Detection of Heterocyclic Amine Content in Different Types of Pork
Products
Ting Li
1,3 a
, Haibin Li
2b
, Jiayun Kang
1,3 c
, Wenhui Liang
1,3 d
, Fang Wang
1,3 e
,
Jinghua Qi
1,3 f
and Meixia Pang
1,3,* g
1
Beijing Key Laboratory of Agricultural, Product Detection and Control of Spoilage Organisms and Pesticide Residue,
Beijing University of Agriculture, Beijing, China
2
Pengcheng food branch of Beijing Shunxin agriculture Co., Ltd, Beijing, China
3
Beijing Innovation Consortium of Swine Research System, Beijing, China
Keywords:
Heterocyclic Amines, Pork Products, LC-MS/MS.
Abstract:
There are different levels of heterocyclic amines in pork products processed by different methods, and the
total heterocyclic amine content was 7.68-64.18 ng/g. Among them, non-polar heterocyclic amines Harman
and Norharman were mostly 1.42-3.69 ng/g and 1.40-40.47 ng/g. The distinction within the content of
heterocyclic amines in meat products with different processing methods was obvious. The total amount of
heterocyclic amines from high to low were cured meat, sauced meat, dried meat, roast meat, fried meat,
canned, sausage and ham.
1 INTRODUCTION
1
Along with our country residents living standards
improve, people produce harmful substances in food
processing is more and more attention, and
heterocyclic amines in a lot of food and different
processing technology produces, has been more and
more researchers focus, but bad amine for
miscellaneous domestic research is still in its infancy,
the related research and less reported, and focused on
the detection method to explore and optimize. Pork is
the main type of meat consumed by Chinese
residents, and there are four main cooking methods in
China: sauce, barbecue, pan frying and deep-frying.
As can be seen from foreign research reports, hcas are
ubiquitous in meat products, and most of the meat
products studied are cattle, sheep, fish and poultry,
while pork is rarely studied. In addition, the
a
https://orcid.org/0000-0002-5644-5768
b
https://orcid.org/0000-0003-0899-5068
c
https://orcid.org/0000-0003-0174-9710
d
https://orcid.org/0000-0001-8982-7693
e
https://orcid.org/0000-0002-0438-6786
f
https://orcid.org/0000-0001-9204-3697
g
https://orcid.org/0000-0003-3025-5305
traditional way of cooking in China and abroad have
very big different, the kinds of heterocyclic amines
and content in the meat products also have bigger
difference, at the same time of heterocyclic amines in
overseas has for meat formation mechanism,
influencing factors, analysis method and suppression
measures have a more in-depth study, and domestic
research is just getting started in recent years, various
aspects are in-depth study, Therefore, this study
focuses on the influencing factors of hCAS content in
pork products.
According to GB/T 26604-2011(2011), meat
products are primarily separated into cured meat,
sauced meat, roast meat, dried meat, fried meat,
sausage, ham, prepared meat products and other meat
products. This study chose 14 sorts of pork items
which are well known and loved by consumers within
the market. This paper analyzed the influence of
Li, T., Li, H., Kang, J., Liang, W., Wang, F., Qi, J. and Pang, M.
Detection of Heterocyclic Amine Content in Different Types of Pork Products.
DOI: 10.5220/0011293600003438
In Proceedings of the 1st International Conference on Health Big Data and Intelligent Healthcare (ICHIH 2022), pages 289-294
ISBN: 978-989-758-596-8
Copyright
c
 2022 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved
289

processing methods on the content of heterocyclic
amines, and provided data reference for future studies
on the population exposure of heterocyclic amines.
2 MATERIALS AND METHODS
2.1 Materials and Reagents
Pork floss, jerky, dried meat crisps, spiced pork,
bacon, preserved pork, grilled tenderloin, braised
pork head, American ham, starch-free ham, wide-
flavored sausage, ham, spam, and toothpick meat
were all purchased from the market.
10 HAAs standards (IQ, MeIQ, 8-MeIQx, 4,8-
DiMeIQx, 4,7,8-TriMeIQx, PhIP, Norharman,
harman, Trp-P-1, AαC), Methanol, Acetonitrile,
Ammonium formate, Formic acid (HPLC, TRC
Canada); Dichloromethane, Sodium hydroxide,
Ammonium acetate, Hydrochloric acid (AR,
Sinopharm reagent Co., Ltd.); Diatomite (Extrelut-
20NT, Merck, Germany); PRS-SPE column, C18-
SPE column (500mg / mL, 300mg / mL, Agilent,
USA).
