Biogenic amines in foods and their role in food safety and public health

1. M.Venkatasami,
M.Tech (Processing and Food Engineering)
Department of Food Process Engineering
AEC&RI, TNAU.
1BIOGENIC AMINES

2. CONTENTS
Introduction
Formation and degradation of biogenic amines
Occurrence in food
Significance in food safety
Quality indicators
Methods of determination
2BIOGENIC AMINES

3. INTRODUCTION
Biogenic amines are,
Basic nitrogenous compounds with low molecular weight
Indispensable components of living cells
Important in physiological and biological activities
Causative agents of food poisoning and various physiological reactions
Histamine - associated mainly with fish consumption
Tyramine - initiator of hypertensive crisis
3BIOGENIC AMINES

4. Cont’d…
Toxicity enhancers of histamine - putrescine, cadaverine, tyramine
Reacts with nitrites to form carcinogenic nitrosamines.
Quality indicators of food spoilage
Biogenic amines formation is due to,
Enzymic activity
 Microbial decarboxylation of amino acids
Amination and transamination of aldehydes and ketones
4
Other amines:
Putrescine, cadaverine, tyramine, tryptamine, phenylethylamine, spermine, spermidine, and agmatine
BIOGENIC AMINES

5. FORMATION OF BIOGENIC AMINES
Decarboxylation of free amino acids
Decarboxylation Removal of the α-carboxyl group Amines
5BIOGENIC AMINES

6. Cont’d…
6
Name Precursors
 Histamine  Histidine
 Tyramine  Tyrosine
 Phenylethylamine  Phenylalanine
 Tryptamine  Tryptophan
 Putrescine  Ornithine, arginine, agmatine
 Cadaverine  Lysine
 Spermidine  Putrescine, spermine
 Spermine  Putrescine, spermidine
 Agmatine  Arginine
 Important biogenic amines in foods
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7. Decarboxylation enzymes
7
Enzyme Amine
 Histidine decarboxylase  Histamine
 Arginase  Ornithine
 Ornithine decarboxylase  Putrescine
 Lysine decarboxylase  Cadaverine
 Arginine decarboxylase  Agmatine
 Agmatinase  Putrescine
 Spermine synthase  Spermidine and spermine
Cont’d…
BIOGENIC AMINES

8. 8
The two mechanism on the formation of biogenic amines based on
Pyridoxal-5- phosphate
Part of the enzyme that catalyzes the decarboxylation reaction
Active site of the decarboxylation enzyme is formed by pyridoxal phosphate
It joined in a schiff base linkage to the amino group of a lysyl residue.
Pyruvoyl residue
Covalently bound to the amino group on the enzyme
Acts in a similar way to pyridoxal phosphate in the decarboxylation reaction
Carbonyl group of pyridoxal phosphate reacts with amino acids to form Schiff base intermediates,
which are then decarboxylated to yield the corresponding amines
TWO SPECIFIC MECHANISMS
BIOGENIC AMINES

9. Decarboxylase enzyme present in,
 The genera Bacillus, Pseudomonas and Photobacterium
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MICROORGANISMS RESPONSIBLE FOR AMINE FORMATION
 Lactic acid bacteria (LAB) including Lactobacillus, Enterococcus, Carnobacterium,
Pediococcus, Lactococcus, and Leuconostoc are able to decarboxylate amino acids
Enterobacteriaceae Micrococcaceae
 Citrobacter
 Klebsiella
 Escherichia
 Proteus,
 Salmonella
 Shigella
 Staphylococcus
 Micrococcus
 Kocuria
BIOGENIC AMINES

10. HISTAMINE-PRODUCING MICROORGANISMS
Products Microorganisms
 Fish
Morganella (proteus) morganii, Klebsiella pneumonia, Hafnia alvei, Proteus vulgaris, Proteus
mirabilis, Clostridium perfringens, Enterobacter aerogenes, Acinetobacter lwoffi , Pseudomonas
putrefaciens, Aeromonas hydrophila, Plesiomonas shigelloides, and Vibrio spp.
 Halotolerant
histamine-forming
strains in fish
Staphylococcus, Vibrio, and Pseudomonas
 Meat products
Enterobacter cloacae, E. aerogenes, and Klebsiella oxytoca, Escherichia coli, Morganella
(proteus) morganii, Lactobacillus bulgaricus and Lactobacillus acidophilus
 Dairy products
Streptococcus lactis, lactobacillus helveticus, Streptococcus faecium, Streptococcus mitis,
lactobacillus bulgaricus, Lactobacillus plantarum, and Propionibacteria
 Swiss cheese &
 Gouda cheese
Lactobacillus buchneri
 Beer Lactobacillus spp. and Pediococcus spp.
 Wine Pediococcus spp. and Oenococcus oeni
10BIOGENIC AMINES

