液液萃取-高分辨气相色谱-高分辨双聚焦磁质谱法测定尿中羟基多环芳烃代谢物

doi:10.3724/SP.J.1123.2019.10031

摘要/Abstract

摘要：

建立了灵敏、准确的液液萃取-高分辨气相色谱-高分辨双聚焦磁质谱（LLC-HRGC-HRMS）同时测定人体尿样中8种多环芳烃（PAHs）代谢物的方法。将2 mL尿液与氘或¹³C标记的同位素内标物进行混合，在抗坏血酸的存在下酶解偶联目标物，再将游离化合物用甲苯-戊烷（1：4，v/v）萃取后浓缩至近干，再用甲苯复溶，衍生后经HRGC-HRMS分离和测定。1-羟基萘在0.14~41.6 μg/L、1-羟基菲在0.05~8.33 μg/L、2-羟基菲在0.04~8.33 μg/L、其他5种代谢产物在0.02~8.33 μg/L范围内线性关系良好，相关系数＞0.99，方法的检出限为0.006~0.042 μg/L，平均回收率为81.4%~127.0%，日内和日间精密度分别为2.7%~6.9%和5.1%~10.9%（n=6）。应用该方法测定330份人群尿液样本，3-羟基䓛和6-羟基䓛未检出，其他6种PAHs代谢物检出率为100%。该方法灵敏、准确、稳定，适用于人尿中8种PAHs代谢物的定量分析。

关键词: 高分辨气相色谱-高分辨双聚焦磁质谱, 液液萃取, 生物监测, 羟基多环芳烃代谢物, 尿

Abstract:

A sensitive and accurate method was developed to quantify eight polycyclic aromatic hydrocarbon (PAH) metabolites in human urine by liquid-liquid extraction-high resolution gas chromatography-high resolution dual-focus magnetic mass spectrometry (LLC-HRGC-HRMS). About 2 mL urine was mixed with a deuterium- or ¹³C-labeled isotopic internal standard, and the conjugated targets were enzymatically digested in the presence of ascorbic acid. The free compounds were extracted with toluene-pentane (1:4, v/v) and condensed to near dryness. Then, the target compounds were redissolved in toluene. After derivatization, they were separated and quantified by HRGC-HRMS. The linear ranges of the 1-hydroxynaphthalene, 1-hydroxyphenanthrene, and 2-hydroxyphenanthrene were 0.14-41.6 μg/L, 0.05-8.33 μg/L, and 0.04-8.33 μg/L, respectively, and those for the other five PAH metabolites were 0.02-8.33 μg/L. The correlation coefficients were greater than 0.99. The limits of detection were in the range of 0.006-0.042 μg/L, and the recoveries were 81.4%-127.0%. The intra-day and inter-day relative standard deviations (RSDs) were less than 6.9% and 10.9%, respectively. This method was utilized for the determination of 330 human urine samples. The results showed that 3-hydroxychrysene and 6-hydroxychrysene were not detected, and the detection rate of the other six PAH metabolites was 100%. This method is sensitive, accurate and stable, and it is suitable for the determination of the eight PAH metabolites in human urine.

Key words: high resolution gas chromatography-high resolution dual-focus magnetic mass spectrometry (HRGC-HRMS), liquid-liquid extraction (LLE), biological monitoring, hydroxyl polycyclic aromatic metabolites, urine

付慧, 陆一夫, 胡小键, 张续, 杨艳伟, 朱英. 液液萃取-高分辨气相色谱-高分辨双聚焦磁质谱法测定尿中羟基多环芳烃代谢物[J]. 色谱, 2020, 38(6): 715-721.

FU Hui, LU Yifu, HU Xiaojian, ZHANG Xu, YANG Yanwei, ZHU Ying. Determination of polycyclic aromatic hydrocarbon metabolites in urine by liquid-liquid extraction-high resolution gas chromatography-high resolution dual-focus magnetic mass spectrometry[J]. Chinese Journal of Chromatography, 2020, 38(6): 715-721.

