9
Toxicants Resulting from Food Processing
9.1 INTRODUCTION
The primary objective of food processing operations is to
improve the quality of foodstuffs to make them palatable.
Nevertheless, some processing operations do induce the
formation of materials that are potentially toxic and harm-
ful to humans. Thus, a toxic substance may be formed by
interaction between any endogenous and/or exogenous
food components or their derivatives, or between these
substances and outside agents, such as oxygen. Chemical
degradation can also occur as a result of exposure to heat,
light, enzymes, and other agents and in turn may result in
the formation of toxic compounds.
Thermal processing of foods is probably the most
commonly used unit operation in the food industry. Heat-
ing operations are associated with cooking, frying, toast-
ing, evaporation, sterilization, and similar processes. Even
milder thermal operations, such as pasteurization, may
bring about a variety of changes in the treated foods. The
most noticeable adverse effects generally observed are a
loss of heat-susceptible nutrients, especially vitamins and
amino acids. Consequently, the nutritional quality of the
food is often lowered. It is now well known that normal
cooking of foods can induce the formation of many non-
pyrolytic toxicants derived from amino acids and proteins.
When foods are subjected to still higher temperatures
(>200°C–300°C), pyrogenic compounds are produced.
Many of these compounds are now known to be potent
mutagens and carcinogens. Heating of fats and oils to
high temperatures can result in oxidation and polymeriza-
tion reactions, the products of which appear to be harmful
to humans. Other processing operations, such as irradia-
tion, used for food preservation, can form radiolysis prod-
Copyright 2002 by Marcel Dekker. All Rights Reserved.
ucts, many of which are known to be potent toxicants to
humans.
The very large number of reactive endogenous
and/or exogenous food components can readily lead to the
formation of a number of derivatives. Several such derived
toxicants have been identified and their toxicological prop-
erties studied. Yet, many more have not been assessed tox-
icologically. Similarly, many toxicological studies have
been done with individual compounds. It is not only essen-
tial that such compounds be identified and their toxicity
assessed; the toxicological properties of mixtures of such
compounds must also be tested. In all likelihood, the toxi-
cological profile of a foodstuff may be different in the
presence of such mixtures; for additive, synergistic and/or
antagonistic effects may come into play.
In this chapter, some of the more important toxicants
derived as a result of food processing operations are dis-
cussed. The categories of derived toxicants discussed in-
clude pyroorganic toxicants, nonpyrolytic toxicants
derived from amino acids and proteins, Maillard reaction
products, toxicants produced in rancid fats and oils as well
as during their thermal degradation, toxic amino acids
formed during alkali processing of proteins, toxicants such
as nitrosamines produced by degradation or reaction with
contaminants, and radiolysis products formed during food
irradiation operations.
9.2 POLYCYCLIC AROMATIC
HYDROCARBONS
The polycyclic aromatic hydrocarbons (PAHs) are formed
from the incomplete combustion of organic materials.
 There are two major sources of PAHs in the human food
chain. The most important source entails the deposition
and uptake of PAHs from polluted air on food crops. As a
result, cereals, vegetables, fruits, and seed oils are impor-
tant contributors to human intake of PAHs. This aspect of
PAHs as industrial or environmental contaminants is dis-
cussed at greater length in Chapter 17. The other signifi-
cant source is derived from the formation and deposition
of PAHs on foods during heat processing using methods
such as roasting, smoking, and grilling. The formation of
PAHs is only significant at high temperatures. At tempera-
tures below 400°C, only small amounts are formed;
amounts increase linearly in the range 400°C–1000°C
(Toth and Potthast, 1984; Concon, 1988). At these tem-
peratures, a significant amount of charred or tarry products
is formed. Benzo[a]pyrene (3,4-benzpyrene) has been
identified as the active compound in charred material
(Figure 9.1). Other carcinogenic PAHs identified as con-
stituents of charred materials include dibenzo[a,h]an-
thracene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, and
benzo[b]fluroanthene (Badger, 1962). All are potent car-
cinogens. However, benzo[a]pyrene is probably the most
potent and consequently has received the most attention.
9.2.1 Synthesis
The formation of pyrogenic compounds such as PAHs is a
complex process. This differs from other heat-induced
processes in that the former is preceded by an initial, ex-
tensive breakdown of the molecular structures of organic
compounds to simpler, reactive fragments. Combinations
of these fragments to more stable compounds follow,
provided the conditions preclude rapid formation of CO
or CO2.
The PAHs are most likely pyrosynthesized from
degradative products consisting of four- or two-carbon
units, such as butadiene or ethylene radicals (Badger,
1962; Hoffmann and Wynder, 1972). The ease of their for-
mation at elevated temperatures follows from their ther-
modynamic stability. Some possible pathways showing the
formation of benzo[a]pyrene are shown in Figure 9.2.

