In addition, S- phenyl mercapturic acid and 3OH-benzoapyrene were measured in urine to monitor exposure to benzene and benzoapyrene. Individual smoking.
Epidemiology • Volume 22, Number 1, January Supplement 2011
concern, time-weighted air nicotine concentrations measured in this study were markedly higher than concentrations measured in bars and nightclubs from several African, American, Asian, and European countries. Implementing a comprehensive smoke-free legislation that protects workers and customers from exposure to secondhand smoke is urgently needed in Mongolia.
PP-30-059 Biological Monitoring of Aromatic Amines, Benzene, and Benzo关A兴Pyrene in Workers of a Modern European Coke Oven Plant Tobias Weiss,1 Holger M. Koch,1 Heiko U. Ka¨fferlein,1 Jana Henry,1 Volker Harth,1 Kai Su¨sselbeck,2 and Thomas Bru¨ning1 1Institute for Prevention and Occupational Medicine of the German Social Accident Insurance (IPA) Institute of the Ruhr-Universita¨t Bochum, Bochum, Germany; and 2IGF – Institut fu¨r Gefahrstoff-Forschung der Berufsgenossenschaft Rohstoffe und Chemische Industrie an der RuhrUniversita¨t Bochum, Bochum, Germany. Background/Aims: Coke oven emissions are complex mixtures of coal and coke particles, vapors, gases, and tars containing metals, polycyclic aromatic hydrocarbons (PAH), and aromatic amino and nitro compounds. Some epidemiologic literature suggests that coke oven workers may bear an elevated bladder cancer risk. Occupational exposure to aromatic amino and nitro compounds, and PAH is discussed being responsible for that observation. Methods: In this study, we examined 47 workers from a modern coke oven plant regarding their external and internal exposure to 24 aromatic amino and nitro compounds, including the known human bladder carcinogens o-toluidine, 2-naphthylamine, benzidine, and 4aminobiphenyl. Internal exposure was assessed by arylamines in pre- and postshift urine and the corresponding hemoglobin adducts. In addition, Sphenyl mercapturic acid and 3OH-benzo关a兴pyrene were measured in urine to monitor exposure to benzene and benzo关a兴pyrene. Individual smoking behavior was assessed by questionnaire and urinary cotinine. Results: Neither urinary arylamines nor hemoglobin adducts correlated with the individual external exposure, no significant increases during shift could be observed. Oven-charging operations and leakage around poorly sealed coke oven doors are expected the major sources of gaseous emissions from coke ovens. However, workers engaged in working tasks near these sources did not show higher internal exposure to arylamines. Internal exposure to arylamines was within the range of environmental exposures observed in smokers and nonsmokers of the general population. But internal exposure to benzene and benzo关a兴pyrene was elevated to some extent. Conclusion: The results of biological monitoring reveal that internal exposure to arylamines was primarily influenced by individual smoking habits. On the other hand, workers were occupationally exposed to benzene and PAH although its exposure was on a comparatively low level. If coke oven workers in this modern plant bear an elevated bladder cancer risk, it could hardly be explained by occupational exposure to those aromatic amino compounds known to be bladder carcinogens in human beings.
PP-30-060 Inflammation, Oxidative Stress, and PAHs Exposure Biomarkers in Children With Atopic Dermatitis Sanghwan Song, Wonjun Heo, Young-Mi Lee, Choonghee Park, and Seung-Do Yu, and Daesun Kim National Institute of Environmental Research, Incheon, Republic of Korea. Background/Aims: Particulate-bounded polycyclic aromatic hydrocarbons (PAHs) are common air pollutants generated from automobile exhaust in urban area. The traffic-related pollutants can exaggerate existing atopic
© 2010 Lippincott Williams & Wilkins
Abstracts
symptoms such as asthma and atopic dermatitis. This study was to investigate urinary eosinophil cationic protein (ECP) as an inflammation biomarker and 8-hydroxydeoxyguanosine (8-OHdG) as an oxidative stress in children exposed to PAHs. Methods: We recruited 43 children with physician-diagnosed atopic dermatitis who attended elementary school located in Incheon city, Korea. With subjects’ agreements, their urine samples were repeatedly obtained. The levels of urinary ECP, 8-OHdG, 1-OHP, and 2-napthol were determined. Results: Of 228 urine samples, the level of ECP, 8-OHdG, 2-napthol, and 1-hydroxypyrene were 0.2 ⫾ 0.36, 13.9 ⫾ 6.8, 3.4 ⫾ 4.6, and 0.2 ⫾ 0.12 g/g creatinine in mean ⫾ standard deviation, respectively. The urinary 1OHP was positive correlate with 8-OHdG but not significant. The urinary 2-napthol showed positive and significant correlation with ECP and 8OHdG concentrations. There was also significant correlation between urinary ECP and 8-OHdG in children. Conclusion: In atopic children, exposure to PAHs may increase urinary inflammation and oxidative stress markers.
