Science of the Total Environment 374 (2007) 183 198 www.elsevier.

com/locate/scitotenv

Review

A critique of benzene exposure in the general population

Eric S. Johnson a,, Sverre Langrd b , Yu-Sheng Lin a
a

Department of Environmental and Occupational Health, School of Public Health, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX 76107, United States b Center for Occupational and Environmental Medicine, Rikshospitalet-Radiumhospitalet, Oslo, Norway Received 3 August 2006; received in revised form 21 November 2006; accepted 28 November 2006 Available online 29 January 2007

Abstract Benzene risk assessment indicates that exposure to a time-weighted average (TWA) of 15 parts per million (ppm) benzene in ambient air for 40 years is associated with an increased risk of acute myeloid leukemia. Decreased white blood cell count, platelet count and other hematological indices have also been observed in persons exposed to as low as 1 ppm airborne benzene. Evidence from studies worldwide consistently shows elevated levels of benzene biomarkers that are equivalent to 0.12 ppm benzene in ambient air, or even higher in the general population without occupational exposure to benzene (including children). The public health significance of these observations depends on to what extent these levels reflect actual benzene exposure, and whether such exposures are life-long or at least occur frequently enough to pose a possible health threat. We reviewed the evidence and discussed possible explanations for these observations. It was concluded that while there is reason to suspect that benzene contributes significantly to elevated levels of biomarkers in the general population, there is growing concern that this cannot be definitively ascertained without concomitant consideration of the role of other factors such as metabolic polymorphisms and sources of biomarkers other than benzene, which have been insufficiently studied to date. Such studies are urgently needed for valid assessment of this potential public health problem. 2006 Elsevier B.V. All rights reserved.
Keywords: Benzene; Metabolites; Background; Non-occupational; Leukemia; Hematotoxicity

Contents 1. 2. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . Overview of background benzene exposure assessment . . . . . 2.1. Benzene in ambient air . . . . . . . . . . . . . . . . . . 2.2. Benzene in breath, blood, and urine . . . . . . . . . . . 2.3. Urinary metabolites of benzene . . . . . . . . . . . . . . 2.4. Blood albumin adducts of benzene . . . . . . . . . . . . 2.5. Benzene in adipose tissues . . . . . . . . . . . . . . . . Factors that determine background levels of benzene biomarkers 3.1. Petrochemical-related sources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184 186 186 189 189 190 190 191 191

3.

Corresponding author. Tel.: +1 817 735 0327. E-mail address: ejohnson@hsc.unt.edu (E.S. Johnson). 0048-9697/$ - see front matter 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.scitotenv.2006.11.045

184

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

3.2. Tobacco smoking . . . . . . . . . . . . . . 3.3. Ingestion and skin absorption of benzene . . 3.4. Dietary sources of benzene . . . . . . . . . 3.5. Sources of metabolites other than benzene . 3.6. Genetic susceptibility in benzene metabolism 4. Conclusion . . . . . . . . . . . . . . . . . . . . . Acknowledgements . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

. . . . . . . .

191 192 192 192 193 194 195 195

1. Introduction It is generally accepted that benzene is a risk factor for acute myeloid leukemia in humans (Infante et al., 1977; International Agency for Research on Cancer, 1982; Crump and Allen, 1984; Rinsky et al., 1987; Paustenbach et al., 1992; Utterback and Rinsky, 1995; Crump, 1996). Rinsky et al. (1987) in a study of the Pliofilm rubber cohort estimated that exposure to ambient benzene at the level of an 8-hour timeweighted average (TWA) of 1 to 5 parts per million (ppm) over a period of 40 years is associated with a 3-fold risk of myeloid leukemia. In comparison, Crump (1996) estimated a 2-fold relative risk for the same cohort. Likewise, Hayes and his colleagues (Hayes et al., 1997, 2000) reported a relative risk of 6.0 for cases of acute myeloid leukemia combined with myelodysplastic syndrome in subjects with a cumulative exposure of 40 99 ppm-years (i.e. exposed to 1 to 2.5 ppm benzene over 40 years). A recently published study further found that decreased white blood cell count, platelet count, and other hematological values could be observed in persons exposed to 1 ppm benzene (Lan et al., 2004). Thus, these data consistently indicate that persons with 40 or more years exposure to 12 ppm benzene in ambient air and possibly lower, may be at increased risk of developing benzene toxicity, including hematological cancer. It is to be noted that an increased risk of childhood leukemia (odds ratio = 4.0, 95% confidence interval = 1.5 to 10.3) was reported among children living next to petrol stations or a repair garages with exposure duration of 1 to 35 months (no direct environmental measurement was available) (Steffen et al., 2004), and meanwhile about 30,000 new cases of leukemia are diagnosed in the United States each year, but the causes of these leukemia cases are still largely unknown (Sandler and Ross, 1997). Although the mechanism of benzene hematotoxicity and carcinogenicity are not completely understood, it is accepted that one or more of the reactive

metabolites are involved, including benzene oxide (BO), 1,2- and 1,4-benzoquinone (1,2- and 1,4-BQ), and muconaldehyde, as shown in Fig. 1 (Snyder, 2000, 2002). Benzene oxide first spontaneously rearranges to phenol, or undergoes further metabolism to catechol via dihydrodiol dehydrogenases. Two other minor BO metabolites include S-phenylmercapturic acid (SPMA) derived from the conjugation of glutathione with BO (Henderson et al., 2005), and muconaldehyde produced as a result of a second CYP oxidation of oxepin (not shown) that ultimately gives rise to t,t-muconic acid (MA). Phenol could be further converted by CYP2E1 to produce hydroquinone (HQ). Subsequent oxidation of HQ and catechol, either spontaneously or via peroxidases, produces 1,4-BQ and 1,2-BQ, respectively. These reactive electrophiles, including BO, 1,2- and 1,4-BQ, can ultimately form covalent bonds with a wide variety of macromolecules including DNA and proteins (Waidyanatha et al., 1998), and presumably involve in the cancer process by inducing cell toxicity in bone marrow cells (Snyder, 2000, 2002). Evidence from studies of benzene biomarkers in blood, urine, or adipose tissue samples (especially urinary metabolites of benzene) indicates that some subjects in the general population may be, at least periodically, exposed to elevated levels of benzene (Tables 14). Cross-sectional studies of benzene biomarkers in subjects without known exposure to benzene have consistently recorded biomarker levels equivalent to 12 ppm 8-hour TWA ambient air benzene in some subjects, and even up to 3 ppm (Ong et al., 1995; Lin et al., 2006). These high levels have also been recorded in children (Weaver et al., 1996; Amodio-Cocchieri et al., 2001). In the study by Weaver et al. (1996), for example, the maximum urinary MA concentration for a group of children was 2001.2 ng/ml (median = 59.5 ng/ml, the concentration of 1000 ng/ml urinary MA is approximately equivalent to 8-hour TWA of 1 ppm benzene). However, the sources or duration of sustenance of these elevated levels of background

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

185

Fig. 1. Major metabolic pathway of benzene (legend: CYP, cytochrome P450; GSH, glutathione; MPO, myeloperoxidase; NQO1, NAD(P)H: quinone oxidoreductase).

biomarkers are not well known, thus limiting the assessment of the public health significance of these findings. Given the known benzene hematoxicity, a better understanding of benzene exposure in the general environment may help determine the extent to which

benzene exposure in the general population contributes to the occurrence of myeloid leukemia and other outcomes in the general population. The purpose of this paper is to examine the evidence for the occurrence of elevated background levels of benzene in the general population, and to draw attention to the need for

186

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

Table 1 Benzene in breath, blood, and urinary samples in the general populations without occupational or known exposure to benzene a Country Belgium China China China and Malaysia Estonia Analyte Blood benzene Exhaled benzene Urinary benzene Urinary benzene Urinary benzene Blood benzene Exhaled benzene Urinary benzene Blood benzene Urinary benzene Blood benzene Minimum/ lower 3 STD b d 7 ng/l 27 ng/l 0.64 ng/l 2 nmol/l <3 nmol/l 0.1 nmol/l 15 nmol/l (NS) 28 ng/l (S) 85 ng/l 0.26 g/L 0.20 g/L 0.12 g/L Singapore Thailand Blood benzene Urine benzene Blood benzene 1.21 nmol/l 18.8 ppt Median/ mean 120 ng/l 69 ng/l 1.49 ng/l 12 nmol/l 7 nmol/l 0.1 nmol/l 110 ng/l (NS) 219 ng/l(S) 1155 ng/l 0.63 g/L 0.30 g/L 0.17 g/L 1.27 nmol/l 1.29 nmol/l 65.6 ppt Maximum/ upper 3 STD b ND16.1 g/l e ND0.41 ppm e 10,142 ng/l 2060 ng/l 10.4 nmol/l 45 nmol/l 22 nmol/l 0.2 nmol/l 462 ng/l (NS) 940 ng/l 1978 ng/l 2.30 g/L 0.68 g/L 0.23 g/L 1.33 nmol/l 2.61 nmol/l 471 ppt Comments c ND0.29 e, f [1.14] g ppm 0.53 f ppm 0.11 f [0.09] g ppm 0.48 g ppm 0.009 0.009 f (mean STD) [1.11] g ppm 0.10 g ppm (NS) 0.210.26 ppm g (S) 0.31 f [0.08] g ppm (bus drivers) 0.24 f [0.71] g ppm (service attendants) 0.06 f [0.18] g ppm (street vendors) 0.02 f [0.06] g ppm (office workers) 0.014 f [0.007] g ppm 12.3 f ppb (children) Reference Hotz et al. (1997) Kim et al. (2006) Waidyanatha et al. (2001) Ong et al. (1995) Kivisto et al. (1997)

