Food Sci. Biotechnol.

24(5): 1907-1913 (2015)

DOI 10.1007/s10068-015-0251-4

Determination of Biogenic Amines in Fish Products

Bilge Bilgin* and Hüseyin Gençcelep
Department of Food Engineering, Faculty of Engineering, Ondokuz Mayis University, 55200 Samsun, Turkey

Received November 24 2014

Abstract Concentrations of 5 biogenic amines were simultaneously determined in 63 samples of fish
Revised April 28 2015
Accepted May 5 2015 products using HPLC. Fish products were purchased from different cities in Turkey. Fish samples
Published online October 31 2015 included canned sardines, canned mackerel, canned tuna, and marinated anchovies. The total level of
biogenic amines ranged from 26.58 to 406.55 mg/kg. Histamine (HIS) and cadaverine (CAD) were the
*Corresponding Author
Tel: +903623121919 dominant biogenic amines. Concentrations of tyramine (TYR) and HIS were less than 50 and 100 mg/
Fax: +903624576094 kg, respectively. The relationship between formation of biogenic amines in fish products and
E-mail: [email protected] thiobarbituric acid (TBA), total volatile basic nitrogen (TVB-N), and pH values was also investigated.
pISSN 1226-7708
There were significant negative correlation (p<0.01) between TBA, and TVB-N, pH respectively.
eISSN 2092-6456 Significant positive correlation was found between TVB-N, and pH, protein, and CAD, TYR, and HIS.
There were significant (p<0.01) positive correlations between TBA, TYR, tryptamine, pH, and TVB-N
© KoSFoST and Springer 2015 values in fish samples.

Keywords: biogenic amine, putrescine, histamine, tyramine, fish product

Introduction regulations, the level of histamine in fish should not exceed 200 mg/
kg (Turkish Fisheries Products Regulation no 2008/27004).
Biogenic amines are nitrogenous organic bases produced during Biogenic amines are biologically active compounds and consumption
decarboxylation of amino acids or transamination of aldehydes and of food containing high amine levels may lead to different food-born
ketones in foods (1-3). The most important biogenic amines are diseases. Biogenic amines lead to several health issues due to toxicity.
histamine (HIS), tyramine (TYR), tryptamine (TRP), putrescine (PUT), Therefore, it is critical to monitor these compounds in foods (7).
and cadaverine (CAD), which are formed from the free amino acids Histamine, tyramine, β-phenylethylamine, and tryptamine may
histidine, tyrosine, tryptophane, ornithine, and lysine, respectively affect the nervous and vascular systems. Common symptoms of
(1). intoxication are nausea, respiratory distress, flushing, sweating, heart
High biogenic amine concentrations are randomly detected in fish palpitations, headache, prupitus, oral burning, and either hypo or
and fish products, matured cheese, sausages, wine, and fermented hypertension (8), which are mainly caused by histamine, tyramine,
vegetables like pickles. Formation of biogenic amines in aquatic and β-phenylethylamine. Histamine intoxication, known as histamine
products depends on different factors, such as the free amino acid poisoning, exhibits symptoms of reddening of the skin, edema, rash,
contents in fish, the presence of bacterial biogenic amine decarboxylases, and dilatation of peripheral blood vessels resulting in hypotension,
the quality parameters of raw materials, and environmental conditions headache, and contraction of intestinal smooth muscle causing
(1). diarrhoea and vomiting (9). Tyramine levels of 100-800 and 30 mg/
Many countries and international organizations have established kg of β-phenylethylamine have been reported to be toxic in foods (1).
regulations and legal requirements for maximum levels of biogenic A total biogenic amine level of 1,000 mg/kg in food is considered
amines (4). A histamine level lower than 50 mg/kg has been suggested harmful to human health. Histamine intake from 8-40, 40-100 mg/
by the U.S. Food and Drug Administration (FDA) as a guidance level in kg, and higher than 100 mg/kg may cause slight, intermediate, and
fish (5). The FDA has also recommended that 100 mg/kg of tyramine intensive poisoning, respectively (10). Nout (11) pointed out that
and 1,000 mg/kg of total biogenic amines should be tolerance levels maximum allowable levels of histamine and tyramine in foods
in fish (5). In the European Union (EU), the acceptable level of should range from 50-100 and 100-800 mg/kg, respectively. This
histamine in Scombridae fish is suggested to be less than 100 mg/kg, study was conducted to determine levels of biogenic amines and the
and the maximum level of total biogenic amines should be less than chemical quality of fish products sold in Turkish markets.
200 mg/kg in fish and fish products (6). According to Turkish
1908 Bilgin and Gençcelep

