Upload
vuanh
View
237
Download
1
Embed Size (px)
Citation preview
ANALYSIS OF BIOGENIC AMINES IN FISH BY USING
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
NUR IDATUL DIANA BT AHMAD ZAKI (35120)
Bachelor of Science with Honours
(Resource Chemistry)
2015
ANALYSIS OF BIOGENIC AMINES IN FISH BY USING
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
NUR IDATUL DIANA BT AHMAD ZAKI
A Final Year Project Report is submitted in partial fulfillment of the requirements for the
award of the degree of Bachelor of Science with Honours
(Resource Chemistry)
Faculty of Resource Science and Technology
UNIVERSITI MALAYSIA SARAWAK
2015
i
ACKNOWLEDGEMENT
Firstly I would to express my special thanks to my research supervisor Professor Dr
Zaini Bin Asim for his guidance in helping me to finish my research project and this degree
thesis. Besides, the useful critiques, valuable suggestion during the development of this
research project and also willingness to spend his time has been much appreciated. I also
would like to dedicate special thanks to Mr Leo Bulin Unting, Science Officer Faculty
Resource Science and Technology whose guide and educating me in the usage of High
Performance Liquid Chromatography (HPLC), to my family, senior and my friends that give
moral support to me for completing this project and report.
ii
UNIVERSITI MALAYSIA SARAWAK
Grade:____________
Please tick (√)
Final Year Project Report
Masters
PhD
DECLARATION OF ORIGINAL WORK
This declaration is made on the ……………..day of……………..2015.
Student’s Declaration:
I NUR IDATUL DIANA BT AHMAD ZAKI, 35120, FACULTY OF RESOURCE SCIENCE
AND TECHNOLOGY hereby declare that the work entitled, ANALYSIS OF BIOGENIC
AMINES IN FISH BY USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY is
my original work. I have not copied from any other students’ work or from any other sources
except where due reference or acknowledgement is made explicitly in the text, nor has any
part been written for me by another person.
Date submitted NUR IDATUL DIANA BT AHMAD ZAKI (35120)
Supervisor’s Declaration:
I, PROF. DR.ZAINI BIN ASSIM, hereby certify that the work entitled, ANALYSIS OF
BIOGENIC AMINES IN FISH BY USING HIGH PERFORMANCE LIQUID
CHROMATOGRAPHY was prepared by the above named student, and was submitted to the
“FACULTY OF RESOURCES SCIENCE AND TECHNOLOGY as a full fulfillment for the
conferment of BACHELOR DEGREE OF SCIENE (CHEMISTRY) WITH HONORS, and
the aforementioned work, to the best of my knowledge, is the said student’s work.
Received for examination by: _____________________ Date:___________________
PROF. DR. ZAINI BIN ASSIM
iii
I declare this Project/Thesis is classified as (Please tick (√)):
CONFIDENTIAL(Contains confidential information under the Official Secret Act 1972)*
RESTRICTED (Contains restricted information as specified by the organisation where research was done)*
OPEN ACCESS
Validation of Project/Thesis
I therefore duly affirmed with free consent and willingness declared that this said
Project/Thesis shall be placed officially in the Centre for Academic Information Services with
the abide interest and rights as follows:
This Project/Thesis is the sole legal property of Universiti Malaysia Sarawak
(UNIMAS).
The Centre for Academic Information Services has the lawful right to make copies for the purpose of academic and research only and not for other purpose.
The Centre for Academic Information Services has the lawful right to digitise the
content to for the Local Content Database.
The Centre for Academic Information Services has the lawful right to make copies of the Project/Thesis for academic exchange between Higher Learning Institute.
No dispute or any claim shall arise from the student himself / herself neither third party
on this Project/Thesis once it becomes sole property of UNIMAS.
This Project/Thesis or any material, data and information related to it shall not be
distributed, published or disclosed to any party by the student except with UNIMAS
permission.
