Special issue

Current issues and perspectives in

food safety and risk assessment

Human and Experimental Toxicology

2015, Vol. 34(12) 12861290
The Author(s) 2015
Reprints and permission:
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DOI: 10.1177/0960327115598408
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G Eisenbrand

Abstract
In this review, current issues and opportunities in food safety assessment are discussed. Food safety is considered an essential element inherent in global food security. Hazard characterization is pivotal within the continuum of risk assessment, but it may be conceived only within a very limited frame as a true alternative to
risk assessment. Elucidation of the mode of action underlying a given hazard is vital to create a plausible basis
for human toxicology evaluation. Risk assessment, to convey meaningful risk communication, must be based on
appropriate and reliable consideration of both exposure and mode of action. New perspectives, provided
by monitoring human exogenous and endogenous exposure biomarkers, are considered of great promise to
support classical risk extrapolation from animal toxicology.
Keywords
Food safety, risk assessment, exposure, biomarkers

Preface
Since ancient times food has played a pivotal role in
the development of human society. Food security,
synonymous for a reliable and adequate supply of
healthy and safe foods, not only means freedom from
hunger. By relieving individuals, families and populations from the constraints of laborious daily food procurement, it can be seen as a main catalyst, allowing
for individual and societal development and education. Another beneficial aspect relates to the continuing technologic evolution inherent in modern food
production, processing, storage, and distribution. This
provides the consumer with an ever increasing multiplicity of high-quality and innovative food items. This
freedom of choice, something never previously
achieved, allows for individualized nutrition and
brings about additional hedonistic value. Thus, food
security in fact may be seen as a prerequisite for the
development of society.
Food safety is an inherent element of food security.
However, the latter term more specifically means that
food as normally consumed should not pose health
risks to the consumer. On a global scale, inadequate
techniques of food production, storage, processing,
and distribution today pose substantial risks to both
food security and food safety. Food security is still

at risk in underdeveloped regions of the world and

starvation is far from being eradicated even though
in theory conventional food production should suffice to feed todays world. On the other side, inadequate distribution of resources governing societal
food supply has led to major detrimental public
health effects. Adiposity-associated diseases as a
consequence of overnutrition of large parts of populations in developed (Western) countries today represent major public health risks, unrelated to food
safety.

Hazard versus risk-based approaches

The basic methodology of safety evaluation on the
basis of animal experiments has changed little during
the past decades. Repeat dose toxicity is studied in
laboratory animals, in compliance with established
Division of Food Chemistry and Toxicology, Department of
Chemistry, University of Kaiserslautern, Kaiserslautern, Germany
Corresponding author:
G Eisenbrand, Division of Food Chemistry and Toxicology,
Department of Chemistry, University of Kaiserslautern, ErwinSchoedinger-Str. 52, 67663 Kaiserslautern, Germany.
Email: eisenbra@rhrk.uni-kl.de

Eisenbrand

guidelines, mostly within the frame of 90-day subchronic or 2-year chronic toxicity studies. They
inform us about a dose level associated with the
non-observed adverse effect level (NOAEL).
By application of canonical uncertainty (safety)
factors (mostly 100), an acceptable/tolerable intake
level is derived from the NOAEL. The response in
experimental animals is evaluated by using general,
clinical chemistry, hematological, and histological
parameters as indicators of organ damage and further
parameters when applicable. The continuing progress in genomics, transcriptomics, proteomics, and
metabolomics in combination with novel tools in
bioinformatics and system biology has brought about
promising new avenues toward improved toxic
hazards characterization. Very often, the decision for
in vivo testing is based on previously obtained in
vitro evidence that calls for in vivo substantiation.
However, in vitro systems provide much more information, especially with respect to mechanistic elucidation of toxicity outcomes. In this respect, the
pivotal importance of in vitro systems for hazard
characterization cannot be overemphasized. Complementary to in vivo toxicity testing, appropriate
in vitro toxicological systems have eminent value
and not only provide enhanced predictivity for
hazards posed by food-associated chemicals.1 They
are also uniquely important for characterizing key
(bio)molecular events governing toxic outcomes
in animals and/or humans. Such in vitro mechanistic studies are also of great value when there is
suspicion or a structural alert with respect to
genotoxicity.
In case there is a genotoxic carcinogen present in
food, the European Food Safety Agency (EFSA)
recommends implementation of appropriate mitigation measures to minimize consumers exposure down
to a level in food resulting in a margin of exposure
(MOE) of not less than 10,000. This MOE is deemed
of low concern with respect to human health.2 The
MOE is calculated by dividing the benchmark dose
(BMD) derived from an appropriate carcinogenicity
study associated with a benchmark response, e.g.
10% tumor incidence at the lower bound confidence
interval (BMDL10), through the estimated human
exposure level.
Thus, safety assessment of food constituents and/or
contaminants not only requires the evaluation of a
hazard potentially exerted by a specific compound but
also needs to take into account the exposure of the
consumer.