Materials and Methods 50% solvent B and ended at 90% solvent B after 25 min. The system
was equilibrated before subsequent analyses. The flow rate was 1.0
Materials Fish products were purchased at local department stores and the column temperature was 40oC. A 20 µL sample was injected
and retail markets from different cities in Turkey in June of 2012. All into the column. Peaks were controlled at 254 nm using the HPLC
fish products were produced in Turkey. Fish products included system with a Spherisorb ODS2 150A column (150×4.60 mm)
canned tuna (18), chunk canned tuna (21), marinated anchovies (12), (Waters, Milford, MA, USA), an HPLC apparatus (Shimadzu Corporation),
canned mackerel (6), and canned sardines (6). Properties of fish a CBM-20A system control unit, an LC-20AT pump unit, an SIL-20A35
products are shown in Table 1. autosampler, an SPD-M20A PDA detector, and a computer (all from
Shimadzu Corporation). Quantitative determinations were carried
Proximate chemical analysis Protein contents were analyzed out following the internal standard of 1.7-diaminoheptane method
following the Kjeldahl method. (12). The crude protein content was using peak areas (15). Biogenic amine contents were expressed as
calculated based on multiplication of the total Kjeldhal nitrogen mg/kg. The limit of detection was between 0.005-0.050 µg/mL and
amount by 6.25. The ash content was determined from the weight of limits of quantification were between 0.010-0.100 µg/mL for
a fish product sample burned at 550oC for 4 h. The moisture content different biogenic amines.
was calculated from the weight loss of a fish product sample of
approximately 5 g after oven-drying at 105oC for 2 h, or until a Statistical analysis Data were subjected to a one-way analysis of
constant weight was obtained. variance (ANOVA). All statistical analyses were performed using SPSS
Version 20.0 for Windows (SPSS Inc., Chicago, IL, USA). A value of
Determination of pH, TBA, and TVB-N values All pH values were p<0.05 was used to identify significant differences. Contents of
measured using a pH meter equipped with an electrode after biogenic amines in different samples were expressed as mean±
homogenization of 10 g of fish sample in 100 of distilled water. The standard deviation (SD) of at least 3 determinations for each sample.
level of total volatile basic nitrogen (TVB-N) was determined Principal component analysis (PCA) was used to study relationships
following the distillation procedure reported by Antonacopoulos and between quality parameters and biogenic amines.
Vyncke (13). Thiobarbituric acid (TBA) level was determined according
to the method proposed by Tarladgis et al. (14) and oxidation
products were quantified as malondialdehyde (mg of MA/kg) Results and Discussion
equivalents.
Chemical analysis of fish product samples Sample names and
Biogenic amine analysis their physical properties are given in Table 1. The chemical
Standards and reagents: Six aqueous standard solutions containing compositions of fish product samples are shown in Table 2 and 3. The
cadaverine dihydrochloride, putrescine dihydrochloride, tyramine water contents of marinated anchovy and canned tuna samples
hydrochloride, tryptamine hydrochloride, histamine dihydrochloride, were significantly (p<0.05) different (Table 2 and 3). Also, significant
and 1.7-diaminoheptane (as an internal standard) were purchased (p<0.05) differences were identified between TBA values of different
from Sigma-Aldrich (St. Louis, MO, USA). All reagents were of canned sardine samples (Table 2) because S1 samples did not contain
analytical grade, except reagents used in HPLC analysis, which were sunflower oil while S2 samples did.
chromatographic grade (Sigma-Aldrich). Deionized water was produced The pH value is an important factor influencing aminodecarboxylase
using a Milli-Q system (Millipore Corp, Molsheim, France). Mobile activity, which is stronger in an acidic environment with an optimum
phases were filtered through PVDF membranes (0.45 mm pore size; pH between 4.0 and 5.5 (15). Marinated anchovy samples exhibited
ISOLAB GmbH, Wertheim, Germany). lower pH values than other fish product samples resulting from use
Determination of biogenic amine levels: Biogenic amine contents of of organic acid in production of marinated anchovies. Generally, the
samples were determined following the method of Eerola et al. (15). pH value was high in samples that had low biogenic amine levels (S2,
Biogenic amines were extracted from 2.0 g samples using 0.4 M M1, M2, and CT2 and CT4). There was a significant (p<0.01) negative
perchloric acid and centrifugation (Hettich, Tuttlingen, Germany) at correlation between TBA and TVB-N values (r= −0.355) and a positive
795×g for 10 min performed at 20oC. The supernatant was collected correlation between pH and TVB-N values (r=0.813) in samples
and brought to 25 mL using 0.4 M perchloric acid. The resulting (Table 4).
solution was filtered through Whatman No 1 filter paper. Derivitazation The TVB-N value is well documented as an index of the quality of
was performed using 2 mL of dansyl chloride. Then, 20 µL of a fresh fish because an increase is related to bacterial spoilage. The
standard (0.4 mg/kg) and sample derivates were injected into an TVB-N value is mainly determined by ammonia and primary, secondary,
HPLC apparatus (Shimadzu Corporation, Kyoto, Japan). The gradient and tertiary amines. TVB-N and free amino acid (precursors of
elution system was 0.1 M ammonium acetate as solvent A and biogenic amines) contents both increase during manufacture and
acetonitrile as solvent B. The gradient elution program was started at storage of fish (16). TVB-N values ranged from 5.57 to 47.23 mg/100