Italy Italy Mexico

Brugnone et al. (1998) Gobba et al. (1997) Romieu et al. (1999)

Ong et al. (1996) Navasumrit et al. (2005)

Abbreviations: Cr, creatinine; ln, natural logarithm; S = smokers; ND = non-detectable; NS = nonsmoker; STD, standard deviation. a Including control workers. b 99% confidence limit. c Values (maximum) in the table represent our estimates based on data using the equation(s) published and/or the values actually measured when available in each study. When the equations were not available, we used the following equations to estimate the airborne benzene concentration(s) [molecular weight of benzene 78 g/mol]: For blood benzene BBz: airbrone benzene ppm BBz; ng=l 251 Brugnone et al:; 1998; 0:913:21000

EBz; ppm Yu and Weisel; 1996; 1 0:64   lnUBz; nmol=l 5:42 Kim et al:; 2006: For urinary benzene UBz: airborne benzene ppm exp 0:886 For exhaled benzene EBz: airborne benzene ppm
d e f g

No data available (including not detectable). Results from different job categories. Measured airborne level (maximum) given by the author(s). Estimated airborne level (maximum) using the equation(s) given by the author(s) or the working equation(s) in this study.

determining whether these occurrences pose a public health threat. 2. Overview of background benzene exposure assessment 2.1. Benzene in ambient air A wide range of benzene concentrations in ambient air, both personal and area sampling, have been reported (Wallace et al., 1987; IARC, 1989). Although the mean levels are usually reported as not exceeding 5 parts per

billion (ppb) (Hricko, 1994; Wallace, 1996), and therefore appear not to be cause for concern, these studies have periodically reported much higher maximum levels of airborne benzene that can be up to 0.2 ppm in both area and personal samples (IARC, 1982, 1989; Wallace et al., 1985, 1987; Texas Commission on Environmental Quality, 2006), and occasionally up to 1 ppm (Berlin, 1985), or even higher. For instance, daily average benzene concentrations of 0.03 to 8.92 ppm in ambient air were recorded at different roadside locations in Calcutta, India, in 1993, primarily due to the use of coal burning stoves (Chattopadhyay et al., 1997).

Table 2 Urinary t,t-muconic acid in the general populations without occupational or known benzene exposure a Median/mean 410500 ng/mg Cr (NS + S) 0.4 ppm (NS + S) Maximum/upper 3 STD b Reference Lauwerys et al. (1994) Bechtold et al. (1991) Inoue et al. (1989) Qu et al. (2000) Waidyanatha et al. (2004) Ducos et al. (1992) Ruppert et al. (1997) Amodio-Cocchieri et al. (2001) Ghittori et al. (1995) Gobba et al. (1997) Lee et al. (1993) Ong et al. (1996) Navasumrit et al. (2005) Johnson et al. (1999) Melikian et al. (1994) Weaver et al. (1996) Yu and Weisel (1996) Boogaard and van Sittert (1995) Comments c

Country

Minimum/lower 3 STD b

Belgium

d 60 ng/mg Cr (NS) 130 ng/mg Cr (S) 270 ng/mg Cr (NS)

China China

China China France Germany

Italy

<100 ng/ml (male) <100 ng/ml (female) 0.02 mg/l (NS + S) <40 ng/ml 20 ng/mgCr (NS) 60 ng/mg Cr (S) 28 g/g Cr 500 g/g Cr 0.09 mg/l (NS + S) 130 ng/ml 65 ng/mg Cr (NS) 130 ng/mg Cr (S) 141 g/g Cr

1470 ng/mg Cr (NS) 2000 ng/ml (male) 1600 ng/ml (female) 2600 g/g Cr 0.34 mg/l (NS + S) 660 ng/ml 590 ng/mg Cr (NS) 390 ng/mg Cr (S) 840 g/g Cr

Italy

Italy Singapore

Singapore Thailand

USA USA

10 ng/mg Cr (NS) 93 ng/mg Cr (S) 10 g/g Cr (NS) 30 ng/ml (NS) 30 ng/ml (S) 0.010.03 mg/g Cr (adults) 0.010.02 mg/g Cr (children)

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

USA USA West Europe

7.1 ng/mg Cr (children)

114 ng/mg Cr (NS) 255 ng/mg Cr (S) 115 g/g Cr (NS) 130 ng/ml (NS) 250 ng/ml (S) 140 ng/ml Cr 0.040.11 mg/g Cr (adults) 0.030.13 mg/g Cr (children) 116 ng/mg Cr 5060 ng/mg Cr (NS) 220270 ng/mg Cr (S) 177 ng/mg Cr (children) 124 ng/mg Cr (NS) 0.04 mg/g Cr (NS) 0.06 mg/g Cr (S)

235 ng/mg Cr (NS) 604 ng/mg Cr (S) 637 g/g Cr (NS) 330 ng/ml (NS) 770 ng/ml (S) 350 ng/ml Cr 0.100.30 mg/g Cr (adults) 0.110.66 mg/g Cr (children) 1570 ng/mg Cr 71210 ng/mg Cr (NS) 10002700 ng/mg Cr (S) 2579 ng/mg Cr (children) 824 ng/mg Cr (NS) 0.71 mg/g Cr (NS + S)

1.2 ppm (NS) 2.0 ppm (male) 1.6 ppm (females) 2.0 ppm 0.3 ppm (NS + S) 0.6 ppm 0.5 ppm (NS) 0.3 ppm (S) 0.7 ppm (children 1114 years) 0.2 ppm (NS) 0.4 ppm (S) 0.5 ppm (NS) 0.3 ppm (NS) 0.8 ppm (S) 0.3 ppm 0.10.3 ppm (adults) 0.10.6 ppm (children) 1.3 ppm 0.10.2 ppm (NS) 0.82.3 ppm (S) 2.1 ppm (children) 0.7 ppm (NS) 0.4 ppm (NS + S)

Abbreviations: Cr, creatinine; MA, t,t-muconic acid; S = smokers; NS = nonsmoker; STD, standard deviation. a Including control workers. b 99% confidence limit. c Values (maximum) in the table represent our estimates based on data using the equation(s) published and/or the values actually measured when available in each study. When the equations were not available, we estimated the airborne benzene concentration based on the assumption that urinary muconic acid concentration of 1000 ng/ml urine or 1200 ng/mg Cr is roughly equivalent to exposure to 1 ppm 8-hour TWA benzene in ambient air (Inoue et al., 1989). But various studies have reported MA levels for 1 ppm benzene to vary from 500 g/g Cr to 1880 g/g Cr (Waidyanatha et al., 2004). d No data available (including not detectable).

187

188

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

Table 3 Urinary S-phenylmercapturic acid in the general populations without occupational or known benzene exposure a Country Belgium China China China Europe and Saudi Arabia Europe and North America Germany Germany Netherlands West Europe Minimum/ lower 3 STD b <0.120.28 g/g Cr d 0.002 mg/l (NS + S) <2 g/g Cr 0.9 g/l (NS) 2.2 g/l (S) Median/mean 841.53 g/g Cr 1.87 g/g Cr 1.2 g/g Cr 0.018 mg/l (NS + S) 0.9 mol/mol Cr (NS) 1.7 g/l (NS) 6.1 g/l (S) 4.0 g/g Cr 0.94 mol/mol Cr (NS) 1.71 mol/mol Cr (S) 1.99 g/g Cr (NS) 3.61 g/g Cr (S) Maximum/ Upper 3 STD b 2.5919.86 g/g Cr 5.59 g/g 8.1 g/g Cr 0.079 mg/l (NS + S) 6 g/g Cr 1.9 mol/mol Cr (NS) 2.4 g/l (NS) 11.3 g/l (S) 16 g/g Cr 1.39 mol/mol Cr (NS) 2.52 mol/mol Cr (S) 2.86 g/g Cr (NS) 5.32 g/g Cr (S) Comments c 0.4 ppm 0.54 ppm 0.2 ppm 2.0 ppm (NS +S) 0.1 ppm 0.1 ppm (NS) 0.1 ppm (NS) 0.3 ppm (S) 0.3 ppm 0.07 ppm (NS) e 0.12 ppm (S) 0.1 ppm (NS) 0.1 ppm (S) Reference Hotz et al. (1997) Melikian et al. (2002) Qu et al. (2000) Waidyanatha et al. (2004) van Sittert et al. (1993) Aston et al. (2002) Einig et al. (1996) Stommel et al. (1989) Boogaard and van Sittert (1996) Boogaard and van Sittert (1995)

Abbreviations: Cr, creatinine; ELISA, enzyme-linked immunosorbent assay; S = smokers; NS = nonsmoker; SPMA, S-phenylmercapturic acid; STD, standard deviation. a Including control workers. b 99% confidence limit. c Values (maximum) in the table represent our estimates based on data using the equation(s) published and/or the values actually measured when available in each study. When the equations were not available, we estimated the airborne benzene concentration based on the assumption that urinary SPMA concentration of 46 g/g Cr is equivalent to exposure to 1 ppm 8-hour TWA benzene in ambient air (van Sittert et al., 1993). d No data available (including not detectable). e Note that 1 ppm benzene is equivalent to 0.04 mg SPMA/l or 21 mol SPMA/mol Cr in this study.

