International Journal of Science & Technology Volume 3, No 1, 59-65, 2008
Investigation of Histamine Level and Some Chemical Quality Parameters in Fish Consumed in Elazig City* 1
2
Özlem EMİR ÇOBAN1 and Bahri PATIR2
Department of Fish Processing Technology, Faculty of Fisheries, Firat Uni., Elazig, TURKIYE Department of Food Hygiene and Tech., Faculty of Veterinary Medicine, Firat Uni., Elazig, TURKIYE
[email protected] (Received: 03.12.2007; Accepted: 20.06.2008)
Abstract: In this study, histamine level and some other quality parameters have been examined in anchovy, horse mackerel, mackerel, rainbow trout, mirror carp and barbel consumed as fresh in Elazig. For this purpose, fish samples were obtained 15 day intervals between September 2003 and April 2004 and they were filleted. Histamine, TMA-N, TVB-N and pH values were examined in these fillets.Mean histamine level were determined as 24.24 ppm for anchovy, 26.49 ppm for horse mackerel, 25.81 ppm for mackerel, 12.21 ppm for rainbow trout, 11.27 ppm for barbel and 10.33 ppm for mirror carp. Histamine level was in marine fish in freshwater fish (p<0.05). In addition, the differences in histamin levels between the monts were significant (p<0.05).In the result of the statistical analyses showed that the TMA-N values determined from fish species were not important (p>0.05). An important difference was observed between TVB-N amount and fish species (p<0.05). An important differences was recorded between pH values of fish species (p<0.05).In conclusion, histamine level and other quality parameters were found under the suggested limits for various fish species (anchovy, horse mackerel, mackerel, rainbow trout, barbel and mirror carp) consumed as fresh in Elazig. So they have not any risk for human healty at this moment, but it is necessary to keep them under cold preservation conditions. Key Words: Histamine, TMA-N, TVB-N, pH, Fresh Fish
Elazığ’da Tüketime Sunulan Balıklarda Histamin Düzeyi İle Diğer Bazı Kimyasal Kalite Parametrelerinin Araştırılması Özet:Bu çalışmada, Elazığ’da taze olarak tüketime sunulan hamsi, istavrit, uskumru, gökkuşağı alabalığı, aynalı sazan ve bıyıklı balıklarda histamin düzeyleri ile diğer kalite parametreleri araştırıldı. Bu amaçla balık örnekleri Eylül 2003 ile Nisan 2004 tarihleri arasında onbeş günlük aralıklarla elde edildi. Çalışmada balıkların filetoları çıkarıldı ve Histamin, TMA-N, TVB-N, pH değerleri yönünden analiz edildi.Ortalama histamin miktarları, hamside 24,24 ppm, istavritte 26,49 ppm, uskumruda 25,81 ppm, gökkuşağı alabalığında 12,21 ppm, bıyıklı balıkda 11,27 ppm ve aynalı sazanda 10,33 ppm olarak tesbit edildi. Yapılan istatistiki analizde histamin değerlerinin deniz balıklarında daha yüksek olduğu (p<0,05) belirlendi. Ayrıca aylar arasındaki histamine ait belirlenen farklılıklar önemli bulundu (p<0,05). Balık türlerinde saptanan farklı TMA-N değerlerinin önemli olmadığı (p>0,05) görüldü. TVB-N miktarı bakımından ise balık türleri arasında önemli farklılıkların olduğu (p<0,05) tespit edildi. Balıklarda pH değerleri bakımından belirlenen farklılıkların önem arz ettiği (p<0,05) gözlemlendi. Sonuç olarak; Elazığ’da taze olarak tüketime sunulan farklı balık türlerinde (hamsi,istavrit, uskumru, aynalı sazan, gökkuşağı alabalığı ve bıyıklı balık) histamin miktarları ile diğer kalite parametrelerinin önerilen limitleri aşmadığı ve bu aşamada insan sağlığı için risk taşımadığı, ancak avlamadan sonra soğuk muhafaza şartlarının gerekli olduğu kanısına varıldı. Anahtar Kelimeler: Histamin, TMA-N, TVB-N, pH, Taze Balık
*This study was abstracted from thesis called Investigation of Histamine Level and Some Chemical Quality Parameters in Fish Consumed in Elazığ City .This study was supported by Fırat University Scientific Research Projects Coordination Office (FUBAP) as the Project number 786
O. Emir Coban and B.Patir
1.Introduction OFA (orthophtaldialdehyde) and fluorometrically determined [7]. Trimethylamine amount analysis was made according to Dyer[8]. It was extracted from the sample with TCA and all other bases except trimethylamine were fixed by formaldehyde. Then trimethylamine was extracted from the basic environment by toluene and was reacted by picric acid and result was read in spectrophotometer at 410 nm . TVB-N analysis was made according to Lucke and Gedel [8]. Homogenized sample was added magnesium oxide, volatile vapors were separated by water vapor distillation and distillate was titration was made with HCl in order to determine the amount of TVB-N. All statistical analyses were performed with Duncan’Multiple Range Test by using SPSS 12.0 computer program (SPSS Inc. Chicago,ıllinois).