Food Sci. Biotechnol.

 Determination of Biogenic Amines in Fish Products 1909

Table 1. Properties of fish product samples

Sample code Sample type Ingredients and Properties
S1 Sardine Fillet with sunflower oil, canned
S2 Sardine Fillet with salt, canned
M1 Mackerel In tomato sauce, filet, canned
M2 Mackerel Fillet with seasoning, canned
A1 Anchovy Marinated fillets with sauce
A2 Anchovy Marinated fillets
A3 Anchovy Marinated fillets with sauce
A4 Anchovy Marinated fillets
T1 Tuna In Sunflower oil, solid, canned
T2 Tuna Light, solid, in water and salt, canned
T3 Tuna In sunflower oil, solid, canned
T4 Tuna Light, solid, in water and salt, canned
T5 Tuna In olive oil, solid, canned
CT1 Tuna In sunflower oil, chunk, canned
CT2 Tuna In sunflower oil, chunk, canned
CT3 Tuna In sunflower oil, chunk, canned
CT4 Tuna In sunflower oil, chunk, canned
CT5 Tuna With peas and corn, in water and salt, chunk, canned
CT6 Tuna In tomato sauce, chunk, canned
CT7 Tuna In olive oil, chunk, canned
CT8 Tuna In sunflower oil, minced, canned