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By definition, a hazard in food means a biological,

chemical, or physical agent present in food that may
have an adverse health effect. The term also encompasses an inherent property of an agent or situation
having the potential to cause adverse effects. By contrast, the term risk describes the probability of an
adverse effect and its magnitude in an exposed system
or (sub)population.
At present, legislation in the European Union and
elsewhere often includes a mix of both hazard-based
and risk-based approaches for safety assessment.
Under specific circumstances, for example, when
immediate estimates of potential health concern are
required, a hazard-based approach can be of value.
This may for instance apply to emergency situations
created by cases of accidental contamination or food
adulteration, especially when human exposure is estimated to be close to thresholds for adverse effects.
Sporadic examples may be mentioned here, for example, historic examples of adulteration of wine/spirits
with diethylene glycol/methanol, adulteration of milk
with melamines or, currently coming into focus, the
illegal presence of drugs in food supplements.3
Hazard-based approaches also may apply to agents
exerting potent non-thresholded effects, such as certain
strong genotoxic carcinogens. Of note, a hazard-based
element also is intrinsic to the so-called threshold of
toxicological concern (TTC) concept. This concept has
evolved over the years, in regulatory authorities and
elsewhere, as an attempt to develop a generic approach
to the safety assessment of groups or individual chemicals of unknown toxicity, that is, with no or insufficient
toxicological data.4 The concept provides guidance
about achieving acceptable risks by deriving toxicologically insignificant exposures according to hazard and
chemical structure.57 This approach was introduced
by the US Food and Drug Administration as threshold
of regulation (1995) for compounds migrating into
food from packaging materials.8 It is based on
structure-guided grouping of the compounds in question with respect to their expected toxicological properties in comparison with well-defined and studied
reference compounds (Read across). In brief, on the
basis of grouping well-investigated compounds into
different structural classes with respect to noobserved toxicological effect dose levels, three socalled Cramer classes (I, II, and III)9 were generated.
Toxicologically insignificant exposure levels were
derived using the 5th percentile of each class, divided
by a conventional uncertainty factor of 100. For
hazards in foods associated with chemicals bearing

1288

structural alerts of genotoxic activity, the toxicologically insignificant exposure level was set at 0.15 mg/kg.
Zero tolerance of risks is not feasible for the majority of foods and their constituents/contaminants. In
general, therefore, for food safety assessment, riskbased approaches are adequate. Such risk assessment
of food has to consider man-made chemicals, as well
as naturally occurring compounds, which might
influence human health. The first prerequisite for
risk-based approaches is a reliable exposure estimate, taking into proper consideration uncertainties
in exposure assessment. The second important prerequisite comprises elucidation and appropriate consideration of the mode of action (MOA). Importantly,
the MOA is required to be put into perspective with
an appropriate estimate of consumers exposure.
Furthermore, there is need to establish a continuous
dialogue with the consumer in order to ensure appropriate risk communication, thereby creating societal
confidence in the risk assessment. Moreover, any
risk assessment appropriate for society should also
take account of the benefits, including potential consequences for food security, sustainability, nutritional value, and others. The aim of such risk
benefit analysis should be to arrive at a sciencebased societal consensus concerning the acceptability of a given risk. This is quite important, given that
consumers usually are risk averse, especially about
exposure to chemicals in food.

Exposure assessment
Reliable metrics relating to the amount of an agent
present in food; its bioavailability from the food
matrix; and the duration, magnitude, and frequency
of exposures are major determinants of the knowledge
needed for risk assessment.1 Concerning food additives and ingredients, there may be substantial variation in formulations from standard recipes taken as
basis for daily average dietary exposure estimates.
This is a shortcoming recognized by most regulatory
agencies, and it is also of relevance for exposure
assessment of contaminants or residues. There is
therefore a need for sustained development of methodology, encompassing aggregate exposure from all
routes.
Exposure assessment is undergoing a fundamental
change, as new tools become available, based on
methodology of extreme specificity and sensitivity.
Significant progress has been made toward monitoring biomarkers of exposure in humans. Presently,

