Regulatory Toxicology and Pharmacology 62 (2012) 62–73

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Regulatory Toxicology and Pharmacology

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Biomonitoring Equivalents for benzene

Sean M. Hays a,b,⇑, David W. Pyatt b,c, Chris R. Kirman d, Lesa L. Aylward e
a
Summit Toxicology, L.L.P., Allenspark, CO, United States
b
University of Colorado, Denver CO, United States
c
Summit Toxicology, L.L.P., Superior, CO, United States
d
Summit Toxicology, L.L.P., Cleveland, OH, United States
e
Summit Toxicology, L.L.P., Falls Church, VA, United States

a r t i c l e i n f o a b s t r a c t

Article history: Biomonitoring Equivalents (BEs) are defined as the concentration or range of concentrations of a chemical
Received 14 September 2011 or its metabolite in a biological medium (blood, urine, or other medium) that is consistent with an exist-
Available online 9 December 2011 ing health-based exposure guideline such as a reference dose (RfD) or tolerable daily intake (TDI). BE val-
ues can be used as a screening tool for the evaluation of population-based biomonitoring data in the
Keywords: context of existing risk assessments. This study reviews available health based risk assessments and
Biomonitoring exposure guidance values for benzene from the United States Environmental Protection Agency (US
Biomonitoring Equivalent
EPA), Texas Commission on Environmental Quality (TCEQ), California’s Office of Environmental Health
Risk assessment
Pharmacokinetics
Hazard Assessment (OEHHA) and the Agency for Toxic Substances and Disease Registry (ATSDR) to derive
Benzene BE values for benzene in blood and urine. No BE values were derived for any of the numerous benzene
metabolites or hemoglobin and albumin adducts. Using existing physiologically based pharmacokinetic
(PBPK) models, government risk assessment values were translated into corresponding benzene levels
in blood assuming chronic steady-state exposures. BEs for benzene in urine were derived using measured
correlations between benzene in urine with benzene in blood. The BE values for benzene in blood range
from 0.04 to 1.29 lg/L, depending upon the underlying non-cancer risk assessment used in deriving the
BE. Sources of uncertainty relating to both the basis for the BE values and their use in evaluation of bio-
monitoring data, including the transience of the biomarkers relative to exposure frequency, are discussed.
The BE values derived here can be used as screening tools for evaluation of population biomonitoring data
for benzene in the context of the existing risk assessment and can assist in prioritization of the potential
need for additional risk assessment efforts for benzene relative to other chemicals.
Ó 2011 Elsevier Inc. All rights reserved.

1. Introduction (NHANES) and other national biomonitoring efforts such as those

Large biomonitoring studies such as those conducted as part of
the US National Health and Nutrition Examination Survey
Abbreviations: ADI, acceptable daily intake; AEGL, acute exposure guideline
level; AML, acute myologenous leukemia; ATSDR, Agency for Toxic Substances and
Disease Registry; BE, Biomonitoring Equivalent; BEPOD, Biomonitoring Equivalent
point of departure; BMDL, benchmark dose lower limit; BO, benzeneoxide; Bz,
benzene; ESL, effects screening level; MRL, Minimal Risk Level; MW, molecular
weight; NOAEL, no observed adverse effect level; 1,4-BQ, 1,4-benzoquinone; POD,
point of departure; RfD, reference dose; SPMA, s-phenyl mercapturic acid; TCEQ,
Texas Commission of Environmental Quality; TD05, The tumorigenic dose05, the
dose level that causes a 5% increase in tumor incidence over background; TDI,
tolerable daily intake; tt-MA, trans, trans-muconic acid; UF, uncertainty factor;
USEPA, United States Environmental Protection Agency; WHO, World Health
Organization.
⇑ Corresponding author at: Summit Toxicology, P.O. Box 427, Allenspark, CO
80510, United States.
E-mail address: shays@summittoxicology.com (S.M. Hays).
conducted in Canada and in Germany are providing valuable data
on the prevalence and concentrations of chemicals in biological
media such as blood or urine from individuals in the general pop-
ulation. These measured concentrations provide an integrated
reflection of exposures that may occur via multiple routes and
pathways, and biomonitoring is increasingly being relied upon as
a state-of-the-art tool for exposure assessment of environmental
chemicals (Sexton et al., 2004). The potential significance of the
measured concentrations of chemicals in the context of existing
toxicology data and risk assessments can be assessed if chemical-
specific quantitative screening criteria are available. Such
screening criteria would ideally be based on robust datasets relat-
ing potential adverse effects to biomarker concentrations in human
populations (see, for example, the US Centers for Disease Control
and Prevention (CDC) blood lead level of concern; see http://
www.cdc.gov/nceh/lead/). However, development of such epide-
miologically-based screening criteria is a resource- and time-

