Fact sheet:

Biogenic amines

Version: 09/01/2015

GMP+ International B.V.

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 Content

General Summary

Summary of GMP+ products standards for the animal feed sector

More Facts
1. Nature, history and prevalence of biogenic amines
2. Transmission to the environment, plants, animals and humans
3. Diagnose of poisoning
4. Potential hazards and adverse effects
5. Severity of the hazard
6. Standards
7. Analysis methods
8. Control measures
9. References
10. Websites

APPENDIX / APPENDICES

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 General Summary

Name: Biogenic amines

code: C05

Description: Substances which are created through the degradation of amino acids as a result of
the spoilage (protein decay) of protein-rich animal feed materials.
Type: chemical

Severity: high

Control measures: - (Locked) closed storage

- (Locked) closed transport
- Check of odour and appearance
- Climate/temperature control
- Establish the time between reception and processing
- Maximum pH
- Prevent condensation

The control measures specified in this fact sheet are all control measures which can be used depending on the
product and/or process step.

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 SUMMARY OF GMP+ SPECIFIC FEED SAFETY LIMITS FOR THE ANIMAL FEED SECTOR
Contaminant Product Action limit(1) Rejection limit(1) Source Supplementary Analysis method7
requirements
Chemical: Other undesirable substances and products

C5 Biogenic amines* - Animal feeds> 30% crude protein - 4,000 mg/kg GMP+ * In as far as these are OZM Part 2; OSP-
protein created by decay due to 23
protein decay and are not
the consequence of the
normal processing of the
product

[1 ] Action limit: A feasible limit agreed in consultation with the sector, supplier or customer. If this limit is exceeded then an investigation into the cause should be undertaken and corrective measures should be
taken to remove or control that cause. Maximum levels in mg/kg (ppm) of the feed materials or compound feeds, derived to a moisture content of 12% unless mentioned differently.
Rejection limit: A feasible limit agreed in consultation with the sector, supplier or customer. If this limit is exceeded then the product is not suitable for use as feed material or animal feed. Maximum levels in mg/kg
(ppm) of the feed materials or compound feeds, derived to a moisture content of 12% unless mentioned differently.
[7] The research methods (OZM) can be found via the PDV website (www.pdv.nl ; quality; research methods)

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 More facts

Chemical name
Cadaverine : pentane-1,5-diamine
Histamine : 2-(1H-imidazol-5-yl)ethanamine
Putrescine : butane-1,4-diamine
Spermidine : N-(3-aminopropyl)butane-1,4-diamine
Spermine : N,N'-bis(3-aminopropyl)butane-1,4-diamine

CAS-number
Cadaverine : 462-94-2
Histamine : 51-45-6
Putrescine : 110-60-1
Spermidine : 124-20-9
Spermine : 71-44-3

Synonyms (non exhaustive)

Cadaverine : 1,5-pentanediamine; pentamethylenediamine
Histamine : 1H-imidazole-4-ethanamine; 5-imidazoleethylamine
Putrescine : 1,4-butanediamine; 1,4-diaminobutane; tetramethylenediamine
Spermidine : N-( -aminopropyl)tetramethylenediamine; spermidin
Spermine : N,N'-bis(3-aminopropyl)tetramethylenediamine; gerontine; musculamine
In this fact sheet will be referred to biogenic amines as BAs.

1. Nature, history and prevalence of biogenic amines

Spoiled foodstuffs and especially fermented foods tend to contain elevated levels of biogenic
amines, although their concentrations vary extensively not only between different food
varieties but also within the varieties themselves (Bodmer et al., 1999). BAs are generated in
course microbial, vegetable, and animal metabolisms and are nitrogenous compounds
formed mainly by decarboxylation of amino acids or by amination and transamination of
aldehydes and ketones (precursors) (Karoviov and Kohajdov, 2005) as shown in table1.
The names of many biogenic amines correspond to the names of their originating amino
acids.

Table 1. Several BAs and their amino acid precursors (Karoviov and Kohajdov, 2005).
Biogenic amine Amino acid precursor
Cadaverine Lysine
Histamine Histidine
Putrescine Ornithine
Spermidine Putrescine
Spermine Spermidine

In this fact sheet is focussed on five BAs: cadaverine, histamine, putrescine, spermidine and
spermine because of their importance in the food and feed industry.

As shown in figure 1, the chemical structure of BA can either be: aliphatic (putrescine,
cadaverine, spermine, spermidine), aromatic (tyramine, phenylethylamine), heterocyclic
(histamine, tryptamine).

Figure 1. Chemical structure cadaverine, histamine, putrescine, spermidine and spermine (Karoviov and
Kohajdov, 2005).

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 Cadaverine Histamine Putrescine

Spermidine Spermine

Some BAs are classified as polyamines, e.g. putrescine, spermidine and spermine (Kala,
2009). Some authors also classify cadaverine as a polyamine (Karoviov and Kohajdov,
2005).