In spite of the rapid population increase in the world, animal protein sources not increasing properly, causes the animal protein deficit per person to reach to serious levels. Production of fishery products should be increased because of their cost being low and supply being easy. For this reason, fishery products are relied on for solution of the hunger problem bound to food insufficiency [1]. In defection of fishery products, like in other foods, bacterial, autolitic and oxidative effects take place. Various chemical compounds form due to fish hunting, handling and marketing conditions [2]. Biogenic amines and bio-toxins form as a result of microbial activities in fish meat. Histamine which is very important among biogenic amines forms by an enzymatic decarboxilation catalyzed by histidine decarboxilase enzyme and L-histidine amino acid [3]. Histamine poisoning occurs when fishes are consumed which include high level of free histidine in their muscle tissues [4]. Poisoning symptoms occur between the first few minutes and a few hours after the histamine including food. The illness period is short and disappears within a few hours. However sometimes it may last for a few days [5]. In our country where 80-85% of the fishery products are consumed fresh, studies according to determine the level of histamine started recently. In this study made with the fishes that are consumed in Elazig, the histamine levels and also some chemical quality parameters such as TMA-N, TVB-N, pH were observed and determined if they comply to standards or not.
3.Results Histamine level and other chemical quality parameters of the consumed fish in Elazig are shown in table 3.1-3.2 and figure 3.1-3.4
2.Materials and Method The fishes that are used as materials in the study, were obtained from the fish bazaars in 15 day 7 intervals between September 2003 and April 2004. Skins, organs and head of the fishes that were brought to the laboratory were taken out and histamine, TMA-N, TVB-N and pH analyses were made with triple parallel. Histamine amount analysis was made according to the method mentioned by Lerke and Bell [6].Samples were extracted with TCA (Trichloraceticacid) and the extract was purified by weak acid cation changer and then derived by
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Investigation of Histamine Level and Some Chemical Quality Parameters in Fish Consumed in Elazig City
Table 3.1. The amount of histamine obtained from some marine and freshwater fishes (ppm) Months
Anchovy
Horse mackarel
Mackarel
Mirror carp
Rainbow trout
Barbel
September
31,94cy
33,17dz
32,48cdx
15,31bx
16,37br
13,20ar
October
30,10bcxy
27,72by
31,68cx
13,99ax
15,84ar
13,99ar
November
27,98crx
26,67cy
31,41dx
11,35ar
16,11br
10,83apr
December
24,82cr
22,97crx
27,46dr
9,77ar
13,99br
7,66ap
January
18,22cp
21,39epr
19,82dp
7,68ap
10,29bp
10,05bpr
February
17,82cp
21,12dpr
21,12dp
7,41ap
7,66ap
14,52br
March
24,55er
20,60cdp
21,12dp
7,39ap
9,24bp
19,82cx
April
25,08brx
24,03bx
21,37bp
10,30ar
8,20ap
13,20ar
a,b,c,d,e: The average values shown with different letters in same lines are statisticaly important (P<0,05). p,r,x,y,z:The average values shown with different letters in same columns are statisticaly important (P<0,05. Table 3.2. The amount of TMA –N, TVB – N and pH obtained some marine and freshwater fishes n Fish species Anchovy Horse mackarel
48 48
TMA-N(mg/100g)
TVB-N(mg/100g)
pH
x±Sx
x±Sx
x±Sx
2,29±0,53x
29,23±1,78yz
6,53±0,077y
(1,30-4,71)
(25,2-36,0)
(6,40-6,71)
2,18±0,69
x
(1,00-4,67) Mackerel
48
Mirror carp
48
Rainbow thout
48
1,98±0,68
x
29,23±2,35
yz
(25,2-35,0) 28,00±1,63
(6,31-6,78)
yz
48
6,44±0,054x
(0,90-3,60)
(26,6-33,6)
(6,23-6,56)
-
27,90±1,69y
6,45±0,056x
(25,2-36,0)
(6,38-6,74)
-
x
26,25±1,24
(23,0-33,6) Barbel
6,55±0,110y
-
29,89±,1,72 (25,2-34,2)
6,58±0,105y (6,31-6,78)
x
6,52±0,087y (6,27-6,75)
n: Analysis number ( ): Minimum and maximum values were showed in parahanthesiz x,y,z: The average values shown with different letters in same columns are statisticaly important (P<0,05).