0273-2300/$ - see front matter Ó 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.yrtph.2011.12.001
 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73
intensive effort, and in practice, data to support such assessments
exist for only a few chemicals. As an interim approach, the concept
of Biomonitoring Equivalents (BEs) has been developed (Hays et al.,
2007), and guidelines for the derivation and communication of
these values have been prepared (Hays et al., 2008; LaKind et al.,
2008).
A Biomonitoring Equivalent (BE) is defined as the concentration
or range of concentrations of a chemical or its metabolites in a bio-
logical medium (blood, urine, or other medium) that is consistent
with an existing health-based exposure guidance value such as a
reference dose (RfD) or tolerable daily intake (TDI). Existing chem-
ical-specific pharmacokinetic data are used to estimate biomarker
concentrations that are consistent with the point of departure
(POD) used in the derivation of an exposure guidance value (such
as the RfD or TDI), and with the exposure guidance value itself.
BEs can be estimated using available human or animal pharmaco-
kinetic data (Hays et al., 2008), and BEs have been derived for
numerous compounds including acrylamide, cadmium, 2,4-dichlo-
rophenoxyacetic acid, toluene, and others (reviewed in Hays and
Aylward, 2009; available at www.biomonitoringequivalents.net).
BEs are intended to be used as screening tools to provide an assess-
ment of which chemicals have large, small, or no margins of safety
compared to existing risk assessments and exposure guidance val-
ues. BE values are only as robust as are the underlying exposure
guidance values and pharmacokinetic data used to derive the val-
ues. BE values are not intended to be diagnostic for potential health
effects in humans, either individually or for a population.
This manuscript presents the background and derivation of BE
values for benzene (CAS #71-43-2). Benzene is a ubiquitous envi-
ronmental chemical that is also present in cigarette smoke and var-
ious petroleum derived products including gasoline. Cigarette
smoke, and second hand smoke, are the largest sources of expo-
sures among the general population, and are estimated to be an or-
der of magnitude greater than exposures from all other sources
(Wallace, 1996). However, exposures to benzene can occur via
multiple routes (inhalation, oral and dermal) and from many dif-
ferent sources. As a result, biomonitoring has been used as a poten-
tial tool for better assessing integrated exposures to benzene. The
BEs derived here can be used as a tool to help regulators and risk
managers interpret benzene biomonitoring data in a public health
risk context.
2. Methods
2.1. Exposure guidance values, critical effects and mode of action
The hematopoietic toxicity of benzene exposure has been rec-
ognized for decades and has been consistently observed in both
experimental studies and epidemiological evaluations of worker
populations (ATSDR, 2007). Chronic, high-level occupational expo-
sures to benzene have been positively associated with bone mar-
row failure and aplastic anemia as well as an increased risk of
various subtypes of acute myeloid leukemia (AML) and myelodys-
plastic syndromes (MDS) (Pyatt, 2004). Chronic occupational expo-
sure to high levels of benzene is also an established cause of
various cytopenias observed in the peripheral circulation (reduc-
tions in specific cell lineages) including pancytopenia (a reduction
in all three major cell lines) (Green et al., 1981; Rozen et al., 1984;
Snyder et al., 1978). Peripheral cytopenias, in particular lymphocy-
topenia, are generally believed to be the most sensitive manifesta-
tion of benzene toxicity. As such, lymphocytopenia forms the basis
for most non-cancer regulatory limits set for benzene (Rothman
et al., 1996).
Exposures to benzene may occur due to: (1) the production of
benzene in combustion processes (e.g., forest fires, cigarette smok-
63
ing), (2) the natural presence of benzene in foods, (3) proximity to
industrial sources (e.g., refineries, petrochemical manufacturing,
service stations), (4) pumping and potentially storing gasoline
and (5) emissions from gasoline and other types of engines. Expo-
sure to benzene can occur via the oral, dermal and/or inhalation
routes, although due to benzene’s high vapor pressure, inhalation
is the most important exposure pathway, especially in occupa-
tional environments.
A robust body of experimental evidence supports the hypothe-
sis that benzene must be metabolized to be hematotoxic (Gad-El
Karim et al., 1985; Ross, 1996; Snyder et al., 1981, 1983; Valentine
et al., 1996). The primary step in benzene metabolism is oxidation
to benzene oxide (see Fig. 1) via the 2E1 isozyme of the cytochrome
P-450 system (CYP2E1). Benzene oxide undergoes enzymatic or
non-enzymatic rearrangement to phenol or is metabolized by
epoxide hydrolase (Parke and Williams, 1950, 1953). Although
the primary site of benzene’s oxidation by CYP2E1 occurs mainly
in the liver, additional metabolism also occurs in the bone marrow
(Andrews et al., 1979). Human bone marrow lacks detectable levels
of CYP2E1, but is a rich source of myeloperoxidase (MPO) (Ross,
1996; Ross et al., 1996). MPO, which has been measured in high
concentrations in granulocytic precursor cells, can further oxidize
various polyphenolic metabolites of benzene and is thought to play
an important role in the hematopoietic toxicity of benzene (Gad-El
Karim et al., 1985; Kalf, 1987; Rana and Verma, 2005; Ross, 1996).
The mechanism of action for benzene’s hematopoietic toxicity is
largely unknown. However, there are a few well-defined steps that
seem to be required for benzene to exert its toxicity. These steps
can be used to describe the more general mode of action (MOA)
(Meek and Klaunig, 2010). Benzene must be absorbed into the sys-
temic circulation and metabolized into its various phenolic metab-
olites. These metabolites must be stable enough to persist into the
peripheral circulation and possibly to the bone marrow, where
they likely undergo additional metabolic conversion. As discussed
above, the exact metabolite or combination responsible for hemat-
opoetic toxicity has not been determined with certainty. However,
hydroquinone and p-benzoquinone are likely candidates. Once in
the bone marrow, and perhaps in the peripheral circulation as well,
it appears that the various metabolites interact with either mature
blood cells or early precursor cells that would ultimately produce
mature blood cells (Baarson et al., 1984). The toxicity of benzene
has been shown to be greater in rapidly dividing cells, where more
actively dividing cells are targeted but quiescent hematopoietic
stem cells are spared (Dempster and Snyder, 1990, 1991; Green
et al., 1981; Irons, 1981; Irons et al., 1979a; Keller and Snyder,
1988; Longacre et al., 1980). Additional evidence indicates that
benzene metabolites can induce a block in cell cycle as well as in-
duce apoptosis in precursor cells (Ross et al., 1996; Yoon et al.,
2001). Other studies have shown that benzene metabolites alter
important signaling pathways in hematopoietic cells that may also
result in cytotoxicity (Irons et al., 1992; Pyatt et al., 1998, 1999a,b;
Renz and Kalf, 1991, 1992).
As with peripheral cytopenias, the precise molecular mechanism
underlying benzene’s ability to induce AML has not been identified.
Hypotheses include topoisomerase inhibition and/or hyper-respon-
siveness to various growth factors (Dempster and Snyder, 1990,
1991; Huff et al., 1989; Irons et al., 1992; Lindsey et al., 2005;
Mondrala and Eastmond, 2010). The MOA on the other hand, has
generally been described and there are several steps that have gen-
eral acceptance (Meek and Klaunig, 2010). As with the hematopoi-
etic toxicity described above, the first steps in the MOA for AML are
absorption, metabolism and distribution to the bone marrow. Once
in the marrow, the various benzene metabolites either create or
provide a selective advantage for initiated target cells. These cells
harboring specific mutations (perhaps evidenced by common cyto-
genetic markers) likely achieve a selective growth advantage over
64 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73
Fig. 1. Overview of metabolic scheme for benzene.
their normal counterparts (hematopoeitic progenitor cell [HPC]) in
the bone marrow. With the possibility of requiring additional geno-
toxic events or the formation of a permissive microenvironment,
these cells possess an abnormal ability for proliferation. This results
in the formation of a dangerous population of immature, aberrant
precursor cells that crowd out the normal hematopoiesis typically
occurring in a healthy bone marrow.
Benzene is one of the most widely studied chemicals, with a ro-
bust literature base describing both human and non-human toxic-
ity (ATSDR, 2007; Irons, 1997; Pyatt, 2004). As briefly described,
there is a relative abundance of high quality human data (ATSDR,
2007; USEPA, 2000). As such, human data forms the basis for most
of the toxicity and reference values established by various scien-
tific agencies and regulatory bodies. The one exception is the acute
Minimal Risk Level (MRL) derived by the Agency for Toxic Sub-
stances and Disease Registry (ATSDR), which is based on a study
in mice (ATSDR, 2007).
The various non-cancer and cancer reference values established
by governmental agencies and were readily available via searches
of government websites and publications are summarized in
Table 1. In addition to chronic exposure guidance values, two
recently-derived acute exposure guidance values are presented as
well for context and comparison. As discussed above, the most
important route of exposure is inhalation. Therefore, most refer-
ence values for benzene are for inhalation exposures. The few oral
reference values in existence are typically derived from the inhala-
tion values, using standard assumptions regarding absorption dif-
ferences via the two routes of exposure.
2.2. Potential biomarkers
Numerous biomarkers have been employed to assess exposures
to benzene (see Table 2), some more commonly employed for
occupational exposures (tt-MA, SPMA, benzene in exhaled breath)
or for environmental exposures (benzene in blood or urine). The
advantages and disadvantages of each, in the context of Biomoni-
toring Equivalents, are described below and in Table 2.
Biomarkers fall within one or more of several categories. Histor-
ically, biomarkers have been classified as markers of exposure, ef-
fect or susceptibility (NRC, 2006). For development of BE values,
biomarkers are assessed with respect to their ease of interpretation
in a risk assessment context. This assessment focuses on consider-
ations of specificity, sensitivity, mode of action for the effect(s) of
most relevance for humans, and stability. The ideal biomarker for
interpretation in a risk assessment context is:
 Related to the mode of action (i.e., is the proximate toxicant,
parent compound or metabolite, or a compound upstream of
the proximate toxicant in the metabolic scheme),
 Is specific to the compound of interest (i.e., levels measured in
biological fluids cannot arise from exposures to other
compounds),
 Is sensitive enough to detect typical exposures encountered in
the environment, and
 Has a sufficiently long half-life, in relation to the frequency of
exposures, so as to avoid highly transient biomarkers (when
possible).
For many compounds, not all of these criteria may be met, in
which case the advantages and disadvantages of the various bio-
markers must be weighed against these criteria to select the best
overall compound-specific biomarker(s) in the context of the BE.
Most research on benzene biomonitoring has focused on bio-
markers of exposure in the occupational environment. Efforts have
focused on correlation of a biomarker with a measure of external
exposures (typically time-weighted average benzene in air via
 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73 65