Student’s signature ___________________ Supervisor’s signature: _____________ ( /5/2015 ) ( /5/2015)
Current Address:
21 JALAN SAGA 2 TAMAN SAGA 28400 MENTAKAB PAHANG Notes: * If the Project/Thesis is CONFIDENTIAL or RESTRICTED, please attach
together as annexure a letter from the organisation with the period and reasons of
confidentiality and restriction.
[The instrument was duly prepared by The Centre for Academic Information Services]
iv
TABLE OF CONTENTS
Content Page
ACKNOWLEDGEMENT i
DECLARATION ii-iii
TABLE OF CONTENT iv-vi
LIST OF ABBREVIATIONS vii
LIST OF TABLES viii
LIST OF FIGURES ix
ABSTRACT x
ABSTRAK Xi
1. INTRODUCTION
1.1 General Introduction 1-2
1.2 Problem Statement 3
1.3 Objectives of the Project 3
2. LITERATURE REVIEW
2.1 Biogenic Amine 4-5
2.2 Hazard Characteristics
2.2.1 Histamine 6
2.2.2 Cadaverine and Putriscine 6-7
2.3 Microorganism Involved in Biogenic Amine Production 7
2.4 Condition Supporting the Formation of Biogenic Amines
v
2.4.1 Substrate availability 7-8
2.4.2 Temperature 8
2.4.3 Effect of pH 8
2.4.4 Others factors 9
2.5 Toxicology Aspects of Biogenic Amine in Response of Human
2.5.1 Histamine 9-10
2.5.2 Cadaverine and Putrescine 10
2.5.3 Tyramine and Phenylethylamine and Tryptamine 10-11
2.6 Biogenic Amine in Fish 11-12
2.7 Biogenic Amine as Quality Index in Meat and Meat Products 12-13
2.8 Analytical Methods for Biogenic Amine 13-14
3. MATERIALS AND METHODS
3.1 Chemical and Reagent 15
3.2 Chromatographic Condition 15
3.3 Application of Validated Method for Biogenic Amines in Fish
3.3.1 Sample of Raw Fish 16
3.3.2 Analysis of Biogenic Amines in Fish 16
3.4 Method Validation for Analysis of Biogenic Amines
3.4.1 Preparation of Biogenic Amine Standards 16
3.4.2 HPLC Analysis of Biogenic Amine Standards 17
3.4.3 Validation Parameters 17
vi
3.5 Statistical Analysis using One Way ANOVA 18
4. RESULTS AND DISCUSSION
4.1 Method Validation 19-23
4.2 Linearity and Working Range 24-27
4.3 Limit of Detection(LOD) and Limit of Quantification (LOQ) 27-28
4.4 Accuracy
4.4.1 Repeatability Precision 29-30
4.4.2 Reproducibility Precision 31-32
4.5 Precision 33-35
5. CONCLUSION 36
REFERENCES 37-44
APPENDICES 45-47
vii
LIST OF ABBREVIATIONS
Cad Cadaverine
CE Capillary Electrophoresis
DAO Diamine oxides
HMT Histamine-N-methyltransferase
GC Gas Chromatography
Him Histamine
HOC Histidine decarboxylase
HPLC High Performance Liquid Chromatography
LOD Limit of detection
LOQ Limit of quantification
MAO Monoamine oxidase
ppm Part per million
Put Putrescine
RT Retention time
RSD Relative standard deviation
Spm Spermine
SD Standard deviation
Spd Spermidine
SDr Standard deviation repeatability
SDR Standard deviation reproducibility
TLC Thin Layer Chromatography
viii
LIST OF TABLES
Table Page
Table 2.1 Structure and molecular weight and boiling point of six standard biogenic
amines
4
Table 2.2 Concentration of histamine in fish related to histamine fish poisoning 12
Table 4.1 Parameters in method validation 19
Table 4.2 Comparison of different columns used in the identification of biogenic
amines
20
Table 4.3 Determined retention time of Biogenic Amines on Purospher® STAR
column
22
Table 4.4 Result of SD & RSD of area and retention time of biogenic amines 23
Table 4.5 Correlation coefficients of calibration curves of biogenic amines 27
Table 4.6 LOD and LOQ values for each investigation of biogenic amines 28
Table 4.7 SDr and RSD value of biogenic amines in repeatability condition in muscle
fish.
