A STUDY ON HISTAMINE LEVEL IN SOME KINDS OF LOCAL AND IMPORTED CHEESE

Dept of Food Hygiene,

Port-Said Laboratory, Animal Health Research Institute, Dokki, Giza.

A STUDY ON HISTAMINE LEVEL IN SOME

KINDS OF LOCAL AND IMPORTED CHEESE

(With 2 Tables)

By

Marcel F. Galab

(Received at 2/4/2009)

دراسة عن مستوى الهستامين فى بعض انواع من الجبن المحلى والمستورد

مارسيل فخرى جلاب

تم فى هذا البحث تجميع 120 عينة عشوائية من الجبن ( ايدام , شيدر, ريکفورت) محلى ومستورد على التوالي من السوبر مارکت فى مدينة بورسعيد لقياس نسبة الهستامين فيها. وبلغت نسبة متوسطات الهستامين 3.84 مجم نسبة مئوية و5.19 مجم نسبة مئوية و6.50  مجم نسبة مئوية فى الجبن محلى الصنع (ايدام وشيدر وريکفورت) على التوالى. بينما بلغت نسبة الهستامين فى الجبن المستورد 3.23 مجم نسبة مئوية و4.33 مجم نسبة مئوية و6.14 مجم نسبة مئوية فى الجبن الايدام والشيدر والريکفورت على الترتيب. ويمکن القول ان نسبة الهستامين فى عينات الجبن المصنع محليا تزيد زيادة طفيفة عن نسبة الهستامين فى عينات الجبن المستورد وتعتبر نسبة الهستامين احد الدلائل على سوء تخزين هذه الأطعمة. هذا وقد تم مناقشة أهم التوصيات لمنع تکوين الهستامين.

SUMMARY

A total of 120 random samples of local and imported cheese(Edam,` Cheddar, and Roquefort cheese) were screened for determination of histamine level. The obtained results revealed that the mean values of histamine content in these samples were 3.84 ± 0.58 mg %, 5.19 ± 0.80 mg% and 6.50 ± 0.97 mg% in local Edam, Cheddar and Roquefort cheese, respectively. Meanwhile, the mean values of histamine were 3.23 ± 0.48 mg%, 4.33 ± 0.72 mg %, and 6.14 ± 0.94 mg %, in imported Edam, Cheddar and Roquefort cheese samples, respectively. It was noticed that the examined local Edam, Cheddar and Roquefort cheese samples contained level of histamine higher than imported cheeses. The public health significance and recommendation for prevention of the formation of histamine and other biogenic amines were discussed.                            .                                                               

Key words: Biogenic amines; histamine; cheese; E dam, Cheddar and Roquefort cheese

INTRODUCTION

Histamine is one of the biogenic amines that belongs among the naturally occurring substances (Stratton et al., 1991). Biogenic amines such as histamine, tyramine, putrescine, and others can be formed in foods as a result of metabolic processes of microorganisms (Veciana-no-gues et al., 1997). Histamine is one of them which is formed in foods largely from growth of microorganisms that possess the histidine decarboxylase enzyme (Arnold and Brown, 1978). This enzyme converts histidine to histamine via an enzymatic decarboxylation reaction (Taylor, 1986 and Fletcher et al., 1998). Numerous genera of bacteria have been reported as capable for amine production (Tyramine, Histamine and Tryptoamine). These are Escherichia, Enterobacter, Salmonella, Shigella, Acromobacter, Pseudomonas, Alcaligenes, Proteus, Clostridium perfringes, Micrococcus, Streptococcus, Lactobacillus and Leuconostoc (Rice et al., 1976 and Voigt and Eitenmiller, 1977).

Histamine and other amines are formed by the growth of certain bacteria and the subsequent action of their enzymes on certain amino acids in foods, either during the production of a product such as Swiss cheese or by bacterial spoilage of foods. Raw milk has very little free histidine but milk protein may contain 9.69 gm of histidine per 100ml (Jenness and Patton, 1976).Likewise, Taylor (1985) suspected that proteolysis plays an important role in histamine formation in cheese, since milk does not contain large quantities of free histidine. Proteolysis either autolytic or bacterial may play a role in the release of free histidine from tissue protein.                                        

Histaminepoisoningsometimes called Scombroid poisoning.                      Histamine poisoning is manifested by a wide variety of symptoms.     The characteristic symptoms affecting the cutaneous system include rash, urticaria, edema and localized inflammation (Taylor, 1986).

Several outbreaks of histamine poisoning have occurred after the consumption of cheese containing high levels of histamine(Chambers and Strusskiewiez, 1978 and Edwards and Sandine, 1981). The toxicity of histamine appears to be enhanced by the presence of other biogenic amines found in foods that can inhibit histamine enzymes in the small intestine (Stratton et al., 1991).

        The histamine is of great importance as health hazard especially with sensitive human and children. Therefore, the present work was conducted to observe levels of histamine content in some kinds of local and imported cheese.                                                                   

MATERIALS and METHODS

1- Collection of samples:

A total of 120 random samples (20 each) of Edam, Cheddar and Roquefort cheese (local and imported) were collected from different Port- Said city markets in an ice box and were directly transferred to the laboratory as possible for estimation of histamine level  in these samples.                            