PP-30-061 Hemoglobin Adducts of Alkylating Substances—-Long-term Parameters of Smoking During Pregnancy Heiko Ka¨fferlein,1 Ricarda Kopp,1 Julia Latzin,1 Stephan Koslitz,1 Michael Kumbartski,2 and Thomas Bru¨ning1 1IPA Center of Toxicology, Ruhr University Bochum, Bochum, Germany; and 2Department of Gynaecology, University Duisburg-Essen, Essen, Germany. Background/Aims: Few data exist on the transplacental passage of carcinogenic and mutagenic compounds of tobacco smoke. In our study, exposure to alkylating substances was studied in maternal venous and umbilical cord blood samples from 47 mother/child pairs including maternal smokers (15) and nonsmokers (32). Methods: Hemoglobin adducts at the N-terminal valine were analyzed in terms of N-2-carbamoylethylvaline (AAV), N-2-cyanoethylvaline (CEV), and N-2-hydroxyethylvaline (HEV). An N-alkyl-Edman degradation was used for sample preparation. Maternal smoking status was assessed by questionnaire. Results: Nearly identical median levels of AAV were found in maternal and fetal samples of nonsmoking mothers (41.9 and 41.4 pmol/g globin, maternal/fetal ratio ⬃1). Concentrations were about 50% higher in mother/child pairs of maternal smokers. Maternal AAV was associated to their corresponding fetal levels (P ⫽ 0.0003, r ⫽ 0.51). CEV could be detected in maternal and fetal blood of smoking mothers only. The median levels were about twice as high in maternal compared with the fetal samples (70.9 vs. 39.4 pmol/g, maternal/fetal ratio of ⬃2). No association for maternal and fetal CEV was found in smoking mothers. Median concentratons of HEV were 19.8 and 3.3 pmol/g in nonsmoking mother/child pairs, whereas 121.8 and 48.7 pmol/g could be determined in smoking mother/child pairs. The maternal/fetal ratio was ⬃7 in nonsmoking but only 2.5 in smoking mother/child pairs. Maternal HEV was associated with their corresponding fetal levels with borderline significance (P ⫽ 0.0424, r ⫽ 0.30). Conclusion: The results show that alkylating substances are either fully (AAV) or partly transported to the fetus (CEV and HEV), leading to similar or lower concentrations in the fetus. However, no active transport to the fetal unit was observed. Smoking leads to higher adduct levels in the maternal/fetal unit and unequivocally, similar to adults, pose a mutagenic and carcinogenic hazard to the fetus. Overall, our results contribute to the risk assessment of alkylating substances in the most vulnerable population, the fetus.
www.epidem.com | S235
Biological Monitoring of aromatic amines, benzene and benzo[a]pyrene in workers of a modern European coke oven plant Tobias Weiss1, Holger M. Koch1, Heiko U. Käfferlein1, Jana Henry1, Volker Harth1, Kai Süsselbeck2 and Thomas Brüning1
Introduction
Results • Aniline, o-, m- und p-toluidine were found in all air samples • Aniline: Median 8.8 ng/m3 (maximum 78 ng/m3) • o-Toluidine: Median 1.3 ng/m3 (maximum 6.4 ng/m3) • m-Toluidine: Median 1.1 ng/m3 (maximum 9.2 ng/m3) • p-Toluidine: Median 2.7 ng/m3 (maximum 118 ng/m3) • 1 and 2-Naphthylamine, 4-aminodiphenyl, benzidine and o-anisidine were only found in few samples (LOD = 0.1 ng/m3)
Coke oven plant Prosper in Bottrop 7 Number of participants N= 47 Recruiting and logistics 12 • 05:00 Urine sampling pre-shift • 06:00 – 13:00 Air monitoring • 13:00 Questionnaire, urine sampling post-shift
Coke production Coke handling Maintenance
500 400
200 100 0
600 550
NH2
500
NH 2
CH3
400
Non-smoking coke oven workers N=21
Smoking coke oven workers N=26
(Weiss 2005)
(Weiss 2005)
(This study)
(This study)
Biological Monitoring of benzene: Pre- vs. post-shift
N= 47
350
10
300 250 200 150 100 50 0 o-T preshift
o-T postshift
2-NA preshift
2-NA postshift
4-ADP 4-ADP Benzidine Benzidine postprepostpreshift shift shift shift
9 8 7 6 5 4 3 2
No differences between pre- and post-shift specimen
1
Biological Monitoring of aromatic amines: Workplaces near vs. far from ovens
0
Pre-shift
600 NH2
550
NH 2
CH3
500
NH 2