These reports raise some concern when the following are considered: 1) Given that typically environmental data are positively skewed, it may not be suitable to use

mean levels of contaminants for assessing the public health importance of environmental exposures, as some members of the general population with high exposures

Table 4 Protein (albumin and hemoglobin) adduct of benzene in the general populations without occupational or known benzene exposure a Albumin adduct BO-Alb Country Range (median) China Comments b Reference Rappaport et al. (2002a)

BO-Alb

China

81.6398 (170) pmol/g Alb (M) Reference group (benzene-exposed workers): 90.6544 (176) pmol/g Alb (F) Median adduct levels was 357 pmol/g Alb for males (median benzene exposure = 4.61 ppm), and 393 pmol/g Alb for females (median benzene exposure = 2.93 ppm) 44.7248 (115) pmol/g Alb <0.0160.110 ppm 2064716 (550) pmol/g Alb 69.3160 (107) pmol/g Alb 20.671.1 (34.2) pmol/g Hb

1,4-BQ-Alb USA BO-Alb China BO-Hb

BO-Alb China 1,4-BQ-Alb BO-Hb

6.9248 (106) pmol/g alb 9499410 (2110) pmol/g alb 23.362.5 (37.1) pmol/g alb

1.083.64 ppm Reference group (benzene-exposed workers): BO-Alb For exposure to >31 ppm, range: 339 to 3780 pmol/g Alb For exposure to <31 ppm, range: 238 to 220 pmol/g Alb BO-Hb For exposure to >31 ppm, range: 25.3 to 241 pmol/g Hb For exposure to <31 ppm, range: 21.9 to 111 pmol/g Hb Reference group (benzene-exposed workers exposed to <31 ppm) Yeowell-O'Connell Range of BO-Alb was 1601520 pmol/g alb et al. (2001) Range of 1,4-BQ-Alb was 241015,020 pmol/g alb Range of BO-Hb was 27.4157 pmol/g alb

Rappaport et al. (2002b) Lin et al. (2006) Yeowell-O'Connell et al. (1998)

Abbreviations: BO-Alb, benzene oxidealbumin adduct; 1,4-BQ-Alb, 1,4-benzoquinone albumin adduct. a Including control workers. b The levels of benzene exposure were reported in the papers.

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

189

may be masked and not identified. More importantly, since the reported values could be significantly lower than actual peak exposure that far exceeds the exposure thresholds for toxicity, it may not provide appropriate information in protecting the population, especially for sensitive subgroups (e.g., pregnant women, children, etc.) who are likely at increased risk from unidentified environmental exposure. Presentation of the full or interquartile range, together with the proportions of subjects exposed at various exposure ranges, should be included in the reports of environmental measurements. 2) Reported mean and maximum values usually represent concentrations averaged over an 8- or 12hour period, thus the actual peak exposure concentrations of benzene for some individuals would be expected to be even significantly higher than the reported interval values. For example, brief exposure to high levels of benzene in air (up to nearly 10 ppm) may occur at least once weekly for subjects in industrialized countries during refueling automobiles, indicating that significant non-occupational benzene exposures may occur over much of a life-time (Egeghy et al., 2002). Also, when unscheduled monitoring of the Port Arthur and Beaumont communities adjacent to petrochemical plants in Texas was carried out for other volatile organics, such as 1,3-butadiene and methyl-tertbutyl ether, maximum instantaneous levels of 0.65 ppm and 1.6 ppm were recorded, respectively (Morris et al., 2004). On the other hand, mean levels were usually reported at ppb levels in these same areas (Texas Natural Resource Conservation Commission, 2000). 3) Air emissions from industrial sources are usually monitored by area air sampling. However, this approach may be limited for the following reasons: First, monitors do not always cover the area of interest, and there is difficulty in determining the optimal sampling locations. In addition, area sampling is not truly representative of individual exposures given the fact that people don't always stay at the same location. Use of personal air sampling overcomes most of the disadvantages of area air sampling, but it still does not account for exposure pathways other than inhalation exposure, such as dermal absorption or ingestion food contaminated with benzene. 2.2. Benzene in breath, blood, and urine Although inhalation exposure is the primary route for benzene exposure (Rinsky et al., 1981), the amounts contributed by other sources may become increasingly significant at low environmental levels. For example, maximum levels of 0.02 to 1 ppm benzene in drinking

water have been reported (Westrick et al., 1994). Thus, exposures through routes other than inhalation, such as through skin or ingestion, are expected to play an important role in determining the total body burden from environmental exposures. Accordingly, the measurement of unmetabolized benzene in breath, blood, and urine has been increasingly used to assess benzene exposure (US, EPA 1987; Weaver et al., 1996; Brugnone et al., 1998). A wide range of blood, breath, and urinary benzene levels have been observed in the general populations worldwide (Table 1). One of the main advantages of using benzene in breath, blood, or urine, as an exposure surrogate is that it accounts for all known and unknown possible exposure routes (i.e., inhalation, ingestion, and dermal absorption) and unexpected exposures (e.g., accidents). Also, it represents the amount of the pollutant actually absorbed into the body by reflecting individual differences in uptake or genetic heterogeneity (Droz et al., 1991; Lin et al., 2005). In general, there is a good linear relationship of ambient benzene concentrations to benzene levels in biological matrices, namely, blood, exhaled breath and urine (Brugnone et al., 1989; Waidyanatha et al., 2001), especially in occupational settings. In contrast, it is less clear whether benzene in biological matrices (blood, urine, and breath) is useful in non-occupational settings due to inconclusive findings. For instance, a reasonably good correlation (r = 0.77) was observed between benzene exposure and benzene in expired breath in the exposure range of 3.8 10 3 to 11.3 ppm in the study by Egeghy et al. (2002). Another study, however, did not recommend the analysis of exhaled breath when benzene concentrations are below about 1 ppm (Money and Gray, 1989). Also, given the relatively short biologic half-life of benzene (approximately a couple of hours) in the human body (Brugnone et al., 1989; Waidyanatha et al., 2001; Lin et al., 2005), the levels of benzene in breath, blood, and urine only reflect exposures during the preceding hours, and may not be able to reflect cumulative body burden. Moreover, since benzene is ubiquitous, contamination of samples from extraneous sources including the collection tubes, may result in falsely elevated measured levels, and this may be especially critical at the low levels of background exposure. 2.3. Urinary metabolites of benzene Urinary metabolites of benzene can be reliably used to monitor above background air benzene levels of as low as 0.10.3 ppm (Boogaard and van Sittert, 1995;

190

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

Hotz et al., 1997; Kim et al., 2006). In general, the halflives of urinary benzene metabolites are less than 24 h (Qu et al., 2000). While there is concern that the other matrix compounds in urine could interfere with the measurement of low concentrations of urinary metabolites (e.g., MA) (Boogaard and van Sittert, 1996), studies have consistently reported that the levels of the benzene biomarkers, SPMA and MA, are equivalent to 0.052 ppm 8-hour TWA ambient air benzene, and even up to 3 ppm in the general population without known or occupational benzene exposure (Tables 2 and 3). Moreover, in one of these studies, structural confirmation of the high levels of MA measured by the high performance liquid chromatography (HPLC) was also provided by the gas chromatography/mass spectometry (GC/MS) analysis (Weaver et al., 1996). Additionally, several of these studies have reported that as much as 5 to 22% of the studied general population at any one time may have metabolite levels equivalent to exposure to more than an 8-hour TWA of 1 ppm benzene in ambient air (Johnson and Lucier, 1992; Gobba et al., 1997). It is important to note that there are no other known sources of urinary SPMA in humans other than benzene. Of particular interest is that exposure assessment studies of children also show that the levels of urinary metabolites are comparable to exposure to as much as 1 to 2 ppm 8-hour TWA ambient air benzene (Weaver et al., 1996; Amodio-Cocchieri et al., 2001). In the study reported by Weaver et al. (1996), urinary MA levels were as high as 2000 ng/ml in children with a mean age of 4.3 years, and 7.6% of the children had levels greater than 500 ng/ml (Table 2). Note that the background levels are almost as high as those observed in the workplace (Kim et al., 2006), yet they could pose a significantly greater cancer risk to children than adults because their bodies are growing (increased cell divisions) and children often have less-developed systems to excrete chemicals (Suk et al., 2003; Smith et al., 2005). 2.4. Blood albumin adducts of benzene Considerable levels of background albumin adduct, including benzene oxide albumin adduct (BO-Alb) and 1,4-benzoquinone albumin adduct (1,4-BQ-Alb), have been observed in populations without occupational exposure to benzene (Table 4). For instance, Rappaport et al. (2002a) found that the levels of 1,4-BQ-Alb observed in non-smoking, unexposed workers in Tianjin, China, were equivalent to those of workers exposed to about 6.3 mg m 3 (2 ppm) benzene in air. Similarly, it was found that the predicted airborne level