The molecular formulas are:

Cadaverine : C5H14N2
Histamine : C5H9N3
Putrescine : C4H12N2
Spermidine : C7H19N3
Spermine : C10H26N4

Cadaverine is soluble in water, alcohol and slightly soluble in ether. Histamine is soluble in
water and alcohol, it is practically insoluble in ether. Putrescine is also soluble in water.
Spermidine is soluble in water, ethanol and ether. Spermine is soluble in water and is
practically insoluble in ether (O'Neil et al., 2006). BAs are heat stable (Min et al.1,2007),
however the heat stability of the micro-organisms producing BAs varies.

Some BAs are used commercially, like putrescine which is used as a pesticide, as bait in
traps, and is approved as a food contact material (RIVM-RIKILT, 2009).

BAs can have beneficial effects in the animal and human body. Polyamines, as some BAs
are classified, are important for the growth, renovation, and metabolism of every organ in the
body and essential for maintaining the high metabolic activity of the normal functioning and
immunological system of gut (Karoviov and Kohajdov, 2005). However in food
microbiology they have been related to spoilage and fermentation processes (Santos, 1996).
This is also the case in feed microbiology. These BAs can be also be toxic for animals an
humans.

BAs are important from a hygienic point of view as they have been implicated as the
causative agents in a number of food poisoning episodes (Shalaby, 1996). In non fermented
foods these compounds were found useful as indicators and markers of food decomposition.
Spoiled foods are also rich in BAs and usually contain high levels of putrescine and
cadaverine. So they can be used as indicators of the degree of freshness or spoilage of food
(Karoviov and Kohajdov, 2005). Also Total Volatile Nitrogen (TVN) (Ricque-Marie et al.,
1998) or ammonia (FAO2, 2001) content can also be used as an indicator for freshness,
since they are related to the presence of BAs.

Because the BAs in food and feed products are mainly generated by decarboxylation of the
corresponding amino acids precursors, for the formation of BAs the following conditions are
necessary (Karoviov and Kohajdov, 2005):
Availability of free amino acid;
Presence of decarboxylase-positive micro organisms;

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 Conditions that allow bacterial growth, decarboxylase synthesis and decarboxylase
activity.
So in virtually all foods that contain protein or free amino acids and are subject to conditions
enabling microbial or biochemical activity BAs can be expected (Karoviov and Kohajdov,
2005). Different BAs tend to predominate in different foods and feed, depending on the
amino acids present, the nature of the bacterial population and the nature of the processing
and storage conditions (Lawley et al., 2008).

2. Transmission and likelihood of occurrence

Environment
Although amino acid decarboxylases are not widely distributed among bacteria, species of
many genera such as Bacillus, Citrobacter, Clostridium, Klebsiella, Escherichia, Proteus,
Pseudomonas, Shigella, Photobacterium and the lactic acid bacteria Lactobacillus,
Pediococcus, and Streptococcus are capable of decarboxylating one or more amino acids.
Microbial strains with high proteolytic enzyme activity also potentially increase the risk for
BAs formation in food systems, by increasing the availability of free amino acids (Karoviov
and Kohajdov, 2005).
Amino acid decarboxylase activity is stronger in an acidic environment, the optimum pH
being between 4.0 and 5.5. Furthermore, in such an environment bacteria are more strongly
encouraged to produce these enzymes, as a part of their defence mechanisms against the
acidity. The presence of fermentable carbohydrate, enhances both growth and amino acid
decarboxylase activity in bacteria (Karoviov and Kohajdov, 2005).
Oxygen supply also appears to have a significant effect on the biosynthesis of BAs.
Enterobacter cloacae produces about half the quantity of putrescine in anaerobic compared
with aerobic conditions, and Klebsiella pneumoniae synthesizes significantly less
cadaverine but acquires the ability to produce putrescine under anaerobic conditions. The
redox potential of the medium also influences BAs production. Conditions resulting in a
reduced redox potential stimulate histamine production, and histidine decarboxylase activity
seems to be inactivated or destroyed in the presence of oxygen.
Amine formation by bacteria is decisively influenced by temperature. Temperature between
20C and 37C is optimal for the growth of the most bacteria containing decarboxylases,
decreased temperature stops their growth (Karoviov and Kohajdov, 2005).

Several bacteria have been shown to be heat stable, so heating feed or food products, will
not prevent, depending on the bacteria type, biogenic amine production. Biogenic amines are
very heat stable and once formed, they will not be destroyed even by dramatic heat
treatment such as autoclaving (FAO1, 2003).

Plants
As stated by Karoviov and Kohajdov (2005), BAs are generated in course vegetable
(endogen) metabolisms. In plants, putrescine, spermidine and spermine are implicated in a
number of physiological processes, such as cell division, flowering, fruit development,
response to stress and senescence.
The levels of endogenous polyamines, like putrescine, have been shown to increase in plant
cells challenged with low temperature. the accumulation of putrescine under cold stress is
essential for proper cold acclimation and survival at freezing temperatures (Cuevas et al.,
2008).
Besides the production of endogenous BAs, BAs are also formed during, e.g. fermentation
processes or storage of food or feed products of vegetal origin. Anli et al. (2006) indicated
that both storage temperature and storage time were important factors affecting biogenic
amine content in beer. There are three types of biogenic amine groups in beers based on
their origin. The first group, which occurs naturally in raw materials, includes agmatine,
putrescine, spermidine, and spermine in malt. The next group, associated with mashing and

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 wort boiling, includes tyramine, cadaverine, and agmatine. The last group includes tyramine
and tryptamine and these can form as a result of fermentation.