61
O. Emir Coban and B.Patir
Anchovy
35
Horse mackarel Mackarel
Histamine level (ppm)
30
Mirror carp
25
Rainbow trout Barbel
20 15 10 5 April
March
February
January
December
November
September
October
0
Months
Figure 3.1. The amount of histamine monthly measured in marine and freshwater fishes Anchovy
3,5
Horse mackarel
TMA-N (mg/100g)
3
Mackarel
2,5 2 1,5 1 0,5 April
March
February
January
December
November
October
September
0
Months
Figure 3.2. The amount of TMA - N monthly measured in marine fishes
Anchovy
TVB -N (mg/100g)
35
Horse mackarel
30
Mackarel
25
Mirror carp
20
Rainbow trout
15
Barbel
10 5 April
March
February
January
December
November
October
September
0
Months
Figure 3.3. The amount of TVB -N monthly measured in marine and freshwater fishes
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Investigation of Histamine Level and Some Chemical Quality Parameters in Fish Consumed in Elazig City
Anchovy
6,8
Horse mackarel
6,7
Mackarel Mirror carp
6,6
Rainbow trout
pH
6,5
Barbel
6,4 6,3 6,2 April
March
February
January
December
November
October
September
6,1
Months
Figure 3.4. The amount of pH monthly measured in marine and freshwater fishes
4.Discussion kinds of fishes that have white meat (rainbow trout, barbel and carp), are parallel with this study. Regarding TMA-N, levels that are obtained from anchovy, mackerel and horse mackerel are not over the above mentioned limits. The TMAN levels of the samples were found between 1,3 –3,49mg/100g. In the result of the statistical analyses showed that the TMA-N values determined from fish species were not important (p>0.05). Regarding TMA-N, tolerable limits are vary in the literature. According to Krzymıen and Elias TMA-N limit values for fishery products are good up to 4 mg/400g, sellable up to 10 mg/100g and spoiled up to 12 mg/100g [15]. According to Karnop and coworkers, tolerable limit for TMA-N is 15 mg/100g [16]; according to Stockemer and Nieper tolerable limit for TMA-N is 12 mg/100g [17]. Schormüller, states the TMA-N limit value standard is 10-15 mg/100g [8]. The average TVB-N values of the samples were between 25mg/100g – 34 mg/100g and they were all below 35 mg/100g. An important difference was observed between TVB-N amount and fish species (p<0.05). Fishery products classification according to TVB-N is “very good” up to 25 mg/100g,
In this study the consumed fishes in Elazig between September 2003 and April 2004 were observed according to some quality parameters. Histamine level was found to be 24,24 ppm in anchovy, 26,49 ppm in horse mackerel, 25,81 ppm in mackerel, 12,21 ppm in rainbow trout, 11,27 ppm in barbel and 10,33 ppm in carp. It was observed that histamine level decreases by decreasing temperature in winter.The level of histamine in marine fish was found higher than that of freshwater fish (p<0.05). In addition, the levels histamin in all fish species were different between monts (p<0.05). Average histamine level in anchovy was found 13,32 ppm by Galarini and coworkers [9], 0,50 ± 0,55 ppm by Veciana – Nogues and others [10], 1 ppm by Mahendratta and Schwedt [11], and 24,97 ppm by Vidal – Carou and coworkers [12]. The results of Vidal – Carou and coworkers [12] are suitable with the results in this study; the other study results are different. Histamine level in mackerel was found 16,17 ppm by Galarini and coworkers [12] and 17,93 ppm by Çıtak [13] These levels are much lower than the levels obtained in this study which is 25,81 ppm. The study results of Edmunds and Eitenmiller [14], that are made with various
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O. Emir Coban and B.Patir
3. URL 1, 2002. Trouble times. Histamin. FDA Center For Food Safety And Applied Nutritionid, http : // vm.cfsan.fda.gov / mow / chap 38.html. 4. URL 2, 2002. Scombroid poisoning. Clemson Extension. Home and Garden Information. http : // hgic.clemson.edu / factsheets / HG / C 3362.html. 5. URL 3, 2002. Fish poisoning. General Travel Center, http // www. Travel healt.com / article / ? aid =38 and cid = 1. 6. Lerke, P.A. and Bell, L.D., 1976. A Rapid fluorometric method for the determination of histamine in canned tuna. Journal of Food Sci. Vol. 41. 7. Ata, U., 1999. Deniz Ürünlerinde Histamin Düzeyinin Araştırılması. İstanbul Üniversitesi, Sağlık Bilimleri Enstitüsü, Farmakoloji ve Toksikoloji Anabilim Dalı. Doktora Tezi. İstanbul. 