of benzene based upon observed levels of 1,4-BQ-Alb, was equivalent to occupational exposures between 1 and 3 ppm of benzene in the US general population (Lin et al., 2006). In comparison with blood, breath, and urine benzene and the urinary metabolites mentioned above, albumin adducts of electrophilic benzene metabolites, namely, BO-Alb and 1,4-BQ-Alb have a relatively longer halflife of 21 days and 13.5 days, respectively (Rappaport et al., 2002a). No data to date are available for albumin adducts of 1,2-benzoquinone (1,2-BQ-Alb), but presumably it would be similar to 1,4-BQ-Alb. Thus, albumin adducts of benzene are suitable for assessing long-term exposures to benzene due to the slow elimination in human body. 2.5. Benzene in adipose tissues The partitioning of lipophilic toxic vapors, such as benzene and 1,3-butadiene, into blood or adipose tissues has been shown to be one of the most important physicochemical properties in determining the respiratory kinetics and toxicity of volatile chemicals or vapors in humans (Droz et al., 1989; Lin et al., 2002). In the National Human Adipose Tissue Survey study by the US Environmental Protection Agency (EPA), benzene levels of up to 0.02 ppm were recorded in composite samples of adipose tissue derived from biopsy and autopsy specimens, which were routinely collected from a sample of hospitals nationwide for pollutant analysis (US EPA, 1982). In this study, measurements were made on 100 samples pooled together each time, thus some individual levels could have been substantially higher than the reported maximum of 0.02 ppm. Given the invasiveness of this approach, it is difficult to obtain adipose tissue from live human subjects. Thus, benzene concentrations in adipose tissues are usually determined using pharmacokinetic models (Medinsky et al., 1995; Rappaport et al., 2005a, b). A study of the US general population had shown a good agreement among the levels of benzene in measured personal air, measured breath air, and the simulation-derived adipose tissue (Hattemer-Frey et al., 1990). Traditionally, the population average rather than individual values is used to determine the distribution of airborne pollutants in human tissues. However, this approach would be inappropriate if there was a significant variation in the simulation model parameters (i.e., ventilation or partition coefficients) across people, or even within the same individual across time (Lin et al., 2002).

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

191

3. Factors that determine background levels of benzene biomarkers Elevated background levels of benzene biomarkers observed in some subjects in the general population could be attributed to any of the following reasons, alone or in any combination: 1) these subjects are exposed to much higher levels of benzene than do the rest of the population; 2) all community members have similar background benzene exposure, but certain individuals have higher biomarker levels because of inherent genetic, physiological or anatomical differences; 3) all community members have similar exposures to background benzene, but some of them may also be exposed to sources of the biomarkers other than benzene that could lead to higher levels of the biomarkers. 3.1. Petrochemical-related sources One of the main sources of elevated ambient air benzene is auto exhaust and/or gasoline vapor emission. It is estimated that as much as 60% of air benzene is derived from vaporization and incomplete combustion of gasoline (Perry and Gee, 1994; Wallace, 1996) and it may vary by the benzene content in the gasoline (Verma and des Tombe, 2002). Thus traffic, driving, and refueling at gasoline stations are important sources of benzene exposure in the general population. For example, benzene concentrations at breathing zone have been reported to be as high as 1.1 ppm during refueling at gasoline stations (Tironi et al., 1986). In another study of self-service customers monitored with personal air samplers, benzene exposure was in the range of < 0.02 to 11 ppm (mean standard deviation = 0.91 1.81 ppm) roughly equivalent to 0.001 to 0.4 ppm 8-hour TWA while refueling, and postexposure breath levels varied from < 0.9 ppb to 0.4 ppm (mean standard deviation = 0.08 0.05 ppm) (Egeghy et al., 2002). Several similar studies have reported that in-vehicle concentrations of benzene can be as much as 1.5 to 10 times ambient air background concentrations (Chan et al., 1991; Wallace, 1996). The other important sources include point sources like petrochemical plants or oil refineries. In a study by US EPA, personal air benzene concentrations of up to 0.2 ppm were recorded in the Elizabeth and Bayonne communities adjacent to chemical plants in New Jersey (Wallace et al., 1987). The maximum background benzene concentrations typically ranged from 0.03 to 0.07 ppm with a minimum level of 0.01 ppm in areas away from chemical plants. Maximum levels of

airborne benzene up to 0.2 ppm (interquartile range = 0.130.55 ppb; median = 0.26 ppb) were also recorded for outdoor air measurement in Texas City, Texas, an industrial city with huge oil refineries and petrochemical factories. In comparison, the concentrations of background benzene were significantly lower with the maximum level of 3.5 ppb (interquartile range = 0.100.26 ppb; median = 0.16 ppb) in typical localities without such plants during 20042005 (data not shown, Texas Commission on Environmental Quality, 2006). This indicates that populations living close to chemical plants could be at increased risks from exposure to elevated levels of environmental benzene. To what extent this may occur is not known because information on the periodicity or sustenance of these exposures is not available. 3.2. Tobacco smoking Tobacco smoking is known to be a significant contributor to benzene exposure in the general population (Wallace, 1989; Rappaport et al., 2002a; Lin et al., 2006). It is estimated that smokers receive about 90% of their benzene exposure from smoking (mainstream and environmental tobacco smoke), with an average benzene body burden about 6 to 10 times that of nonsmokers (Wallace, 1996; Gordon et al., 2002). On the other hand, nonsmokers receive only about 10% of their benzene exposure from environmental tobacco smoke, 6% from point sources such as petrochemical plants or refineries, and the majority of their benzene exposure from auto exhaust or gasoline vapor emissions (Wallace, 1996). It should be pointed out that these reports only referred to known sources of benzene, and did not take into account benzene uptake through other routes like skin absorption or ingestion. The levels of urinary metabolites of benzene found in smoking populations worldwide were usually much lower (approximately 510 times) than those derived from exposure to 12 ppm 8-hour TWA airborne benzene. The fact that in these populations, urinary MA levels in smokers are no more than 23 times higher than those in non-smokers (Table 2), strongly suggested that smoking per se cannot explain the findings of high metabolite levels in other populations. Also, Hotz et al. (1997) reported end-of-shift benzene in exhaled air of (0.41) ppm, and personal air levels averaged over the entire shift of 0.25 ppm (95th percentile), in groups of unexposed workers with median daily cigarette use of zero. These types of results may be a reminder of the existence of unidentified significant sources of benzene other than

192

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

smoking in the general environment. This is borne out by the fact that the high levels of benzene metabolites in the general population of concern (comparable to up to 12 ppm benzene in ambient air) have frequently been reported not only in non-smoking workers without occupational exposure, but also in children (Tables 2 and 3). For example, high urinary MA concentrations, ranging from 1567 to 2716 ng/mg creatinine, were observed in unexposed workers who were non-smokers as confirmed by both urinary nicotine and cotinine levels in a study of supermarket workers (Johnson et al., 1999). Furthermore, smoking cannot account for the very high levels of urinary MA (up to 2 ppm) seen in children aged 414 years in the United States (Weaver et al., 1996) and Italy (Amodio-Cocchieri et al., 2001). Also, it was found that MA levels in these children were directly related to time spent outside playing in the street (Weaver et al., 1996). This suggests that emission from automobile exhausts containing benzene was probably one of the main sources of the muconic acid. This finding is in agreement with reports that traffic is the main source of benzene in the environment (Perry and Gee, 1994). 3.3. Ingestion and skin absorption of benzene Usually there is little cause for concern regarding benzene in drinking water. However, maximum levels of 0.02 ppm to 1 ppm have been recorded (Westrick et al., 1994; ATSDR, 1995). Notwithstanding, it has been estimated that the actual level of benzene exposure from drinking water could be 10 times higher than the recorded concentration in the water (Burg and Gist, 1992). In a study of gasoline-contaminated ground water in a home in North Carolina, the benzene concentration in ground water was 0.3 ppm, well above the EPA's maximum contaminant standard of 0.005 ppm (Lindstrom et al., 1994). The authors also found that bathroom air levels reached 0.1 to 0.2 ppm benzene with shower stall concentrations in the range of 0.24 to 0.52 ppm on three consecutive days. Likewise, in a case study of benzene exposure by Blank and McAuliffe (1985), the authors found that benzene could be absorbed at the rate of 7.5 /h from inhalation, 7.0 l/h from direct skin contact with gasoline, and 1.5 l from body exposure to ambient air in a worker simultaneously exposed to ambient air benzene level of 10 ppm and with 100 cm2 of glabrous skin in contact with gasoline containing 5% benzene, and his entire skin of 2 m2 exposed to ambient air. These examples clearly show that in addition to exposure to airborne benzene, the routes and sources of non-