Food substances that have been prepared by a fermentative process, or have been exposed
to microbial contamination during aging or storage, are likely to contain amines. Alcoholic
beverages such as beers can contain BAs, as do some other fermented foods such as
sauerkraut and soy bean products. Some fruits and vegetables were also found to contain
high concentrations of various BAs (Shalaby, 1996). Additionally Lawley et al. (2008) also
mention that wine can contain high levels of BAs. Certain BAs are also found naturally in a
range of fruit juices and fresh fruit and vegetables, including cocoa beans, mushrooms and
lettuce. Dadkov et al. (2009) studied the presence of BAs and polyamines in edible
mushrooms and stated that mushrooms can contain a very high spermidine levels.

Animals
Exposure
No data was found concerning BAs levels of exposure of animals.
Concerning likelihood of occurrence Phuntstok et al. (1998) found substantial amounts of
BAs in silages: alfalfa and maize, as shown in table 1.

Table 1. Levels of BAs in silages and hay (mg/kg of dry matter) (Phuntstok et al., 1998).
Biogenic amine Alfalfa silage Alfalfa hay Maize silage Grass silage
Cadaverine 3987 38 724
Histamine 3078 5 510
Putrescine 2953 20 667

In addition to that, Van Os et al. (1996) found BAs in grass silages. Total amine content of
the grass was low (100-200 mg/kg dry matter). The well preserved silages without additives
(e.g. formic acid, molasses, degrading enzymes etc.) contained up to 7400 mg/kg dry matter.
Tyramine, cadaverine, putrescine and histamine were, in descending order, the principal BAs
formed, representing together 90% of the total biogenic amine content of the silages.
Formation of BAs occurred mainly during the first 10 days of fermentation, and was highest
in silages with a slow acidification rate. Ensiling at high dry matter content, with formic acid or
inoculation with large numbers of lactic acid bacteria significantly reduced the amount of BAs
in the silage. Van Os et al. (1996) concluded that the formation of BAs in grass silage is
related to protein degradation, and that amine formation can be reduced by restriction of
fermentation in the silage, or by achieving rapid acidification during the first phase of ensiling.

Kse et al. (2003) studied BAs in fish meal and investigated the effect of processing and
storage conditions on histamine formation during fish meal production. It was found that most
histamine concentrated in the press liquor (stickwater) meal after processing. Histamine
levels were mainly decreased in mackerel samples but increased in cod samples after
processing into fish meal. No bacterial growth was observed in the press-cake when fish was
cooked and pressed during fish meal production. After drying of solids and the stickwater,
bacterial growth was observed. This is an indication that fish meal is apparently
microbiologically hygienic after cooking process, then recontamination occurs. The majority
of histamine was found in mackerel flesh meal rather than in offal meal (guts, heads and
bones, etc.). Histamine was mainly concentrated in the stickwater meal for all samples. Since
histamine is water soluble, it is natural that histamine will be extracted from raw fish during
processing (cooking and pressing) into press water. The samples that were packed in
polyethylene bags seemed to show a slight increase in histamine levels up to fifth week but a
significant decrease occurred gradually for the samples stored at 0, 20, 30 and 35 C.
However, storage temperature did not have significant effect on histamine levels for the
packed samples. The results of storage trial with unpacked samples showed that histamine
values decreased gradually with time.
In table 2 BA levels in animal feed of animal origin are shown.

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 B
Table 2. Mean biogenic amine levels (mg/ kg product) in feed materials of animal origin (Brinker et al., 2002 ,
C
Radosevich, 2009 ).
Biogenic amine SPD SPN PUT CAD HIM TYM
B
Fish meal - - 546 992 273 -
C
Fish meal 19 19 243 365 77 95
C
Chicken by product meal 25 39 81 125 14 25
C
Chicken meal 35 57 164 249 26 67
C
Meat meal 07 08 68 84 13 10
- = Not Determined; SPD = Spermidine; SPN = spermine; PUT = putrescine; CAD = cadaverine; HIM =
histamine; TYM = tyramine.

Absorption
Excess, not metabolizable, BAs are absorbed. No data was found concerning absorption
rates.

Distribution
BAs can be transferred to the liver and brain. No other data found.

Metabolism
Phuntsok et al. (1998) state that experimental results in ruminants suggest that extensive
metabolism of dietary amines takes place during ruminal fermentation and substantially
lowers the amounts of amines potentially absorbed relative to intake. Apparent passage of
amines in abomasal digesta was several fold less than amounts consumed. Other research
demonstrated that considerable amounts of dietary histamine disappeared intraruminally. So
BAs may be extensively metabolized by ruminal microorganisms. Increased ruminal NH3
after the addition of BAs to in vitro ruminal fluid incubations indicated that these compounds
can be metabolized rapidly. Cadaverine and putrescine are essential membrane components
for Gram-negative ruminal bacteria such as Selenomonas ruminantium, Veillonella purvula,
and Veillonella alcallescens. The uptake of BAs for use in bacterial cell-wall structure may
partly explain their extensive ruminal metabolism and relatively low recovery in abomasal
digesta.
No data was found concerning the metabolism of other animals species.

