利用串聯式液相層析質譜法同時分析丙烯醯胺職業暴露工人尿液代謝物

Simultaneous Determination of Urinary Mercapturic Acids for Acrylamide Exposed Workers by LC-MS/MS

10.7005/JOSH.200909.0385

吳焜裕(Kuen-Yuh Wu)；陳美蓮(Mei-Lien Chen)；劉紹興(Saou-Hsing Liou)；汪禧年(Shi-Nian Uang)；黃鈺芳(Yu-Fang Huang)

丙烯醯胺 ； 尿液代謝物N-acetyl-S-propionamide-cysteine AAMA ； N-acetyl-S-carbamoyl-2-hydroxyethyl-cysteine GAMA2 ； N-acetyl-S-3-amino-2-hydroxy-3-oxopropyl-cysteine GAMA3 ； 液相層析串聯式質譜儀 ； Acrylamide ； N-acetyl-S-propionamide-cysteine AAMA ； N-acetyl-S-carbamoyl-2-hydroxyethyl-cysteine GAMA2 ； N-acetyl-S-3-amino-2-hydroxy-3-oxopropyl-cysteine GAMA3 ； Liquid chromatography-tandem mass spectrometry LC-MS/MS

勞工安全衛生研究季刊

17卷3期（2009 / 09 / 01）

385 - 397

繁體中文

Acrylamide (AA) has been widely used in industry to produce flocculants and grouting agents as well as in biological laboratories for the preparation of polyacrylamide gels for electrophoresis. It is classified as a probable human carcinogen (Group 2A) by the IARC. Workers are at risk of AA exposures. AA can be metabolized into AA N-acetyl-S-(propionamide)-cysteine (AAMA) and of Glycidamide (GA) N-acetyl-S-(carbamoyl-2-hydroxyethyl)-cysteine (GAMA2) and N-acetyl-S-(3- amino-2-hydroxy-3-oxopropyl)-cysteine (GAMA3). Analysis of these mercapturic acids (MAs) can serve as biomarkers for AA exposures and illustrate the metabolism of AA in human. In our study, a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously analyze urinary AAMA, GAMA2 and GAMA3 for the AA-exposed workers. Fifty six AA-exposed workers and 36 controls were recruited and provided a pair of pre- and post-shift urine samples for this study. The results showed the limit of detection (LOD) values were ＜10 ng/mL (GAMA2) , 20 ng/mL (GAMA3) and 5 ng/mL (AAMA) in urine and the detection rate was 34%, 38% and 92% for GAMA2, GAMA3 and AAMA, respectively. The geometric mean (range) level of GAMA2, GAMA3 and AAMA in the whole collective (n=100) was 38.2 (8.1-343) (μg/g creatinine), 94.9(10.8-1754.6) (μg/g creatinine) and 516.7 (4.7-115843) (μg/g creatinine), respectively. There was a significantly positive correlation between naturally log-transformed concentrations of AAMA and naturally log-transformed GAMA2 and GAMA3 (Pearson correlation: r=0.695, p＜0.001). Besides, the pre- and post-shift urinary AAMA concentrations for the AA-exposed workers were higher than those for the controls. These results suggest that AAMA can serve as a sensitive, specific, non-invasive, and easily accessible biomarker for AA exposures.

