Intake of heterocyclic aromatic amines from meat in the European
Prospective Investigation into Cancer and Nutrition (EPIC)-Heidelberg cohort
Sabine Rohrmann*, Dorothee Zoller, Silke Hermann and Jakob Linseisen
Division of Clinical Epidemiology, German Cancer Research Centre, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany
(Received 9 January 2007 – Revised 22 May 2007 – Accepted 24 May 2007)
It was the aim of the present study to estimate the intake of heterocyclic aromatic amines (HCA) from meat, which have been associated with
cancer risk in several epidemiological studies, of 21462 subjects who participated in the European Prospective Investigation into Cancer and
Nutrition (EPIC) in Heidelberg. This was accomplished by using a detailed dietary questionnaire that assessed meat consumption, cooking
methods, and degree of browning of the respective food items. Median total HCA intake from meat was 31ng/d (mean 69ng/d), which was
lower than results observed in previous studies. 2-Amino-1-methyl-6-phenylimidazo[4,5b]pyridine was the most common HCA in this cohort
(median 17; mean 48ng/d). The present study offers the opportunity of a detailed examination of the associations between meat cooking as
well as HCA intake from meat and cancer risk in a prospective way.
Heterocyclic aromatic amines: Meat: Cancer risk: European Prospective Investigation into Cancer and Nutrition
Meat and fish are usually cooked before being consumed.
Cooking methods that induce high temperatures and a direct
exposure to a hot surface, such as grilling or frying, or to
direct flame, for example, barbecuing, are discussed in relation
to carcinogenesis. Results from several epidemiological
studies show associations between meat prepared at high tem-
peratures and the risk of several types of cancer1–5. One of the
reasons for the increased risk is thought to be the heat-depen-
dent formation of heterocyclic aromatic amines (HCA). When
meat is cooked at temperatures over 1308C, for example, when
frying, barbecuing or grilling, these compounds are formed
from amino acids, creatinine and sugar6. The amount of
HCA production depends mainly on cooking method, tem-
perature and the type of meat, with amounts ranging in most
studies from 1 to 80ng/g meat for 2-amino-1-methyl-6-pheny-
limidazo[4,5b]pyridine (PhIP), the most abundant HCA in
the human diet, followed by 2-amino-3,8-dimethyl-3H-imida-
zo[4,5f]quinoxaline (MeIQx), with usual amounts up to 6ng/g
meat and 2-amino-3,4,8-trimethyl-3H-imidazo[4,5f]quinoxa-
line (DiMeIQx), with usually up to 1ng/g meat7. Recent epi-
demiological studies have shown associations between the
estimated intake of HCA from diet and the risk of colorec-
tal1,8, breast4and prostate9cancer, although other studies
did not observe positive associations10,11.
We examined the distribution of HCA intake from meat in a
German cohort of middle-aged men and women. Previous
studies reported a wide range of the daily HCA intake from
as low as 77ng/d in a Swedish study10to more than 1mg/d
in a US study12. The intake of HCA in a pilot project has
been described previously13. This new analysis was conducted
in the entire European Prospective Investigation into Cancer
and Nutrition (EPIC)-Heidelberg cohort.
Material and methods
EPIC is a prospective cohort study conducted in ten countries
that started in the early 1990s14. In Heidelberg (Germany),
25544 subjects, aged 35–65 years (women) and 40–65
years (men), were recruited for participation between 1994
and 1998. During recruitment, information on diet, lifestyle
and health have been collected. All subjects are being con-
tacted in approximately 2-year intervals to collect information
on chronic disease status as well as diet and lifestyle15. During
the second follow-up (2001–3), 25049 participants have been
contacted. Of those, 86% completed a 158-item FFQ that
assessed food consumption during that past 12 months,
which had previously been used in the baseline assessment
(1994–8) (for details, see Bohlscheid-Thomas et al.16). This
FFQ included questions on meat consumption and preparation.
Participants were asked how often they consumed sixteen
types of meat (beef roast, including goulash, roulade; beef
steak, filet or tenderloin; pork roast, including goulash; pork
*Corresponding author: Dr Sabine Rohrmann, fax þ6221 422203, email email@example.com
Abbreviations: DiMeIQx, 2-amino-3,4,8-trimethyl-3H-imidazo[4,5f]quinoxaline; EPIC, European Prospective Investigation into Cancer and Nutrition; HCA,
heterocyclic aromatic amine; MeIQx, 2-amino-3,8-dimethyl-3H-imidazo[4,5f]quinoxaline; PhIP, 2-amino-1-methyl-6-phenylimidazo[4,5b]pyridine.
British Journal of Nutrition (2007), 98, 1112–1115
q The Authors 2007
British Journal of Nutrition
steak, schnitzel, cutlet, filet or tenderloin; hamburgers or meat-
balls; frying sausage; Wieners; bacon; liverloaf; fried chicken,
turkey breast, turkey goulash) and which cooking methods
they prefer for each type of meat (steaming or boiling, pan-
frying, breading and frying, frying or broiling, grilling or bar-
becuing). Additionally, with the help of four pictures, subjects
stated which degree of browning they favoured (lightly
browned, moderately browned, strongly browned, extremely
browned)17. If a subject indicated to vary between two or
more cooking methods per food items these cooking methods
were weighted equally. The same degree of browning was
assumed for each cooking method used for a specific type of
Total HCA concentration and concentration of the most
abundant HCA PhIP, MeIQx and DiMeIQx were estimated
using published data of their content in different types of
meat18–21. HCA intake from steaming or boiling or from
breading and frying was considered to be zero7. In addition
to meat cooking, participants were asked about the use of
meat drippings to prepare gravy. The intake of HCA from
gravy was calculated by multiplying the amount of gravy
with the HCA concentration in gravy of the corresponding
meat item. This was automatically added to a specific meat
item’s HCA intake. By combining information on degree of
browning, cooking method and the amount of meat intake,
the mean daily dietary intake of HCA from meat was esti-
mated. We also calculated HCA intake per MJ to take into
account differences in energy intake that might contribute to
differences in HCA intake. Because HCA intake was not nor-
mally distributed we computed medians and interquartile
ranges of HCA intake and used the Wilcoxon test and Krus-
kal–Wallis test to compare the intake of different subgroups
of our participants. All tests were two-sided; P values
,0·05 were considered to be statistically significant. EPIC-
Heidelberg has been approved by the ethical committee of
the Heidelberg University Medical School.