Assessment of benzene exposure and identification of exposure hot spots: 26 Workplaces near ovens 21 Workplaces far from ovens
400 350 300 250 200
Methods
150 100
• Personal Air Monitoring (GC-HRMS) • o-Toluidine, 2-naphthylamine, 4-aminodiphenyl, benzidine (IARC: all K1) • o-Anisidine (K2B), aniline (K3) • m- and p-Toluidin,1-naphthylamine (not classified) as to its carcinogenicity
NH2
2-NA far
Range of remaining collective
4-ADP Benzidine near 4-ADP near Benzidine far far
Biological Monitoring of benzo[a]pyrene: Post-shift
Biological Monitoring of aromatic amines: Non-smokers vs. smokers
2,0
600 NH2
550
NH 2
CH3
500
H 2N
NH 2
NH 2
450 400 350
21 Non-smokers (NS) 26 Smokers
300 250
CH3
200 NH
150 100
e.g. NAT2
• Suitable parameters for Biological Monitoring • Specific • Elimination into urine within a few hours • Reference values known
50
4-Aminodiphenyl in urine [ng/L]
OH
• Suitable parameter for Biological Monitoring • Specific for B[a]P • Represents PAH with high carcinogenic potency • Elimination into urine within a few hours
70 60 50
Comparable associations for o-toluidine und 2-naphthylamine
2000
4000
6000
8000 10000 12000 3OH-Cotinine in urine [µg/L]
Far from ovens
Near ovens NS
Far from ovens NS
99.9
99
Modern coke oven plant (This study)
95 90
50
20
Near ovens
Biological Monitoring of benzo[a]pyrene: Comparison with data from the literature
40 30
Nonsmokers
Occupational exposure mainly at workplaces near the ovens
75
Internal exposure clearly influenced by smoking habits
Mean Smokers*
Steel convertor Refractory brickwork Another coke oven plant Graphite electrode manufacturing (Data from Förster et al. 2008)
25 10 5 1 0.1
Mean Non-smokers *
1
10 3OH-B[a]P [ng/g creatinine] * Lafontaine et al. 2006
Summary and Conclusions
OH
GSH COOH
COOH S
HN
HN
COOCH 3
COOCH 3
S-Phenyl mercapturic acid
• Reaction product of the benzene metabolite benzene epoxide with glutathion • Suitable parameter for Biological Monitoring • Specific • Elimination into urine within a few hours • Correlation with airborne benzene well known • Sources of internal background burden • Non-smokers: ~ 0.1 – 0.3 µg/g creatinine • Smokers: ~ 1 – 3 µg/g creatinine
y = 0.006x + 5.0578 r = 0.83
80
0,0 0,0
Biological Monitoring of benzene: S-phenyl mercapturic acid in urine
S-Phenyl premercapturic acid
90
10
• Sources of internal background burden • Only few data published in literature • Non-smokers: ~ 0.02 – 0.1 ng/g creatinine • Smokers: ~ 0.02 – 0.2 ng/g creatinine • Diet
Benzene epoxide
Smokers
Internal exposure clearly influenced by smoking habits
O
Benzene
0,5
0,0
Influence of tobacco smoking: Association 3-OH-cotinine vs. 4-aminodiphenyl
S
1,0
o-T o-T 2-NA 2-NA 4-ABP 4-ABP Benzidine NS Smokers NS Smokers NS Smokers NS Benzidine Smokers
Biological Monitoring of PAH: 3-Hydroxy benzo[a]pyrene in urine
O
1,5
0
• Sources of internal background burden • Tobacco smoking • Further sources mainly unknown
50 German EKA correlation 45 (exposure equivalents for carcinogenic materials) 40 35 Former German 30 threshold (TRK) 1 ppm (3.25 mg/m3) 25 20 Maintenance worker 15 10 5 0 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 Benzene in air [ppm; ml/m3]
Higher internal exposure in maintenance workers
No differences between workplaces near and far from ovens
relative frequency [%]
O
NH2
o-T far
2-NA near
B[a]P in urine [ng/g creatinine, post-shift]
Biological Monitoring of aromatic amines: Unmetabolized aromatic amines and its phase-II conjugates in urine
0
ng/L urine
• Biological Monitoring • Aromatic amines in urine (GC-MS/MS) • S-Phenyl mercapturic acid in urine as metabolite of benzene (K1) (LC-MS/MS) • 3-Hydroxybenzo[a]pyrene in urine as metabolite of B[a]P (K1) (LC-MS) • Cotinine und 3OH-cotinine in urine to evaluate smoking habits (LC-MS/MS)
50 o-T near
Post-shift Pre-shift Post-shift Pre-shift Post-shift near near far from far from ovens ovens ovens ovens
Occupational exposure apparent
NH 2
H 2N
450
26 workplaces near ovens 21 workplaces far from ovens
Smokers of the general population N=45
No occupational exposure apparent
NH 2
H 2N