9.2.2 Occurrence

Intensive heat treatment leads to the production of PAH. In

Benzo[a]pyrene
Dibenzo[a,h]anthracene
Benzo[k]fluoranthene
Benzo[b]fluoranthene
Figure 9.1 Polycyclic aromatic hydrocarbons (PAHs) com-
monly found in pyrolytic products.
Benzo[a]anthracene
cooked foods, they are most typically formed from pyroly-
sis of fats at temperatures exceeding 400°C. This can oc-
cur if portions of the food or fat drippings encounter
charcoal or very hot surfaces. Thus, there are a limited
number of cooking applications that promote the forma-
tion of these compounds. The mechanism of formation
suggests that PAHs may be produced from pyrolysis of
other food components. However, in most cases they are
Dibenzo[a,h]pyrene
not likely to result in edible products.
Grilling or broiling of meat and fish on an open fire
leads to PAH contamination in several different ways.
First, the high temperatures lead to endogenous formation
of PAH on the surface of the food. PAHs can also be
formed during the combustion of the fuel used in the grill-
ing. Finally and perhaps most significantly, PAHs are
formed when melted fat drips down on the heat source.
The PAHs so formed during pyrolysis of the fat are spread
to the atmosphere and partially deposed on the surface of
Dibenzo[a,i]pyrene the meat (Larsen and Poulsen, 1987).
The fat content of the meat is an important factor af-
fecting the PAH level in foods (Lijinsky and Ross, 1967).
This is probably most evident when a “clean” fuel, such as
charcoal, is used. For example, hamburgers with high fat
content when broiled close to the flame produce 43 ppb of
PAHs, of which 2.6 ppb is benzo[a]pyrene. In contrast, the

Copyright 2002 by Marcel Dekker. All Rights Reserved.