g in this study. TVB-N levels were lower than 30 mg/100 g in all Significant (p<0.05) differences between biogenic amine contents in
samples, except for S1, T5, and CT3. Thus, almost all fish samples fish product samples were identified. Biogenic amine levels were low
analyzed were fresh. Generally, some amine contents were high in or moderate in most fish products. All biogenic amines were not
samples that had high TVB-N levels (Fig. 1). The rate of unacceptable detected in all fish product samples. HIS (50/63), PUT (28/63), CAD
fish samples was 22.22% (14/63) for TVB-N, based on a decomposition (44/63), TYR (28/63), and TRP (27/63). Most fish product samples
level of 30 mg/100 g for fish quality. showed less than 20 mg/kg of each biogenic amine.
The TBA value is a measure of malondialdehyde (MA), which is an Cadaverine and putrescine are important in foods, especially fish and
end product of lipid oxidation. A decrease in MA levels may be fish products because they have been shown to potentiate the
caused by interactions between MA and amines, nucleosides, nucleic toxicity of histamine. In this study, canned sardines contained low
acids, amino containing phospholipids, proteins, or other aldehydes histamine concentrations (32.65-70.67 mg/kg) (Table 5). Soares and
that are by-products of lipid oxidation. TBA values were between Gloria (18) reported histamine values in canned sardines from Brazil
0.22-5.12 mg of MA/kg in this study. The highest TBA values were ranged from 1.2 to 36 mg/kg. In a Polish study, imported canned
identified in marinated anchovy samples (Table 2). The initial sardines and mackerel had histamine contents exceeding 200 mg/kg
microbiological load of a product, hygiene measures, and the presence (19). Zhai et al. (20) reported that the maximum histamine level was
of bruises are all factors that can influence degradation of anchovies 22.38 mg/kg in canned sardine samples, which was less than levels
(17). The type of anchovy processing has an impact on the quantity reported herein.
of biogenic amines and amine chemical properties because marinated The histamine level of canned mackerel in this study was in the
samples had high concentrations of histamine, cadaverine, and range of 25.70 to 26.34 mg/kg (Table 5), which was more than
tryptamine due to the bacteriostatic action of salt ripening. previous results for histamine reported to be in the range of ND to
Development of lactic flora, potential producers of biogenic amines, 2.4 mg/kg (21). In Spanish mackerel samples, histamine was detected
is still possible under acidic conditions (4). However, the major within the range of 15.74-28.70 mg/kg, whereas the maximum
factors that determine production of biogenic amines are properties histamine level was 26.95 mg/kg in canned anchovies (20). Hwang et
of raw fish. al. (22) reported large amounts of histamine in dried fish products
(63.1-479.0 mg/kg). In Spain, histamine was found in 83.3% of tuna
Biogenic amines in fish product samples Canned sardines, samples from a retail market in Barcelona with an average value of
mackerel, marinated anchovies, and tuna products were analyzed for 8.9 mg/kg (23). Silva et al. (24) reported that canned tuna products
biogenic amine contents measured from 21 different fish products available in the Brazilian market also contained low levels of
currently distributed in Turkey. The concentrations of biogenic histamine with an overall mean value of 4.41 mg/kg. Zarei et al. (25)
amines in 63 fish product samples are shown in Table 5 and 6. reported histamine in 57.5% of canned tuna samples in the range of

October 2015 | Vol. 24 | No. 5

1910 Bilgin and Gençcelep

Table 2. Chemical analysis of canned sardines, canned mackerel, and marinated anchovies (mean±SD)
Sample Total Water Protein Ash TVB-N TBA
pH
code N % % % (mg/100 g) (mg of MA/kg)
S1 3 54.49±1.47a1) 20.32 ±0.37a 3.16±0.37a 6.22±0.06 a 47.23±1.04 a 3.36±0.88a
S2 3 55.79±0.78a 16.08±0.30b 2.87±0.07a 6.03±0.03b 27.08±1.71 b 0.79±0.05b
Mean±SD 6 55.14±1.27 18.20±2.34 3.01±0.29 6.12±0.11 37.16±11.11 2.07±1.51
M1 3 52.93±0.95a 14.39 ±0.03a 1.25±0.64a 5.52±0.05a 19.23±1.91a 0.81±0.02a
M2 3 51.65±1.18a 14.55±0.09 b 1.64±0.16a 6.28±0.03 b 22.60±0.77 b 1.08±0.03b
Mean±SD 6 52.30±1.19 14.47±0.11 1.44±0.47 5.90±0.42 20.91±2.26 0.95±0.15
A1 3 52.53±2.97b 19.43±0.36ab 2.38±0.16c 3.54±0.08a 5.57±0.21c 1.85±0.04d
A2 3 50.67±2.16b 19.65±0.10ab 3.54±0.12a 3.27±0.07c 10.37±0.68a 3.03±0.09c
A3 3 56.38±0.50a 19.17±0.21 b 3.65±0.22a 3.29±0.06bc 10.58±0.77a 3.85±0.17b
A4 3 57.42±1.39a 19.90±0.48a 3.18±0.02b 3.41±0.02b 6.97±0.01b 5.12±0.36a
Mean±SD 12 54.25±0.39 19.54±0.39 3.19±0.54 3.38±0.12 8.37±2.30 3.46±1.26
1)a-b
Different lowercase letters within a sample column and chemical analysis indicate significant differences between fish samples (p<0.05).