Table 1
Health-based exposure guidance values for benzene from various agencies.

Organization, criterion, reference Study description
Inhalation non-cancer chronic exposure guidelines
USEPA RfC (USEPA, 2003) (Rothman et al., 1996): Study of
human population
occupationally exposed
TCEQ Chronic ReV (TCEQ, 2007) Rothman et al. (1996): Study of
human population
occupationally exposed
ATSDR Chronic MRL (ATSDR, Study of human population (2/3
2007) female) occupationally exposed
California REL (CA, 2000) Tsai et al. (1983): Study of male
refinery workers
Inhalation non-cancer acute exposure guidelines
USEPA 8-h AEGL (USEPA, 2006) Srbova et al. (1950): controlled
exposures of volunteers
TCEQ 1-h Acute ReV (TCEQ, 2007) Rozen et al. (1984): Mice
exposed via inhalation
(depressed peripheral
lymphocytes and depressed
mitogen-induced blastogenesis
of femoral B-lymphocytes).
Organization, criterion, reference Study description
Inhalation cancer exposure guidelines
USEPA unit risk levels (USEPA, PlioFilm cohort: study of human All leukemias
2000) population occupationally
exposed
TCEQ ESL cancer (TCEQ, 2007) PlioFilm cohort: study of human Acute myelogenous and
population occupationally
exposed
Critical endpoint and dose
Decreased lymphocyte count
BMCL: 23.0 (mg/m3)
BMCLADJ: 8.2 (mg/m3) (adjusted
for 10/20 m3/d, 5/7 d/wk
exposure)
Decreased lymphocyte count
POD (at 1 SD from control):
23.0 mg/m3 (7.2 ppm)
PODHEC (adjusted by a factor of
10/20 m3 and 5/7 d/wk): 8.2 mg/
m3 (2.6 ppm)
Decreased B cell count
BMCL0.25SD: 0.1 ppm
BMCL0.25SD (adjusted by a factor
of 8/24 for 24-h, 6/7 d/wk
exposure): 0.03 ppm
Hematological effects
NOAEL: 0.53 ppm adjusted by a
factor of 10/20 m3/day, 5/7 d/wk
CNS effects
NOEL: 110 ppm, 2 h duration
adjusted by a factor of 2/8 h
PODadj: 18.5 ppm
Most sensitive endpoint
monocytic leukemia (AMML)
Uncertainty factors Value
300 – total 30 lg/m3 (9 ppb)
10 – interindividual
3 – subchronic to chronic
3 – effect level
extrapolation
3 – database deficiency
30 – total 280 lg/m3 (86 ppb)
10 – interindividual
3 – incomplete database
10 – total 10 lg/m3 (3 ppb)
10 – interindividual
10 – total 60 lg/m3 (20 ppb)
10 – interindividual
3 – intraspecies 29000 lg/m3 (9,000 ppb)
100 – total 590 lg/m3 (185 ppb)
3 – animal to human
3 – LOAEL to NOAEL
10 – interindividual
1E-04 Risk level 1E-06 Risk level
13.0–45.0 lg/m3 (4– 0.13–0.45 lg/m3 (0.04–
14 ppb) 0.14 ppb)
45 lg/m3 (14 ppb) 0.45 lg/m3 (0.14 ppb)

Table 2
Potential biomarkers of exposure to benzene.