Human and Experimental Toxicology 34(12)

we witness the advent of metabolomics, techniques

enabling us to picture the totality of metabolite spectra in a given body fluid or compartment, thus opening
the possibility of collecting comprehensive analytical
information about specific food intakes and their
interaction with the organism.10
As yet however, the monitoring of selected biomarkers is the preferred option. It requires detailed
knowledge of the metabolism of the compound in
focus to be able to use specific metabolites in body
fluids or tissues as quantitative exposure indicators.
For human studies, metabolite monitoring in urine
and blood appears to be the method of choice.
An example is provided by the considerable database already available of human exposure estimates
for the genotoxic carcinogen acrylamide (AA), based
in part on biomarkers of exposure.11 AA is generated
during heat treatment of food from innocuous precursors, namely, asparagine and a reducing sugar during
the course of the Maillard reaction. AA itself is not a
genotoxic agent but is converted in the organism by
CYP450 to the corresponding genotoxic epoxide,
2,3-epoxypropane amide (glycidamide, GA). The latter reacts with DNA, forming DNA base adducts, predominantly at N7 of guanine. AA and GA react
covalently with a spectrum of nucleophilic centers
in biopolymers and plasma constituents and to a major
extent also with glutathione. The glutathione adducts
are terminally excreted in the urine as the corresponding mercapturic acids.1214 Thus, the main biomarkers
for internal exposure to the parent compound and its
genotoxic metabolite, GA are:
i. urinary mercapturic acids (MAs),
ii. hemoglobin (Hb) adducts of AA and GA, and
iii. DNA adducts of GA.
Of note, monitoring these biomarkers not only
allows for aggregate dietary exposure estimates. The
direct comparison of MA metabolites and/or Hb
adducts also indicates relative rates of toxification
and detoxification at diet-related, low-exposure
levels.
Uncertainties associated with assessment of consumer exposure to AA were recently adressed in
detail in an European Food Safety opinion.11
It was recommended that:
1. reporting of AA occurrence data regarding the
mode of preparation the products before analysis be improved;

Eisenbrand

2. duplicate diet studies be undertaken to provide

more accurate indication of AA levels in food
as prepared and consumed; and
3. data on urinary metabolite levels from individuals participating in duplicate diet studies be generated for the purpose of biomarker validation.
Beyond AA, these recommendations paradigmatically identify future research needs for the development and validation of biomarkers as adequate
metrics for aggregate consumers exposure to genotoxic agents.
DNA adducts induced by genotoxic agents today
are amenable to specific quantitation at levels of
around 1 adducted DNA base in 10 8 DNA bases.
This allows direct measurement of DNA damage
(e.g. in peripheral white blood cells) associated with
dietary intake of a genotoxic carcinogen at very low
levels of food contamination (low mg/kg range). New
methods increasingly become available that allow
specific simultaneous detection of multiple DNA
adducts in human tissue samples, including those
formed by endogenous lipid peroxidation.15 A further
development, baptized adductomics, includes
DNA adducts of known and of unknown identity.16,17
This rapid technological progress offers new perspectives for assessment of the risk associated with
dietary exposure to genotoxic contaminants at levels
of present-day consumers dietary intake. The induction of DNA damage can be viewed as the first step
in the sequence of cellular key events leading to fixation of a mutation and eventually to malignant transformation. However, it has to be kept in mind that not
every DNA lesion translates into a mutation since a
spectrum of efficient repair systems are in place
before cell division is achieved and a mutation fixed.
Therefore, doseresponse relationships of DNA damage induction and of mutation induction are not the
same. It may thus be inferred that monitoring of DNA
adducts as an exposure biomarker is a conservative
approach with respect to potential biological consequences. Yet it provides a very significant step forward since reliable dosimetry at low levels of
consumer exposure can now be achieved, obviating
the necessity for high to low (virtually safe) dose
extrapolation mainly used until now in classical risk
assessment of genotoxic agents.
Another aspect largely disregarded until now
comes from our increasing knowledge concerning the
substantial background of DNA damage in human tissues, consistently induced by endogenous genotoxic

1289

agents generated during normal energy/housekeeping

metabolism. It has been shown that a number of such
background DNA adducts are identical to those arising in humans by exposure to exogenous genotoxic
agents, including, for instance, ethylene oxide, vinyl
chloride, formaldehyde, acetaldehyde, acrolein, and
lipid peroxidation products. Risk assessment of exposure to genotoxic agents may therefore in the future
assess the quantitative contribution of exogenous
exposure to the endogenous background DNA damage, especially when the DNA adducts formed are
identical to endogenous adducts.13,18,19 This certainly
will need extended establishment of databases on
background DNA damage in various segments of the
population, including sensitive subpopulations. Once
available, such data will provide promising novel perspectives for risk assessment, enabling assessment of
the significance of DNA damage induced by a given
exposure in relation to the physiological background.
Conflict of interest
The author declared no potential conflicts of interest with
respect to the research, authorship, and/or publication of
this article.

