoring (MRM) mode. The analyte was quantified by matrix-matched external standard curves. The results showed good linear relationship in the range of 0.1-10 ng/mL, and the correlation coefficient (r²) was greater than 0.99. The spiked recoveries of diazepam were 88.2%-101.1% at the spiked levels of 1.5, 3.0 and 15.0 μg/kg, and both the intra-and inter-day precisions were less than 10%. The developed method is simple, rapid and accurate, and it can meet the requirements for diazepam determination in aquatic product samples.

Key words: ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS), pass-through solid phase extraction, diazepam, aquatic products

宿书芳, 孙立臻, 薛霞, 公丕学, 魏莉莉, 李新玲, 祝建华, 刘艳明, 张峰. 通过式固相萃取-超高效液相色谱-串联质谱法测定水产品中地西泮[J]. 色谱, 2020, 38(7): 791-797.

SU Shufang, SUN Lizhen, XUE Xia, GONG Pixue, WEI Lili, LI Xinling, ZHU Jianhua, LIU Yanming, ZHANG Feng. Determination of diazepam in aquatic products by ultra performance liquid chromatography-tandem mass spectrometry with pass-through solid phase extraction[J]. Chinese Journal of Chromatography, 2020, 38(7): 791-797.

图/表 11

图1 地西泮的化学结构式

Fig. 1 Chemical structure of diazepam

表1 地西泮的母离子、子离子及其他质谱参数

Table 1 Precursor ion, daughter ion and other MS parameters of diazepam

Analyte	Precursor ion (m/z)	Daughter ion (m/z)	Declustering potential/V	Collision energy/eV
*Quantitative ion.
Diazepam	285.2	154.0	100	37
		193.0^*	100	38

图2 地西泮标准溶液(1 ng/mL)的提取离子流色谱图

Fig. 2 Extracted ion current chromatograms of diazepam in the standard solution (1 ng/mL)

表2 采用不同提取方式时地西泮的回收率(n=6)

Table 2 Recoveries of diazepam using different extraction methods (n=6)

Pretreatment method	Recoveries/% (RSDs/%)
Pretreatment method	Yellow croaker	Carp	Crucian	Shrimp
Enzymolysis	82.1 (6.9)	79.5 (9.2)	80.6 (7.5)	92.5 (5.7)
Non enzymolysis	103.0 (5.4)	109.0 (4.3)	94.5 (8.3)	101.0 (2.9)

图3 不同提取剂对地西泮回收率的影响

Fig. 3 Effect of different extraction solvents on the recoveries of diazepam Solvent 1: acetonitrile; solvent 2: acetonitrile (containing 1% (v/v) acetic acid); solvent 3: acetonitrile (containing 1% (v/v) ammonium hydroxide); solvent 4: ethyl acetate.

表3 采用不同净化方式时地西泮的基质效应

Table 3 Matrix effects of diazepam using different purification methods

Purification method	Matrix effects/%
Purification method	Yellow croaker	Carp	Crucian	Shrimp 1	Shrimp 2
Prime HLB	6.7	3.9	7.6	10.1	5.5
QuEChERS	11.8	13.5	9.9	16.3	15.0
Non purification	55.2	58.9	37.6	68.0	56.2

图4 不同规格净化柱对地西泮回收率的影响

Fig. 4 Effect of purification columns with different specifications on the recoveries of diazepam

图5 上样溶液中乙腈的体积分数对地西泮回收率的影响

Fig. 5 Effect of volume fractions of acetonitrile on the recoveries of diazepam in sample loading solutions

图6 氮吹对不同样品中目标物回收率的影响(n=3)

Fig. 6 Effect of the nitrogen blowing on the recoveries of the different samples (n=3)

表4 地西泮的线性方程、相关系数、检出限和定量限

Table 4 Linear equations, correlation coefficients (r2), LODs and LOQs of diazepam

Matrix	Linear equation	r²	LOD/(μg/kg)	LOQ/(μg/kg)
Y: peak area; X: mass concentration, ng/mL.
Yellow	Y=1.826×10⁵X+1.463×10⁴	0.9992	0.5	1.5
croaker
Shrimp	Y=2.021×10⁵X+5.873×10³	0.9970	0.5	1.5

表5 水产品中地西泮的回收率及精密度(n=6)

Table 5 Recoveries and precisions of diazepam in aquatic products (n=6)

Matrix	Spiked level/(μg/kg)	Average recovery/%	Intra-day RSD/%	Inter-day RSD/%
Yellow croaker	1.5	95.4	7.0	5.9
	3.0	93.5	4.8	3.3
	15.0	101.1	5.2	5.1
Shrimp	1.5	88.2	2.3	7.7
	3.0	95.6	3.8	5.2
	15.0	97.3	6.4	5.0