8. Schormüller, J., 1968. Handbuch der Lebensmittelchemie Band III/2 Teil. Springer – Verlag Berlin Heidelberg, Newyork. 1482 – 1537 p. 9. Galarini, R., Hoovet, M.N. and Manuali, E., 1996. Heavy metals and histamine content during 1988 – 1995 period. İndustric Alimentari. 35(353): 1194 – 1198 (Abst). 10. Veciana – Nogues, M.T., Hernandez – Jover, T., Marine – Font, A. and Vidal – Carou, M.C., 1995. Liquid chromatographic method for determination of biogenic amines in fish and fish products. Journal of the AOAC İnt., 78(4): 1045 – 1050. 11.Mahendratta, M. and Schwedt, G., 1998. Sample preparation for photometric determination of histamine in foodstuffs compared with capillary electrophoresis. Z Lebensm Unters Forsch A, 206: 246 – 250 12.Vidal – Carou, M.C., Veciana – Nogues, M.T. and Marine – Font, A., 1990. Spectrofluorometric determination of histamine in fish and meat products. J. AOAC., 71(4): 565 – 567. 13.Çıtak, C., 1994. Uskumru (Scomber scombrus) Balığında Histamin Oluşumu Üzerine Depolama Sıcaklığı ve Süresinin Etkisi. Ankara Üniversitesi, Fen Bilimleri Enstitüsü, Gıda Mühendisliği Anabilim Dalı. Yüksek Lisans Tezi. Ankara. 14.Edmuns, W.J. and Eitenmiller, R.R., 1975. Effect of storage time and temperature on histamine content and histidine decarboxylase activity of aquatic species.J. Food Sci., 40: 516 – 519. 15.Krzymıen, M.E. and Elias, L.,1990.Feasibility study on the determinetion of fish freshness by trimethylamine headspace analysis. J. of Food Sci. Vol.55,No: 5. 16.Karnop, G., 1988. Histamine in salzsardellen.Archiv für Lebensmittelhygiene 39:57 – 84.
“good” up to 30 mg/100g, “sellable” up to 35 mg/100g and “spoiled” over 35 mg /100g’dan [16]. Although Karnop and coworkers mentions the tolerable limit values as 40 mg/100g Krzymıen and Elias, assumes the 35 mg/100g TVB-N level as “sellable” and 40 mg/100g TVB-N value as “spoiled” [17]. In a study made in the region, [18] TVB-N average level in the consumed carp fillets were found to be 11.67 mg/100g The reason of such a low level might be the analysis being made right after hunting of the fish. In the study, pH values were found 6,41 – 6,66 mg/l for anchovy, 6,45 – 6,77 mg/l for horse mackerel and 6,37 – 6,51 mg/l for mackerel. Regarding fresh water fishes, the values were found 6,43 – 6,74 for rainbow trout, 6,3 – 6,60 for barbel and 6,39 – 6,54 for mirror carp. An important differences was recorded between pH values of fish species (p<0.05). Although Çıtak determined the average pH value of mackerel as 6,24 – 6,34 mg/l, this value is 6,37 – 6,51 mg/l in this study. The reason of the high pH value in this study may be storing period and season temperature. Histamine, TMA-N and TVB-N values were found to be below the consumable limits for the fishes in Elazig such as anchovy, horse mackerel, mackerel, rainbow trout, mirror carp and barbel. However, hygienic rules must be applied in order to decrease contamination during hunting of anchovy, horse mackerel and mackerel which have tendency to form histamine. Suitable storage conditions should also be provided. 5.Acknowledgements We would like to present many thanks to Fırat University Scientific Research Projects Coordination Office (FÜBAP) supporting this study as the Project number 786. 6. References 1. Yılsay, K., Yücel, A., 1997. Ton konservelerinin kimyasal özelliklerinin belirlenmesi ve TS 355’e uygunluğunun araştırılması. Vet. Hek. Der. Derg., 68 (1),55 – 58. 2. İnal, T., 1992. Besin Hijyeni-Hayvansal Gıdaların Sağlık Kontrolü. Genişletilmiş ikinci baskı. Final Ofset A.Ş. İstanbul.
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Investigation of Histamine Level and Some Chemical Quality Parameters in Fish Consumed in Elazig City
17.Stockemer, J., Nıeper, L., 1984. Parameter zur beurteilung des verderbs von nordsee krabben (Crangan crangon) Archiv lebensmittelhygiene. 35: 5-7. 18.Patır, B., Gürel, A., Ateş, G., Dinçoğlu, A.H., 2001. Potasyum sorbat uygulanmış aynalı sazan
(Cyprinus carpio) filetolarının üretimi ve muhafazası sırasında meydana gelen mikrobiyolojik ve kimyasal değişimler üzerine araştırmalar. Vet. Bil. Derg. 17(2): 31- 44.
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