Non-smokers of the general population N=145
NH 2
450
28
Comparable results for 2-naphthylamine and 4-aminodiphenyl
600
Biological Monitoring of aromatic amines: Pre- vs. post-shift
ng/L urine
Collektive
700
300
• No differences between workplaces near the ovens and other workplaces
• Evaluation of occupational exposure to aromatic amines and additional hazardous substances such as benzene and PAH • Examination of existing technical safety measures • Identification of exposure hot spots • Establishment of protection measures if necessary • Human Biomonitoring is preferred method, because • aromatic amines, benzene and B[a]P penetrate through the skin • of changing working tasks • of outdoor working places
Collective and methods
Biological Monitoring of aromatic amines: Comparison with background burden of the general population
SPMA in urine [µg/g creatinine]
Motivation • Prevention of occupational diseases • Bladder cancer due to aromatic amines • Cancer of the hematopoietic system due to benzene • Lung cancer due to coke oven emissions • Lack of data of external and internal exposure to aromatic amines
Personal Air Monitoring of aromatic amines:
ng/L urine
• Dry distillation of stone coal at 1100 °C for 24 hours under airtight conditions • Coke (~ 75%) • Side products (~ 25%), e.g. • Tar • Gas • Benzene • Ammonium sulfate • Sulfuric acid
o-Toluidine in urine [ng/L]
2
Institute for Prevention and Occupational Medicine (IPA) of the German Social Accident Insurance, Institute of the Ruhr-Universität Bochum Buerkle-de-la-Camp-Platz 1, 44789 Bochum, Germany Institut für Gefahrstoff-Forschung (IGF) der Berufsgenossenschaft Rohstoffe und chemische Industrie at the Ruhr-Universität Bochum Waldring 97, 44789 Bochum, Germany
SPMA in urine [µg/ g creatinine]
1
• Aromatic amines (K1: o-Toluidine, 2-naphthylamine, 4-aminodiphenyl, benzidine) • Internal burden within the range of the general population • Clearly influenced by smoking status (exception: benzidine) • No additional occupational exposure apparent • Benzene (K1) • Occupational internal exposure apparent, comparable to external exposure < 0.05 ppm; for comparison: general population < 0.01 ppm • Maintenance workers had elevated internal exposure maybe due to dermal uptake • PAH: Benzo[a]pyrene (K1) • Occupational internal exposure apparent mainly in workers with working tasks near the ovens • Internal exposure was lower than formerly reported in the literature If coke oven workers in this modern plant bear an elevated bladder cancer risk it could hardly be explained by occupational exposure to aromatic amino compounds known to be bladder carcinogens in humans.
Abbreviations and references
o-T 4-ADP B[a]P SPMA
= o-toluidine; 2-NA = 2-naphthylamine; = 4-aminodiphenyl; = benzo[a]pyrene; = S-phenyl mercapturic acid; NS = non-smokers
Blome H, Lichtenstein N, Kredel P, Goergens U: 2-Naphthylamin. Gefahrstoffe Reinhaltung der Luft (1999) Nr. 11/12, S. 445-446 Förster K, Preuss R, Rossbach B, Brüning T, Angerer J, Simon P: 3-Hydroxybenzo[a]pyrene in the urine of workers with occupational exposure to polycyclic aromatic hydrocarbons in different industries. Occup Environ Med. 2008 Apr;65(4):224-9. Lafontaine M, Champmartin C, Simon P, Delsaut P, Funck-Brentano C: 3-Hydroxybenzo[a]pyrene in the urine of smokers and non-smokers. Toxicol Lett. 2006 Apr 10;162(2-3):181-5. Riedel K, Scherer G, Engl J, Hagedorn HW, Tricker AR: Determination of three carcinogenic aromatic amines in urine of smokers and nonsmokers. J Anal Toxicol. 2006 Apr;30(3):187-95. Weiss T, Angerer J: Simultaneous determination of various aromatic amines and metabolites of aromatic nitro compounds in urine for low level exposure using gas chromatography-mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. (2002) Nr. 5;778(1-2), S. 179-92 Weiss T: Entwicklung & Anwendung analytischer Methoden zum Biologischen Monitoring & Biochemischen Effektmonitoring von aromatischen Aminen im Rahmen arbeits- & umweltmedizinischer Fragestellungen. Erlangen 2005.