30
Table 4.8 Result of accuracy (intermediate precision) for the chromatographic method
evaluated with the same HPLC in three consecutive days
31
Table 4.9 Recovery values in % of spiked fish 34
ix
LIST OF FIGURES
Figure Page
Figure 2.1 Structure of histamine, tyramine, 2-phenylethylamine, putrescine and
cadaverine
5
Figure 2.2 Formation of biogenic amine 9
Figure 4.1 Chromatogram for 100 ppm biogenic amines analyzed using
Purospher® STAR HPLC column
20
Figure 4.2 Chromatogram for 100 ppm biogenic amines analyzed using μ
Porasil HPLC column
21
Figure 4.3 The calibration curve of the biogenic amines standards of tyramine 25
Figure 4.4 The calibration curve of the biogenic amines standards of cadaverine 25
Figure 4.5 The calibration curve of the biogenic amines standards of histamine 26
Figure 4.6 Components of accuracy 29
Figure 4.7 HPLC chromatogram of spiked sample 35
Figure 4.8 HPLC chromatogram of unspiked sample 35
x
Analysis of Biogenic Amines in Fish by using
High Performance Liquid Chromatography
ABSTRACT
Biogenic amines (BAs) are organic basis formed in food by microorganisms through
enzymatic decarboxylation of amino acid, non volatile and heat stable. Biogenic amine can be
found in several of food especially food that inappropriate handle and stored scombroid fish
like tuna. The aim of this study was for the verification of an high performance liquid
chromatography (HPLC) method for the determination of biogenic amines in fish. The
implementation of the method included the optimization of sample preparation, HPLC
analysis and data evaluation. Thereafter, the method performance was verified to demonstrate
that the results obtained were consistent, correct and satisfactory for analysis of biogenic
amines in fish. Separation of biogenic amine was achieved on a Purospher® STAR column
with gradient elution system coupled with a UV detector (198 nm). A mixture of standard
biogenic amines consist of tyramine, cadaverine and histamine were used for the verification
test such as linearity, limit of detection, limit of quantification, accuracy, precison and
recovery. The calibration curves for biogenic amines are linear from the concentration of 10
ppm until 75 ppm and linearity coefficients (r2) >0.900 for tyramine and histamine. Relative
standard deviation (RSD) for biogenic amines <10% which are 4.74%, 5.15% and 5.53% for
tyramine, cadaverine and histamine. The recovery in the concentration of tyramine and
cadaverine are 86% and 95%, respectively.
Keywords: biogenic amine, HPLC, validation, fish
xi
ABSTRAK
Amina biogenik merupakan organik asas yang terbentuk di dalam makanan oleh bakteria
melalui pendekarboksilan enzim asid amino, tidak mudah meruap dan tahan kepada suhu yang
tinggi. Amina biogenik boleh dijumpai di dalam makanan terutamanya makanan yang
dikendalikan dengan cara yang tidak sesuai dan kumpulan ikan scrombroid seperti tuna yang
telah disimpan. Tujuan utama kajian ini adalah bagi menentukan kandungan amina biogenik
di dalam ikan dengan menggunakan kromatografi cecair prestasi tinggi (KCPT). Perlaksanaan
kaedah ini adalah termasuk pengoptimuman penyediaan sampel, analisis KCPT dan proses
pengolahan data. Seterusnya kaedah yang terhasil disahkan bagi menunjukkan keputusan yang
diperolehi konsisten, betul dan memuaskan bagi analisis amina biogenik di dalam ikan.