Table 3. Chemical analysis of canned solid tuna (Mean±SD)

Sample Total Water Protein Ash TVB-N TBA
pH
code N % % % (mg/100 g) (mg of MA/kg)
T1 3 66.53±1.73b 20.95±1.04a1) 1.42±0.21b 5.75±0.02c 27.29±0.66b 0.35±0.02a
T2 3 76.49 ±0.15c 20.73±0.11a 1.25±0.09 b 5.52±0.02a 22.28±0.45a 0.45 ±0.02b
T3 3 61.41 ±0.82a 20.61 ±0.51a 1.71±0.03c 5.53 ±0.05a 27.53±1.71b 1.07 ±0.06c
T4 3 76.86 ±0.59c 22.87 ±0.55b 1.65 ±0.02c 5.63±0.03b 21.39±1.02a 0.39 ±0.09a
T5 3 62.33 ±0.51a 22.37 ±1.15b 1.30±0.20b 5.63±0.05b 30.53±0.63c 0.36 ±0.03a
Mean±SD 15 68.72±7.00 21.50±1.17 1.46±0.22 5.61±0.09 25.80±3.67 0.52±0.29
CT1 3 68.37±0.79c 21.80±0.32def 1.70 ±0.05e 5.71±0.02d 22.57±0.50c 0.55±0.05c
CT2 3 61.40 ±2.20a 21.91±0.24de 1.40±0.11c 5.57±0.03bc 24.71±0.16cd 0.47±0.04b
CT3 3 61.34±0.97a 22.32±1.08f 1.51±0.18cd 5.66±0.03cd 34.75 ±1.74e 0.43 ±0.04b
CT4 3 64.55 ±0.40b 21.19±0.43de 1.64±0.12de 5.52 ±0.01b 26.56±2.48d 0.67 ±0.02d
CT5 3 77.05 ±0.89e 18.02±0.61b 1.36±0.11c 5.10±0.02a 20.27±0.32b 0.22±0.02a
CT6 3 74.00 ±0.18 d 16.91 ±0.54a 1.45±0.01c 5.08 ±0.06a 13.78±0.79a 0.47 ±0.02b
CT7 3 60.66 ±0.44a 20.02 ±0.12e 1.17 ±0.02b 5.69±0.01d 23.23±1.08c 1.22 ±0.02f
CT8 3 67.55 ±0.68 c 20.78 ±0.59cd 0.97 ±0.02a 5.58±0.02bc 26.60±0.25d 0.95 ±0.03e
Mean±SD 24 66.86 ±5.90 20.37 ±1.92 1.40 ±0.24 5.49 ±0.25 23.95 ±5.90 0.62 ±0.30
1)a-h
Different lowercase letters within each sample column and chemical analysis indicate significant differences between fish samples (p<0.05).