11. Cont’d…
Tyramine producing microorganisms
Putrescine and cadaverine-producing microorganisms
Pseudomonads - putrescine (horse mackerel, mahimahi)
Cadaverine - Enterobacteriaceae (beef, pork, lamb, and minced beef)
Fermented meat products- Staphylococcus carnosus and Staphylococcus piscifermentans
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Products Microorganisms
 Meat products L. Divergens and L. Carnis
 Dry sausage Carnobacterium, L. Curvatus, and L. Plantarum,
 Miso Enterococcus faecium and Lactobacillus bulgaricus
 Beer Pediococcus spp., mainly P. damnosus
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12. DEGRADATION OF BIOGENIC AMINES
Physiologically inactivated by amine oxidases present in bacterium, fungus, plant, and animal cells
Amine oxidases can able to catalyze the oxidative deamination of amines with the production of
aldehydes, hydrogen peroxide, and ammonia
Microbial strains in amine degradation belongs to the genera Lactobacillus, Pediococcus, and
Micrococcus, as well as to the species Staphylococcus carnosus and Brevibacterium linens.
S. Xylosus in fermented sausages showed significant reduction in the concentration of tyramine and
putrescine
Histaminase activity has been detected with several types of bacteria, including M. morganii,
Vibrio spp., and Klebsiella spp.
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13. Tyramine degradation is dependent on pH (with an optimum at 7.0), temperature, and NaCl, as
well as glucose, oxygen availability, and hydralazine concentration.
Bacterial histaminase is best produced under alkaline conditions (pH 7.5 to 8) but that moderate
activity occurs under slightly acidic conditions.
The highest degradation rate of this amine was observed at 37°C, ( at 22 and 15°C the
degradation was considerable.)
Histaminase activity has its optimum temperature at 37°C and retains about 50% of its maximum
activity at 20°C.
13
Cont’d…
BIOGENIC AMINES

14. OCCURRENCE OF BIOGENIC AMINES IN FOODS
Level of biogenic amines in foods depends on
 Type of food
Quality of the raw material
Type and extent of contamination
Processing, Distribution and Storage condition
Biogenic amines are mainly present in,
Fish products, meat products, dairy products, wine, beer, vegetables, fruits, nuts, and
chocolate
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BIOGENIC AMINES
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15. SIGNIFICANCE OF BIOGENIC AMINES IN FOOD SAFETY
Important physiological effects are either psychoactive or vasoactive
Psychoactive amines
Affects nervous system (acting on neural transmitters)
Vasoactive amines
Affects vascular system
Detoxification system in the intestinal tract prevents amines from entering into the bloodstream
Detoxification systems are,
Monoamine oxidase (MAO)
Diamine oxidase (DAO)
Polyamine oxidase (PAO)
 Respiratory and coronary problems or those with hypertension or vitamin B12 deficiency are sensitive to
lower doses of biogenic amines
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16. HISTAMINE TOXICITY
Exerts toxicity by binding to three types of receptors, H1, H2, and H3, Cellular membranes in
the respiratory, cardiovascular, gastrointestinal, and hematological/immunological systems.
Causes contraction of the smooth muscle of the uterus, the intestine, and the respiratory tract
(regulated by the H1 receptor)
Controls gastric acid secretion (regulated by the H2 receptor located on the parietal cells);
Affects liberation of adrenaline and noradrenaline from the suprarenal gland; stimulates both
sensory and motor neurons; and causes dilatation of the peripheral blood vessels.
The most significant food-borne disease associated with histamine is Histamine (or scombroid)
fish poisoning (HFP)
16BIOGENIC AMINES

17. Cont’d…
Causes of HFP are eating fish of the families scombridae and scomberesocideae, including tuna,
saury, bonito, seer-fish and butterfly kingfish.
Common symptoms of histamine poisoning are limited to rash, diarrhea, flushing, sweating and
headache
Guideline levels for the histamine content of fish:
<5 mg/100 g (safe for consumption)
5 to 20 mg/100 g (possibly toxic)
20 to 100 mg/100 g (probably toxic)
>100 mg/100 g (toxic and unsafe for human consumption)
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18. TYRAMINE TOXICITY
Vasoactive amines
 Physiological effects are,
Peripheral vasoconstriction
Increased cardiac output
Increased respiration
Elevated blood glucose
Release of norepinephrine
Reacts with MAO inhibitor (MAOI) drugs, giving rise to hypertensive crisis.
The dose of tyramine required for acute oral toxicity was more than 2,000 mg/kg body weight
The no-observed-adverse-effect level of tyramine was 180 mg/kg body weight/day.
18BIOGENIC AMINES
Tyramine