Funding
The author received no financial support for the research,
authorship, and/or publication of this article.

References
1. Eisenbrand G, Pool-Zobel B, Baker V, et al. Methods
of in vitro toxicology. Food Chem Toxicol 2002; 40:
193236.
2. European Food Safety Authority (EFSA). Results on
acrylamide levels in food from monitoring years
2007-2009 and exposure assessment. EFSA J 2011;
9(4): 21332180. Available at: http://www.efsa.
europa.eu/efsajournal (accessed 23 July 2015).
3. Available at: http://www.dfg.de/dfg_profil/gremien/
senat/bewertung_lebensmittel/ (accessed 23 July
2015).
4. Barlow S. Threshold of Toxicological Concern (TTC),
A tool for assessing substances of unknown toxicity
present at low levels in the diet. Belgium: ILSI Press,
2005, p. 32. Available at: http://www.ilsi.org/Pages/
Publications.aspx (accessed 23 July 2015).
5. Kroes R, Renwick AG, Cheeseman M, et al. Structure-based thresholds of toxicological concern (TTC):
guidance for application to substances present at low
levels in the diet. Food Chem Toxicol 2004; 42: 6583.

1290
6. Kroes R, Renwick AG, Feron V, et al. Application of
the threshold of toxicological concern (TTC) to the
safety evaluation of 4 Human and Experimental Toxicology cosmetic ingredients. Food Chem Toxicol
2007; 45: 25332562.
7. Munro IC, Renwick AG and Danielewska-Nikiel B.
The Threshold of Toxicological Concern (TTC) in risk
assessment. Toxicol Lett 2008; 180: 151156.
8. US Food and Drug Administration. Food additives:
threshold of regulation for substances used in
food-contact articles; final rule. Federal Register
60: 3658236596. Available at: https://federalregister.
gov/a/95-17435 (accessed 23 July 2015).
9. Cramer GM, Ford RA and Hall RL. Estimation of toxic
hazard - a decision tree approach. Food Cosm Toxicol
1976; 16: 255276.
10. Scalbert A, Brennan L, Manach C, et al. The food
metabolome: a window over dietary exposure. Am J
Clin Nutrit 2014; 99: 12861308.
11. EFSA CONTAM Panel (EFSA Panel on Contaminants
in the Food Chain). Scientific opinion on acrylamide in
food. EFSA J 2015; 13(6): 41044424. Available at:
http://www.efsa.europa.eu/efsajournal (accessed 23
July 2015).
12. Berger F, Feld J, Bertow D, et al. Biological effects of
acrylamide after daily ingestion of various foods in
comparison to water: a study in rats. Mol Nutr Food
Res 2011; 55: 387399.

Human and Experimental Toxicology 34(12)

13. Watzek N, Bohm N, Feld J, et al. N7-glycidamide-guanine DNA adduct formation by orally ingested acrylamide in rats: a dose-response study encompassing
human diet-related exposure levels. Chem Res Toxicol
2012; 25: 381390.
14. Watzek N, Scherbl D, Feld J, et al. Profiling of mercapturic acids of acrolein and acrylamide in human urine
after consumption of potato crisps. Mol Nutr Food Res
2012; 56: 18251837.
15. Monien BH, Schumacher F, Hermann K, et al. Simultaneous detection of multiple DNA adducts in human
lung samples by isotope-dilution UPLC-MS/MS. Anal
Chem 2014; 87: 641648.
16. Balbo S, Turesky RJ and Villalta PW. DNA adductomics. Chem Res Toxicol 2014; 27: 356366.
17. Balbo S, Hecht SS, Upadhyaya P, et al. Application of
a high-resolution mass-spectrometry-based DNA
adductomics approach for identification of DNA
adducts in complex mixtures. Anal Chem 2014; 86:
17441752.
18. Swenberg JA, Lu K, Moeller B, et al. Endogenous versus exogenous DNA adducts: their role in carcinogenesis, epidemiology, and risk assessment. Toxicol Sci
2011; 120(Suppl 1): S130S145.
19. Lu K, Gul H, Upton PB, et al. Formation of hydroxymethyl DNA adducts in rats orally exposed to stable
isotope labeled methanol. Toxicol Sci 2012; 126:
2838.

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