Non-volatile Amines Research Articles

Monitoring Volatile and Nonvolatile Amines in Dried and Salted Roes of Tuna (Thunnus thynnus L.) during Manufacture and Storage

Dried and salted roe, obtained from the reproductive organs of female tuna (Thunnus tynnus L.), is a typical fish-based food in the Mediterranean area of Spain. In the present study, we monitored the formation of volatile amines (trimethyamine nitrogen [TMA-N] and total basic volatile nitrogen [TBVN]) and nonvolatile amines (biogenic amines) in dried and salted tuna roe after processing and storage for 8 weeks at 4, 20, and 30°C. The salting and drying process significantly increased the TBVN, cadaverine, tyramine, phenylethylamine, and tryptamine contents, and bacteria with histamine decarboxylase activity were detected both in raw and in dried and salted tuna roes. During storage of tuna roe, TMA-N and TBVN levels increased significantly after the fourth week of storage at 30°C, whereas biogenic amine contents remained more or less constant. However, samples stored at 30°C showed histamine formation after the first week of storage, with a concentration of <50 ppm. The volatile and nonvolatile amine concentrations in tuna roe were below the consumer safety limit, with the exception of the total biogenic amine level in roe stored at 30°C, which exceeded the European Community's recommended limit (300 ppm). These results indicate that in properly stored tuna roe, histamine formation will not represent a serious health risk to consumers unless the tuna roe has previously been mishandled.

Production of tyramine in "moromi" mash during soy sauce fermentation

The concentrations of 7 non-volatile amines, tyramine (Tym), histamine (Him), phenethylamine (Phm), putrescine (Put), cadaverine (Cad), spermidine (Spd) and spermine (Spm) in the liquid part of "moromi" mash during soy sauce fermentation were studied. These amines, except for him and Cad, were detected during fermentation by the conventional production method in the laboratory. Put and Spd were detected at the beginning, and Tym, Phm and Spm appeared later; these 5 amines increased gradually during the fermentation. Put, Spd, Spm and Cad were present in the raw starting material for soy sauce; thus, Tym and Phm were produced by the fermentation. When "moromi" mash was added to liquid medium and cultivated, Tym was detected in some "moromi" mash and the other amines were not detected. Tym-producing bacterial strains were isolated from the liquid culture media of Tym-positive "moromi" mash. The Tym-producing strain was a gram-positive coccus. The conditions for production of amines by Tym-producing bacterial strains were examined. These strains grew and produced tyramine under various conditions, which may occur during soy sauce fermentation. Namely, Tym was produced at pH 5-10, at salt concentrations of less than 8%, under either aerobic or anaerobic conditions. During soy sauce fermentation, it is assumed that Tym would be produced by these strains during the early stages of soy sauce aging within a short period when the salt concentration and pH conditions are optimal for growth. Based on the bacteriological properties, the strains were identified as Enterococcus faecium. With the exception of Phm and Him, which did not exist in the starting raw material, non-volatile amines (including Put, Cad, Spd and Spm) were not produced and microorganisms producing them are not believed to be present during "moromi" fermentation.

Content of biogenic amines in a Chardonnay wine obtained through spontaneous and inoculated fermentations.

This paper describes the content of biogenic amines in wines obtained from a Chardonnay must inoculated with different strains of Saccharomyces cerevisiae and in a wine fermented with the indigenous yeasts (control wine). The concentrations of nonvolatile amines and phenethylamine in the wines from the inoculated must were superior to those of the control wine. This was probably due to the fact that consumption of the precursor amino acids of these amines, during fermentation, was also greater in the inoculated samples than in the control sample. Furthermore, from the results obtained it may be said that, at least to some extent, the nonvolatile amines were formed by yeasts during fermentation. The concentrations of dimethylamine, ethylamine, and pyrrolidine (volatile amines) were different for the different wines, although they did not reach concentrations sufficiently high to have any effect on the aroma.

Volatile and Nonvolatile Amines in Mediterranean Hake as Function of their Storage Temperature

ABSTRACT: The main amines in frozen and fresh hake (Merluccius merluccius) are the natural polyamines, spermidine. The levels of volatile and nonvolatile amines during storage of hake were strogly dependent on temperature. In samples stored at ‐20° only changes in dimethylamine and agmatine were found, whereas major changes were observed in samples stored at higher temperatures. Cadaverine was the major amine formed followed by histamine, putrescine and tyramine. The maximum levels of biogenic amines were lower than those reported for other fish species. Trimethylamine and biogenic amines related to hake spollage showed levels higher in fresh than in frozen samples, whereas levels of natural polyamines were not statistically different.