Excretion
BAs are transferred to milk, as stated by Karoviov and Kohajdov (2005).

Humans via animal products

The biogenic amine content of various foods and feed have been widely studied and found in
cheese, fish and meat products and eggs (Shalaby, 1996). Lawley et al. (2008) mention that
fermented meat products can contain having high levels of BAs.

The study of biogenic amine quantities in meat as a function of conservation time, could be a
useful tool to control meat spoilage. The formation of some BAs and concentration increase
of those already existing in meat, are due to degrading processes in food, which are
promoted by enzymatic reactions caused by external microbial activity or by endogenous
tissue activities. In an experiment by Vinci and Antonelli (2002) the results show that in red
meat (adult bovine) the biogenic amine levels (being tryptamine, putrescine, cadaverine,
serotonin, tyramine, spermidine, spermine) were still low until 9 days of storage ( 30 mg/kg)
and that over 36 days only cadaverine and tyramine concentrations became very high ( 120
mg/kg). In white meat (chicken) all the biogenic amine levels remained quite low ( 40 mg/kg)
all over the 36 days, instead of the cadaverine content which gained 50 mg/kg at the seventh
day of storage.

BAs are also present in organs. In an experiment of Krauslov et al. (2006) putrescine,
spermidine and spermine levels were studied in livers of young bulls, cows, pigs and chicken
24 hours after slaughtering. The results are shown in table 3.

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 Table 3. Mean putrescine, spermidine an spermine level (mg/kg) in livers (Krauslov et al., 2006).
Species Putrescine Spermidine Spermine
Cows 25 161 35
Young bulls Negligible 122 43
Pigs Negligible 32 115
Chickens Negligible 57 120

Kalac (2009) states that bovine, porcine and chicken liver, kidney, spleen and heart all
have a high content of spermine; bovine liver also of spermidine.

Paulsen et al., (2008) also studied biogenic amine levels in organs. Studied animals were
hare and roe deer. The results are shown in table 4.

Table 4. Mean biogenic amine levels (mg/kg) in organs of hare and roe deer (Paulsen et al., 2008).
Biogenic amine Spleen Spleen Liver Liver Kidney Kidney Mean
*
(hare) (deer) (hare) (deer) (hare) (deer) organs
Cadaverine <5
Histamine 14.6 11.6
Putrescine < 7.8
Spermidine 52.0 42.9 37.2 8.5 24.4 10.7
Spermine 91.1 102.2 111.2 94.6 82.8 79.9
Tyramine <5
*
= mean of spleen, liver and kidney of hare and roe deer.

Also the presence of BAs in fish products has received a lot of attention. Especially
histamine-producing species which include tuna, mahi mahi, escolar, bonito, yellowtail,
bluefish, sardine, pilchard, abalone, mackerel (FDA1, 2004) and in freshwater fish (Shakila
and Vasundhara, 2002). A fresh product typically has barely detectable levels of histamine.
Histamine can be present in fresh, canned and cooked product as the toxin survives
processing. The formation of histamine is typically associated with decomposed product
(FDA1, 2004) as a result of inadequate handling and preservation (Karoviov and
Kohajdov, 2005).
The formation of high levels of histamine in fish products can be fairly rapid and develops on
the number of micro organisms present. Several bacteria are involved in toxicity, such as
Proteus morganii, Hafnia alvei, Acromonas hydrophila, Vibrio alginolyticus, Pseudomonas
sp., Klebsiella sp., etc. These bacteria are capable of producing hazardous amounts of
histamine in very short period of time when fish are held at elevated temperatures. Low
storage temperatures are used in the fishery industry to control bacterial histamine
production Karoviov and Kohajdov, 2005).
Besides histamine, other BAs can be present in fish products, e.g. putrescine, cadaverine,
spermidine, spermine and tyramine (Lebiedziska et al. 1991).
In table 5 biogenic amine levels are shown in various fish products.
*
Table 5. Mean biogenic amine levels (mg/100 g) in fish and fishproducts (Hui, 2006).
Biogenic amine SPD SPN PUT CAD HIM TYM PHM
Tuna
Good 0.44 0.95 0.12 0.15 0.38 - -
Borderline 0.36 0.67 0.23 1.03 2.36 - -
Decomposed 0.07 0.12 0.25 1.03 25.3 - -
Tuna
Fresh - - 0.04 0.02 nd nd nd
0C / 21 days - - 0.52 2.44 10.8 1.38 nd
8C / 9 days - - 1.11 5.62 368 3.25 0.69
20C / 3 days - - 0.45 10.84 687 1.71 0.81
*
nd. = Not Detectable; - = Not Determined; SPD = Spermidine; SPN = spermine; PUT = putrescine; CAD =
cadaverine; HIM = histamine; TYM = tyramine; PHM = phenylethylamine.