2.2 HAAs Analysis
According to the method of Shen(Shen 2017), the
sample was purified by PRS column and C18 solid
phase extraction, then detected and analyzed by LC-
MS/MS.
2.3 Statistical Analysis
All data was organized using Microsoft Excel, and
Origin 8.0 was used for graphing, and IBM SPSS
Statistics 20 statistical software was used for data
analysis.
3 RESULTS AND DISCUSSION
The samples of pork products collected in this
experiment are all bagged products sold in the market,
except that toothpick meat is currently unpackaged.
Bacon, preserved pork and grilled tenderloinare all
raw meat products, after heating mature meat
products, and then tested.
The contents of polar heterocyclic amines of all
meat products are shown in Table 1, and the contents
of non-polar heterocyclic amines are shown in Table
2
.
Table 1: The content of polar HAAs in different meat products.
Meat products IQ MeIQ 8-MeIQx 4,8-DiMeIQx PhIP Total
Pork floss 0.97±0.05
e
0.66±0.02
d
0.07±0.02
d
0.05±0.01
d
0.04±0.03
e
1.78±0.06
fg
jerky 1.18±0.02
e
0.66±0.01
d
0.06±0.02
d
0.03±0.02
d
0.10±0.03
d
2.03±0.17
fg
dried meat crisps 1.24±0.01
e
0.64±0.01
d
0.15±0.05
c
0.08±0.01
d
0.12±0.04
d
2.23±1.01
f
Spiced Pork 1.84±0.07
e
1.01±0.12
bc
0.41±0.48
b
0.32±0.17
b
0.12±0.04
d
3.70±1.230
e
bacon 6.19±0.32
a
0.83±0.03
c
0.40±0.02
b
nd 0.13±0.02
d
7.55±0.51
b
preserved pork 5.42±0.12
b
0.81±0.04
c
0.76±0.34
a
0.17±0.07
c
1.95±0.16
a
9.10±2.23
a
grilled tenderloin nd 0.75±0.02
cd
0.15±0.04
c
0.17±0.07
c
0.15±0.01
d
1.23±0.15
g
braised pork head nd 0.91±0.02
c
0.12±0.04
cd
0.18±0.03
c
0.20±0.06
d
1.41±0.16
fg
American ham nd 1.08±0.02
bc
0.18±0.09
c
0.27±0.09
b
0.30±0.07
cd
1.83±0.37f
g
starch-free ham 4.17±0.24
c
1.82±0.11
b
0.12±0.01
cd
0.13±0.02
c
0.16±0.01
d
6.41±2.12
cd
wide-flavored sausage 2.69±0.10
d
3.56±0.11
a
0.26±0.02
c
0.29±0.02
b
0.33±0.02
c
7.12±2.43
bc
ham 1.37±0.17
e
1.92±0.45
b
0.18±0.01
cd
0.26±0.02
b
0.33±0.01
c
3.42±1.20
e
spam nd 2.51±0.12
b
0.39±0.08
b
0.50±0.12
a
0.46±0.16
c
3.86±0.58
e
toothpick meat 1.69±0.11
e
1.95±0.24
b
0.81±0.10
a
0.51±0.35
a
1.22±0.10
b
6.16±2.11
d
nd=not detected.
Each value is represented as mean± standard deviation (n= 3). Means with different superscript letters within the
same column are significantly different at P<0.05.
ICHIH 2022 - International Conference on Health Big Data and Intelligent Healthcare
290

It can be seen from Table 1 that the content of IQ
was nd-6.19 ng/g, and the average content was 1.91
ng/g. The highest content was Bacon at 6.19 ng/g.
Among which the highest content is bacon 6.19 ng/g,
which has a significant difference compared with
other meat products (P <0.05), but it is not detected in
grilled tenderloin, braised pork head, American ham
and spam. The IQ content of most meat products was
1-3 ng/g.It should be noted that bacon is sealed
packaging of raw bacon, bought by the canteen after
fried and then tested, because when fried meat and
heat source direct contact, The various precursors
formed by heterocyclic amines in meat constantly
exude from the meat and react on the contact surface
to form more IQ-type heterocyclic amines. The
content of MeIQ ranged from 0.64-3.56ng/g, and the
average content was 1.37 ng/g. The content of the
sausage was the highest. There was no significant
difference in most samples (p> 0.05). Wide-flavored
sausages are processed through pickling, enema,
airing, and baking. The auxiliary materials contain a
relatively high proportion of sugar(Huang 2019).