Pemisahan amina biogenik terhasil baik dengan menggunakan turus Purospher@ STAR dan
UV sebagai pengesan (198 nm). Amina biogenik seperti tiramina. kadaverina dan histamina
digunakan dalam pengesahan analisis seperti kelinearan, had pengesanan, had kuantifikasi,
ketetapan, dan kepresisan. Graf kalibrasi bagi amina biogenik adalah linear pada julat
kepekatan antara 10 hingga 75 ppm dan menunjukkan nilai pekali kelinearan >0.900 bagi
tiramina dan histamina. Sisihan piawai relatif untuk amina biogenik <10% iaitu sebanyak
4.74%, 5.15% and 5.53% untuk tiramina, kadaverina dan histamina. Nilai perolehan semula
bagi amina biogenik tiramina dan kadaverina adalah 86% dan 95%, masing-masingnya.
Kata kunci : amina biogenik, kromatografi cecair prestasi tinggi (KPTC), ikan
1
CHAPTER 1
INTRODUCTION
1.1 General Introduction
High Performance Liquid Chromatography (HLPC) is the instrument that is widely
applicable techniques leading the field of analytical chemistry and the most versatile (McNair
and Miller, 1998). The advantage by using HPLC is the sample does not need to derivatise like
the Gas Chromatography (GC). In HPLC, the mobile phase will carried the analyte through
column or stationary phase. Water, methanol, acetonitrile are the example for the mobile
phase. The analyte will be separate by the stationary phase and the elute enter the detector
which produces an electrical signal. The data system generates an image called
chromatography.
Biogenic amines (BAs) are result of removal the carboxylic acid group on the amino
by enzymic reactions from corresponding amino acid. The bacterial actions cause the
formation of biogenic amine in raw food. Biogenic amine can be found in diverse foods like
meat, fermented foods, fish and fish product and cheese (Eerola et al., 1993). Mishandled
during the storage and processing, can causes breakdown of protein to free amino. Amino acid
can also be naturally found within the food. According to Den Brinker et al. (1995) food that
was contaminated with bacteria that contain the decarboxylase enzymes, produce biogenic
amine because the free amino acids undergo decarboxylation. Biogenic amines may cause
vasoactive or psychoactive effects if too large quantity of biogenic amine is consumed orally
(El-Habib, 2011). Microbial growth, availability of free amino acids, temperature, and high the
presence of decarboxylase enzymes are factor influenced the amount of biogenic amines are
2
conditions (Halasz et al.,1994). Histamine poisoning, which results from intake of foods that
contain significant amounts of histamine, has been reported to be one of the major illness
among foodbrone disease. Other amines like putrescine and cadaverine have been described as
potentiators that enhance the toxicity of histamine (El-Habib, 2011). Histidine decarboxylase
is the enzyme that involved in the production of histamine. Temperaute greater than 15 ºC and
30 ºC as the optimum temperature is needed for the production of histamine. Fish are usually
caught in temperatures exceeding 20ºC in the tropical areas of the world and if the fish are not
refrigerated immediately, conditions are favorable for biogenic amine production, providing
bacteria containing decarboxylase enzymes. According to Ahmed (1991), when the
temperature is around 0-5ºC, bacterial growth will stop, but enzymic activity will still continue
resulting in further amine production. Taylor (1985) reported, histamine is toxic in larger
doses, but in small doses it has little effect. Excess quantity of histamine in the body can cause
dilating blood cells and can result in hypotension. The incubation period of histamine
poisoning is short. According to Taylor et al. (1989), eating of a meal that contain high level
of histamine can cause the poisoning effects occur within several minute to a few hours.
Diamine oxides (DAO) and histamine-N-methyl transferase (HMT) is enzyme in human body,
which convert histamine to harmless degradation products (Den Brinker et al., 1995). These
enzymic reactions will be inhibited by putrescine and cadaverine and therefore potentiate of
histamine toxicity. There are several fish suspected of causing histamine but only one type of
fish will be used as samples which are tenualosa toli (terubuk).
3
1.2 Problem Statements
Biogenic amines are basic nitrogenous compounds found in a variety of foods like fish,
meat, fish product and many others. The formations of biogenic amines were mainly due to the
amino acid decarboxylase activity of certain microorganisms. Availability of free amino acid,
microbial growth and elevated temperature conditions are the factor that influences the
biogenic amines production. They can cause different toxicological effects to humans,
depending on biogenic amines concentration, individual sensitivity and the specific biogenic
amines. Some of them have powerful physiological effect and have an important biological
activity. This study will focus on validation of analytical method using HPLC for detection of
biogenic amine. Information regarding the biogenic amine contains in raw fish and fish
product in Sarawak is still rather scanty and inadequate. Hence, attention should be given to
ensure the safety of this product.