airborne benzene exposure may assume disproportionately even greater importance when low environmental exposures are of concern. 3.4. Dietary sources of benzene Benzene has been reported to be present in eggs (25100 g/egg), irradiated beef (19 g/kg), canned beef (2 g/kg), fish, cooked chicken, roasted nuts, various fruits, vegetables and dairy products (IARC, 1982). The average dietary intakes of benzene were in the range of 0.9 2.4 g/day (UK Ministry of Agriculture, Fisheries and Food, 1995) and could be as high as 250 g/day (Brief et al., 1980). However, it is less likely that dietary sources are the major contributors to elevated levels of benzene metabolites in the general population, since the intake of benzene from the diet is usually about 1000 times less than that derived from heavy cigarette smoking (estimated average = 1.8 mg/day) (Wallace, 1989). 3.5. Sources of metabolites other than benzene It has been suggested that ingestion of sorbic acid present in food, drinks, cosmetics, and certain pharmaceuticals could account for high levels of muconic acid seen in unexposed persons (Gobba et al., 1997; Ruppert et al., 1997; Pezzagno et al., 1999; Weaver et al., 2000). On average, daily intake of sorbic acid is only 25 mg in the United States (Yu and Weisel, 1996). Similarly, the average daily intake of sorbic acid observed in Europe was in the range of 630 mg (Ruppert et al., 1997). Experimental studies indicated that ingestion of very high concentrations of sorbic acid can produce urinary MA levels that are equivalent to 12 ppm 8-hour TWA ambient air benzene exposure. For example, Pezzagno et al. (1999) reported that ingestion of a single dose of 447 mg sorbic acid resulted in a mean concentration of 1313 g/l MA (maximum of 2787 g/l) in nonsmokers. Similarly, Ducos et al. (1990) reported that urinary MA levels increased from non-detectable at baseline to 610 and 660 ng/ml in two volunteers 34 h following ingestion of 200 mg of sorbic acid. Other studies, such as Ruppert et al. (1997) and Renner et al. (1999), also reported a considerable increase in urinary MA levels following ingestion of sorbic acid. These results undoubtedly indicate that the usual daily ingestion of small doses of sorbic acid ( 30 mg) cannot account for the high urinary MA levels in the general population. On the other hand it is evident that extreme consumption of sorbic acid in the diet may produce these high levels of urinary MA. For example, a

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

193

second study by Weaver et al. (2000) observed large increases in urinary MA concentrations in 8 adults and children who consumed food assumed to contain sorbic acid. Individual urinary MA peaks ranged as high as 1673.7 ng/ml in the adults and 1221.3 ng/ml in the children following ingestion. However, interpretation of this study should be tempered with caution for the following reasons: 1) the sorbic acid content if any in the foods was not measured; 2) it appears that the ingestion of foods was carried out at home without supervision, and was based on self-report; 3) although baseline MA levels were available, there was no control group of subjects who did not ingest the foods this is important, as metabolite levels are known to show diurnal variation in the absence of known benzene exposure (Weaver et al., 1996; Johnson et al., 1999); 4) the use of 24-hour personal air benzene monitoring could have missed peak exposures; 5) one of the participants did not show any change in MA levels on ingestion of one of the foods even when the challenge was repeated. Similarly, elevated biomarker levels for catechol, HQ, and phenol have been reported from dietary intake (Maga, 1978; Deisinger et al., 1996). In the case of HQ, sources of the compound other than benzene include diet, medications, metabolism of dietary tyrosine by gut bacteria flora, or tyrosine formed from phenylalanine, smoking, etc. (McDonald et al., 2001). Deisinger et al. (1996) showed that arbutin (the beta-D-glucopyranoside conjugate of HQ) occurs in wheat (110 ppm), pears (415 ppm), and free HQ occurs in a variety of food items including: coffee (0.2 ppm), red wine (0.5 ppm), and wheat cereals (0.20.4 ppm). The average concentrations of total HQ in blood and urine peaked 5 and 12 times background levels, respectively, at approximately 2 h after volunteers ingesting arbutin- and HQrich foods. In comparison, the average ratios of peak to background levels of total HQ were 1.5 and 2.5 in blood and urine, respectively, after smoking four cigarettes. These results also indicate that extremely high intake of HQ from dietary sources could conceivably give rise to background levels of blood and urinary HQ approaching those that are of concern. It is concluded that the contribution of non-benzene dietary sources of biomarkers to the elevated levels of these markers seen in some subjects in the general population is variable and depends on the type of benzene biomarker, varying from negligible in the case of SPMA, to significant for MA and HQ in extreme cases, especially when interest is in low levels of benzene exposure (Ong et al., 1995, 1996; Qu et al., 2000; Weaver et al., 2000; Waidyanatha et al., 2001, 2004).

3.6. Genetic susceptibility in benzene metabolism After entering human bodies, benzene is first transformed by cytochrome P4502E1 (CYP2E1) to benzene oxide (BO)-oxepin (Fig. 1). BO could be further transformed into other reactive species (e.g., 1,4benzoquinone, 1,4-BQ) by activation enzymes such as myeloperoxidase (MPO). Alternatively, theses reactive metabolites of benzene can transformed to less harmful derivatives (e.g., hydroxylbenzenes) by reduction enzymes including microsomal epoxide hydrolase (mEH) and/or NAD(P)H: quinone oxidoreductase (NQO1) (Snyder, 2000; Lan et al., 2004). Genetic polymorphisms of metabolic enzymes have been found to be associated with their functional activities. For instance, a 13- to 50-fold variability has been observed in cytochrome P4502E1 activity in humans (Seaton et al., 1994; Klotz and Ammon, 1998; Bolt et al., 2003). Polymorphisms in metabolic enzymes may determine the concentrations of benzene metabolites at a given benzene exposure (Johnson and Lucier, 1992; Gobba et al., 1997; Johnson et al., 1999). In genetically predisposed individuals, benzene could be preferentially converted along the toxic pathways from which reactive intermediates (i.e., muconaldehyde, or benzoquinone) are generated, eventually leading to elevated levels of the corresponding metabolites observed in the general population. Gobba et al. (1997) provided evidence, for instance, that there is significant variation in individual ability to metabolize benzene to MA, in that higher levels of the urinary metabolite were observed in efficient metabolizers (n = 18), 22% of the study population (n = 80). Large differences also are observed between the minimum and maximum background levels of benzene metabolites and adducts, and these differences can be as much as 80- to 360-fold (Tables 24) (Weaver et al., 1996; Amodio-Cocchieri et al., 2001). Some of this variability presumably could be attributable to genetic differences. For example, MA concentrations were related to glutathione S-transferase (GST) polymorphism with significantly higher values observed in null individuals (GSTM1 and GSTT1 combined) (Verdina et al., 2001), and to the CYP2E1 polymorphism in individuals with variant allele of CYP2E1 (RsaI/DraI), which had an influence on the levels of urinary benzene as well (Fustinoni et al., 2005). It is to be noted that GSTT1 polymorphism is also critical to SPMA formation across individuals (Sorensen et al., 2004; Qu et al., 2005). Thus, as shown in Fig. 1, the influence of genetic polymorphism on benzene metabolite levels may not be a simple one because there are complementary pathways

194

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

involving multiple enzymes, each of which may exhibit polymorphism in the genes coding for them. As a result, depending on the overall genetic makeup they possess, one individual may have lower or higher levels of blood or urinary benzene or benzene metabolites than the other. For benzene toxicity, genetic polymorphisms have also been observed in nearly all of the main enzymes involved in the metabolism of benzene, and genetic variations have been linked to the occurrence of leukemia (Table 5). Low NAD(P)H:quinone oxidoreductase 1 (NQO1) activity resulting from a cysteine-tothreonine substitution polymorphism at nucleotide 609 is associated with increased risk of acute leukemia in adults (Smith et al. 2001; Lan et al., 2004). The finding is consistent with previous research that subjects with rapid CYP2E1 activity and NQO1 609 C > T mutation had a 7.6-fold increased risk of benzene poisoning (Rothman et al., 1997). Similarly, the study by Morgan and Smith (2002) had reported that glutathione Stransferase theta (GSTT1) null genotype and possibly glutathione S-transferase mu (GSTM1) null type are associated with increased risk of adult acute leukemia or myelodysplastic syndrome. Noteworthy is that joint interaction among genetic polymorphisms is also critical in determining benzene toxicity. For instance, a study by Wan et al. (2002) reported increased health risks from benzene exposure among individuals with lower NQO1 and GSTT1 activity and higher CYP2E1 activity, but none of them was significant when examined separately. In addition to the importance of genegene interaction, attention should also be drawn to geneenvironment interaction. Previous studies, for instance, have observed an alcohol-induced 3- to 20-fold increase in CYP2E1 activity (Ingelman-Sundberg et al., 1993;