 H2C CH2
Pyrosynthesis
Pyrodegradation
Organic matter
Pyrodegradation
Benzo[a]pyrene
Figure 9.2 Possible pathways showing the formation of benzo[a]pyrene.
lean product produces only 2.8 ppb PAHs and no
benzo[a]pyrene. Reducing the amount of fat in grilled or
broiled foods thus appears to reduce the level of PAHs.
Thus, the contamination of grilled food with PAHs can be
prevented or minimized by using charcoal as fuel, by
avoiding open flames, and by employing special grill con-
structions that prevent the fat from dripping onto the heat
source.
Smoking is also a major contributory factor to the
PAH levels found in smoked foods. The main purpose of
smoking is to give products a special desirable taste and
palatability. Curing smoke is normally produced from
wood (sawdust) by the initial pyrolytic changes of lignin,
hemicellulose, and cellulose, followed by secondary reac-
tions leading to the formation of a variety of different
chemical compounds. The most important compounds
formed are phenols, carbonyls, acids, furans, alcohols,
esters, lactones, and PAHs (Larsen and Poulsen, 1987).
Low-molecular-weight PAHs, such as phenanthrene, an-
thracene, and pyrene, are frequently found in smoked
foods at levels of 10 ppb. The higher-molecular-weight
PAHs, such as benzo[a]pyrene, benzo[a]anthracene,
benzo[b]fluoranthene, dibenzo[a,c]anthracene, and di-
benzo[a,h]anthracene, are found in much lower concentra-
tions. Smoked fish and meat normally contain less than 1
ppb PAHs, and benzo[a]pyrene is most often found at lev-
els of 0.1–0.5 ppb. Under special conditions, higher levels
Copyright 2002 by Marcel Dekker. All Rights Reserved.
can be found, especially on the outside of heavily smoked
products (Adrian et al., 1984).
In addition to grilled and smoked products, foods
processed by other means can also produce PAHs. For ex-
ample, the soot and skin of coffee beans that had been
roasted by direct contact with the combustion gases were
found to be very rich (15–28 ppb) in benzo[a]pyrene (Yan-
nai, 1980). Similarly, the bread crust, which receives a
more severe heating during baking than the crumb, con-
tains considerably more benzo[a]pyrene than does the lat-
ter. This difference is much greater if the bread is baked in
a wood-fueled oven than in an electric oven.
Several amino acids, especially tryptophan, glutamic
acid, valine, proline, and lysine, may form potent mu-
tagens when heated. Some of the tryptophan derivatives
have been synthesized in sufficient quantities to permit an-
imal testing and have been found to be very active mu-
tagens (Concon, 1988).
Levels of benzo[a]pyrene found in smoked and other
foods are summarized in Table 9.1. The levels of this car-
cinogen vary, depending on the food and the manner of
cooking. Generally, the closer the exposure to the source
of the smoke or heat, the higher the levels of carcinogen
formed. Although there is no clear evidence implicating
foodstuffs containing benzo[a]pyrene and other PAHs as
causes of cancer in humans, these foods may constitute a
real health hazard.
 Table 9.1 Benzo[a]pyrene Content of Selected Foodstuffs

Benzo[a]pyrene Benzo[a]pyrene
Food (ppb) Food (ppb)

Smoked fish Barbecued meats

(charcoal broiled)
Eel 1.0 Hamburgers 11.2
Herring 1.0 Pork chop 7.9
Sturgeon 0.8 Chicken 3.7
Chubs 1.3 Sirloin steak 11.1
White fish 6.6 T-bone steak 57.4
Kippered cod 4.5 T-bone steak 4.4
(flame-broiled)
Ribs 10.5
Smoked meats Other steaks 5.8–8.0
Ham 0.7–55.0
Mutton Miscellaneous foods
Close to stove 107.0 Spinach 7.4
Distant from stove 21.0 Kale 12.6–48.1
Lamb 23.0 Yeast 1.8–40.4
Sausage (with casing) Tea 3.9–21.3
Cold smoked 2.9 Coffee 0–15.0
Hot smoked 0.7 Cereals 0.2–4.1
Salami 0.8 Soybean 3.1
Bacon 3.6 Cheese (Provola) 4.1–6.2
Source: From Concon (1988) and Howard and Fazio (1983).