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 Ramos et al., (2009) studied BAs in eggs. The presence of 11 BAs was studied, among
which putrescine, cadaverine, histamine, tyramine, spermine and spermidine. Two
experiments were conducted to evaluate yolk biogenic amine concentrations of fresh and
stored eggs, and to explain the effect of temperature and time of storage in the levels of BAs
during egg shelf-life. Only five of the 11 BAs under study were detected: putrescine,
cadaverine, propylamine, ethylamine and ethanolamine. Storage time during shelf-life
presented a significant effect on the levels of the five amines. On the contrary, storage
temperature did not presented a significant effect on the levels of the mentioned amines. Min
et al.2 (2004) state that egg yolk had more BAs than egg white, however it is unknown which
BAs were studied.

Occurrence of BAs in the milk is low, about 1 mg/dm3, but in cheese their content can
achieve 1 g/dm3. Cheese contains proteins, enzymes, cofactors, water, salt, and bacteria,
and therefore represents an ideal environment for BAs production from free amino acids by
decarboxylating enzymes of micro organisms during cheese ripening. Large amounts in
cheese could indicate a failure, from a hygienic point of view, in the milk used for cheese
products or during the cheese making (Karoviov and Kohajdov, 2005).
Sumner et al. (1990) studied the effect of heating raw milk, containing the histamine-
producing isolate of L. buchneri, used in producing Swiss cheese. The L. buchneri survived
heating at 49 to 80C for 10 minutes, suggesting that this organism would survive the normal
heating process applied to raw milk used prior to making Swiss cheese.

Humans
Exposure
A mean daily intake by humans of 18.7, 12.6 and 11.0 mg of putrescine, spermidine and
spermine, respectively, was reported for the United Kingdom, Italy, Spain, Finland, Sweden
and the Netherlands. Reported mean daily intake by humans in Japan was 9.9, 12.0 and 7.9
mg and in the USA 14.0, 7.9 and 7.2 mg for putrescine, spermidine and spermine,
respectively (Kala, 2009).
No data was found concerning cadaverine and histamine.

Absorption
Kala (2009) studied the biological role of dietary polyamines in humans. The gastrointestinal
tract can represent a significant source of polyamines originating from intestinal bacteria,
sloughed cells and pancreatic bile and intestinal secretions. Considerable polyamine levels
were observed in the lumen of human gut during the fasting state, which suggests
endogenous secretion. A significantly higher content was determined in the jejunum than in
the ileum, which suggests proximal absorption.
Besides endogenous BAs, humans ingest BAs via food. Normally, during the food intake of
small amounts of BAs, BAs are metabolized to physiologically less active degradation
products. However, upon intake of high loads of biogenic amines with foods, this
detoxification system is unable to eliminate biogenic amines sufficiently (Bodmer et al., 1999)
and absorption will occur.

Distribution
Almost all mammalian tissues contain histamine in amounts ranging from less than 1g up to
100 g/g tissue. Concentrations in plasma and other body fluids are generally very low, but
human cerebrospinal fluid contains significant amounts. The mast cell is the predominant
storage site form histamine in most tissues, especially in the skin, the mucosa of the
bronchial tree and the intestinal mucosa. Mast cells in human cardiac tissue are also
reported to contain high levels of histamine (Rambali et al., 2002).
No data was found concerning the routes of excretion of other BAs.

Metabolism

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 In the intestinal tract of mammals affects a detoxification system which is capable of
metabolizing normal dietary intake of BAs. Under normal conditions in humans exogenous
amines absorbed from food are rapidly detoxified by the action of amine oxidase or
conjugation, but in the case of allergic individuals or if monoamine inhibitors are applied or
when too high levels are consumed the detoxification process is disturbed and BAs
accumulate in the body. The enzymes monoamino oxidase (MAO) and diamino oxidase
(DAO) play an important role in detoxification process. MAO and DAO occur in the gut
epithel and thus oxidation products of BAs are getting into the blood circulation. Polyamines
are usually in the first place acetylated and consequently oxidized by DAO or polyamino
oxidases (Karoviov and Kohajdov, 2005).

Excretion
The human kidney has a considerable capacity for removing histamine from blood. When
healthy individuals were infused intravenously with histamine, a large proportion was
methylated by the kidney and excreted in the urine and a smaller proportion was excreted
unchanged in the urine (Karoviov and Kohajdov, 2005).
No data was found concerning the routes of excretion of other BAs.

3. Diagnose of poisoning

Animals
Diagnosing biogenic amine poisoning should be based upon a combination of history
(possible dietary exposure), symptoms and chemical analysis of feed / food and body tissues
/ fluids or animal product (e.g. eggs). BAs can be analysed in several types of samples, e.g.
in blood, urine, organs, muscles, eggs. Animal feed can also be analysed for BAs, TVN or
ammonia.

Humans
Food can be analysed for BAs, TVN or ammonia. No other data found.

4. Potential adverse effects

Environment
No data found.

Animals
Ruminants potentially receive BAs from both dietary and ruminal microbial sources and thus
have the potential to absorb greater amounts than other species. BAs have been implicated
as being responsible for the depressed dry matter intake of dairy cattle fed silage. BAs have
been implicated as causative factors in ketonemia / acetonemia (Phuntsok et al., 1998).
However Lingaas and Tveit (1992) state that these may be a contributory factor in causing
ketonemia. So the role, either causative or contributory, is not uniformly established and will
therefore in this fact sheet not be included (yet) as an adverse effect caused by BAs.
Putrescine has been shown to decrease feed intake and milk production of dairy cows
(Phuntsok et al., 1998).
In poultry BAs have been implicated in a malabsorption syndrome characterized by
decreases in feed efficiency and enlargement of the proventriculus (Barnes et al., 2001).
Also gizzard erosion has been observed in poultry. This erosion is caused by gizzerosine, a
thermal decomposition product of histamine and lysine. Gizzerosine has been reported to be
a much more potent stimulus of gastric acid secretion in the chicken than histamine (Smith et
al., 2000).
Histamine can promote acid secretion in the stomach and cause ulcers in swine (Smith et
al., 2000).