Ronis et al., 1998). This may explain why alcohol consumption has been associated with benzene toxicity (Verma and Rana, 2001; Wan et al., 2002). 4. Conclusion This review demonstrated that elevated levels of benzene biomarkers, namely, blood, exhaled breath and urinary benzene, urinary metabolites (MA and SPMA), and albumin adducts are constantly observed in the general population without occupational exposure to benzene. The data suggest that background benzene exposure could account for a substantial part of these elevated levels. The main sources could be auto exhaust and gasoline vapor emissions, emissions from chemical plants, or unidentified sources. Although tobacco smoking is also one of the important contributing sources explaining a significant part of inhaled benzene in the general population, it does not fully explain the occurrence of these elevated background benzene biomarkers. Similarly, dietary sources of benzene themselves are not likely to be a major contributor. However, for some of the benzene metabolites such as muconic acid and hydroquinone, sources of these compounds other than benzene, for example diet, could be of importance and may need to be considered. In the case of muconic acid, a daily ingestion of 30 mg or less of sorbic acid is unlikely to contribute significantly to background levels of muconic acid. Nevertheless, it is conceivable that individuals consuming extraordinary amounts of sorbic acid from the diet or other sources could produce high concentrations of muconic acid. These levels may not pose a health threat, as the conversion of sorbic acid to muconic acid does not involve the formation of

Table 5 Relationship between genetic polymorphisms involved in benzene metabolism and benzene biomarkers Genetic Metabolite/ polymorphism adduct/end-point EH GST MPO NQO1 CYP2E1 NQO1 CYP2E1 MPO NQO1 GSTM1 GSTT1 Muconic acid Findings Significant relationship in homozygotes especially those with GST null Reference Bergamaschi et al. (1999)

WBC, platelet MPO and NQO1 influenced susceptibility to benzene hematotoxicity, and Lan et al. (2004) Colony formation benzene exposure of <1 ppm caused reduction in WBC and progenitor cells Benzene poisoning Subjects with rapid CYP2E1 related excretion of chlorzoxazone activity and Rothman et al. (1997) NQO1 609 C > T mutation had a 7.6-fold increased risk of benzene poisoning Benzene poisoning NQO1 genotype was associated with benzene poisoning in smokers and alcohol Wan et al. (2002) drinkers. Also certain alleles of CYP2E1, NQO1 and GSTT1 were more susceptible to benzene toxicity.

Abbreviations: CYP2E1, cytochrome P4502E1; EH, microsomal epoxide hydrolase; GSTM1 and GSTT1, glutathione S-transferase T1 and M1, respectively; MPO, myeloperoxidase; NQO1, NAD(P)H: quinone oxidoreductase (NQO1); WBC, white blood cell.

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198

195

muconaldehyde, the toxic precursor of benzene-induced muconic acid (Westoo, 1964), but they may confound the assessment of background benzene exposure. On the other hand, it is not clear whether ingestion of hydroquinone from dietary and non-dietary sources poses a health threat. Ingested hydroquinone is rapidly conjugated in the liver, and much reduced amounts of free hydroquinone are expected to reach the bone marrow. Thus the amount that will be converted to benzoquinone will be similarly much reduced (Smith, 1996), but HQ from these sources may also confound background exposure assessment of benzene. Another significant influential factor is genetic susceptibility in benzene metabolism. The role of genetic polymorphisms on urinary levels of various benzene metabolites is complex and not easily predicted. It is important to remember that disproportionately high levels of metabolites can be observed in some individuals even when benzene exposure is not increased. Also, genetically predisposed individuals could be at increased health risks due to preferential conversion of benzene to reactive, presumably more toxic, metabolites (i.e., muconaldehyde or benzoquinone). Given the complicated genegene and gene environmental interactions, further investigations that consider multiple genes from different pathways simultaneously, including toxicokinetics and sources of inter- and intra-individual variation, are needed to establish a clear link between benzene biomarkers and public health risks. The public health significance of high background levels of benzene or benzene-induced biomarkers in the general population rests on the periodicity or sustenance of these levels and the number of individuals affected. For example, if these exposures occur daily over the lifetime of a significant number of individuals, their occurrence would clearly constitute a major public health problem. On the other hand, this issue would be less of a concern if such exposure scenarios are rare. Unfortunately, only limited information on the background exposure scenario is available. Thus, there is an urgent need to carry out comprehensive longitudinal studies of background benzene exposure that will incorporate the issues discussed above, in order to throw light on this subject. Acknowledgements We are grateful to Professor S.M. Rappaport, Dr. S. Waidyanatha, and the Texas Commission for Environmental Quality for providing valuable comments. The authors have no competing interests.

This work is part of the community health study supported by G67702 Seed Research Program from the University of North Texas Health Science Center at Fort Worth. References
Amodio-Cocchieri R, Del Prete U, Cirillo T, Agozzino E, Scarano G. Evaluation of benzene exposure in children living in Campania (Italy) by urinary trans,trans-muconic acid assay. J Toxicol Environ Health A 2001;63:7987. Aston JP, Ball RL, Pople JE, Jones K, Cocker J. Development and validation of a competitive immunoassay for urinary S-phenylmercapturic acid and its application in benzene biological monitoring. Biomarkers 2002;7:10312. ATSDR. National exposure registry: benzene subregistry. Baseline technical report. Atlanta, GA: Agency for Toxic Substances and Disease Registry; 1995. Bechtold WE, Lucier G, Birnbaum LS, Yin SN, Li GL, Henderson RF. Muconic acid determinations in urine as a biological exposure index for workers occupationally exposed to benzene. Am In Hyg Assoc J 1991;52:4738. Bergamaschi E, Brustolin A, De Palma G, Manini P, Mozzoni P, Andreoli R, et al. Biomarkers of dose and susceptibility in cyclists exposed to monoaromatic hydrocarbons. Toxicol Lett 1999;108: 2417. Berlin M. Low level benzene exposure in Sweden: effect on blood elements and body burden of benzene. Am J Ind Med 1985;7: 36573. Blank IH, McAuliffe DJ. Penetration of benzene through human skin. J Invest Dermatol 1985;85:5226. Bolt HM, Roos PH, Thier R. The cytochrome P-450 isoenzyme CYP2E1 in the biological processing of industrial chemicals: consequences for occupational and environmental medicine. Int Arch Occup Environ Health 2003;76:17485. Boogaard PJ, van Sittert N. Biological monitoring of exposure to benzene: a comparison between S-phenylmercapturic acid, trans, trans,-muconic acid, and phenol. Occup Environ Med 1995;52: 61120. Boogaard PJ, van Sittert NJ. Suitability of S-phenyl mercapturic acid and trans-trans-muconic acid as biomarkers for exposure to low concentrations of benzene. Environ Health Perspect 1996;104 (Suppl 6):11517. Brief RS, Lynch J, Bernath T, Scala RA. Benzene in the workplace. Am Ind Hyg Assoc J 1980;41:61623. Brugnone F, Perbellini L, Faccini GB, Pasini F, Maranelli G, Romeo L, et al. Breath and blood levels of benzene, toluene, cumene and styrene in non-occupational exposure. Int Arch Occup Environ Health 1989;61:30311. Brugnone F, Perbellini L, Romeo L, Bianchin M, Tonello A, Pianalto G, et al. Benzene in environmental air and human blood. Int Arch Occup Environ Health 1998;71:5549. Burg J, Gist GL. Chronic low-level exposure to benzene. J Occup Med 1992;34:6812. Chan CC, Spengler JD, Ozkaynak H, Lefkopoulou M. Commuter exposures to VOCs in Boston, Massachusetts. J Air Waste Manage Assoc 1991;41:1594600. Chattopadhyay G, Samanta G, Chatterjee S, Chakraborti D. Determination of benzene, toluene, and xylene in ambient air of Calcutta for three years during winter. Environ Technol 1997;18: 2118.