19. TOXICITY OF SPERMIDINE, SPERMINE,
PUTRESCINE, AND CADAVERINE
Spermine was the most toxic, leading to emaciation, aggressiveness, convulsions, paralysis of the hind legs,
slight anemia, and changes in plasma clinical chemistry
Slight increases in packed cell volume, hemoglobin concentration, and thrombocytes occurred with
cadaverine.
The no observed- adverse-effect level was,
2,000 ppm (180 mg/kg body weight/day) of cadaverine and putrescine
1,000 ppm (83 mg/kg body weight/day) of spermidine
200 ppm (19 mg/kg body weight/day) of spermine
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20. BIOGENIC AMINES AS SPOILAGE
INDICATORS OF FOODS
Potential indicators of food spoilage
The quality of canned tuna monitored by the biogenic amine-based index (BAI)
BAI comprises levels of histamine, putrescine, cadaverine, spermidine,and spermine
An index value of fish acceptability is 10
BAI limit of acceptability established for tyramine included BAI limit is in the range of
15 to 20 μg/g for Mediterranean hake
Useful indicator for the freshness of common squid is agmatine
20BIOGENIC AMINES

21. Cont’d…
The most effective spoilage index of sardine and saury pike is cadaverine
Cadaverine and putrescine could be used to assess freshness of chilled-stored rainbow trout (Salmo irideus)
BAI value (sum of putrescine, cadaverine, histamine, and tyramine) for bologna sausage, minced beef and pork is
500 mg/kg
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Cadaverine value, mg/100g Indication
 15  Good quality
 15 and 20  Initial decomposition
 > 20  Advanced decomposition
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22. Cont’d…
BAI value (sum of putrescine, cadaverine, histamine, and tyramine) for fresh pork are,
22
BAI value, mg/kg Indication
 <5  Good quality
 5 and 20  Acceptable quality
 20 and 50  Low meat quality
 >50  Spoiled meat
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23. DETERMINATION OF BIOGENIC AMINES IN FOODS
The determination of biogenic amines in foods based on,
Thin-layer chromatography (TLC)
Liquid chromatography
Gas chromatography
Biochemical assays
Capillary electrophoresis
Crucial step in analytical procedure: Extraction of amines from the samples.
Solvents used: hydrochloric acid, trichloroacetic acid, perchloric acid, methanesulfonic
acid, petroleum ether and other organic solvents
23BIOGENIC AMINES

24. Cont’d…
The simplest method: TLC
Disadvantages:
Excessive time needed for analysis
Semi-quantitative nature of the obtained results.
The most frequently reported technique: High-pressure liquid chromatography (HPLC)
Advantage: Simultaneous separation and quantification of multiple biogenic amines in foods
Gas chromatography (GC) for the analysis of biogenic amines is not very common
Methods based on GC alone or in combination with mass spectrometry have been developed for the
determination of histamine and other amines
BIOGENIC AMINES 24

25. Capillary electrophoresis (CE) is another method which has been used widely in recent years
Advantage: High sensitivity and short analysis time.
Biogenic amines were separated in less than 9 min by CE compared to the 20 min achieved by
HPLC
CE methods used for the determination of biogenic amines are based on,
 Indirect UV
Lamp-induced fluorescence
Laser-induced fluorescence
Conductometric
Pulsed amperometric detection
BIOGENIC AMINES 25
Cont’d…

26. Cont’d…
Very promising techniques for histamine detection: enzymaticmethods, such as radioimmunoassays
and the enzyme-linked immunosorbent assay system
Advantage: Rapid and not requiring expensive instrumentation
Analytical methods for the determination of biogenic amines require a derivatization procedure (low
volatility and lack of chromophores of most biogenic amines)
Chemical reagents used in derivatization procedure:
 Dansyl and dabsyl chloride
 Benzoyl chloride
Fluoresceine
9-fluorenylmethyl chloroformate
O-phthalaldehyde (OPA)
Naphthalene-2, 3-dicarboxaldehyde
BIOGENIC AMINES 26

27. Juneja, VK, and JN Sofos. 2010. Pathogens and toxins in foods: ASM Press.
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REFERENCE
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