图/表 6

参考文献

1	Duan X L , Tao S , Xu D Q , et al. Exposure Measurement and Health Risk Assessment of Human Exposure to Polycyclic Aromatic Hydrocarbons Beijing China Environmental Science Press 2011
1	段小丽, 陶澍, 徐东群, et al. 多环芳烃污染的人体暴露和健康风险评价方法北京中国环境科学出版社 2011
2	Amirdivani S , Khorshidian N , Dana M G , et al. Int J Dairy Technol, 2019, 72 (1): 120
3	Wang C X , Zhang R R , Xun Z Q , et al. Journal of Instrumental Analysis, 2018, 37 (4): 459
3	汪晨霞, 张瑞瑞, 寻知庆, et al. 分析测试学报, 2018, 37 (4): 459
4	Song X J , Li H Y , Yin M M , et al. Chinese Journal of Chromatography, 2018, 36 (1): 51
4	宋晓娟, 李海燕, 尹明明, et al. 色谱, 2018, 36 (1): 51
5	Zhang H Q , Rao Z , Wang X C , et al. Journal of Instrumental Analysis, 2017, 36 (10): 1197
5	张红庆, 饶竹, 王晓春, et al. 分析测试学报, 2017, 36 (10): 1197
6	Li Z , Romanoff L C , Trinidad D A , et al. Anal Chem, 2006, 78 (16): 5744
7	CDATA[Centers for Disease Control and Prevention. Monohydroxy-Polycyclic Aromatic Hydrocarbons (OH-PAHs) Laboratory Procedure Manual. (2013-03-12). https://www.cdc.gov/nchs/data/nhanes/nhanes_11_12/pahs_g_met.pdf
8	CDATA[Centers for Disease Control and Prevention. Fourth National Report on Human Exposure to Environmental Chemicals Update Tables. (2017-01-01)[2019-10-28]. https://www.cdc.gov/exposurereport/pdf/FourthReport_UpdatedTables_Volume1_Jan2017.pdf
9	CDATA[Government of Canada. Fourth Report on Human Biomonitoring of Environmental Chemicals in Canada. (2017-08-23)[2019-10-28]. https://www.canada.ca/en/health-canada/services/environmental-workplace-health/reports-publications/environmental-contaminants/fourth-report-human-biomonitoring-environmental-chemicals-canada.html
10	Health Canada. Second Report on Human Biomonitoring of Environmental Chemicals in Canada. Hu G C, Yu Y J, Zhang L J, et al. transl. Beijing: China Environment Press, 2015
10	加拿大卫生部.加拿大第二次环境污染物生物监测报告.胡国成, 于云江, 张丽娟, 等译.北京: 中国环境出版社, 2015
11	Nian Q X , Liu Y M , Sun B , et al. Chinese Journal of Chromatography, 2019, 37 (3): 252
11	念琪循, 刘园满, 孙冰, et al. 色谱, 2019, 37 (3): 252
12	Li P , Cui J T , Guo X X , et al. Journal of Instrumental Analysis, 2018, 37 (3): 282
12	李佩, 崔君涛, 郭溪香, et al. 分析测试学报, 2018, 37 (3): 282
13	Hu G Y , Wang M M , Nian Q X , et al. Chinese Journal of Chromatography, 2018, 36 (4): 370
13	胡桂羽, 王曼曼, 念琪循, et al. 色谱, 2018, 36 (4): 370
14	Fu H , Hu X J , Chen X , et al. Chinese Journal of Chromatography, 2018, 36 (5): 487
14	付慧, 胡小键, 陈曦, et al. 色谱, 2018, 36 (5): 487
15	Chen X L , Li D M , Xu H J , et al. Journal of Instrumental Analysis, 2019, 38 (3): 270
15	陈雪蕾, 李冬梅, 徐厚君, et al. 分析测试学报, 2019, 38 (3): 270
16	Ding C M , Jin Y L , Lin S B . Chinese Journal of Analytical Chemistry, 2012, 40 (3): 397
16	丁昌明, 金银龙, 林少彬. 分析化学, 2012, 40 (3): 397
17	Zheng L , Ding C M , Hu X J , et al. Journal of Environment and Health, 2011, 28 (4): 348
17	郑磊, 丁昌明, 胡小键, et al. 环境与健康杂志, 2011, 28 (4): 348
18	Zhao M T. Hoodbook of Analytical Chemistry, 9B, Inorganic Mass Spectrometry. 3rd ed. Beijing: Chemical Industry Press, 2016
18	赵墨田.分析化学手册, 9B, 无机质谱分析. 3版.北京: 化学工业出版社, 2016
19	Li M P , Li R , Wang Z J , et al. Chinese Journal of Analytical Chemistry, 2019, 47 (2): 288
19	李美萍, 李蓉, 王志娟, et al. 分析化学, 2019, 47 (2): 288
20	Huang C Q , Chen Q K , Chen L , et al. Chinese Journal of Chromatography, 2019, 37 (10): 1048
20	黄超群, 陈钦可, 陈丽, et al. 色谱, 2019, 37 (10): 1048
21	Li X X , Cui S W , He M F , et al. Chinese Preventive Medicine, 2012, 13 (4): 246
21	李欣欣, 崔师伟, 何民富, et al. 中国预防医学杂志, 2012, 13 (4): 246
22	Li Z , Romanoff L C , Trinidad D A , et al. Anal Bioanal Chem, 2014, 406 (13): 3119
23	CDATA[U S Environmental Protection Agency. Definition and Procedure for the Determination of the Method Detection Limit, Revision 2. (2016-12-01)[2019-10-28]. https://www.epa.gov/cwa-methods.