Analyte Medium Advantages Disadvantages

Benzene Blood Sensitive and specific; highly relevant to Invasive, short half-life
target tissue concentrations.
Urine Sample easily obtained; specific biomarker Not directly relevant to target tissue concentrations
Exhaled Sample easily obtained; specific biomarker Difficult to obtain reproducible results
air
Trans, trans-muconic acid Urine Sample easily obtained Non-specific
S-phenyl mercapturic acid Urine Sample easily obtained Potential downstream/de-activation metabolite. Extremely
small percentage of metabolites.
Phenol Urine Sample easily obtained Non-specific, potential downstream/de-activation metabolite
Hydroquinone Urine Sample easily obtained Non-specific, Potential downstream/de-activation metabolite
Benzene oxide (BO) hemoglobin or Blood Potentially reflects integrated exposures Invasive, background sources unresolved
albumin adducts
1,4-benzoquinone (1,4-BQ) hemoglobin Blood Potentially reflects integrated exposures Non-specific; Invasive; perhaps not as stable as BO-Alb adducts,
or albumin adducts background sources unresolved

either area monitors or personal badges). However, many of the
benzene biomarkers have very short half-lives. As a result, correla-
tions between the transient biomarker concentrations and time-
weighted measures of external air concentration reported in the
literature have been highly variable, and the relationships ob-
served are likely influenced by a number of factors. For example,
the timing of the measured concentrations in air (and the time per-
iod of exposure and integration of those measured concentrations)
compared to that of sample collection would influence the degree
to which the sample reflects very recent air concentrations versus
concentrations earlier in the monitoring period (e.g., a biomarker
only reflects the previous four half-lives of exposure, with the
exposures most recent influencing the biomarker concentration
most). Because the half-life of benzene in blood is less than one
hour, the last 1–2 h of exposures will have the greatest influence
on benzene in blood. If the concentration of benzene in air is
66 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73
variable, the correlation of benzene in blood with benzene in air
will be highly influenced by the timing of blood collection in rela-
tion to only the previous few hours of exposure (i.e., the first 4–6 h
of exposure will have little effect on benzene levels at the end of
the 8-h exposure period – the typical length of time used for
TWA measurements). The type of biological sample (blood, urine,
or exhaled air) also will influence the air vs. biomarker concentra-
tion relationship, as urine will reflect a more averaged exposure le-
vel compared to blood or air. Finally, because of the high volatility,
accurate sample collection, handling, and analytical measurement
of low levels of benzene is likely to be challenging.
Table 2 lists many of the candidates previously assessed or used
as biomarkers of benzene exposure. As noted above, many of the
benzene metabolite biomarkers have mostly been assessed and
used in the context of occupational benzene exposures, which
are likely to be many-fold higher than environmental exposures.
As a result, some of the biomarkers of benzene exposure that are
useful in an occupational context may not be useful to evaluate
environmental exposures due to the presence of low-level alterna-
tive sources of these biomarkers. In the occupational environment,
the relative contribution of benzene to the measured levels of
these metabolites is large, so the biomarker is more reliable. How-
ever, in the environmental arena, the alternative sources become
substantial and complicate interpretation of these metabolites as
specific to benzene exposure.
For example, trans, trans-muconic acid (tt-MA) has been a pop-
ular biomarker, in particular in the workplace (ACGIH, 2001). How-
ever, at background levels of benzene exposures, tt-MA
confounding from sorbic acid is problematic (Fustinoni et al.,
2005). Phenol and various phenolic metabolites downstream have
also been used over the years, but since all of these are non-specific
metabolites of benzene (they can arise from exposures to phenol,
including endogenous production of phenol in humans), interpre-
tation in a risk assessment context is difficult, especially at low
environmental exposures to benzene (Lawrie and Renwick, 1987;
Renwick et al., 1988).
More recently, hemoglobin and albumin adducts of benzene
oxide and 1,4-benzoquinone have been used as biomarkers of
exposure to benzene (Lin et al., 2007; Yeowell-O’Connell et al.,
1998, 2001; Rappaport et al., 2002a,b). A benzene-related adduct
has a potentially huge advantage for assessing exposures to ben-
zene in that it has a much longer half-life than benzene or any
other metabolite of benzene, thereby giving a better indicator of
integrated exposures and be less variable within and across days.
Several attempts have been made to relate these adducts to ben-
zene exposures in animals (Sun et al., 1990; McDonald et al.,
1993, 1994) and humans (Lin et al., 2007; Yeowell-O’Connell
et al., 1998, 2001; Rappaport et al., 2002a,b). In controlled labora-
tory experiments using rodents, a clear dose–response relationship
has been found (Sun et al., 1990; McDonald et al., 1993, 1994).
Among workers exposed to high levels of benzene, a relationship
between benzene oxide (BO) hemoglobin and albumin adducts
and 1,4-benzoquinone (1,4-BQ) albumin adducts and workplace
air benzene levels have also been found (Yeowell-O’Connell et al.,
1998, 2001; Rappaport et al., 2002a,b). However, every human
study has found high levels of BO hemoglobin and albumin adducts
among control populations (presumably non-exposed or low-ex-
posed) individuals. Currently, the source of the observed elevated
background levels of these adducts is unclear (Yeowell-O’Connell
et al., 1998, 2001; Rappaport et al., 2002a,b). Several researchers
have even suggested a potential endogenous source of BO produc-
tion (Yeowell-O’Connell et al., 1998). The issues with background
sources of benzene-related adducts precludes their utility as a BE
at environmentally relevant exposure levels. Nonetheless, ben-
zene-related adducts may be useful for a BE related to occupational
exposures. This BE dossier is not deriving occupational BEs,
therefore the large source of adducts among general population
exposures preclude the utility of these benzene-related adducts
for BE derivation for general population exposures. Further re-
search on this potentially useful biomarker is warranted.
S-phenyl mercapturic acid (SPMA) is a specific biomarker for
benzene, however it is a very minor metabolite (it has been esti-
mated that less than one percent of absorbed benzene is metabo-
lized into SPMA), with high inter-individual variability (Boogaard
and van Sittert, 1996). It also most likely represents a deactivated
metabolite. Thus, SPMA is of limited utility either as a quantitative
biomarker of exposure or as a marker for measures of internal
exposure to the proximate toxicant.
Measures of benzene in blood, urine and exhaled air have the
advantage of being specific to benzene exposures, but have the dis-
advantage of being highly transient. In measuring benzene in ex-
haled breath it has been difficult for investigators to obtain
consistent results (Ong and Lee, 1994), and special collection meth-
ods are required to assure that consistent end alveolar air is col-
lected. Similarly, stringent sample collection and storage
methods are required to ensure that any benzene excreted in the
urine doesn’t volatilize during collection.
Benzene in blood has the advantage of being relevant to the
health effects related to benzene exposures, and is the only avail-
able analyte that is specific to benzene exposures at environmental
levels. Because benzene in urine can be correlated to benzene in
blood, benzene in blood and urine are the biomarkers selected
for BE calculations in this analysis.
2.3. Pharmacokinetic data and models potentially useful for deriving
BEs
Following inhalation, absorption of benzene is very efficient.
Following absorption, benzene and/or its metabolites are widely
distributed throughout the body via the systemic circulation. Be-
cause benzene and many of its main metabolites are highly lipo-
philic, these various metabolites distribute in the bone marrow
and other tissues with fat stores (Irons et al., 1979b; Srbova
et al., 1950). Distribution to the bone marrow has been suggested
an important step in benzene toxicity, as the bone marrow is the
site of most hematopoietic activity in humans.
Given most exposure guidance values for benzene have been
derived using human epidemiology data, the pharmacokinetics of
benzene in humans is required for deriving BE values. The possible