196

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198 Hricko A. Rings of controversy around benzene. Environ Health Perspect 1994;102:27681. IARC. Monographs on the evaluation of the carcinogenic risk of chemicals to humans. Some Industrial Chemicals and Dyestuffs, vol. 29. Lyon, France: International Agency for Research on Cancer; 1982. IARC. Monographs on the evaluation of the carcinogenic risk of chemicals to humans. Benzene, vol. 45. Lyon, France: International Agency for Research on Cancer; 1989. Infante PF, Rinsky RA, Wagoner JK, Young RJ. Leukaemia in benzene workers. Lancet 1977;2:768. Ingelman-Sundberg M, Johansson I, Yin H, Terelius Y, Eliasson E, Clot P, et al. Ethanol-inducible cytochrome P4502E1: Genetic polymorphism, regulation, and possible role in the etiology of alcohol-induced liver disease. Alcohol 1993;10:44752. Inoue O, Seiji K, Nakatsuka H, Watanabe T, Yin SN, Li GL, et al. Urinary t,t-muconic acid as an indicator of exposure to benzene. Br J Ind Med 1989;46:1227. Johnson ES, Lucier G. Perspectives on risk assessment impact of recent reports on benzene. Am J Ind Med 1992;21:74957. Johnson ES, S.H., Netto G, Lucier G, Bechtold W, Henderson R. Detection of low level benzene exposure in supermarket wrappers by urinary muconic acid. Biomarkers 1999;4:10617. Kim S, Vermeulen R, Waidyanatha S, Johnson BA, Lan Q, Rothman N, et al. Using urinary biomarkers to elucidate dose-related patterns of human benzene metabolism. Carcinogenesis 2006;27: 77281. Kivisto H, Pekari K, Peltonen K, Svinhufvud J, Veidebaum T, Sorsa M, et al. Biological monitoring of exposure to benzene in the production of benzene and in a cokery. Sci Total Environ 1997;199:4963. Klotz U, Ammon E. Clinical and toxicological consequences of the inductive potential of ethanol. Eur J Clin Pharmacol 1998;54: 712. Lan Q, Zhang L, Li G, Vermeulen R, Weinberg RS, Dosemeci M, et al. Hematotoxicity in workers exposed to low levels of benzene. Science 2004;306:17746. Lauwerys RR, Buchet JP, Andrien F. Muconic acid in urine: a reliable indicator of occupational exposure to benzene. Am J Ind Med 1994;25:297300. Lee BL, Ong HY, Shi CY, Ong CN. Simultaneous determination of hydroquinone, catechol and phenol in urine using high-performance liquid chromatography with fluorimetric detection. J Chromatogr 1993;619:25966. Lin YS, Smith TJ, Wypij D, Kelsey KT, Sacks FM. Association of the blood/air partition coefficient of 1,3-butadiene with blood lipids and albumin. Environ Health Perspect 2002;110:1658. Lin YS, Kupper LL, Rappaport SM. Air samples versus biomarkers for epidemiology. Occup Environ Med 2005;62:75060. Lin YS, McKelvey W, Waidyanatha S, Rappaport SM. Variability of albumin adducts of 1,4-benzoquinone, a toxic metabolite of benzene, in human volunteers. Biomarkers 2006;11:1427. Lindstrom AB, Highsmith VR, Buckley TJ, Pate WJ, Michael LC. Gasoline-contaminated ground water as a source of residential benzene exposure: a case study. J Expo Anal Environ Epidemiol 1994;4:18395. Maga JA. Simple phenol and phenolic compounds in food flavor. CRC Crit Rev Food Sci Nutr 1978;10:32372. McDonald TA, Holland NT, Skibola C, Duramad P, Smith MT. Hypothesis: phenol and hydroquinone derived mainly from diet and gastrointestinal flora activity are causal factors in leukemia. Leukemia 2001;15:1020.

Crump KS. Risk of benzene-induced leukemia predicted from the Pliofilm cohort. Environ Health Perspect 1996;104(Suppl. 6):143741. Crump KS, Allen BC. Quantitative estimates of risk of leukemia form occupational exposure to benzene. OSHA Docket H.059b, Exhibit 152 (Appendix B). Washington DC: U.S. Occupational Safety and Health Administration; 1984. Deisinger PJ, Hill TS, English JC. Human exposure to naturally occurring hydroquinone. J Toxicol Environ Health 1996;47:3146. Droz PO, Wu MM, Cumberland WG, Berode M. Variability in biological monitoring of solvent exposure. I. Development of a population physiological model. Br J Ind Med 1989;46:44760. Droz PO, Berode M, Wu MM. Evaluation of concomitant biological and air monitoring results. Appl Occup Environ Hyg 1991;6: 46574. Ducos P, Gaudin R, Robert A, Francin JM, Maire C. Improvement in HPLC analysis of urinary trans,trans-muconic acid, a promising substitute for phenol in the assessment of benzene exposure. Int Arch Occup Environ Health 1990;62:52934. Ducos P, Gaudin R, Bel J, Maire C, Francin JM, Robert A, et al. trans, trans-muconic acid, a reliable biological indicator for the detection of individual benzene exposure down to the ppm level. Int Arch Occup Environ Health 1992;64:30913. Egeghy PP, Nylander-French L, Gwin KK, Hertz-Picciotto I, Rappaport SM. Self-collected breath sampling for monitoring low-level benzene exposures among automobile mechanics. Ann Occup Hyg 2002;46:489500. Einig T, Dunemann L, Dehnen W. Sensitive gas chromatographic method for determination of mercapturic acids in human urine. J Chromatogr B Biomed Appl 1996;687:37985. Fustinoni S, Consonni D, Campo L, Buratti M, Colombi A, Pesatori AC, et al. Monitoring low benzene exposure: comparative evaluation of urinary biomarkers, influence of cigarette smoking, and genetic polymorphisms. Cancer Epidemiol Biomarkers Prev 2005;14:223744. Ghittori S, Maestri L, Fiorentino ML, Imbriani M. Evaluation of occupational exposure to benzene by urinalysis. Int Arch Occup Environ Health 1995;67:195200. Gobba F, Rovesti S, Borella P, Vivoli R, Caselgrandi E, Vivoli G. Interindividual variability of benzene metabolism to trans,transmuconic acid and its implications in the biological monitoring of occupational exposure. Sci Total Environ 1997;199:418. Gordon SM, Wallace LA, Brinkman MC, Callahan PJ, Kenny DV. Volatile organic compounds as breath biomarkers for active and passive smoking. Environ Health Perspect 2002;110:68998. Hattemer-Frey HA, Travis CC, Land ML. Benzene: environmental partitioning and human exposure. Environ Res 1990;53:22132. Hayes RB, Yin SN, Dosemeci M, Li GL, Wacholder S, Travis LB, et al. Benzene and the dose-related incidence of hematologic neoplasms in China. Chinese Academy of Preventive Medicine National Cancer Institute Benzene Study Group. J Natl Cancer Inst 1997;89:106571. Hayes RB, Yin S, Rothman N, Dosemeci M, Li G, Travis LT, et al. Benzene and lymphohematopoietic malignancies in China. J Toxicol Environ Health A 2000;61:41932 [In process citation]. Henderson AP, Barnes ML, Bleasdale C, Cameron R, Clegg W, Heath SL, et al. Reactions of benzene oxide with thiols including glutathione. Chem Res Toxicol 2005;18:26570. Hotz P, Carbonnelle P, Haufroid V, Tschopp A, Buchet JP, Lauwerys R. Biological monitoring of vehicle mechanics and other workers exposed to low concentrations of benzene. Int Arch Occup Environ Health 1997;70:2940.

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198 Medinsky MA, Kenyon EM, Schlosser PM. Benzene: a case study in parent chemical and metabolite interactions. Toxicology 1995;105: 22533. Melikian AA, Prahalad AK, Secker-Walker RH. Comparison of the levels of the urinary benzene metabolite trans,trans-muconic acid in smokers and nonsmokers, and the effects of pregnancy. Cancer Epidemiol Biomarkers Prev 1994;3:23944. Melikian AA, Qu Q, Shore R, Li G, Li H, Jin X, et al. Personal exposure to different levels of benzene and its relationships to the urinary metabolites S-phenylmercapturic acid and trans,transmuconic acid. J Chromatogr B Analyt Technol Biomed Life Sci 2002;778:21121. Money CD, Gray CN. Exhaled breath analysis as a measure of workplace exposure to benzene ppm. Ann Occup Hyg 1989;33: 25762. Morgan GJ, Smith MT. Metabolic enzyme polymorphisms and susceptibility to acute leukemia in adults. Am J Pharmacogenomics 2002;2:7992. Morris DL, Barker PJ, Legator MS. Symptoms of adverse health effects among residents from communities surrounding chemicalindustrial complexes in southeast Texas. Arch Environ Health 2004;59:1605. Navasumrit P, Chanvaivit S, Intarasunanont P, Arayasiri M, Lauhareungpanya N, Parnlob V, et al. Environmental and occupational exposure to benzene in Thailand. Chem Biol Interact 2005;153154:7583. Ong CN, Kok PW, Lee BL, Shi CY, Ong HY, Chia KS, et al. Evaluation of biomarkers for occupational exposure to benzene. Occup Environ Med 1995;52:52833. Ong CN, Kok PW, Ong HY, Shi CY, Lee BL, Phoon WH, et al. Biomarkers of exposure to low concentrations of benzene: a field assessment. Occup Environ Med 1996;53:32833. Paustenbach DJ, Price PS, Ollison W, Blank C, Jernigan JD, Bass RD, et al. Reevaluation of benzene exposure for the Pliofilm (rubberworker) cohort (19361976). J Toxicol Environ Health 1992;36:177231. Perry R, Gee IL. Vehicle emissions and effects on air quality: indoors and outdoors. Indoor Environ 1994;3:22436. Pezzagno G, Maestri L, Fiorentino ML. Trans,trans-muconic acid, a biological indicator to low levels of environmental benzene: some aspects of its specificity. Am J Ind Med 1999;35:5118. Qu Q, Melikian AA, Li G, Shore R, Chen L, Cohen B, et al. Validation of biomarkers in humans exposed to benzene: urine metabolites. Am J Ind Med 2000;37:52231. Qu Q, Shore R, Li G, Su L, Jin X, Melikian AA, et al. Biomarkers of benzene: urinary metabolites in relation to individual genotype and personal exposure. Chem Biol Interact 2005;153154: 8595. Rappaport SM, Waidyanatha S, Qu Q, Shore R, Jin X, Cohen B, et al. Albumin adducts of benzene oxide and 1,4-benzoquinone as measures of human benzene metabolism. Cancer Res 2002a;62:13307. Rappaport SM, Yeowell-O'Connell K, Smith MT, Dosemeci M, Hayes RB, Zhang L, et al. Non-linear production of benzene oxide albumin adducts with human exposure to benzene. J Chromatogr B Analyt Technol Biomed Life Sci 2002b;778:36774. Rappaport SM, Waidyanatha S, Yeowell-O'connell K, Rothman N, Smith MT, Zhang L, et al. Protein adducts as biomarkers of human benzene metabolism. Chem Biol Interact 2005a;153154:1039. Rappaport SM, Kupper LL, Lin YS. On the importance of exposure variability to the doses of volatile organic compounds. Toxicol Sci 2005b;83:22436.