编辑推荐

Metrics

阅读次数

全文

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	8	0	0	63

来源	本网站	其他网站

次数	70	1
比例	99%	1%

摘要

112

最新录用	在线预览	正式出版

0	0	112

来源	本网站	其他网站

次数	111	1
比例	99%	1%

Compound	Abbreviation	Retention time/min	Monitoring ion [M⁺] (m/z)	IS
1-Hydroxynaphthalene	1-OHN	9.40	216.0970	1-OHN-d₇
2-Hydroxyfluorene	2-OHF	16.34	254.1127	2-OHF-d₉
3-Hydroxyfluorene	3-OHF	15.94	254.1127	3-OHF-d₉
1-Hydroxyphenanthrene	1-OHP	20.30	266.1127	1-OHP-d₉
2-Hydroxyphenanthrene	2-OHP	21.32	266.1127	2-OHP-d₉
1-Hydroxypyrene	1-OHPyr	25.73	290.1127	1-OHPyr-d₉
3-Hydroxychrysene	3-OHC	27.52	316.1283	3-OHC-d₁₁
6-Hydroxychrysene	6-OHC	27.03	316.1283	6-OHC-¹³C₆
1-Hydroxynaphthalene-d₇	1-OHN-d₇	9.35	223.1410
2-Hydroxyfluorene-d₉	2-OHF-d₉	16.24	263.1692
3-Hydroxyfluorene-d₉	3-OHF-d₉	15.86	263.1692
1-Hydroxyphenanthrene-d₉	1-OHP-d₉	20.17	275.1692
2-Hydroxyphenanthrene-d₉	2-OHP-d₉	21.19	275.1692
1-Hydroxypyrene-d₉	1-OHPyr-d₉	25.70	299.1692
3-Hydroxychrysene-d₁₁	3-OHC-d₁₁	27.48	327.1975
6-Hydroxychrysene-¹³C₆	6-OHC-¹³C₆	27.03	322.1485