DETERIORATIVE CHANGES DURING ICE STORAGE OF IRRADIATED BLUE JACK MACKEREL (TRACHURUS PICTURATUS)

Changes in the content of histamine (Him), tyramine (Tyr), agmatine (Agm), spermidine (Spd), putrescine (Put), cadaverine (Cad) and volatile basic nitrogen (VBN) were examined during the ice storage of blue jack mackerel irradiated at different levels (0, 1, 2 and 3 kGy). The variation of these amines in white and dark muscle offish irradiated with 0 and 3 kGy was also investigated. The levels of Him, Tyr, Put, Cad and VBN increased with storage time and their production was significantly reduced by irradiation of fish. Him content in the control lot exceeded the maximum allowed in fresh fish (100 g/kg) after 7 days, while in the irradiated lots a maximum value of 54.16 mg/kg of Him was determined after 23 days. No significant differences were determined in nonvolatile amine production in samples irradiated with 1, 2 and 3 kGy. Although there was a correlation between fish quality deterioration and both Him and other amine production in irradiated fish, the highest values detected in spoiled fish were considerably lower than the maximum levels allowed. Thus, since the production level of Him and other nonvolatile amines is irradiation dependent the use of these compounds as quality indices may not be appropriate. Nevertheless, it is evident that irradiation, even at the lowest level used, extends the ice storage shelf-life of blue jack mackerel.

Use of carbon dioxide enriched atmospheres in the refrigerated storage (2 °C) of salmon (Salmo salar ) steaks

Selected microbial counts (total viable microbiota, Brochothrix thermosphacta, lactic acid bacteria and Enterobacteriaceae), biochemical parameters [pH, total volatile nitrogen, nucleotide breakdown products, non-volatile amines, d(–) and l(+)-lactic acids and short-chain fatty acids] and sensory attributes (colour and odour) of refrigerated (2 °C) salmon (Salmo salar) steaks stored under CO2-enriched [CO2/air (20/80, v/v), CO2/air (40/60, v/v)] and air atmospheres were determined. When compared with air, sensory results showed shelf-life extension of 6 days and 15 days for 20% and 40% CO2-enriched atmospheres, respectively. Microbial and biochemical results also revealed that the 40% CO2-enriched atmosphere was the most effective packaging type for refrigerated salmon.

Microbial and physicochemical modifications of hake (Merluccius merluccius) steaks stored under carbon dioxide enriched atmospheres

Microbial counts (total viable, Brochothrix thermosphacta, lactic acid bacteria and Enterobacteriaceae), biochemical parameters [pH, total volatile nitrogen, nucleotide breakdown products, non-volatile amines, D(−) and L(+) lactic acid and short-chain fatty acids] and sensory attributes (colour and odour) of refrigerated (2 °C) hake (Merluccius merluccius) steaks under carbon dioxide enriched air (CO2 /air, 20/80 v/v and CO2 /air, 40/60 v/v) and air atmospheres were determined. When compared with air, sensory results showed shelf-life extension of 4 and 11 days for 20% and 40% CO2-enriched atmospheres, respectively. Microbial and biochemical results also revealed that the 40% CO2-enriched atmosphere was the most effective packaging type for refrigerated hake. © 2000 Society of Chemical Industry

The formation of non-volatile amines in relation to concentrations of free basic amino acids during postmortem storage of the muscle of scallop ( Pecten maximus), herring ( Clupea harengus) and mackerel ( Scomber scombrus)

The concentrations of the non-volatile biogenic amines that form during storage of the adductor muscle of scallop ( Pecten maximus ) and the skeletal muscles of mackerel (Scomber scombrus) and herring ( Clupea harengus ) were determined by high-performance liquid chromatography, using postcolumn derivatisation with o-phthalaldehyde. Initially, concentrations of amines were low in all muscles but, as time of storage progressed, the concentration of agmatine increased steadily in scallop muscle reaching a level of 100 mg per 100 g wet weight after 10 days of storage in ice or 3 days at 10°C. In mackerel and herring only slight increases were observed, with levels of less than 1.0 mg after 10 days in ice or 2 days at 10°C. In scallop, arginine, the precursor of agmatine, decreased steadily in concentration. In herring muscle, histidine, the precursor of histamine, decreased considerably in concentration, whereas in mackerel it changed only slightly. © 1997 Published by Elsevier Science Ltd