参考文献

1	Sun L , Zhang L , Xu Q , et al. Chinese Journal of Chromatography, 2010, 28 (1): 38
1	孙雷, 张骊, 徐倩, et al. 色谱, 2010, 28 (1): 38
2	Zhang S , Zhou S , Chen D W , et al. Journal of Instrumental Analysis, 2014, 33 (11): 1213
2	张烁, 周爽, 陈达炜, et al. 分析测试学报, 2014, 33 (11): 1213
3	Qu Y , Lu Y , Feng N , et al. Food Science, 2012, 33 (8): 252
3	渠岩, 路勇, 冯楠, et al. 食品科学, 2012, 33 (8): 252
4	Zhang Q M . Journal of Anhui Agricultural Sciences, 2016, 44 (25): 71
4	张倩勉. 安徽农业科学, 2016, 44 (25): 71
5	Wang L P , Li X , Sun Y , et al. Chinese Journal of Analytical Chemistry, 2005, 33 (7): 951
5	汪丽萍, 李翔, 孙英, et al. 分析化学, 2005, 33 (7): 951
6	Xu C B , Suo R , Zhang F , et al. Chinese Journal of Chromatography, 2012, 30 (5): 457
6	许成保, 锁然, 张峰, et al. 色谱, 2012, 30 (5): 457
7	Sun S S , Zhu L J , Hu Y X , et al. Chinese Journal of Chromatography, 2018, 36 (2): 150
7	孙珊珊, 朱丽君, 胡延喜, et al. 色谱, 2018, 36 (2): 150
8	Wang L , Liu C Y , Wang Z J , et al. Journal of Instrumental Analysis, 2016, 35 (2): 219
8	王璐, 刘成雁, 王志嘉, et al. 分析测试学报, 2016, 35 (2): 219
9	Zhang X Y , Cai X X , Zhang X Y . Chinese Journal of Chromatography, 2010, 28 (1): 23
9	张秀尧, 蔡欣欣, 张晓艺. 色谱, 2010, 28 (1): 23
10	Chang Q , Ma H Y , Wang F J , et al. Chinese Journal of Chromatography, 2011, 29 (11): 1082
10	常青, 马虹英, 王方杰, et al. 色谱, 2011, 29 (11): 1082
11	Li R , Yang L Q , Zhang P Y , et al. Chinese Journal of Chromatography, 2018, 36 (2): 125
11	李蓉, 杨璐齐, 张鹏云, et al. 色谱, 2018, 36 (2): 125
12	Yu H J , Qian B L , Huang D M , et al. Chinese Fishery Quality and Standards, 2011, 1 (1): 54
12	于慧娟, 钱蓓蕾, 黄冬梅, et al. 中国渔业质量与标准, 2011, 1 (1): 54
13	Qian X D , Yu H J , Hui Y H , et al. Hunan Agricultural Sciences, 2010 (19): 134
13	钱晓东, 于慧娟, 惠芸华, et al. 湖南农业科学, 2010 (19): 134
14	Wei J M , Luo Y Z , Bai Y X . Science and Technology of Food Industry, 2014, 35 (10): 95
14	魏晋梅, 罗玉柱, 白云旭. 食品工业科技, 2014, 35 (10): 95
15	Li P D , Han H R , Zhai X H , et al. J Chromatogr Sci, 2012, 50 (2): 108
16	Guo D H , Deng X J , Zhao S Z , et al. Chinese Journal of Analytical Chemistry, 2010, 38 (3): 318
16	郭德华, 邓晓军, 赵善贞, et al. 分析化学, 2010, 38 (3): 318
17	Zhang X , Zhang J , Shao B . Chinese Journal of Food Hygiene, 2017, 29 (6): 684
17	张昕, 张晶, 邵兵. 中国食品卫生杂志, 2017, 29 (6): 684
18	Wei J M , Luo Y Z , Zhang L , et al. J Sci Food Agric, 2015, 95 (3): 598
19	Lee S , Han E , In S , et al. J Anal Toxicol, 2011, 35 (5): 312
20	Leandro C C , Hancock P , Fussell B J , et al. J Chromatogr A, 2007, 1144 (2): 161
21	Cheng L L , Zhang Y J , Shen J Z , et al. J Chromatogr Sci, 2010, 71 (1): 155
22	SN/T 3235-2012
23	Liu J Y , Huang X , Jia H X . Journal of Instrumental Analysis, 2017, 36 (3): 305
23	刘家阳, 黄旭, 贾宏新. 分析测试学报, 2017, 36 (3): 305
24	Zhang Y , Chen Y N , Zheng X Z , et al. Chinese Journal of Hospital Pharmacy, 2002, 22 (6): 337
24	张吟, 陈一农, 郑兴中, et al. 中国医院药学杂志, 2002, 22 (6): 337
25	Zou Y , Shao L Z , Chen S M , et al. Chinese Journal of Chromatography, 2017, 35 (8): 801
25	邹游, 邵琳智, 陈思敏, et al. 色谱, 2017, 35 (8): 801
26	Zhang H , Wu Y L , Zhang Y W , et al. Chinese Journal of Chromatography, 2019, 37 (12): 1314
26	章豪, 吴银良, 张宜文, et al. 色谱, 2019, 37 (12): 1314
27	Zhang Y X , Xue X , Su S F , et al. Food Anal Methods, 2018, 11: 2865
28	Jiang D Z , Xin L N , Tan X M , et al. Science and Technology of Food Industry, 2019, 40 (22): 259
28	蒋定之, 辛丽娜, 谭喜梅, et al. 食品工业科技, 2019, 40 (22): 259