2- Determination of histamine contents:

The histamine analysis was carried by Thin Layer Chromatography (TLC) method according to Su et al. (2000). 10gm of cheese were homogenized in the homogenizer vessel with approximately 70 ml of methanol for 1 min., decanted into 100 ml conical flask and placed in 60 °C water bath for 15 min. The flask was removed, allowed to cool and made up with methanol to a final volume of 100 ml, mixed thoroughly and filtered. The filtrate was used to spot directly on the TLC plate. The plates were activated at 105°C for 30 min to 1 hr and allowed to cool before spotting with standards histamine dihydrochloride and unknown. A pencil line was first drown along the TLCplates at least 20 mm from the bottom. The position of the unknowns and standards were then marked at approximately 20 mm intervals on the line. To each mark, the appropriate aliquot of unknown and standards (e.g 20ul) was carefully added in small amounts, to ensure keeping the spot to a minimum size. The prepared plate was carefully placed in the equilibrated tank and left to run until the solvent front reached to within 20 mm of the top edge. The plate is then removed and dried thoroughly in an oven at 100°C. It is then cooled and sprayed with ninhydrin. The spots can be developed by heating at 105°Cfor 2-3min. but the best results are obtained by leaving the color to develop slowly in a dark place for overnight. Presence of histamine could be detected by matching the unknown and standards color spots. Quantitative determination of histamine was done according to(EOS, 1990) by eluting the spot from the plate by spatula, dissolving in known amount of methanol and filter by means of spectrophotometer (at 570 nm). A standard curve of various concentrations of histamine was drawn. From the standard curve, histamine levels of the samples were calculated.                

RESULTS

Table 1: Frequency distribution of histamine contents in the examined local and imported cheese samples. n=20                        

NB: The value in (%) are related to number of samples.

Table 2: Statistical analytical results of histamine contents in the examined local and imported cheese samples.                       

Max. Maximum values                      Min. Minimum values        

S.E. Standard Error                            S.D. Standard deviation

DISCUSSION

             Table 1 illustrates the frequency distribution of histamine contents in examined local and imported cheese samples. The results revealed that 7 (35%) and 6 (30%) samples of local and imported Edam cheese, respectively had histamine contents at frequency range of 0-2 mg/100gm cheese. 10(50%) + 1(5%), local Edam and Cheddar cheese samples and 13(65%) + 6 (30%) imported Edam and Cheddar cheese samples had histamine contents at frequency range of >2-4 mg/100gm cheese, respectively. While 3 (15%), 15 (75%), 5 (25%) samples of local Edam, Cheddar and Roquefort cheese and 1 (5%), 14 (70%), 8 (40%) samples of imported Edam, Cheddar and Roquefort cheese, respectively, had frequency range of histamine levels at range of 5mg %. 4(20%), 15(75%) of local Cheddar and Roquefort cheese samples, respectively and 12(60%) of imported Roquefort cheese samples had histamine contents at frequency range of >5-10 mg %. The present results reflected that all examined samples had histamine levels. Likewise, Mohamed and Thakeb (2004) could be detected histamine in examined samples. In this respect, Swiateck and Kisza (1959)stated that, histamine amount of 50-100 mg/100 gm Roquefort cheese is considered harmful level. On the other hand the present results in Table 2 showed that, the mean value of histamine content of local Cheddar cheese was slightly higher than the level of histamine in imported Cheddar cheese (5.19 mg /100 gm cheese ± 0.80 and 4.33 mg /100 gm cheese ± 0.72, respectively). The relatively high concentration of histamine in the local Cheddar cheese than imported one may be attributed either due to processing technology faults or due to the problem of storage at supermarkets and at custom level. However, lower content of histamine level in Cheddar cheese was reported by Silverman and Kosikowski (1956). While the histamine contents of local and imported Edam cheese samples had nearly similar mean values which were 3.84 mg% ± 0.58, and 3.23 mg% ± 0.48, respectively. Finally local and imported Roquefort cheese samples had also similar mean counts of histamine contents which were 6.50 mg% ±0.97 and 6.14 mg% ± 0.94, respectively which had agreed with the results recorded byMohamed and Thakeb (2004). In this concern Taylor et al. (1982) stated that the histamine poisoning occurred in Swiss cheese in one case contained 187 mg% histamine; the symptoms associated with the outbreak occurred in 30 minutes to one hour after ingestion.Graf (1992) reported that the formation of histamine correlated closely with the microbiological quality of the original milk especially with the content of Gram- negative bacteria and Enterobacteriaceae. Moreover Edwards and Sandine (1981) mentioned that higher histidine content in marked cheese ranged from 1.5-12 mg/100gm cheese may be attributed important factor which may be contributed to elevated biogenic amines as histamine. This concentration of substrate would provide non-toxic quantities of corresponding amines indicating that cheese proteolysis is important when toxic amount occurs. Other factors may be present as presence of capable bacteria or enzymes availability of substrate, presence of co-factor, or existence of a proper environment in the cheese as influenced by pH, temperature, salt and water availability.

              It is extremely difficult to determine the exact threshold of toxicity for amines because most cheeses do not contain histidine-decarboxylating bacteria and because the free histidine concentration in cheese is usually low, histamine levels mostly do not exceed 100mg /kg(Joosten, 1988) since the toxic dose will depend in large measure on the efficiency of each individuals all detoxification mechanisms and the presence of potentiators (such as other biogenic amines like cadaverine and putrescine). Legislation in this area is practically non-existent. However, data obtained from the reported cases of food poisoning indicate that a level of 500 to 1000 mg /kg is potentially dangerous (Han et al. 1996). The best way to control of histamine concentration in the cheese probably is to decrease or to limit the numbers of histamine producing bacteria present in raw milk (Reinbold, 1972). Furthermore the recommendation for prevention the formation of the histamine and other biogenic toxic amines may be summarized in the following steps: the application of HACCP system in dairy products processing plants, good storage condition at custom level & marketing and personal hygiene of customer.                                                                              

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Chambers, T.L. and Strusskiewiez, W.F. (1978): Fluorometeric              determination of histamine in cheese. J. Assoc. of Anal. Chem. 61: 1092-1097.                                                                                                                                                                      

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