1.3 Objectives of the Project
The objectives of this study are as following:
a. to validate the analytical method for biogenic amines analysis using high
performance liquid chromatography (HPLC),
b. to evaluate the recovery efficiency of extract method,
c. to isolate biogenic amines from fish.
4
CHAPTER 2
LITERATURE REVIEWS
2.1 Biogenic Amines
Biogenic amines such as spermine, cadaverine, tryptamine, histamine, putrescine,
heptylamine, and tyramine (Table 2.1) are organic bases formed mainly by the
decarboxylation of amino acid by enzymic reaction.
Table 2.1: Structure and molecular weight and boiling point of six standard biogenic amines
(Nakovich, 2003)
Amines Molecular
Weight
Boiling Point
( ˚C)
Structure Precursor
Histamine 111.15 167
Histidine
putrescine 88.15 158-160
Orthine,
agmatine
cadaverine 102.18 178-180
Lysine
Tyramine 137.18 175-181
Tyrosine
Spermidine 145.25 128-130
Arginine
5
According to Fathi et al. (2013), biogenic amine may be present in various foods that
rich in protein like fish and meat. The chemical structure of biogenic amines are heterocyclic
(tryptamine and histamine) or aliphatic (cadaverin, putrescine, espermine and espermidine) or
as aromatic (phenylethylamine and tyramine) (Figure 2.1).
Heterocyclic amines Aromatic amines
Aliphatic amine
Figure 2.1: Structure of putrescine, tyramine, cadaverine, histamine and 2-phenylethylamine
(EFSA, 2011).
Various bacteria will be produce decarboxylase enzyme in high temperature and kept its
activity to slightly lower 5°C (Fathi et al., 2013). Improper storage of fish can cause quality
degradation and the accumulation of biogenic amine in a short period. Technological
processes for example fermentation, ripening, salting or marination can increase the possibility
formation of biogenic amine (Pons-Sanchez-Cascado et al. 2005a). A study conducted by
Fathi et al. (2013), at low pH around 4.0–5.5, is favorable for enhanced amino acid
decarboxylase activity which can be achieved in salted anchovies. Presence of histamine in
raw fish or fish products is the most important quality indicator as chemical indicators for
spoilage of fish and is considered as a threat to public health bodies.
6
2.2 Hazard Characteristics
2.2.1 Histamine
L-histidine that was contained in Pyridoxal phosphate decarboxylase from the amino
acid histidine in enterochromaffine cells, histaminergic neurons, basophils, mast cells, and
platelets, synthesized the histamine (Maintz and Novak, 2007). Histidine to histamine is the
decarboxylation of the amino acid that is naturally occurring substance in the human body.
Foods that contain free histidine can be present with histamine. Histamine is generated by
certain bacteria during spoilage and fermentation of fish. Endogenous histamine has vital
physiological functions related to local neuromodulation, immune responses and gastric acid
secretion. Sensitive individuals cannot tolerance with histamine-rich foods and food poisoning
may cause because of histamine contamination in fish and fish products (Taylor, 1985).
2.2.2 Putrescine and Cadaverine
Cadaverine and putrescine are example of biogenic amines that may found in fish.
Gram-negative bacteria has mainly been to related cadaverine and putrescine production,
especially in the Pseudomonadaceae, Shewanellaceae and Enterobacteriaceae,family’s
normally related with spoilage (Lopez-Caballero et al., 2001). In the fermented product,
putrescine is one of the most common biogenic amine that can be found. Bacteria like
lactobacilli mainly, staphylococci and lactic acid bacteria have also been reported to be able to
generate cadaverine or putrescine (Arena and Manca de Nadra, 2001; Beneduce et al.,2010;
Coton et al., 2010b). Like histamine, they are produced from amino acids by bacteria during
spoilage and fermentation. The precursors of putrescine and cadaverine are ornithine and
lysine respectively. Putrescine and cadaverine are both found frequently in improperly handled
7
fish and have been studied as spoilage indicators. In some fish spoilage studies, cadaverine
appeared to be formed and increased earlier than histamine (Pons-Sanchez-Cascado et al.,
2005b; Rossi et al., 2002).