Acrylamide (AA) has been widely used in industry to produce flocculants and grouting agents as well as in biological laboratories for the preparation of polyacrylamide gels for electrophoresis. It is classified as a probable human carcinogen (Group 2A) by the IARC. Workers are at risk of AA exposures. AA can be metabolized into AA N-acetyl-S-(propionamide)-cysteine (AAMA) and of Glycidamide (GA) N-acetyl-S-(carbamoyl-2-hydroxyethyl)-cysteine (GAMA2) and N-acetyl-S-(3- amino-2-hydroxy-3-oxopropyl)-cysteine (GAMA3). Analysis of these mercapturic acids (MAs) can serve as biomarkers for AA exposures and illustrate the metabolism of AA in human. In our study, a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously analyze urinary AAMA, GAMA2 and GAMA3 for the AA-exposed workers. Fifty six AA-exposed workers and 36 controls were recruited and provided a pair of pre- and post-shift urine samples for this study. The results showed the limit of detection (LOD) values were ＜10 ng/mL (GAMA2) , 20 ng/mL (GAMA3) and 5 ng/mL (AAMA) in urine and the detection rate was 34%, 38% and 92% for GAMA2, GAMA3 and AAMA, respectively. The geometric mean (range) level of GAMA2, GAMA3 and AAMA in the whole collective (n=100) was 38.2 (8.1-343) (μg/g creatinine), 94.9(10.8-1754.6) (μg/g creatinine) and 516.7 (4.7-115843) (μg/g creatinine), respectively. There was a significantly positive correlation between naturally log-transformed concentrations of AAMA and naturally log-transformed GAMA2 and GAMA3 (Pearson correlation: r=0.695, p＜0.001). Besides, the pre- and post-shift urinary AAMA concentrations for the AA-exposed workers were higher than those for the controls. These results suggest that AAMA can serve as a sensitive, specific, non-invasive, and easily accessible biomarker for AA exposures.