0.12-648.20 mg/kg with an overall mean value of 64.61 mg/kg. (Table 4). Lower histamine concentrations observed in some fish
In this study, HPLC analysis was used to quantify histamine products in this study can be attributed to use of hazard analysis
contents in 63 canned fish samples. Seven tuna samples exhibited critical control point systems (HACCP) by the Turkish canning
histamine levels higher than 100 mg/kg. The histamine content of 44 industry.
samples were in accordance with the FDA toxicity level of 50 mg/100 The presence of cadaverine and putrescine can synergistically
g. Canned sardines and tuna exhibited a considerably higher enhance histamine toxicity due to inhibition of the histamine
histamine level than other samples. Based on an analysis of poisoning metabolizing enzymes diamine oxidase and histamine methyl
episodes, Shalaby (2) suggested guideline levels for histamine transferase (16). Putrescine was detected in approximately 65.21% of
contents of fish of (1) <5 mg/100 g (safe for consumption); (2) 5-20 all samples with values ranging from ND to 116.53 mg/kg. Cadaverine
mg/100 g (possibly toxic), (3) 20-100 mg (probably toxic), and (4) was the major biogenic amine in barramundi fillets kept at 8oC (26),
>100 mg/100 g (toxic and unsafe for human consumption). Significant and was detected in all samples in this study with values ranging
(p<0.05) positive correlations were identified between pH and the from ND to 122.18 mg/kg. Also, sample S1 had a high TVB-N value of
tyramine content (r=0.262) and between pH and the putrescine 47.23 mg/kg.
content (r=0.244) in samples in this study. (Table 4). Also, there was a High cadaverine levels may have been due to the use of poor
significant (p<0.05) positive correlation between putrescine and quality raw fish. Yamanaka et al. (27) suggested that cadaverine
cadaverine contents (r=0.295) and a significant (p<0.01) positive (upper acceptable limit of 100 mg/kg) might be used as an indicator
correlation between histamine and tyramine contents (r=0.348) of freshness in salmonoids. In 9.5% of samples in this study, high

Food Sci. Biotechnol.

 Determination of Biogenic Amines in Fish Products 1911

Table 4. Descriptive statistics and the Pearson correlation matrix (n=63)

TBA TVB-N pH Water Protein Ash Tryptamine Tyramine Histamine Cadaverine Putrescine
1)
TBA 1 -.355** -.655** -.521** .009** .677** -.006 -.211** -.210** .205** -.067**
TVB-N 1 .813** .100** .246** -.197** .025 .214** .038** .197** .303**
pH 1 .262*2) -.024** -.519** .025 .262** .114** .013** .244**
Water 1 .381** -.451** -.028 .068** .011** .158** .232**
Protein 1 -.044** .145 .261** -.128** .313** .366**
Ash 1 -.113 -.312** -.120** .195** -.021**
Tryptamine 1 .095** .184** -.056** -.073**
Tyramine 1 .348** -.147** -.080**
Histamine 1 -.213** -.251**
Cadaverine 1 .295**
Putrescine 1
1)
**Correlation is significant at p<0.01 level (2-tailed).
2)
*Correlation is significant at p<0.05 level (2-tailed).

Table 5. Biogenic amine levels in canned sardines, canned mackerel, and marinated anchovies (mg/kg, Mean±SD)
Sample Histamine Putrescine Cadaverine Tyramine Tryptamine
Total N
Code (HIS) (PUT) (CAD) (TYR) (TRP)
S1 3 32.65±4.06b 58.18±57.30a1) 122,18±68.0a 5.50±4.83a 36.05±1.09a
S2 3 70.67±9.31a ND2) ND ND ND
Mean±SD 6 51.66±21.77 29.09±48.26 61.09±79.57 2.78±4.31 18.03±19.76
M1 3 26.34±8.85a 8.72±0.61a ND ND 29.03±25.14a
M2 3 25.70±6.24a 8.82±1.01a 8.75±7.87a 28.40±2.95a 53.28±78.87a
Mean±SD 6 26.02±6.86 8.77±0.75 4.37±6.91 14.20±15.67 41.15±54.02
A1 3 73.46±10.83a ND ND 8.13±14.07b ND
A2 3 ND ND 47.60±17.61b ND 19.87±34.42c
A3 3 11.43±7.24b 4.28±7.41b 95.69±6.13c ND 190.61±7.67a
A4 3 8.61±4.41b N.D 39.54±7.88b ND 106.56±33.0b
Mean±SD 12 23.37±31.08 1.07±3.71 45.70±35.56 2.03±7.04 79.26±81.78
1)a-b
Different lowercase letters within each sample column and chemical analysis indicate significant differences between fish samples (p<0.05).
2)
ND: Not detected.