Compound	Abbreviation	Retention time/min	Monitoring ion [M⁺] (m/z)	IS
1-Hydroxynaphthalene	1-OHN	9.40	216.0970	1-OHN-d₇
2-Hydroxyfluorene	2-OHF	16.34	254.1127	2-OHF-d₉
3-Hydroxyfluorene	3-OHF	15.94	254.1127	3-OHF-d₉
1-Hydroxyphenanthrene	1-OHP	20.30	266.1127	1-OHP-d₉
2-Hydroxyphenanthrene	2-OHP	21.32	266.1127	2-OHP-d₉
1-Hydroxypyrene	1-OHPyr	25.73	290.1127	1-OHPyr-d₉
3-Hydroxychrysene	3-OHC	27.52	316.1283	3-OHC-d₁₁
6-Hydroxychrysene	6-OHC	27.03	316.1283	6-OHC-¹³C₆
1-Hydroxynaphthalene-d₇	1-OHN-d₇	9.35	223.1410
2-Hydroxyfluorene-d₉	2-OHF-d₉	16.24	263.1692
3-Hydroxyfluorene-d₉	3-OHF-d₉	15.86	263.1692
1-Hydroxyphenanthrene-d₉	1-OHP-d₉	20.17	275.1692
2-Hydroxyphenanthrene-d₉	2-OHP-d₉	21.19	275.1692
1-Hydroxypyrene-d₉	1-OHPyr-d₉	25.70	299.1692
3-Hydroxychrysene-d₁₁	3-OHC-d₁₁	27.48	327.1975
6-Hydroxychrysene-¹³C₆	6-OHC-¹³C₆	27.03	322.1485

Compound	Linear range/(μg/L)	Regression equation	r	LOD/(μg/L)	LOQ/(μg/L)
y: peak area ratio of quantitative ion of the analyte to IS; x: mass concentration ratio of the analyte to IS.
1-OHN	0.14-41.6	y=0.9321x+0.3733	0.994	0.042	0.139
2-OHF	0.02-8.33	y=0.9665x+0.1833	0.992	0.006	0.020
3-OHF	0.02-8.33	y=0.9796x-0.0449	0.997	0.006	0.020
1-OHP	0.05-8.33	y=0.9605x+0.0383	0.993	0.015	0.050
2-OHP	0.04-8.33	y=0.9154x-0.0010	0.992	0.012	0.040
1-OHPyr	0.02-8.33	y=0.9762x+0.0291	0.997	0.006	0.020
3-OHC	0.02-8.33	y=0.9580x-0.0698	0.997	0.006	0.020
6-OHC	0.02-8.33	y=0.9955x+0.0732	0.998	0.006	0.020

Compound	Linear range/(μg/L)	Regression equation	r	LOD/(μg/L)	LOQ/(μg/L)
y: peak area ratio of quantitative ion of the analyte to IS; x: mass concentration ratio of the analyte to IS.
1-OHN	0.14-41.6	y=0.9321x+0.3733	0.994	0.042	0.139
2-OHF	0.02-8.33	y=0.9665x+0.1833	0.992	0.006	0.020
3-OHF	0.02-8.33	y=0.9796x-0.0449	0.997	0.006	0.020
1-OHP	0.05-8.33	y=0.9605x+0.0383	0.993	0.015	0.050
2-OHP	0.04-8.33	y=0.9154x-0.0010	0.992	0.012	0.040
1-OHPyr	0.02-8.33	y=0.9762x+0.0291	0.997	0.006	0.020
3-OHC	0.02-8.33	y=0.9580x-0.0698	0.997	0.006	0.020
6-OHC	0.02-8.33	y=0.9955x+0.0732	0.998	0.006	0.020

Compound	0.125 μg/L		1.25 μg/L		6.25 μg/L		Intra-day RSD/%	Inter-day RSD/%
Compound	Recovery/%	RSD/%	Recovery/%	RSD/%	Recovery/%	RSD/%	Intra-day RSD/%	Inter-day RSD/%
1-OHN	94.3	2.9	82.9	6.1	100.0	8.3	5.8	9.1
2-OHF	127.0	3.1	110.5	1.7	111.4	8.2	4.4	9.7
3-OHF	120.2	2.2	117.8	2.8	117.1	9.5	4.8	6.8
1-OHP	115.3	9.0	98.6	4.1	107.1	7.8	6.9	8.6
2-OHP	121.4	0.2	98.8	5.2	104.8	4.9	3.4	9.7
1-OHPyr	121.9	4.2	84.9	2.1	96.2	1.9	2.7	10.3
3-OHC	95.4	5.6	123.7	7.4	126.1	7.6	6.9	5.1
6-OHC	115.7	6.8	99.2	7.5	81.4	2.2	5.5	10.9