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 In an experiment by Til et al. (1997) studying the subacute toxicity of BAs in rats, neural
effects were noted, like aggressiveness, convulsions and paralysis of the hind legs. Impaired
kidney function occurred with spermine.
In Appendix I the potential adverse effects of BAs in animals are shown. For toxicity data,
see Appendix II.

Humans
BAs do not usually represent health hazards to individuals unless excessive amounts are
ingested, or the natural mechanism for their catabolism is deficient or impaired.
All humans are susceptible to scombroid poisoning; however, the symptoms can be severe
for the elderly and for those taking medications (FDA2, 2009) with inhibiting effect to MAO
and DAO such as antihistamines, antimalaria agents, psychopharmaceutics, might have a
changed metabolism of BAs. Some amines, especially putrescine and cadaverine, inhibit
histamine detoxifying enzymes and thus act as potentiators of histamine toxicity. These
amines in the intestinal tract preferably react with MAO and DAO that tend to increase the
level of histamine in blood. Also, injuries of intestinal mucosa can reduce the function of
biogenic amine detoxification enzymes. Upon intake of high loads of BAs in food, the
detoxification system in the human body is unable to eliminate BAs sufficiently. Furthermore
people with gastrointestinal problems are also at risk because the activity of oxidases in their
intestines is usually lower than that in healthy individuals (Karoviov and Kohajdov, 2005).

The International Agency for Research on Cancer has not studied the carcinogenicity of BAs
as a group or of specific BAs. Polyamines, participate in cell growth and proliferation and as
a consequence in tumour growth. They accumulate in cancerous tissues and their content is
elevated in the body fluids of cancer patients (Kala, 2009). BAs are also potential
precursors for the formation of carcinogenic N-nitroso compounds (Karoviov and
Kohajdov, 2005).

Histamine exerts its effects by binding to receptors on cellular membranes in the respiratory,
cardiovascular, gastrointestinal, and haematological/immunological systems and the skin
(Karoviov and Kohajdov, 2005). Possibly also neurological effect occur. However the
duration of exposure to exhibit these effects is not known. These effects are taken into
account in this fact sheet.
Scromboid poisoning (allergic reaction) is also a well known adverse effect of histamine.
However since this is an acute effect, this effect will not be addressed to in this fact sheet.

Hui (2006) states that spermine can cause kidney toxicity and can affect blood coagulation
and pressure, heat beat and respiration.

No data concerning specific adverse effect of other BAs were found.

In Appendix I the potential adverse effects of BAs in humans are shown. For toxicity data,
see Appendix II.

5. Severity of the potential adverse effects

The severity of contamination of food and / or feed materials with BAs is based on the worst
case scenario, as shown in table 6, and is based upon the potential adverse effects stated in
chapter 4 and Appendix I.

The severity of BA toxicosis in animals is classified as medium because the observed

effects in literature are not classified as high for animals.

The severity of PA toxicosis in humans is classified as high because:

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 BAs has effect on cellular membranes in the respiratory, cardiovascular, gastrointestinal,
and haematological/immunological systems and the skin (Karoviov and Kohajdov,
2005);
Spermine causes kidney toxicity (Hui, 2006).

Table 6. Severity of BAs

Severity
Low Medium High
Animals x
Humans x

6. Legislation and standards

No data found.

7. Method of analysis
For determination of BAs numbers of analytical methods are developed. The complex matrix
sample, the presence of potentially interfering compounds, and the occurrence of several
BAs simultaneously are typical problems encountered in the analysis (Karoviov and
Kohajdov, 2005).
Preclean-up includes extraction of sample with suitable extracting reagent. The analytical
methods used for separation and quantification of BAs are mainly based on chromatographic
methods: gas chromatography (GC), thinlayer chromatography (TLC), and high-performance
liquid chromatography (HPLC) with precolumn or postcolumn derivatization techniques
(Karoviov and Kohajdov, 2005).
The ELISA technique has been stated as a suitable detection method for the determination
of histamine in cheese by Aygn et al. (1999).
Determination of the Total Volatile Nitrogen (TVN) content can also be used to assess
freshness of fish meal (Ricque-Marie et al., 1998). Determination of ammonia can also be
used as an indicator for freshness (FAO2, 2001).

8. Possible control measures

Products with a higher risk to contain hazardous levels or exceeding maximum limits of
biogenic amines, are:
Fermented products, or by-products originating from fermentation processes and
high nitrogen containing products, like:
o Products of vegetal origin and products derived from these products, like maize,
alfalfa or grass silages, soy products, products originating from beer
brewing or originating from other processes producing alcoholic
beverages;
o Products of animal origin and products derived from these products, like fish
meal, cheese;
o Product containing biogenic amines by nature (not by fermentation), like:
o Products of vegetal origin, like mushrooms;

Cultivation:
Conditions should be unfavourable for microbiological contamination and BA production
(see storage and transport).