2.3 Microorganism Involved in Biogenic Amine Production
Amino acids and amino acid decarboxylases, synthesized by bacteria required in the
formation of biogenic amine (EFSA, 2011). Some microorganisms that have enzyme histidine
decarboxylase (HDC) can lead the formation of histamine in fish. At the highest level of
temperature, the naturally present of free histidine in the muscle of some fish will be converted
to histamine by HDC. HDC can be form by both Gram-negative and Gram-positive bacteria
but, the forms of the enzymes differ (EFSA, 2011; Bjornsdottir-Butler et al.,2010).
FAO/WHO (2013), reported that in fish, biogenic amine-producing bacteria can be likely to be
found on the gills, skin, or in the gastrointestinal tract. Transfer of these bacteria to the flesh of
the fish, where free amino acids may be present, leads to development of biogenic amines.
Transfer can occur from the gastrointestinal tract after harvest through migration, or via
rupture or spillage of gastric contents during gutting.
2.4 Conditions Supporting the Formation of Biogenic Amines
2.4.1 Substrate availability
The amino acids substrate is one example of the prerequisites for biogenic amines
synthesis. Proteolysis is one of the factors that are directly related to availability of free amino
8
acids which provide a substrate for biogenic amines formation. Biogenic amines formation
increases based on conditions of enhanced proteolysis or accelerated (Leuschner et al.,1998a;
Innocente and D`Agostin, 2002; Fernandez et al., 2007a; Komprda et al., 2008).
2.4.2 Temperature
Temperature is the major factor of the quantity production of biogenic amines (Zaman
et al. 2009). As the temperature increase, the production of amine also increases. On the
contrary, at low temperatures microbial growth and the reduction of enzyme activity is
inhibited, thus the accumulation of the biogenic amine is minimized. 20 to 37 °C is the
optimum temperature for the formation of biogenic amine but at the temperature of 5 °C or
above 40 °C the production of biogenic amine is decrease (EFSA, 2011). In seafood
Morganella morganii is a powerful histamine producer, although at temperatures above 7-10
ºC (Kim et al. 2002; Lehane and Olley, 2000). At 20ºC rather than 10ºC, Klebsiella
pneumoniae was produce cadaverine while an Enterobacter cloaca was able to produce
putrescine (Halaz et al., 1994). Very low temperature is needed for refrigerated foods because
even at 5 ºC, psychro tolerant bacteria can give to the amine accumulation. To reduce the
growth of bacteria and proteolytic and decarboxylase activities, low temperatures should be
applied during the storage (Hernandez-Orte et al. 2008; Rezaei et al., 2007)
2.4.3 Effect of pH
Other factors that influence the amino acid decarboxylase activity are pH (Silla Santos,
1996). In low pH environment, bacteria are more stimulated to produce decarboxylase as a
part of their defence mechanisms (Fernandez et al. 2007a; Bover-Cid et al. 2006b; Molenaar
et al. 1993).
9
2.4.4 Other Factors
Biogenic amines are produces by enzymatic decarboxylation of certain free amino acid
(Figure 2.2). According to Bardocz (1995), bacterial decarboxylases are not very specific, but
their activity varies according to the bacterial species and strain. Thus, the presence of free
amino acids, the decarboxylase enzyme and suitable environmental conditions are required for
biogenic amine formation (Ruiz-Capillas and Jimenez-Colmenero, 2004).