sources of data and/or approaches for deriving BEs for the recom-
mended benzene biomarkers include:
 Correlations between benzene in air (workplace or environmen-
tal) and benzene in blood or urine.
 Controlled human exposure studies (preferably of sufficient
length to achieve steady-state).
 Classical pharmacokinetic (PK) or physiologically-based phar-
macokinetic (PBPK) models for benzene in humans, which have
been developed using data from the controlled human exposure
studies.
One of the main difficulties in using many of the correlation
analyses is the disconnect between the half-life of benzene in bio-
logical media and the duration of measures of benzene
concentration in air. Most correlation studies measure some form
of a time-weighted estimate of benzene in air (e.g., time-weighted
average (TWA) over an 8-h work shift, 24-h composite, etc.). When
a biological sample is retrieved at the end of the time period cov-
ered by the integrated air measure, the benzene in that biological
sample is influenced most by the most recent air concentrations
and then by air exposures over the past 4–5 half-lives. Benzene
in blood and exhaled breath has a half-life of less than about
 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73
30 min (Sato et al., 1975), therefore, only air concentrations during
the previous two hours before collection of the biological sample
have any impact on the measured benzene level in blood and
breath, with the previous1-h time period immediately preceding
measurements having the most impact, and each previous hour
of exposure prior to collection successively having less impact. If
substantial fluctuations have occurred in benzene air concentra-
tions over the period of collection particularly over the most recent
2–4 h periods, the measured benzene concentration in blood or ex-
haled breath will be difficult to accurately correlate with the time-
weighted air concentrations.
2.3.1. Benzene in blood
Benzene is a data-rich compound, for which many biomonitor-
ing studies have been conducted. Concentrations of benzene in
post work-shift blood samples have been found to be reasonably
well correlated with work-shift benzene air exposures (assessed
via 8-h TWA personal air samples) from workers in petrol stations
and refineries (Brugnone et al., 1999). Exposures in the Brugnone
et al. (1999) study cover a wide range, including some that are rel-
atively low in the context of occupational exposures (5–1500 lg/
m3: 1.5–470 ppb). The equation relating measured blood concen-
tration at the end of the work shift to the 8-h TWA concentration
derived by Brugnone et al. (1999) is:
Bzblood ðng=LÞ ¼ 0:9227ðng=L per lg=m3 Þ  Bzair ðlg=m3 Þ
þ 107ðlg=LÞ
The intercept of the equation is substantial in the context of
environmental exposures; it predicts unrealistic levels of benzene
in blood at environmental background air benzene concentrations.
Controlled human exposure studies have been conducted with
few volunteers (Sato et al., 1975; Pekari et al., 1992). Sato et al.
(1975) exposed five men and women to 25 ppm benzene for two
hours and collected blood and breath samples during and for five
hours post exposure. Pekari et al. (1992) exposed three volunteers
to 10 or 1.7 ppm benzene for four hours and collected blood and
breath samples during and for 21 h post exposure. Unfortunately,
it is not clear that either of these studies involved exposures long
enough to develop a relationship between airborne benzene and
steady-state blood levels. Furthermore, the experimental air con-
centrations employed were very high compared to environmental
exposure levels, limiting the ability to extrapolate the observed
correlations to environmentally relevant air concentrations (less
than 10 ppb, in general) (Wallace, 1996).
Numerous physiologically-based pharmacokinetic (PBPK) mod-
els have been developed for benzene in rodents (Cole et al., 2001;
McMahon et al., 1994; Medinsky et al., 1989; Cox, 1996; Watanabe
and Bois, 1996; Travis et al., 1990) and humans (Travis et al., 1990;
Bois et al., 1996; Brown et al., 1998; Sinclair et al., 1999; Yokley
et al., 2006). Most PBPK models have been developed with the goal
of describing blood and breath benzene (Travis et al., 1990; Bois
et al., 1996; Brown et al., 1998; Sinclair et al., 1999). A recently
developed model by Cole et al. (2001) focused on the quantitative
evaluation and prediction of parent and metabolite concentrations
in target tissues in rodents. This model was further developed and
parameterized for humans by Yokley et al. (2006) using Bayesian
analyses and multiple datasets on urinary metabolite elimination
following occupational exposures in air. Brown et al. (1998) devel-
oped their PBPK model to be gender specific, in an attempt to
quantitatively account for gender specific differences in body fat
levels. The Brown et al. model is the most simplistic that is also
consistent with the existing controlled human exposure kinetic
data. Because the focus here is on development of BE values based
on estimation of steady-state benzene concentrations in blood
(with corresponding estimated urine concentrations), the simple
67
PBPK model from Brown et al. (1998) was used for BE derivation.
The Brown et al. (1998) and Yokley et al. (2006) models give con-
sistent results for predicted benzene concentrations in blood in the
concentration range of interest for this derivation.
2.3.2. Benzene in urine
Benzene in urine has the advantage of requiring a non-invasive
sample collection, but issues associated with volatilization during
collection have posed some technical challenges (Ikeda, 1999).
None of the available PBPK models predict urinary concentrations
of benzene; however, a number of studies have attempted to cor-
relate benzene in air with benzene in urine among workers and
environmentally exposed individuals (Kim et al., 2006; Ghittori
et al., 1993; Ong et al., 1995, 1996; Waidyanatha et al., 2001;
Tolentino et al., 2003; Ghittori et al., 1995; Perbellini et al.,
2003). Fig. 2 shows the relationships identified in the literature.
The reported correlations between benzene in urine and benzene
in air vary widely. The variations likely reflect differing efforts
undertaken to prevent volatilization of benzene in collected sam-
ples. The variations may also reflect factors that govern variations
associated with the kinetics of benzene in urine, including time
since last void, time since exposure, time of urinary sample collec-
tion compared to the period of air concentration measurement,
and variations in hydration status leading to variable urine output
rates. Of the available datasets, those from Ghittori et al. (1993,
1995) and Ong et al. (1996) appear to be consistent with each other
ð1Þ
and the most central among the various studies included in this
assessment.
An alternative approach is to correlate benzene concentrations
in urine to those in blood. Perbellini et al. (2003) sampled benzene
in blood, urine and exhaled breath among non-occupationally ex-
posed volunteers to find correlations among the three biomarkers.
Benzene in urine was found to be related to benzene in blood with
the following relationship:
Bzurine ðng=LÞ ¼ 1:1ðng=L urine per ng=L bloodÞ
 Bzblood ðng=LÞ þ 60:8 ðng=L urineÞ
r ¼ 0:4709 p < 0:001 ð2Þ
This relationship was established over a range of benzene in
blood concentrations of 33.5–487.2 ng/L, which correspond to air
concentrations of 0.002–0.03 ppm [2–30 ppb] (based on the PBPK
model of Brown et al., 1998). These air concentrations are relevant
for environmental exposures.
To determine how the relationship between benzene in urine
and benzene in blood compares with the benzene in urine versus
air correlations above (Fig. 2), the Perbellini relationship (Eq. (2))
had to be converted to benzene in urine versus benzene in air. To
achieve this conversion, the following steps were taken:
 Benzene in air was converted to benzene in blood using the
Brown et al. (1998) PBPK model with an assumption of
steady-state exposure conditions.
 Benzene in blood was converted to benzene in urine using the
relationship identified by Perbellini et al. (2003).
Fig. 2 compares the Perbellini relationship (converted to be a
function of benzene in air using the approach outlined above) to
the results of Ghittori et al. (1993, 1995) and Ong et al. (1996).
The observed relationships are remarkably consistent over the
range of air concentrations encompassed by the Ghittori et al.
(1993, 1995) and Ong et al. (1996) and Perbellini et al. (2003) stud-
ies. Thus, the relationship from Perbellini et al. (2003) (Eq. (2)) pro-
vides a basis for estimating steady-state urinary benzene
concentrations from steady-state blood benzene concentrations,
68 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73