编辑推荐

Metrics

阅读次数

全文

132

HTML			PDF

最新录用	在线预览	正式出版	最新录用	在线预览	正式出版
0	0	12	0	0	120

来源	本网站	其他网站

次数	131	1
比例	99%	1%

摘要

193

最新录用	在线预览	正式出版

0	0	193

	来源	本网站

	次数	193
	比例	100%

[1]	赵芮, 黄晴雯, 余智颖, 韩铮, 范楷, 赵志辉, 聂冬霞. QuEChERS-超高效液相色谱-串联质谱法同时测定水果中36种真菌毒素[J]. 色谱, 2023, 41(9): 760-770.
[2]	岳超, 赵超群, 毛思浩, 王展华, 施贝, 徐欣丰, 梁晶晶. 通过式固相萃取净化-超高效液相色谱-串联质谱法测定液体乳中10种氨基甲酸酯类农药残留[J]. 色谱, 2023, 41(9): 807-813.
[3]	孟二琼, 念琪循, 李峰, 张秋萍, 许茜, 王春民. 磺酸化磁性氮化碳固相萃取-超高液相色谱-串联质谱筛检淡水鱼中孔雀石绿和隐色孔雀石绿[J]. 色谱, 2023, 41(8): 673-682.
[4]	王秋旭, 冯启言, 朱雪强. 加速溶剂萃取-固相萃取净化结合超高效液相色谱-串联质谱法测定沉积物中双酚类化合物[J]. 色谱, 2023, 41(7): 582-590.
[5]	夏宝林, 汪仕韬, 殷晶晶, 张维益, 杨娜, 刘强, 吴海晶. 自动上样固相萃取-超高效液相色谱-串联质谱法同时测定水中9类43种抗菌药物残留[J]. 色谱, 2023, 41(7): 591-601.
[6]	陈东洋, 张昊, 张磊, 王一红, 王小丹, 冯家力, 梁静, 钟旋. 固相萃取-超高效液相色谱-串联质谱法测定发酵食品中的曲酸[J]. 色谱, 2023, 41(7): 632-639.
[7]	童学智, 陈东洋, 冯家力, 范翔, 张昊, 杨胜园. QuEChERS-超高效液相色谱-串联质谱法快速测定水产品中5种卤代苯醌[J]. 色谱, 2023, 41(6): 490-496.
[8]	李鑫玉, 赵芳, 平华, 马智宏, 李冰茹, 马挺军, 李成. 稳定同位素稀释气相色谱-三重四极杆质谱法测定水产品中15种卤代多环芳烃[J]. 色谱, 2023, 41(6): 527-534.
[9]	曾永福, 陈美芳, 邵雨, 闫永欢, 张海超, 王敬, 艾连峰, 康维钧. 植物中27种典型药品及个人护理品多残留检测方法的建立及其在芽苗菜中迁移规律的分析[J]. 色谱, 2023, 41(5): 386-396.
[10]	李振环, 胡小键, 陆一夫, 谢琳娜, 朱英. 基于高通量全自动固相萃取的超高效液相色谱-串联质谱法测定人尿中16种抗生素和4种β-受体激动剂[J]. 色谱, 2023, 41(5): 397-408.
[11]	吴少明, 欧阳立群, 孟鹏, 何孟杭, 林钦, 陈言凯, 刘文菁, 苏晓明, 戴明. 固相萃取净化-超高效液相色谱-串联质谱法测定畜肉中18种卡因类麻醉剂[J]. 色谱, 2023, 41(5): 434-442.
[12]	孙佳林, 牛宇敏, 高群, 张晶, 邵兵. 超高效液相色谱-串联质谱法测定灰尘中26种双酚类化合物[J]. 色谱, 2023, 41(5): 417-425.
[13]	朱卫红, 王超, 张霖琳, 袁懋. 超高效液相色谱-串联质谱法直接测定水中5种黄原酸[J]. 色谱, 2023, 41(4): 339-347.
[14]	韩林学, 张续, 胡小键, 张海婧, 邱天, 林潇, 朱英. 超高效液相色谱-串联质谱法测定人尿中12种典型个人护理品[J]. 色谱, 2023, 41(4): 312-322.
[15]	邱天, 张续, 杨艳伟, 胡小键, 罗嵩, 朱英. 超高效液相色谱-串联质谱法测定尿液中7种苯系物代谢物[J]. 色谱, 2023, 41(4): 366-375.