Sugar is the precursor of polar heterocyclic amines.
The high content of MeIQ may be due to the high
sugar content in Wide-flavored sausages. The
measured 8-MeIQx in samples was 0.06-0.81ng/g,
and the average content was 0.29 ng/g. The content of
toothpick meat was the highest. This heterocyclic
amine may be easily formed due to the higher frying
temperature. The content of PhIP ranged from 0.04-
1.95ng/g, the average content was 0.40 ng/g, the
content was low, and most samples were not
significantly different (p>0.05). The total content of
the five polar heterocyclic amines measured in the
surveyed meat samples was 0.96-7.28ng/g. The
highest content of polar heterocyclic amines was
preserved pork, and the lowest content was roasted
tenderloin.
Table 2 The content of non-polar HAAs in different meat products
Meat products Norharman Harman Trp-P-1 AaC Total
Pork floss 3.72±0.21
e
18.74±3.08
b
nd nd 22.46±1.2
c
jerky 4.06±0.95
e
14.25±0.62
b
nd nd 18.31±0.56
d
dried meat crisps 10.35±0.14
c
5.98±0.20
c
nd nd 16.33±0.78
d
Spiced Pork 21.02±0.64
b
32.69±0.34
a
nd nd 53.71±1.39
a
bacon 8.85±0.46
cd
3.49±0.13
cd
nd nd 12.34±0.65
ef
preserved pork 40.47±0.95
a
11.58±0.65
b
nd nd 52.05±2.45
a
grilled tenderloin 4.69±0.13
e
4.89±0.19
c
nd 0.02±0.01
c
9.60±0.34
g
braised pork head 9.37±0.19
c
26.00±0.69
a
nd 0.04±0.01
bc
35.41±0.89
b
American ham 2.35±0.91
f
2.86±0.43
d
nd 0.02±0.01
c
5.24±1.12
h
starch-free ham 2.17±0.15
f
4.79±0.15
c
nd 0.02±0.01
c
6.98±0.91
h
wide-flavored sausage 1.40±0.16
g
1.42±0.15
e
0.26±0.01
a
0.01±0.01
c
3.08±0.89
i
ham 3.73±0.01
e
5.79±0.64
c
0.14±0.03
b
0.02±0.01
c
9.68±0.81
g
spam 5.53±1.00
d
4.62±0.93
c
nd 0.11±0.02
a
10.26±0.65
fg
toothpick meat 5.53±0.26
d
8.10±1.18
c
nd 0.07±0.01
b
13.70±1.11
e
nd=not detected.
Each value is represented as mean± standard deviation (n= 3). Means with different superscript letters within the
same column are significantly different at P<0.05.
The non-polar heterocyclic amine content and
total heterocyclic amine content are shown in Table
2. The content of Norharman ranges from 1.40-40.47
ng/g, with an average content of 8.80 ng/g. The
highest content was Bacon. The bacon is made by
curing, air-drying, smoking and roasting. Although
the temperature of the whole process is low, because
the smoke contains heterocyclic amines, it adheres to
the surface of meat products during smoking. After
heating treatment before eating, Heterocyclic amines
are immersed in the meat from the surface, resulting
in an increase in the content of heterocyclic amines.
Detection of Heterocyclic Amine Content in Different Types of Pork Products
291

It should be noted that bacon is a sealed package of
cured bacon, bought from the supermarket and
cooked in water before being tested.
The content of Harman is in the range of 1.42-
32.69 ng/g, with an average content of 10.37 ng/g,
and the highest content was Sauced meat. Although
the processing method of sauced meat was relatively
mild, the heating temperature was not high at 100℃,
but the cooking time was more than 1h, and the
resulting Harman content was indeed high. The
Norharman and Harman content of wide-flavored
sausage was the lowest. Both Norharman and Harman
belong to β-carboline heterocyclic amines. Regarding
the formation of β-carboline, it is generally believed
that it is directly generated by pyrolysis of a single
amino acid or protein, and the formation temperature
is generally higher than 300 ℃ (MLC 2004), But later
studies have shown that β-carboline can also be
formed at temperatures below 100°C (Liu 2020).
Because Norharman and Harman can enhance the
mutagenicity of other heterocyclic amines in the
Ames/Salmonella experiment, they are called
auxiliary mutagens (Kanithaporn 2011). At the same
time, studies have shown that it is related to
neurotoxins and enzyme inhibitors.