1. (2003).Standards of Permissible Exposure Limits of Airborne Hazardous Substances in Workplace Lao. An-3-Tzu No. 0920073294.Labor Commlssion, Ministry of the Interior on Dec 31.
2. Bjellaas T,Janak K,Lundanes E,Kronberg L,Becher G.(2005).Determination and quantification of urinary metabolites after dietary exposure to acrylamide.Xenobiotica,35,1003-1018.
3. Bjellaas T,Stolen-LH,Haugen M(2008).Urinary acrylamide metabolites as biomarkers for short-term dietary exposure to acrylamide.Food and Chemical Toxicology In Press, Corrected Proof.
4. Boettcher MI,Bolt HM,Drexler H,Angerer J.(2006).Excretion of mercap-tune acids of acrylamide and glycidamide in human urine after single oral administration of deuteriumlabelled acrylamide.Arch Toxicol,80(2),55-61.
5. Bull PJ,Brooke RE,Cocker J,Jones K(2005).An occupational hygiene investigation of exposure to acrylamide and the role for urinary S-Carboxyethyl-Cysteine (CEC) as a biological marker.Ann Occup Hyg,49,683-690.
6. Bull RT,Robinson M,Laurie RD,Stoner GD,Greisiger E,Meier JR(1984).Carcinogenic effects of acrylamide in sencar and A/J mice.Cancer Res,44,107-111.
7. Calleman CJ,Bergmark E,Costa LG.(1990).Acrylamide is metabolized to glycidamide in the rat: evidence from hemoglobin adduct formation.Chem Res Toxicol,3,406-412.
8. Calleman CJ,Wu Y,He F,Tian G.,Bergmark B,Zhang S(1994).Relationships between biomarkers of exposure and neurological effects in a group of workers exposed to acrylamide.Toxicol Appl Pharmacol,126,361-371.
9. Chevolleau S,Jacques C,Canlet C,Tulliez I,Debrauwer L.(1964).Analysis of hemoglobin adducts of acrylamide and glycidamide by liquid chroma tography-electrospray ionization tandem mass spectrometry Toxicology of Acrylamide.Toxicol Appl Pharmacol,103,172-181.
10. Fennell TR,Burgess J,Friedman MA,Snyder RW,Spicer R,Sumner SCJ(2005).Metabolism and hemoglobin adduct formation of acrylamide in humans.Toxicol Sci,85,447-459.
11. Fennell TR,Sumner SCJ,Snyder RW(2006).Kinetics of Elimination of Urinary Metabolites of Acrylamide in Humans.Toxicol Sci,93,256-267.
12. Fuhr U,Boettcher MI,Martina KS,Alexandra W,Alexander J,Andreas L(2006).Toxicokinetics of Acrylamide in Humans after Ingestion of a Defined Dose in a Test Meal to Improve Risk Assessment for Acrylamide Carcinogenicity.Cancer Epidem Biomar,15,266-271.
13. Hagmar L,Tornqvist M,Nordander C,Rosen I,Bruze M,Kautiainen A(2001).Health effects of occupational exposure to acryl amble using hemoglobin adducts as biomarkers of internal dose.Scand J Work Environ Health,27,219-226.
14. Huang CCJ,Li CM,Wu CF,Jao SP,Wu KY.(2007).Analysis of urinary N-acetyl-S-(propionamide)-cysteine as a biomarker for the assessment of acrylamide exposure in smokers.Environ Res,104,346-351.
15. Kellert M,Scholz K,Wagner S,Dekant W,Volkel W.(2006).Quantitation of mercapturic acids from acrylamide and glycidamide in human urine using a column switching tool with two trap columns and electrospray tandem mass spectrometry.J Chromatogr A,1131,58-66.
16. Li CM,Wu KY,Hu CW.(2005).Quantif-ication of urinary N-acctyl-S-(propionamide) cysteine using an on-line clean-up system coupled with liquid chromatography/tandem mass spectrometry.J Mass Spectrom,40,511-515.
17. Mottram DS,Wedieha BL,Dodson AT.(2002).Acrylamide is formed in the Maillard reaction.Nature,419,448-449.
18. Segerback D,Calleman CJ,Echo-eder JL,Costa LG.,Faustman EM.(1995).Formation of N-7-(2-carb-amoyl-2-hydroxyethyl) guanine in DNA of the mouse and the rat following intraperitoneal administration of [14C] acrylamide.Carcin,16,1161-1165.
19. Smith CJ,Perfetti TA,Rumple MA,Rodgman A,Doolittle DJ.(2000)."IARC Group 2A Carcinogens" reported in cigarette mainstream smoke.Food Chem Toxicol,38,371-383.
20. Stadler RH,Blank I,Varga N,Robert F,Hau J,Guy PA(2002).Acrylamide from Maillard reaction products.Nature,419,449-450.
21. Sumner SC,Fennel TR,Selvaraj L,Nauhaus SK.(1997).Urinary metabolites from F344 rats and B6C3FI mice coadministered acrylamide and acrylonitrile for 1 or 5 days.Chem Res Toxicol,10,1152-1160.
22. Sumner SC,Fennell TR,Moore TA,Chanas B,Gonzalez F,Ghanayem BI.(1999).Role of cytochrome P450 2E1 in the metabolism of acrylamide and acrylonitrile in mice.Chem ResToxicol,12,1110-1116.
23. Sumner SC,MacNeela JP,Fennell, TR.(1992).Characterization and quantit-ation of urinary metabolites of [1,2,3-l3C] acrylamide in rats and mice using l3C nuclear Magnetic resonance spcctroscopy.Chem Res Toxicol,5,81-89.
24. Sumner SC,Williams CC,Snyder RW,Krol WL,Asgharian B,Fennell TR.(2003).Acrylamide: A comparison of metabolism and hemoglobin adducts in rodents following dermal, intraperitoneal, oral, or inhalation exposure.Toxicol Sci,75,260-270.
25. Twaddle NC,McDaniel LP,da Costa GG,Churchwell MI,Beland FA,Doerge DR.(2004).Determination of acrylamide and glycidamide serum toxicokinetics in B6C3F1 mice using LC-ES/MS/MS.Cancer Letter,207,9-17.
26. Urban M,Kavvadias D,Riedel K,Scherer G.(2006).Urinary Mercaptunic Acids and a Hemoglobin Adduct for the Dosimetry of Acrylamide Exposure in Smokers and Nonsmokers.Inhal Toxicol,18,831-839.
27. Aerylamide 2006-6-15. OSHA 2004
28. World Health Organization, International Agency for Research on Cancer, and National Cancer Institute(1994).IARC Working Group on the Evaluation of Carcinogenic Risks to Humans, Some industrial chemicals.Geneva:
29. Wu YQ,Zhang J,Cui T,Yu AR,Tang XY.(1994).Determination of acrylamide metabolite, mercapturic acid by high performance liquid chromatography.Biomed. Environ Sci,6,273-280.