The median intake of total HCA from meat was 30·6ng/d
(mean 69·4ng/d), with PhIP contributing most to total HCA
intake frommeat (median
(Table 1). Intake was highest from roast beef, followed by
chicken or turkey, hamburgers or meatballs, and beef steak.
Statistically significant differences in HCA intake were
observed by sex, age, education, BMI and smoking status.
Men had a higher HCA intake than women and smokers a
higher intake than non-smokers (Table 2). Intake decreased
with age and subjects with a higher educational level had a
lower HCA intake than those with a lower educational level.
Intake also differed by BMI, with more obese participants
having a higher HCA intake from meat (Table 2). These
differences were similar for HCA intake per MJ.
This is the first large European cohort study that attempts to
assess the intake of HCA from meat using a detailed ques-
tionnaire on food intake and food preparation methods. We
have previously estimated the intake of HCA in a smaller
group of EPIC-Heidelberg participants13, which revealed a
higher intake of total HCA (median 103ng/d) compared with
the present investigation (median 30·6ng/d). In a Swedish
study that used a similar approach to assess HCA intake,
median total HCA intake was 77ng/d, which also included
HCA intake from fish10. Similar amounts were calculated
in a large Japanese study22and slightly lower levels in a Sin-
gapore study (mean intake 49·95ng/d)23. In US studies, esti-
mated HCA intake is generally higher (mean PhIP intake
78·1ng/d; mean MeIQx intake 21·9ng/d) than in European
studies24. This might be explained by larger portions of
meat consumed in the USA than in Germany, but also by
differences in cooking method preferences, for example, a
preference for HCA-forming methods such as grilling in
US cohorts and for non-HCA-forming methods such as boil-
ing in Germany. Also, meat is usually consumed at a higher
degree of browning in US cohorts12than in our cohort, lead-
ing to a higher intake of HCA. In addition, different con-
sumption habits contribute to the observed differences.
‘Roast beef, roulade and goulash’ contributed most to the
intake of total HCA in our cohort; however, the contribution
of roast beef to HCA intake was negligible in three US
cohorts25. The difference in HCA intake between the pilot
study13and the present evaluation can, at least in part, be
explained by some changes in questionnaire design. We
added the possibility to mark the preparation method ‘boil-
ing’ that does not contribute to HCA intake. Second, in con-
trast to the pilot study, we did not consider fish in the
present study because HCA intake from fish varies widely
depending on the type of fish and its preparation20,26.
Third, HCA intake differed between subgroups mainly due
to higher meat consumption, for example, in men, but
also due to the preference of cooking methods or degrees
of browning. Men tended to consume meat darker than
Table 1. Intake of total heterocyclic aromatic amines (HCA), 2-amino-1-
methyl-6-phenylimidazo[4,5b]pyridine (PhIP), 2-amino-3,8-dimethyl-3H-
imidazo[4,5f]quinoxaline (MeIQx) and 2-amino-3,4,8-trimethyl-3H-imida-
zo[4,5f]quinoxaline (DiMeIQx) as well as total HCA by meat type in
European Prospective Investigation into Cancer and Nutrition (EPIC)-
Heidelberg (n 21462)
(Medians and interquartile ranges)
Median Interquartile range
Total HCA from meat
Intake of single HCA
Total HCA intake by meat type
Beef roast, including
Fried chicken, turkey breast,
Beef steak, filet or tenderloin
Pork steak, schnitzel, cutlet,
filet or tenderloin
Pork roast, including goulash
Bacon, pork belly
Intake of heterocyclic aromatic amines1113
British Journal of Nutrition
women and younger participants tended to prepare meat
more often by frying or grilling than older participants
(data not shown).
In conclusion, we estimated an HCA intake from meat
that was lower than observed in previous studies in
Europe or the USA. Statistically significant differences
were seen for age, sex, BMI, education and smoking
status. EPIC-Heidelberg offers the opportunity to examine
the association of meat consumption and HCA intake from
meat with cancer risk in a prospective way. Further, future
studiescan takeinto account
especially genetic variation in metabolic pathways of HCA
as well as secondary plant products such as phenolic
acids that are well known to have an impact on HCA metab-
olism and cancer risk.
The authors thank the volunteers who participated in EPIC-
Heidelberg. This study was supported by the German
Cancer Research Centre and grants from the Kurt-Eber-
Risk, Nutrition and Individual Susceptibility), a network
of excellence operating within the European Union 6th
Framework Program, Priority 5: ‘Food Quality and Safety’
(Contract No 513943).
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Table 2. Total heterocyclic aromatic amines (HCA) intake by sex, age, education, smoking status and body mass index in European Prospective
Investigation into Cancer and Nutrition (EPIC)-Heidelberg (n 21462)
(Medians and interquartile ranges)
Total HCA intake from meat
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