Storage and transport:

Conditions should be unfavourable for microbiological contamination and BA production:

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 o Feed materials should be stored and transported dry, closed and / or covered;
o Temperature during storage and transport should be unfavourable for biogenic
amine generation. Include processes starting from e.g. fishing at sea, since
during storage conditions at the boat BAs can be generated;
o Avoid deviations in temperature, causing condensation;
o Feed materials should be stored hygienically: controlling the presence of BA
producing micro-organisms.
Preferably no use of BA-based baits in traps, like putrescine based baits. In case of use
of BA-based baits, contact with feed an food material is not possible. The latter should be
verifiable.

Processing feed or food material:

Use the HACCP system to assess the risk of BAs in the feed material.
Processing conditions should be unfavourable for microbiological contamination and BA
production (see storage and transport).
Implement a BAs monitoring program, or analyse for TVN or ammonia, on feed materials.
Also pay special attention to feed material of which a long storage period is known /
suspected;
Preferably no use of BA-based baits in traps, like putrescine, in the processing plant. In
case of use of BA-based baits, contact with feed an food material is not possible. The
latter should be verifiable.

Processing animal feed:

Processing conditions should be unfavourable for microbiological contamination and BA
production(see storage and transport).
Preferably no use of BA-based baits in traps, like putrescine, in the processing plant. In
case of use of BA-based baits, contact with feed an food material is not possible. The
latter should be verifiable.
Implement a BAs monitoring program, or analyse for TVN or ammonia, on feed materials
of risk. Also pay special attention to feed material of which a long storage period is known
/ suspected.
Implement an BAs monitoring program, or analyse for TVN or ammonia, on animal feed,
e.g. high nitrogen containing animal feed or animal feed containing risk products as
mentioned above.

9. References

1 Anli et al., Biogenic Amine Content of Beers Consumed in Turkey and Influence of
Storage Conditions on Biogenic Amine Formation, J. Inst. Brew. 112(3), 2006,
pages 267274
2 Aygn et al., Comparison of ELISA and HPLC for the Determination of Histamine in
Cheese, J. Agric. Food Chem., Volume 47 (5), 1999, pages 1961 1964
3 Barnes et al., Effects of Biogenic Amines on Growth and the Incidence of
Proventricular Lesions in Broiler Chickens, Poultry Science 80, 2001, pages 906 -
911
4 Bodmer et al., Biogenic amines in foods: Histamine and food processing,
Inflammation Research, Volume 48, 1999, pages 296-300
5 Brinker et al., Biogenic amines in fish and fish products, 2002
6 Cuevas et al., Putrescine Is Involved in Arabidopsis Freezing Tolerance and Cold
Acclimation by Regulating Abscisic Acid Levels in Response to Low Temperature,
Journal of Plant Physiology, Vol. 148, 2008, pages 1094 1105
7 Dadkov et al., Content of biogenic amines and polyamines in some species of
European wild-growing edible mushrooms, European food research and