Fig. 2. Correlations between urinary benzene concentrations and air benzene concentrations observed from several studies under a variety of exposure conditions.
Relationship for Perbellini et al. (2003) was calculated from the Brown et al. (1998). PBPK model based on the reported urine to blood concentration in that study with an
assumption that blood concentrations represented steady-state blood concentrations (see text for further discussion). Key: A, Kim et al. (2006, n = 228); B, nonsmokers,
Ghittori et al. (1993, n = 63); C, smokers + nonsmokers, Ghittori et al. (1993, n = 110); D, Ong et al. (1995, n = 78); E, Waidyanatha et al. (2001, n = 37); F, Tolentino et al. (2003,
n = 33); G, Ghittori et al. (1995, n = 124); H, Ong et al. (1996, n = 131); I, Perbellini et al. (2001, n = 61).

and will be used in the calculation of BE values for benzene in deriving a BE for benzene in blood consistent with non-cancer
urine. guidance values are outlined below and illustrated in Fig. 3 and
Table 4.

3. Results: BE derivation
1. Points of departure (PODs) from each exposure guidance value
were adjusted from workplace to continuous exposure (where
3.1. Chronic exposure guidance values
applicable) and sub-chronic to chronic adjustments were made
(where applicable). This value was used as the adjusted POD.
3.1.1. BE values for benzene in blood
2. The gender specific PBPK model of Brown et al. (1998) was used
As discussed above, benzene in blood is the preferred and most
to predict steady-state concentrations of benzene in blood asso-
reliable of the available biomarkers for a BE derivation. The PBPK
ciated with exposures at this adjusted POD. The average of the
model developed by Brown et al. (1998) is used to derive BEs for
predictions for males and females is the BEPOD.
benzene in blood (see Table 3 for PBPK model parameters). Since
3. Uncertainty factors (UFs) associated with within human vari-
all of the available exposure guidance values for benzene are based
ability and database deficiency (where applicable) were applied
on human data, no species extrapolation is required. The steps in
to the BEPOD to yield the BE for benzene in blood.

Table 3
BEs associated with cancer guidance values from USEPA and
PBPK Model Parameters from Brown et al. (1998).
TCEQ were derived by using the PBPK model to simulate continu-
Parameter Male Female ous exposures to the air concentrations associated with 1E-06 and
Physiological parameters 1E-04 risk levels.
Body weight (kg) 70 60 Estimates of steady-state blood benzene concentrations were
Tissue volumes (L) calculated using parameter sets from Brown et al. (1998) for both
Liver 1.82 1.38 males and females. The results varied by less than 15% from one
Fat 14 18
another. Based on this small difference, the average of results from
Richly perfused 4.2 3
Poorly perfused 44.8 33 the male and female models was reported for benzene in blood in
Cardiac output (L/hr) 336 288 both Tables 4 and 5.
Alveolar ventilation (L/hr) 450 363
Tissue blood flow rates (L/hr) 3.1.2. Urinary BE values
Liver 84 72 BE values for benzene in urine were derived by estimating uri-
Fat 26.9 23.0
nary concentrations consistent with the blood BE values using the
Richly perfused 129.4 110.9
Poorly perfused 95.7 82.1 urine to blood benzene relationship identified by Perbellini et al.
(2003); Equation 2 above. Each BEPOD and BE for benzene in blood
Partition coefficients
Blood:air 7.8 8.2 was converted to the urine equivalent using the Perbellini et al.
Liver:blood 2.95 2.8 (2003) relationship (see Table 4 and Fig. 3). The same approach
Fat:blood 54.5 51.8 was used for deriving BEs for benzene in urine for cancer guidance
Richly perfused:blood 1.92 1.8 values (Table 5). However, some of the blood benzene BE values
Poorly perfused:blood 2.05 2
associated with 1E-06 risk levels were below the range of blood
Metabolic constants benzene concentrations for which the Perbellini et al. (2003) rela-
Vmax (mg/hr) 13.9 19.5
Km (mg/L) 0.35 0.35
tionship was established. In such cases, the BEs for benzene in ur-
ine were not calculated.
 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73 69

Fig. 3. Schematic of BE derivation method. The BMCLAdj is the BMCL with applied uncertainty factors for LOAEL to NOAEL and subchronic to chronic exposure duration. As
discussed in the text, the PBPK model was used to estimate average blood concentrations of benzene at the BMCLAdj, which is the BEPOD. Uncertainty factors for intraindividual
variability and database uncertainty factors, if any, are applied (UFH and UFD, respectively) to derive the BE value. Urinary BEs were then estimated using the correlation
between urine and blood concentrations reported by Perbellini et al. (2003) (see text).