The content of Trp-p-1 was nd-0.26 ng/g, and the
average content was 0.03 ng/g. Most samples were
not detected, only wide-flavored sausage and
common ham detected low levels of this heterocyclic
amine, which were 0.26ng/g and 0.14ng/g,
respectively.
The content of AaC was nd-0.11ng/g, and the
average content was 0.02 ng/g. The content was very
low and almost undetectable.
Figure 1: The total HAAs content of pork products in different processing methods.
In order to reflect the influence of different
processing methods on the content of heterocyclic
amines in pork products, the meat products with the
highest content of heterocyclic amines among various
types were selected to compare 8 types of meat
products. It can be seen from Figure1 that the total
amount of heterocyclic amines in descending order
were Cured meat, Sauced meat, Dried meat, Roast
meat, Fried meat, Canned meat, Sausage and Ham.
The total amount of heterocyclic amines in cured
meat products and braised meat products was much
higher than other meat products, at 64.18ng/g and
58.64ng/g, which were almost 2-3 times the content
of other meat products.
ICHIH 2022 - International Conference on Health Big Data and Intelligent Healthcare
292

Figure 2: The polar and nonpolar HAAs content of pork products in different processing methods.
It can be seen from Figure 2 that the level of polar
heterocyclic amines was very low. The content of all
meat products surveyed was less than 10ng/g. Except
for ham, the content of non-polar heterocyclic amines
was almost above 10ng/g, the polar and non-polar
heterocyclic amines of cured meat products were the
highest, reaching 9.10ng/g and 52.05ng/g
respectively. However, only non-polar heterocyclic
amines were higher in cured and sauced meat
products, while polar heterocyclic amines are lower.
4 CONCLUSIONS
The established LC-MS/MS method was used to
detect the content of heterocyclic amines in common
commercial pork products. All kinds of meat products
have heterocyclic amines to varying degrees, and the
total heterocyclic amine content was 7.68-64.18ng/g.
Among them, the most common non-polar
heterocyclic amines Harman and Norharman were
1.42-32.69 ng/g and 1.40-40.47 ng/g. The content of
heterocyclic amines in meat products of different
processing methods was significantly different, and
the content of heterocyclic amines in cured meat
products was the largest. The total amount of
heterocyclic amines in descending order were Cured
meat products, Sauced meat products, Dried meat
products, Smoked and Roasted meat products, Fried
products, Canned meat products, sausage and ham. It
is inevitable to take in different levels of heterocyclic
amines in the daily diet. This study is to investigate
and evaluate the safety of pork products, and to
further establish the exposure and prevention of
heterocyclic amines in meat products.
ACKNOWLEDGEMENTS
This work was supported by Beijing Key Laboratory
of Agricultural, Product Detection and Control of
Spoilage Organisms and Pesticide. The authors
express their thanks to Beijing Innovation
Consortium of Swine Research System and
Pengcheng food branch of Beijing Shunxin
Agricultural Limited Company for providing the
primary sample.
REFERENCES
(2011). Meat product classification. GB/T 26604-2011: 7.
Huang Yuanxiang, Zhao Shenglan, Ma Yage, et al (2019).
Study on the formulation optimization and texture
characteristics of low-calorie, low-sodium, wide-flavor
sausage[J]. Meat Industry, (08):16-24
Kanithaporn P, Wannee J, Scott S J.(2011).Inhibitory
activity of Asian spices on heterocyclic amines
formation in cooked beef patties.[J]. Journal of food
Detection of Heterocyclic Amine Content in Different Types of Pork Products
293

science,76(8).
Liu Tiantian, Yao Yao, Wang Wei, et al.(2020).Formation
and inhibition of three heterocyclic amines, PhIP,
Norharman, and Harman in the model system[J]. Meat
Research,34(03):1-7.
M, L. C. and K. T. H, et al. (2004).Effects of heating time
and antioxidants on the formation of heterocyclic
amines in marinated foods. Journal of chromatography.
B, Analytical technologies in the biomedical and life
sciences 802 (1).
Shen X C, Zhang Y L, Cui Y Q, et al.(2017). Simultaneous
determination of 9 heterocyclic aromatic amines in pork
products by liquid chromatography coupled with triple
quadrupole tandem mass spectrometry[J]. IOP
Conference Series: Earth and Environmental Science,
77(1).
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