利用串聯式液相層析質譜法分析尿液中二鄰氯苯胺甲烷及其代謝物以作為職業暴露之評估

Analysis of 4,4'-Methylene-bis (2-Chloroaniline) (MOCA) and acetyl-MOCA in urine by LC/MS/MS for MOCA Exposure Assessment

施偉仲(Wei-Chung Shih) ； 陳明烽(Ming-Fong Chen) ； 汪禧年(Shi-Nian Uang) ； 吳焜裕(Kuen-Yu Wu)

《勞工安全衛生研究季刊》 15卷1期 (2007/03) Pp. 55-66

https://doi.org/10.7005/JOSH.200703.0055

摘要

二鄰氯苯胺甲烷(4,4'-Methylene-bis(2-Chloroaniline), MOCA)為一致突變物質與動物致癌物，因為MOCA與其代謝物會和生物大分子如DNA和蛋白質反應而形成其共價鍵結物。在工業上廣泛作為固化劑之用，從事生產與使用的勞工潛在暴露於MOCA環境下對其健康的危害已受到重視。本實驗藉由分析MOCA及acetyl-MOCA，來作為MOCA暴露之生物指標，目前已成功利用固相萃取法做尿液樣品之前處理，並配合液相層析串聯式質譜儀(LC/MS/MS)建立分析MOCA與acetyl-MOCA之方法。此分析方法之偵測極限對MOCA及acetyl-MOCA分別是2ppb及0.02 ppb。此外，實驗收集並分析MOCA暴露之勞工尿液樣本結果發現一般不從事MOCA作業者，尿液中並無法分析到MOCA與acetyl-MOCA；而從事與MOCA製造或使用相關工作之作業勞工，每一尿液樣本中都可以分析到MOCA與acetyl-MOCA之含量。其中兩者的相關性則視當日是否暴露MOCA而定。數據顯示有暴露者其MOCA與acetyl-MOCA兩種指標之相關係數R^2為0.68，而他們工作前採樣所得兩種指標之相關性在0.8~0.36。結果顯示此一新發展的LC/MS/MS方法在樣品經固相萃取前處理後，對分析MOCA和acetyl-MOCA具有高度的靈敏度和選擇性。

關鍵字

二鄰氯苯胺甲烷 ； 乙醯基二鄰氯苯胺甲烷 ； 液相層析串聯式質譜

並列摘要

MOCA is a mutagen and animal carcinogen. MOCA and its metabolites can react with biological macromolecules such as DNA and protein to form DNA and protein adducts. Since MOCA is used as a hardness reagent in several industries, there is the potential for negative health effects for workers in MOCA-related workplaces. Analysis of MOCA and acetyl-MOCA in biomarkers may serve as a good indicator of exposure and risk. In our study, a new liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) method after solid-phase extraction was developed to analyze MOCA and acetyl-MOCA in the urine of workers exposed to MOCA. This method gave a low detection limit of 2 ppb for MOCA and 0.02 ppb for acetyl-MOCA. Our results showed that MOCA and acetyl-MOCA were detectable in urine collected from MOCA-exposed workers, but were not detectable in urine from unexposed workers. The correlation coefficient R^2 between urinary MOCA and acetyl-MOCA was 0.68 for urine collected after the shift, but 0.8~0.36 before the shift. These results demonstrate that the newly developed LC/MS/MS technique is highly sensitive and has good selectivity for analyzing MOCA and acetyl-MOCA after solid-phase extraction.

並列關鍵字

4,4'-Methylenebis 2-Chloroaniline ； MOCA ； Acetyl-MOCA ； LC/MS/MS

參考文獻

Cheever KL,DeBord DG,Swearengin TF(1991).4,4`-Methylenebis(2-chloroaniline) (MOCA): the effect of multiple oral administration, route, and phenobarbital induction on macromolecular adduct formation in the rat.Fundam Appl Toxicol.16,71-80.

Cheever KL,DeBord DG,Swearengin TF(1991).4,4`-Methylenebis(2-chloroaniline) (MOCA): the effect of multiple oral administration, route, and phenobarbit al induct ion on macromolecular adduct formation in the rat.Fundam Appl Toxicol.16,71-80.

Chen TH,Kuslikis BI,Braselton WE(1991).Un labeled hemoglobin adducts of 4,4`-methylenebis (2-chloroaniline) in rats and guinea pigs.Arch Toxicol.65,177-185.

Cocker J,Boobis AR,Davies DS(1988).Determination of the N-acetyl metabolites of 4,4`-methylene dianiline and 4,4`-methylenebis(2-choroaniline) in urine.Biomed. Envir. Mass Spectro.17,161-167.

Farmer PB,Rickard J,Robertson S(1981).The metabolism and distribution of 4,4`-methylenebis(2-chloroaniline) (MBOCA) in rats.J Appl Toxicol.1,317-322.