Table 4
Biomonitoring Equivalents for chronic non-cancer exposure guidance values.

BE derivation step USEPA Chronic RfC TCEQ ReV CA REL ATSDR chronic
inhalation MRL
Target organ Decreased lymphocyte Decreased lymphocyte Hematological effects Decreased B cell counts
count count
POD 7.2 ppm (23 mg/m3) 7.2 ppm (23 mg/m3) NOAEL: 0.53 ppm (1.5 mg/m3) 0.1 ppm (0.3 mg/m3)
(BMCL, 1 SD) (BMCL, 1 SD)
LOAEL to NOAEL adj. 3 1 1 1
Duration adjustment (5/7 days/wk, 10/20 m3/ (5/7 days/wk, 10/20 m3/ adjusted by a factor of 10/20 m3/day, (6/7 days/wk, 8/24 h/
day) day) 5/7 d/wk day)
Subchronic to chronic adjustment 3 1 1 1
Adjusted POD, mg/m3 continuous 0.3 ppm (0.9 mg/m3) 2.6 ppm (8.3 mg/m3) 0.2 ppm 0.03 ppm (0.1 mg/m3)
Human equivalent BEPOD, Benzene in 4.4 38.8 2.9 0.44
blood (lg/L):
Benzene in urine (lg/L): 4.9 42.7 3.2 0.54
Intraspecies uncertainty factors
0.5 0.5 0.5
Pharmacodynamic 10 10 10 100.5
Pharmacokinetic 100.5 100.5 100.5 100.5
Incomplete database UF 3 3 1 1
BE value, Benzene in blood (lg/L): 0.15 1.29 0.29 0.04
Benzene in urine (lg/L): 0.16 1.42 0.33 0.05

Table 5
Biomonitoring Equivalents corresponding to cancer risk-specific exposure levels.

Agency, criteria 1E-04 risk 1E-06 risk

USEPA, risk-specific concentrations
Air concentration lg/m3, (ppb) 13.0–45.0 lg/m3 (4–14 ppb) 0.13–0.45 lg/m3 (0.04–0.14 ppb)
BE – Benzene in blood (ng/L) 58.3–204.4 0.58–2.04
BE – Benzene in urine (ng/L) 125–286 NC
TCEQ, ESL cancer
Air concentration lg/m3, (ppb) 44.6 lg/m3 (14 ppb) 0.446 lg/m3 (0.14 ppb)
BE – Benzene in blood (ng/L) 204.4 2.04
BE – Benzene in urine (ng/L) 286 NC

NC – Not calculated since blood concentration was outside the observed range for which this blood-urine correlation was established (33.5–487.2 ng/L) (Perbellini et al.,
2003).