Biogenic Amines in Fish and Shellfish

This chapter focuses on biogenic amines in fish and shellfish. The term “biogenic amines” refers to the nonvolatile amines, such as cadaverine, putrescine, spermidine, spermine, tyramine, tryptamine, and histamine, produced post mortem in fish and shellfish products. The compounds are reported to originate from the decarboxylation of specific free amino acids in fish or shellfish tissue. Chemically, the biogenic amines are defined as low-molecular-weight aliphatic, alicyclic, or heterocyclic organic bases. The use of chemical compounds as objective product standards or the indices of quality has long been suggested because these tests are rapid when compared to traditional microbiological analyses and less subject to individual interpretation than sensory analyses. Moreover, clinical studies have considered the activity of diamine oxidases to be a useful parameter, thus, many different assays to determine the activity of diamine oxidases have been proposed. Procedures using radioactive substrate achieve high sensitivity and most recently the use of high-performance liquid chromatography has been described.

Effect of Carbon Dioxide and Oxygen Enriched Atmospheres on Microbiological and Chemical Changes in Refrigerated Tuna (Thunnus alalunga) Steaks

Bacterial populations and several physicochemical parameters (metmyoglobin, pH, total volatile nitrogen, nucleotide breakdown products, nonvolatile amines, lipid autoxidation, and short-chain fatty acids) and sensory attributes (odor and color) were measured in refrigerated tuna steaks stored at 2 o C in air, CO 2 /air (20/80) (v/v), CO 2 /air (40/60) (v/v), and CO 2 /O 2 (40/60) (v/v) atmospheres. The effects of modified atmospheres on bacterial growth and physicochemical parameters are discussed. From a microbiological point of view, the shelf life of refrigerated tuna steaks was extended about 1 week when CO 2 enriched atmospheres were used. Metmyoglobin formation was minimum when the atmosphere was enriched with O 2 . Therefore, the most effective atmosphere was CO 2 /O 2 (40/60) (v/v), in which the shelf life of tuna may be doubled with respect to both microbial condition and color appearance

Changes in Nitrogenous Components and Bacterial Counts of Fermented Sardine with Rice-Bran Produced by Warmed Brewing.

FSR, “Iwashinukazuke,” is a fermented fish product which takes over 6 months for ripening. The warmed brewing method is able to shorten the ripening period to 100 days. This study aims at obtaining fundamental qualitative knowledge of FSR using warmed brewing (30°C, relative humidity; 80%). The initial moisture (55.8%), NaCl content (31.2%) and pH (5.79) of FSR decreased to 51.0%, 26.3% and 5.20 after 100-days warmed brewing, respectively. TMA-N, VBN, NPN/TN and Amino-N/TN increased and the non-volatile amines of the FSR also increased during the warmed brewing. TBC, MYC and LAC at 100 days reached 8.7×107, 1.7×105 and 3.0×106, respectively. A sensory evaluation indicated that there was no particular difference except for color (rice-bran and meat) and flavor between the warmed FSR and the commercial FSR.

Source of Putrefactive Non-Volatile Amines and Volatile Basic Nitrogen Detected in Kusaya Products

Source of Putrefactive Non-Volatile Amines and Volatile Basic Nitrogen Detected in Kusaya Products

Studies on Halotolerant and Halophilic Histamine-Forming Bacteria. Part 4. Changes in the Number of Halotolerant Histamine-Forming Bacteria and Contents of Non-Volatile Amines in Meat during Processing of Fermented Sardine with Rice-Bran.

This study was carried out to determine whether or not halotolerant and halophilic histamine (Hm)-forming bacteria (HH bacteria) produce non-volatile amines in meat during the traditional processing of fermented sardines with rice-bran (FSR). HH bacteria in FSR for 6 months increased in a number to more than 104/g. Putrescine, Hm and tyramine content markedly increased in the meat of FSR after 6 months. The isolates of HH bacteria from the raw sardine were identified as Staphylococcus, Micrococcus, Vibrio, Pseudomonas III/IV-NH and Pseudomonas III/IV-H. The isolates from FSR after 6 months were identified as Staphylococcus, Micrococcus and Vibrio.