cadaverine levels of approximately 100 mg/kg were identified. Zarei value of 0.7 mg/kg. Nout (11) pointed out that the maximum
et al. (25) found that putrescine was detected in all canned tuna allowable level of tyramine in foods should be in the range of 100-
samples in a range of 0.29-52.83 mg/kg. Köse et al. (28) reported 800 mg/kg.
that putrescine was detected in marinated anchovies in a range of Tryptamine was detected in approximately 42.85% of all samples
3.6-24.9 mg/kg. Zhai et al. (20) reported that putrescine was detected in this study with values ranging from ND to 190.61 mg/kg. High
within a range of ND-25.01 mg/kg, all similar to results reported tryptamine levels may have been related to use of poor quality raw
herein. The maximum level of cadaverine detected in Spanish fish. Zhai et al. (20) reported that tryptamine was detected in the
mackerel was reported to be in the range of 230.14-259.95 mg/kg by range of 10.12-20.15 mg/kg in fish samples. Köse et al. (28) reported
ten Brink et al. (1). Hu et al. (29) detected putrescine in 97% of 74 that tryptamine was detected in marinated anchovies at 55.8 mg/kg.
samples with values ranging between 1.0 and 94.1 mg/kg, and In this study, tryptamine levels were generally higher than 20 mg/kg,
tyramine in 82% of all samples ranging from 2.6 to 24.3 mg/kg. especially in marinated anchovies. Some technological processes,
Tyramine was detected in approximately 42.85% of all samples in such as marination, fermentation, and salting can increase the
this study with values ranging from ND to 48.63 mg/kg. Having toxic possibility of biogenic amine formation. A low pH of 4.0-5.5, which
effects for human health, it is very crucial to underline the presence can be achieved in salted anchovies, is favorable for enhanced amino
of tyramine in fish products. Tyramine levels in all samples in this acid decarboxylase activity (31).
study were lower than 100 mg/kg (1). In octopus and squid samples For all fish product samples tested in this study, maximum levels of
low amounts of tyramine have been observed whereas higher HIS, PUT, CAD, TYR, and TRP were 110.33, 116.53, 122.18, 48.63, and
amounts of tyraminedetected in the scale of 101-222 mg/kg in 190.61mg/kg, respectively. The HIS content was within the range 0-
mackerel, saury, Spanish mackerel, and amberjack samples (30). 10.0 mg/kg in 46.38% of samples, 10.0-50.0 mg/kg in 23.19% of
Zarei et al. (25) reported that tyramine was detected in 55% of samples, and more than 100 mg/kg in 10.14% of samples. Only 2
samples tested in the range of 0.46-9.7 mg/kg with an overall mean samples exhibited values that were not close to acceptable limits.

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1912 Bilgin and Gençcelep

Table 6. Biogenic amine analysis of canned solid tuna (mg/kg, Mean±SD)