3.2. Acute exposure guidance values
As noted in Table 1, a number of acute exposure guidance val-
ues have been derived for benzene. These guidance values
represent exposure limits deemed tolerable over individual,
short-term exposure periods. Derivation of BE values associated
with these guidance values is problematic due to the rapid changes
in blood concentrations expected during the short term exposure
period and immediately following such exposures. Fig. 4 illustrates
the expected time course of blood concentrations of benzene
70 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73
Fig. 4. Simulated blood concentration vs. time curves for the acute exposure
conditions specified by the USEPA AEGL (8-h) and the TCEQ ReV (1-h) (see Table 1
for descriptions). If exposure timing relative to blood sampling is known, the curves
here can be used to compare blood concentrations to those consistent with these
exposure guidance values.
during an 8-h exposure at the AEGL-1 of 9 ppm, or during a 1-h
exposure at the TCEQ ReV of 0.185 ppm. Under these exposure
conditions, peak blood concentrations of approximately 100 lg/L
(AEGL-1) or 1 lg/L (TCEQ ReV) would be attained. If blood samples
are collected during or at a known time following an acute expo-
sure event, the measured concentrations can be compared to the
concentrations in Fig. 4 to evaluate whether exposures likely ex-
ceeded the acute guidance values or not. However, detailed infor-
mation on the time of exposure and the timing of sample collection
relative to exposure are needed in order to make this evaluation.
4. Discussion
The BE values presented here represent estimates of the
steady-state concentrations of benzene in blood or urine that are
consistent with the existing exposure guidance values from vari-
ous governmental agencies as listed in Table 1. These BE values
were derived based on current understanding of the pharmacoki-
netic properties and toxicity of benzene in humans. The derived
values vary due to differences in the underlying exposure guidance
values. These differences stem from alternative interpretations of
the available human exposure–response datasets, selected points
of departure, and applied uncertainty factors. The BEs described
here for benzene should be regarded as screening values that can
be updated or replaced if; (1) the exposure guidance values are up-
dated, (2) if the scientific and regulatory communities develop
additional data on acceptable or tolerable concentrations in human
biological media, or (3) if available PBPK models are updated.
A screening BE value for benzene of 0.1 lg/L corresponding to
the EPA RfC was recently published as part of a broader, cross-
VOC evaluation that relied upon steady-state solutions to the gen-
eric PBPK model for VOCs (Aylward et al. 2010). The BERfC value
presented here (0.15 lg/L) is consistent with the earlier publica-
tion, which presented values to 1 significant figure only.
4.1. Sources of variability and uncertainty
Benzene has a short half-life in blood, exhaled breath, and urine,
and thus biomarker concentrations are transient in nature. Rapid
changes in benzene exposures are quickly reflected in changes in
benzene in blood and exhaled breath, while urinary concentrations
fluctuate more slowly and are more representative of exposures
over a slightly longer averaging time. Therefore, the levels of ben-
zene in biological samples are likely to be highly variable within an
individual throughout a day, across days and through the years of
their exposure (Sexton et al., 2005). Since chronic exposure
guidance values for benzene are estimated based on an assumed
exposure scenario of long-term or lifetime average exposures,
interpreting concentrations of biomarkers with short half-lives col-
lected in spot cross-sectional studies poses challenges. In the case
of such studies, the variations in measured concentrations across
individuals may be a surrogate, in part, of temporal variations
within individuals (Sexton et al., 2005). In this context, the central
tendency of measured concentrations in a population may provide
a general indication of the levels of exposure, but extremes in the
distributions observed may only reflect short-term, high exposures
or results from taking a blood or urine sample close in time to a re-
cent exposure event. Since BEs for chronic exposure guidance val-
ues (e.g., RfDs, RfCs, etc.) assume steady-state chronic exposures,
the BE is best used to interpret measures of longer-term average
exposures. In the case of benzene population biomonitoring, this
is better reflected in the central tendencies of the distribution. As
a result, BEs should only be used to interpret central tendencies
in population levels, and the limitations in ability to interpret the
extremes in the distributions should be acknowledged.
Other sources of variability and uncertainty include variations
due to inter-individual differences in the pharmacokinetics of ben-
zene. The model relied upon here (Brown et al., 1998) presents a
central tendency estimate of the steady-state blood concentration
predicted to result from steady-state chronic exposure at the RfC or
other exposure guidance value. Variations in physiology and met-
abolic capability will result in individual variation in the actual
blood concentration that would be attained in various individuals
exposed at a given air concentration, as well as in the relative pro-
duction of metabolites of interest. The Brown et al. (1998) PBPK
model used here also included parameterizations for both male
and female adults, and as discussed above, the two parameter sets
resulted in predictions of steady-state blood concentrations that
differed by approximately 15%. For the purposes of the BE deriva-
tion, this was considered to be a minimal difference and the results
from the gender-specific models were averaged for the BE deriva-
tion. In addition, biomonitoring data directly reflect the differences
in concentration of benzene attained in blood, and so such varia-
tions will be reflected in higher or lower concentrations measured
in an individual blood sample. Variations in rates of production of
various metabolites (which may be important for development of
toxicological responses) are addressed in the derivation of the BE
values through the preservation of an uncertainty factor of 100.5
to address such toxicokinetic variability among the population
(as included in the EPA RfC derivation).
The correlation between urine and blood concentration from
Perbellini et al. (2003) which was relied upon here for the deriva-
tion of the urinary BE values was strong, but individual variation
from the best-fit correlation of approximately two- to fourfold
were common. This is consistent with expected variation in urine
volume due to variations in hydration status.
Because benzene is a volatile compound, special care must be
taken in the process of sample collection and analysis in order to
produce valid results. Variations in sample handling procedures
may introduce uncertainty or variation in the measured blood or
urine sample concentrations (IUPAC, 2000).
4.2. Confidence assessment
The guidelines for derivation of BE values (Hays et al., 2008)
specify consideration of two main elements in the assessment of
confidence in the derived BE values: robustness of the available
pharmacokinetic data and models, and understanding of the rela-
tionship between the measured biomarker and the critical or rele-
vant target tissue dose metric.
The datasets and models relied upon here for derivation of the
blood-based BE values are relatively robust, and the available PBPK
 S.M. Hays et al. / Regulatory Toxicology and Pharmacology 62 (2012) 62–73
models have been parameterized based on data from several con-
trolled human exposure studies. However, as discussed above,
while benzene in blood is a reliable biomarker of exposure to ben-
zene, benzene itself is unlikely to be the proximate toxicant for the
adverse effects of interest for both the cancer and non-cancer
exposure guidance values. Based on these considerations, the con-
fidence ratings for BE values for benzene in blood are as follows:
 Robustness of pharmacokinetic data: HIGH.
 Relevance of biomarker to relevant dose metrics: MEDIUM.
BE values derived for assessment of benzene in urine are based
on studies that correlate urinary and blood benzene concentra-
tions; none of the available controlled human exposure studies
measured urinary benzene concentrations, nor do any of the avail-
able PBPK models predict urinary benzene excretion. Further, the
relative proportion of inhaled benzene that is ultimately excreted
in urine is relatively low, resulting in the potential for substantial
variability in urinary concentrations associated with a given expo-
sure or blood level. In addition, sample collection and handling of
urinary samples for accurate measurement of benzene is challeng-
ing due to the volatility of benzene. Based on these considerations,
the confidence ratings for BE values for benzene in urine are as
follows:
 Robustness of pharmacokinetic data: LOW.
 Relevance of biomarker to relevant dose metrics: LOW.
4.3. Interpretation of biomonitoring data using BE values
The appropriate uses and limitations of BE values have been dis-
cussed previously (Aylward and Hays 2008; Hays and Aylward
2008; Hays et al., 2008). These BE values can be used as a screening
tool to evaluate population- or cohort-based biomonitoring data in
the context of existing risk assessments. Concentrations in excess
of the BE values, but less than the BEPOD values represent medium
priority for risk assessment follow-up, while those in excess of the
BEPOD indicate high priority for risk assessment follow-up. Based
on the results of such comparisons, an evaluation can be made of
the need for additional studies on exposure pathways, potential
health effects, other aspects affecting exposure or risk, or other risk
management activities.
BE values do not represent diagnostic criteria and cannot be
used to evaluate the likelihood of an adverse health effect occur-
ring in an individual or even among a population. Rather, like the
underlying guidance values from which BEs are derived, BEs are
best used as risk management tools. Measured values in excess
of the identified BE values may indicate exposures at or above
the current exposure guidance values that are the basis of the BE
derivations. However, as discussed above, measured concentra-
tions above the BE values, which are based on steady-state average
biomarker concentrations, would be expected even if exposures do
not exceed the exposure guidance values due to the transient con-
centration profiles in blood and urine expected for benzene, varia-
tions in hydration status, and other factors discussed further above.
Thus, interpretation of data for individuals or of extremes of the
distribution (e.g., the upper or lower percentiles) in population-
monitoring studies using these steady-state BE values is not
appropriate.
BE values, consistent with the underlying risk assessments BEs
are derived from, are not ‘‘bright lines’’ that distinguish safe from
unsafe exposure levels. Chronic exposure guidance values are set
at exposure levels that are expected to be protective over a lifetime
of exposure. For short-lived compounds such as benzene, an
exceedance of the corresponding BE value in a single blood or urine
sample may or may not reflect continuing elevated exposure. In
71
other words, it may reflect on a very recent, short-term excursion
above a certain guidance value. As demonstrated in the limited
available datasets and based on the kinetics of benzene in blood
and urinary elimination, spot blood or urine sample concentrations
may vary substantially both within and across days in an individ-
ual. Thus, occasional exceedances of the BE value in individuals in
cross-sectional studies do not imply that adverse health effects are
likely to occur, but can serve as an indicator of relative priority for
further risk assessment follow-up. Further discussion of interpreta-
tion and communication aspects of the BE values is presented in
LaKind et al. (2008) and at www.biomonitoringequivalents.net.
Conflict of interest statement
The authors received funding from the American Petroleum
Institute to prepare this analysis and manuscript. The authors
had complete freedom to design, carry out, and report the results
of their analyses.
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