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 technology, Volume 32, No. 1, 2009, pages 163-171
8 FAO1, Assessment and Management of Seafood Safety and Quality, FAO
Fisheries Technical Paper 444, 2003
9 FAO2, Non-Sensory Assessment of Fish Quality, 2001
10 FDA1, Seafood Chemistry, ORA Laboratory Manual, Volume IV, Section 9, 2004
11 FDA2, Bad Bug Book: Food borne Pathogenic Micro organisms and Natural Toxins
Handbook: Scombrotoxin, 2009
12 Hui (ed.), Handbook of Food Science, Technology and Engineering, Chapter 13,
Volume 1, 2006
13 Kala, Recent advances in the research on biological roles of dietary
polyamines in man, Journal of Applied Biomedicine, Volume 7, 2009, pages 65 - 74
14 Karoviov and Kohajdov, Biogenic amines in food, Chem. Pap. 59 (1), 2005,
pages 70 - 79
15 Kse et al., Changes in the levels of histamine during processing and storage of
fish meal, Animal Feed Science and Technology 107, 2003, pages 161 172
16 Krausov et al., Content of biologically active polyamines in livers of cattle, pigs
and chickens after animal slaughter , Meat Science, Volume 73, Issue 4, 2006,
pages 640 - 644
17 Lawley et al., Food Safety Hazard Guidebook, 2008
18 Lebiedziska et al., Differences in biogenic amine patterns in fish obtained from
commercial sources, Zeitschrift fr Lebensmitteluntersuchung und -Forschung A,
Volume 192, Number 3, 1991, pages 240 - 243
19 Lingaas and Tveit, Etiology of Acetonemia of Butyric Acid, Valeric in Norwegian
Cattle. 2. Effect Acid, and Putrescine, Journal of Dairy Science, Volume 75, 1992,
pages 2433-2439
20 Kahn (ed.), The Merck Veterinary Manual, Ninth edition, 2005
21 Min et al.1, Control of Micro-organisms and Reduction of Biogenic Amines in
Chicken Breast and Thigh by Irradiation and Organic Acids, Poultry Science,
Volume 86, 2007, pages 2034-2041
22 Min et al.2, Quantitative analysis of biogenic amines in raw and processed foods of
animal origin on Korean domestic market, Asian-australasian journal of animal
sciences , Volume 17, Number 12, 2004, pages 1764 - 1768
23 O'Neil (ed.) et al., The Merck index: An encyclopedia of chemicals, drugs, and
biologicals, fourteenth edition, 2006
24 Paulsen et al., Biogenic amines and polyamines in liver, kidney and spleen of roe
deer and European brown hare, European Food Research and Technology,
Volume 227, Number 1, 2008, pages 209 - 213
25 Phuntsok et al., Biogenic Amines in Silage, Apparent Postruminal Passage,
and the Relationship Between Biogenic Amines and Digestive Function and Intake
by Steers, Journal of Dairy Science, Volume 81, 1998, pages 21932203
26 Radosevich, Raw ingredients freshness: new perspectives on biogenic amines,
2009
27 Rambali et al., The contribution of cocoa additive to cigarette smoking addiction,
RIVM report 650270002/2002, 2002
28 Ramos et al., Changes of yolk biogenic amine concentrations during storage of
shell hen eggs, Food Chemistry, Volume 116, Issue 1, 2009, pages 340 - 344
29 Ricque-Marie et al., Raw material freshness, a quality criterion for fish meal fed to
shrimp, Aquaculture, Volume 165, Issues 1-2, 1998, pages 95 - 109
30 RIVM-RIKILT, Beoordeling inzake biogene amines in kaas, 2009
31 Santos, Biogenic amines: their importance in Foods, International Journal of Food
Microbiology, Volume 29, Issue 2 -3 , 1996, pages 213 - 231
32 Shakila and Vasundhara, Formation of Histamine and Other Biogenic Amines
During Storage of Freshwater Fish Chunks, Asian Fisheries Science 15, 2002,
pages 1 - 6
33 Shalaby, Significance of biogenic amines to food safety and human health, Food

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 Research International, Volume 29, Issue 7, 1996, Pages 675-690
34 Smith et al., Feed-borne biogenic amines: Natural toxicants or growth promotors?,
Avances en Nutricin Acucola V. Memorias del V Simposium Internacional de
Nutricin Acucola. 19-22 Noviembre, 2000, pages 24 - 32
35 Stratton et al., Biogenic amines in cheese and other fermented foods: a review,
Journal of food protection (USA), Volume 54(6), 1991, pages 460 - 470
36 Sumner et al., Factors Controlling Histamine Production in Swiss Cheese
Inoculated with Lactobacillus buchner, Journal of Dairy Science 73, 1990, pages
3050 - 3058
37 Til et al., Acute and subacute toxicity of tyramine, spermidine, spermine, putrescine
and cadaverine in rats, Food and Chemical Toxicology, Volume 35, Issues 3 4,
1997, pages 337-348
38 Van Os et al., Formation of biogenic amines in well fermented grass silages,
Journal of Agricultural Science , Volume 127, 1996, pages 97 - 107
39 Vinci and Antonelli, Biogenic amines: quality index of freshness in red and white
meat, Food Control, Volume 13, Issue 8, 2002, pages 519 - 524
40 VWA, Histamine, Kennisbank Voedselveiligheid, 2008

10. Websites

1 http://www.gmpplus.org
2 http://www.fda.gov/Food/FoodSafety/FoodborneIllness/FoodborneIllnessFoodborn
ePathogensNaturalToxins/BadBugBook/ucm070823.htm
3 http://www.fao.org/DOCREP/006/Y4743E/y4743e00.htm#Contents

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 APPENDIX I Potential adverse effect of biogenic amines

1,2 1,2 1,2 1,2 2

biogenic Death Carcinogen Teratogen Mutagen Internal injury Neurological Immunological Reproductive
2 2
amines (physical effect effects
contamination)
1,2

Animals x

Humans x x

biogenic Effects on Dermal and Respiratory Musculo- Cardiovascular Gastrointestin Hematological Endocrine Body weight
2 2 2 2 2 2 2
amines organs ocular effect effect skeletal effect effect al effect effect effect effect
Animals x x

Humans x x x x x x

1
This potential adverse effect is classified as high severity for animals
2
This potential adverse effect is classified as high severity for humans

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 APPENDIX II Toxicity data several biogenic amines

NOEL (mg/kg BW) ADI (mg/kg BW) LD50 (mg/kg BW)

Cadaverine 180 (Til et al., 1997)1
Histamine 220 (mouse) (Rambali et
al., 2002)
1
Putrescine 180 (Til et al., 1997) TDI=0.6 (RIVM-RIKILT, 740A (RIVM-RIKILT,
2009) 2009)
Spermidine 83 (Til et al., 1997)1
Spermidine trichloride Estimated: 600 (rat)
(RIVM-RIKILT, 2009)
Spermine 19 (Til et al., 1997)1
Spermine Estimated: 600 (rat)
tetrahydrochloride (RIVM-RIKILT, 2009)
1
= during 6 weeks, not known for which effect NOEL was determine
A
= species tested unknown

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