Histamine Putrescine Cadaverine Tyramine Tryptamine
Sample code Total N
(HIS) (PUT) (CAD) (TYR) (TRP)
T1 3 84.00±73.10b 2.36±4.09a1) 103.28±94.05a 33.79±9.43b ND
T2 3 4.20±1.55a 12.59±0.20ab 78.81±33.05a ND ND
T3 3 4.92±1.35a 40.86±24.45b 82.24±4.71a ND ND
T4 3 ND2) 116.53±2.90c 38.09±22.13a 10.24±17.74a ND
T5 3 5.34±4.70a 35.82±31.39ab 86.76±6.09a ND 179.78±63.09a
Mean±SD 15 19.63±43.25 41.63±44.16 79.93±43.58 8.80±15.55 35.96±78.16
CT1 3 93.94±9.26cd 12.96±22.44a 72.36±120.94a 23.43±7.52abc 182.98±158.58b
CT2 3 5.17±4.48a 9.90±8.57a 20.12±17.88a 33.13±4.20bc 19.13±33.12a
CT3 3 10.28±1.98a 13.61±23.56a 10.76±9.62a 28.62±12.63b ND
CT4 3 7.17±6.41a 18.93±4.18a ND 7.71±7.78ab ND
CT5 3 6.21±10.75a ND 64.69±12.20a ND ND
CT6 3 66.07±1.98bc ND 1.69±2.93a 3.74±6.48ab 15.37±26.61a
CT7 3 110.33±9.87d ND 22.49±5.81a 48.63±43.09c 82.33±4.37a
CT8 3 52.53±52.46b ND 7.99±0.43a 23.24±10.33abc ND
Mean±SD 24 34.48±38.47 6.92±12.47 25.01±45.20 21.06±21.15 37.48±78.80
1) a-d
Different lowercase letters within each sample column and chemical analysis indicate significant differences between fish samples (p<0.05).
2)
ND: Not detected.

Levels of other biogenic amines were less than 100 mg/kg, with the
exception of few samples.
Use of poor quality fish as a raw material for canned fish products
or inappropriate handling techniques for fish during processing are
the main reasons for unacceptable levels of biogenic amine in canned
fish. Lopez-Sabater et al. (23) reported that bacterial populations can
rise during thawing and handling of fish. Furthermore, contamination
of fish with histamine-producing bacteria can also occur from the
environment and from equipment used in processing plants.
Equipment used for skinning, scaling, beheading, gutting, and
filleting can cause contamination if not properly cleaned.
PCA results for mean values of biogenic amines, protein, water, Fig. 1. A principal component analysis plot of biogenic amines and
ash, pH, TVB-N, and TBA of fish samples are depicted in a 3- chemical properties of fish products.
dimensional plot (Fig. 1). Principal components (PC) 1, 2, 3, and 4
described approximately 72.89% of total sample variation with the fish product samples contained biogenic amines lower than
32.25% for PC1, 16.28% for PC2, 12.57% for PC3, and 11.77% for PC4. accepted safe values. Therefore, these products were regarded to be
PC 1 was heavily loaded on TBA, water, and ash, component 2 was harmless for human consumption. 45.85% of fish product samples
heavily loaded on TVB-N, pH, and PUT, component 3 was heavily showed high level of biogenic amines exceeding the level of 100 mg/
loaded on HIS, CAD, and TYR, while component 4 was loaded on TRP kg. Some of the analysed tuna samples exhibited increased level of
and protein. Cadaverine, putrescine, tryptamine, protein, and TVB-N HIS, CAD, and TRP and a total biogenic amine level of 300 mg/kg.
values were positively correlated with each other. Positive low Variations in the biogenic amine levels of fish samples could be due
correlation values between histamine, tyramine, and pH are shown to the hygienic qualities of raw materials, manufacturing techniques, or
in Fig. 1. A significant negative relationship between water and specific flora. Instrumental analysis in quality control systems of
cadaverine attributes was also identified (Fig. 1). In this study, no Turkish producers of canned fish is needed.
significant (p>0.05) correlation was identified between amounts of A significant positive correlation (p<0.01) was identified between
HIS, TYR, and other biogenic amines in fish product samples (Fig. 1). biogenic amine levels and chemical properties of fish products (Table
A significantly (p<0.05) high positive correlation between TVB-N and 4). The quality of some products has to be improved, perhaps based
PUT values was identified. No correlations between pH and PUT, or on cooperation between official food control agencies and quality
between CAD and TRP values were identified. assurance systems of producers. Use of good hygienic practices
Histamine and cadaverine are the most important biogenic during manufacture of fish products is necessary for prevention of
amines for fish products. However, there were significant (p<0.05) high amine levels, together with good quality raw materials with low
variations in the biogenic amine contents of fish products. Most of total counts of contaminant microorganisms. Efforts should be made

Food Sci. Biotechnol.

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