197

Renner T, Baer-Koetzle M, Scherer G. Determination of sorbic acid in urine by gas chromatography-mass spectrometry. J Chromatogr A 1999;847:12733. Rinsky RA, Young RJ, Smith AB. Leukemia in benzene workers. Am J Ind Med 1981;2:21745. Rinsky RA, Smith AB, Hornung R, Filloon TG, Young RJ, Okun AH, et al. Benzene and leukemia. An epidemiologic risk assessment. N Engl J Med 1987;316:104450. Romieu I, Ramirez M, Meneses F, Ashley D, Lemire S, Colome S, et al. Environmental exposure to volatile organic compounds among workers in Mexico City as assessed by personal monitors and blood concentrations. Environ Health Perspect 1999;107:5115. Ronis MJ, Huang J, Longo V, Tindberg N, Ingelman-Sundberg M, Badger TM. Expression and distribution of cytochrome P450 enzymes in male rat kidney: effects of ethanol, acetone and dietary conditions. Biochem Pharmacol 1998;55:1239. Rothman N, Smith MT, Hayes RB, Traver RD, Hoener B, Campleman S, et al. Benzene poisoning, a risk factor for hematological malignancy, is associated with the NQO1 609C> T mutation and rapid fractional excretion of chlorzoxazone. Cancer Res 1997;57: 283942. Ruppert T, Scherer G, Tricker AR, Adlkofer F. trans,trans-muconic acid as a biomarker of non-occupational environmental exposure to benzene. Int Arch Occup Environ Health 1997;69:24751. Sandler DP, Ross JA. Epidemiology of acute leukemia in children and adults. Semin Oncol 1997;24:316. Seaton MJ, Schlosser PM, Bond JA, Medinsky MA. Benzene metabolism by human liver microsomes in relation to cytochrome P450 2E1 activity. Carcinogenesis 1994;15:1799806. Smith MT. The mechanism of benzene-induced leukemia: a hypothesis and speculations on the causes of leukemia. Environ Health Perspect 1996;104(Suppl. 6):121925. Smith MT, Wang Y, Kane E, Rollinson S, Wiemels JL, Roman E, et al. Low NAD(P)H: quinone oxidoreductase 1 activity is associated with increased risk of acute leukemia in adults. Blood 2001;97: 14226. Smith MT, McHale CM, Wiemels JL, Zhang L, Wiencke JK, Zheng S, et al. Molecular biomarkers for the study of childhood leukemia. Toxicol Appl Pharmacol 2005;206(2):23745. Snyder R. Overview of the toxicology of benzene. J Toxicol Environ Health A 2000;61:33946. Snyder R. Benzene and leukemia. Crit Rev Toxicol 2002;32:155210. Sorensen M, Poole J, Autrup H, Muzyka V, Jensen A, Loft S, et al. Benzene exposure assessed by metabolite excretion in Estonian oil shale mineworkers: influence of glutathione s-transferase polymorphisms. Cancer Epidemiol Biomarkers Prev 2004;13: 172935. Steffen C, Auclerc MF, Auvrignon A, Baruchel A, Kebaili K, Lambilliotte A, et al. Acute childhood leukaemia and environmental exposure to potential sources of benzene and other hydrocarbons; a case-control study. Occup Environ Med 2004;61:7738. Stommel P, Muller G, Stucker W, Verkoyen C, Schobel S, Norpoth K. Determination of S-phenylmercapturic acid in the urinean improvement in the biological monitoring of benzene exposure. Carcinogenesis 1989;10:27982. Suk WA, Murray K, Avakian MD. Environmental hazards to children's health in the modern world. Mutat Res 2003;544 (23):23542. Texas Commission on Environmental Quality. Hourly Air Quality Data. Austin, TX. 2006. Available: http://www.tceq.state.tx.us/ nav/data/aq_data.html [accessed 1 March, 2006].

198

E.S. Johnson et al. / Science of the Total Environment 374 (2007) 183198 Wallace LA. Major sources of benzene exposure. Environ Health Perspect 1989;82:1659. Wallace LA. Environmental exposure to benzene: an update. Environ Health Perspect 1996;104(Suppl. 6):112936. Wallace LA, Pellizzari ED, Hartwell TD, Sparacino CM, Sheldon LS, Zelon H. Personal exposures, indoor-outdoor relationships, and breath levels of toxic air pollutants measured for 355 persons in New Jersey. Atmos Environ 1985;19:165161. Wallace LA, Pellizzari ED, Hartwell TD, Sparacino C, Whitmore R, Sheldon L, et al. The TEAM (Total Exposure Assessment Methodology) Study: personal exposures to toxic substances in air, drinking water, and breath of 400 residents of New Jersey, North Carolina, and North Dakota. Environ Res 1987;43:290307. Wan J, Shi J, Hui L, Wu D, Jin X, Zhao N, et al. Association of genetic polymorphisms in CYP2E1, MPO, NQO1, GSTM1, and GSTT1 genes with benzene poisoning. Environ Health Perspect 2002;110: 12138. Weaver VM, Davoli CT, Heller PJ, Fitzwilliam A, Peters HL, Sunyer J, et al. Benzene exposure, assessed by urinary trans,trans-muconic acid, in urban children with elevated blood lead levels. Environ Health Perspect 1996;104:31823. Weaver VM, Buckley T, Groopman JD. Lack of specificity of trans, trans-muconic acid as a benzene biomarker after ingestion of sorbic acid-preserved foods. Cancer Epidemiol Biomark Prev 2000;9:74955. Westoo G. On the metabolism of sorbic acid in the mouse. Acta Chem Scand 1964;18:13738. Westrick JJ, Mello JW, Thomas RF. The groundwater supply survey. J AWWA 1994:529. Yeowell-O'Connell K, Rothman N, Smith MT, Hayes RB, Li G, Waidyanatha S, et al. Hemoglobin and albumin adducts of benzene oxide among workers exposed to high levels of benzene. Carcinogenesis 1998;19:156571. Yeowell-O'Connell K, Rothman N, Waidyanatha S, Smith MT, Hayes RB, Li G, et al. Protein adducts of 1,4-benzoquinone and benzene oxide among smokers and nonsmokers exposed to benzene in China. Cancer Epidemiol Biomarkers Prev 2001;10:8318. Yu R, Weisel CP. Measurement of benzene in human breath associated with an environmental exposure. J Expo Anal Environ Epidemiol 1996;6:26177.

Texas Natural Resource Conservation Commission. Interoffice memorandum. Austin, TX. 2000, December 1. Tironi G, Nebel G, Williams R. Measurement of vapor pressure during gasoline refueling. SAE paper, vol. 860087. Detroit, MI: International Congress of the Society of Automotive Engineers; 1986. UK Ministry of Agriculture, Fisheries and Food. Benzene and other aromatic hydrocarbons in food-average UK dietary intakes. Food surveillance information sheet, vol. 58. UK Ministry of Agriculture; 1995. US EPA. NHATS broad scan analysis: Population estimates from fiscal year 1982 specimens. EPA, vol. 560/5-90-001. Washington, DC: Office of Pesticides and Toxic Substances, U.S. Environmental Protection Agency; 1982. US EPA. The Total Exposure Assessment Methodology (TEAM) Study. Washington, DC: U.S. Environmental Protection Agency; 1987. Utterback DF, Rinsky RA. Benzene exposure assessment in rubber hydrochloride workers: a critical evaluation of previous estimates. Am J Ind Med 1995;27:66176. van Sittert NJ, Boogaard PJ, Beulink GDJ. Application of the urinary S-phenylmercapturic acid test as a biomarker for low levels of exposure to benzene in industry. Br J Ind Med 1993;50:4609. Verdina A, Galati R, Falasca G, Ghittori S, Imbriani M, Tomei F, et al. Metabolic polymorphisms and urinary biomarkers in subjects with low benzene exposure. J Toxicol Environ Health A 2001;64: 60718. Verma Y, Rana SV. Biological monitoring of exposure to benzene in petrol pump workers and dry cleaners. Ind Health 2001;39:3303. Verma DK, des Tombe K. Benzene in gasoline and crude oil: occupational and environmental implications. AIHA J (Fairfax, Va) 2002;63:22530. Waidyanatha S, Yeowell-O'Connell K, Rappaport SM. A new assay for albumin and hemoglobin adducts of 1,2- and 1,4-benzoquinones. Chem Biol Interact 1998;115:11739. Waidyanatha S, Rothman N, Fustinoni S, Smith MT, Hayes RB, Bechtold W, et al. Urinary benzene as a biomarker of exposure among occupationally exposed and unexposed subjects. Carcinogenesis 2001;22:27986. Waidyanatha S, Rothman N, Li G, Smith MT, Yin S, Rappaport SM. Rapid determination of six urinary benzene metabolites in occupationally exposed and unexposed subjects. Anal Biochem 2004;327:18499.