9.2.3 Metabolism
The PAHs are oxidized by the cytochrome P-450 system in
the liver to a complex mixture of phenols, dihydrodiols,
and quinines. Arene oxides are formed as intermediate
metabolites, which are spontaneously converted into phe-
nols or further metabolized by epoxide hydrolase to dihy-
drodiols. These primary metabolites undergo further
oxidative metabolism and conjugation reactions (Levin et
al., 1982; Concon, 1988; Larsen and Poulsen, 1987).
As with most known carcinogens, the PAHs are
secondary carcinogens or procarcinogens. Their carcino-
genic activity depends on specific metabolic transforma-
tion. The metabolism of PAHs can be exemplified by
benzo[a]pyrene, whose metabolic transformation is rather
complex, involving epoxidation and hydroxylation (Sel-
kirk, 1977). A part of the metabolic pathways involved in
proximate carcinogen formation is shown in Figure 9.3.
The diol-epoxide I is the major metabolite of benzo[a]-
pyrene. It also binds readily with DNA and RNA (Concon,
1988). It is also the most mutagenic to mammalian cells as
compared to the diol-epoxide II and all 13 other known
benzo[a]pyrene metabolites and derivatives (Huberman et
al., 1976; Concon, 1988). The evidence so far points to
diol-epoxide I as the major proximate carcinogen of
benzo[a]pyrene.
The enzymes known collectively as aryl hydrocar-
bon hydroxylase (AHH) that are responsible for the carci-
nogenic or noncarcinogenic metabolic transformation of
the PAH are affected by several factors. These enzymes,
consisting of both constitutive and inducible components,
may increase or decrease in activity depending on prior
exposure to these carcinogens (Schlede et al., 1970). This
relationship is significant because it has been estimated
that in 1 year the average person in the United States may
be exposed to about 6 g of benzo[a]pyrene (NAS, 1972), a
dose that is several thousand times the amount needed to
induce cancer in mice.
There is also evidence that suggests that conjugation
of the benzo[a]pyrene metabolites with glucuronic acid,
presumably as a detoxification process, does not necessar-
ily render the conjugates inactive (Kinoshita and Gelboin,
1978). β-Glucuronidase, which is abundant in the liver,
spleen, kidney, and other secretory tissues, catalyzes the
hydrolysis of this glucuronide. In the process, an active
intermediate that can bind to DNA molecules with strong-

Copyright 2002 by Marcel Dekker. All Rights Reserved.

 A
Mixed-function oxidase (MFO)
B
D C
O
O MFO O

HO
E MFO F
OH
HO HO

OH OH
G
HO HO HO J
HO
NADPH NADPH
HO HO
HO
OH OH OH
HOH OH
OH HOH HOH OH OH HOH OH
HO HO HO HO

HO HO HO HO
OH H I OH K OH L OH

Figure 9.3 Metabolism of benzo[a]pyrene (BP) showing the formation of possible metabolites that may act as the proximate carcino-
gens or mutagens A, benzo[a]pyrene; B, (-) BP-trans-7,8-epoxide; C, epoxide hydratase; D, (-) BP-trans-7,8-diol; E, BP-diolepoxide I; F,
BP-diolepoxide II; G, 7/8,9-BP-triol; H, 7,10/8,9-BP-tetrol; I, 7/8,9,10-BP-tetrol; J, 7,9/8-BP-triol; K, 7,9,10/8-BP-tetrol; L, 7,9/8,10-BP-
tetrol. The black box represents deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and protein binding that result in mutation and
carcinogenesis.

er affinity than the parent molecule is formed. Thus, car-
cinogenesis distant from the site of contact may be
mediated by glucuronide conjugates, which are well dis-
tributed throughout the body because of their greater water
solubility.
9.2.4 Toxicity
Fifteen PAHs have been identified as carcinogenic in ani-
mals through various routes of exposure (USDHHS,
1991). Some produce skin tumors in the two-stage mouse
skin carcinogenesis system and produce local sarcomas at
the injection site, and others have the ability to produce
lung tumors after either intravenous injection or intratra-
cheal instillation or inhalation (IARC, 1983). In almost all
the studies, benzo[a]pyrene appeared to be the most potent
carcinogen. Mammary tumors induced by intragastric
dosed dimethylbenzo[a]anthracene have become an im-
portant model in the study of breast cancer.
Only four PAHs (benzo[a]pyrene, benzo[a]an-
thracene, dibenzoanthracene, and methylcholanthrene)
have been tested through the oral route and have shown
positive indications of carcinogenicity (Nawrot et al.,
1999). The target organs for these compounds were shown
to be the rodent forestomach; mouse lung, liver, and blood
vessels; and rat and mouse mammary glands. Oral admin-
istration of PAHs has also been found to result in repro-
ductive, hematopoietic, and other systemic toxicity;
however, the carcinogenic effects are considered the most
significant (JECFA, 1991). The carcinogenic PAHs de-
tected in foods have all been found to be genotoxic (US-
DHHS, 1993) and may be reasonably considered to be
human carcinogens.

Copyright 2002 by Marcel Dekker. All Rights Reserved.