Protein
Proteinases
Endopeptidases
Peptides
Tyrosine Histidine Trytophan Lysine Glutamine Ornithine Arginine
Amino acid
decarboxylases
Tyramine Histamine Tryptamine Cadaverine Spermine Putrescine Spermidine
Figure 2.2 Biogenic amine Formation (Ruiz-Capillas and Jimenez-Colmenero, 2004)
2.5 Toxicological Aspects of Biogenic Amine in Responses of Human
2.5.1 Histamine
Intestinal problem, headaches, pseudo-allergic, gastric and migraine are several effect
if consume foods that contain high concentration of biogenic amine which is brought by the
toxic action of tyramine and histamine, known as cheese reaction and histamine poisoning
(Stratton et al., 1991; Taylor, 1985; Smit, 1980). Even though endogenous concentrations of
histamine are necessary and are required for normal physiological function, histamine is toxic
10
when large doses enter the circulatory system. This results in symptoms of poisoning, which
involve a wide range of organs include vascular, neurological, heart and gastrointestinal
(Taylor, 1985).
2.5.2 Cadaverine and Putrescine
Putrescine and cadaverine, can enhance the effect of tyramine and histamine by
interacting with the amino oxidase and interfering with the detoxifying mechanism and also
know as histamine potentiators and both are not considered toxic individually (Sattler et al.,
1988; Taylor, 1985; Rice et al., 1976). The minimum concentration of putrescine or
cadaverine that potentiates histamine toxicity is unknown. The ratio of putrescine or
cadaverine to histamine may need to be high to produce an effect, and it is not clear whether
the levels present in spoiled fish are sufficient to enhance the toxicity of histamine in humans
(FAO/WHO, 2013).
2.5.3 Tyramine, Phenylethylamine and Tryptamine
According to EFSA (2011), phenylethylamine, tryptamine and tyramine are
synthesised in humans from their corresponding amino acids (phenylalanine, tryptophan, and
tyrosine respectively) by decarboxylation and also known as ‘trace amines’. Monoamine
oxidase (MAO) is their main catabolism there are two MAO enzyme exist (A and B), with
different substrate specificity and locations. MAO-A predominates in the placenta, stomach
and intestine has polar aromatic amines (as octopamine and noradrenalin) as preferred
substrates. In the brain MAO-B was predominates and selectively deaminates non-polar
aromatic amines (as dopamine and phenylethylamine) (EFSA, 2011). In humans, tyramine
acts as a catecholamine (including norepinephrine [noradrenaline], dopamine, epinephrine
11
[adrenaline]) releasing agent, resulting in increased blood pressure (FAO/WHO, 2013).
According to Joosten (1987), headaches, increased blood pressure, and migraines are typical
symptoms of tyramine poisoning. Tyramine also causes the release of noradrenaline from the
sympathetic nervous system (Bardocz, 1995).
2.6 Biogenic Amine in Fish
A majority incident of histamine poisoning was related with the fish like tuna,
skipjack, and mackerel (Taylor, 1985). Scombroid fish poisoning, also known as histamine
fish poisoning is because of the histamine as the main toxin involved in. That poisoning
generally associated with high levels of histamine that is higher or equal to 500 mg/kg in fish.
There is not a straightforward dose-response relationship even though there is compelling
evidence to implicate histamine as the causative agent in histamine fish poisoning, because
spoiled fish containing histamine tends to be more toxic than the equivalent amount of pure
histamine dosed orally (Lehane and Olley, 2000). The main evidences supporting the
involvement of histamine in histamine fish poisoning are the identical symptoms of histamine
poisoning to those of intravenous histamine administration and the efficacy of antihistamines
in histamine fish poisoning therapy (Koutsoumanis et al. 2010). Concentration of histamine
that is lower than 500 mg/kg, was also reported as histamine poisoning. For example
scrombroid poisoning was linked to canned anchovies containing 365 mg/kg histamine (NSW
Food Authority, 2010) and tuna burgers with histamine levels of 213 mg/kg were implicated
(Becker et al. 2001). Various histamine concentrations in fish with different range sub-
samples of the same sample, 102 – 180 mg/kg, 27 – 152 mg/kg, 50 – 500 mg/kg of 147 mg/kg,
below 10 to 1000 mg/kg and 5 to 208 were recorder in the European Rapid Alert system for