Determination of Putrefactive Non-Volatile Amines in Foods Following Sample Cleanup by Solid Phase Extraction.

An analytical method for the determination of putrescine (Put), cadaverine (Cad), histamine (Him), tyramine (Tym), and spermidine (Spd) in foods by high performance liquid chromatography was described. The five amines were extracted from fish, fish products, miso, soy sauce, wine and sake with 1% trichloroacetic acid. After 0.1 M sodium octanesulfonate was added to the extract, the solution was passed through a Sep-Pak Vac C18 cartridge column. The column was washed with water, and the amines were eluted with a mixture of methanol - water (6 : 4). The amines in the eluate were converted into the corresponding dansyl amines by reaction with dansyl chloride under alkaline conditions. The dansyl amines were separated on a Cosmosil 5C18-AR column with a mobile phase of acetonitrile-water (65 : 35) and detected with fluorescence detector (Ex. 325 nm, Em. 525 nm). Recoveries of the five amines were more than 79.4%. The detection limit of Put, Cad and Spd in samples were 1 μg/g, Tym was 5 μg/g, and Him was 20 μg/g.

Studies on Halotolerant and Halophilic Histamine-Forming Bacteria-III. Changes in the Number of Halotolerant Histamine-Forming Bacteria and Contents of Non-Volatile Amines in Sardine Meat with Addition of NaCl.

This study was carried out to investigate whether or not halotolerant histamine (Hm)-forming bacteria could produce Hm and some non-volatile amines in sardine products containing a high concentration of NaCl. The total number of Hm-forming and halotolerant bacteria and the contents of non-volatile amines of each sample were measured in composite samples of sardine with the addition of 2 or 12% NaCl. Hm, cadaverine (Cad), and tyramine were not detected in any of the samples before storage. The contents of Cad and Hm increased rapidly during storage at 30°C in the samples with 2% NaCl added. Hm, putrescine, and Cad were formed during storage at 30°C in the sample with 12% NaCl added, in which the number of halotolerant Hm-forming bacteria varied from less than 10/g to 104/g. It is suggested that halotolerant Hm-forming bacteria producenon-volatile amines in sardine meat with 12% NaCl added.

海産魚における耐塩性及び好塩性ヒスタミン生成菌の分布ならびに同菌による不揮発性アミン生成

海産魚における耐塩性及び好塩性ヒスタミン生成菌の分布ならびに同菌による不揮発性アミン生成

Non-volatile Amine Production in Mackerel Muscle during Growth of Different Bacterial Species

The production of various non-volatile amines by Morganella morganii, Klebsiella planticola, Enterobacter aerogenes and Hafnia alvei in the dark and white muscle extracts of mackerel and in intact muscle was investigated during storage at 20°C. The amines were extracted with 2% perchloric acid and quantified by an HPLC method. The major amines produced were putrescine, cadaverine and histamine. The amine levels in the white muscle extracts were higher than those in the dark muscle extracts. All the bacterial species tested produced histamine in large quantities except H. alvei, which mainly produced cadaverine in the muscle extracts together with a little histamine. M. morganii did not produce any cadaverine in either of the extracts during storage. In the intact muscles, inoculated with M. morganii and/or E. aerogenes, there was an increase in amine levels followed by a gradual decrease. The results show that the strong histamine producers are also capable of producing the other amines in considerable amounts, and the rate and the type of the amine produced depend on the bacterial species, some of which show amine-degrading ability, especially during prolonged storage of the fish.

Formation of Tyramine and Histamine during Soybean Paste (Miso) Fermentation.

The concentrations of seven non-volatile amines, tyramine (Tym), histamine (Him), phenethylamine (Phm), putrescine (Put), cadaverine (Cad), spermidine (Spd) and spermine (Spm), during miso fermentation were studied. Tym of these amines which is generally known to be present in miso was detected during fermentation by the conventional production method. Him and Phm were not detected, and concentrations of Put, Cad, Spd and Spm, which were present in the raw starting material for miso, did not change significantly during fermentation. However, Tym and Him were produced when the miso starting materials, i. e., cooked soybeans, rice koji and mashed miso, were added to liquid medium and allowed to stand. Tym- and Him- producing bacterial strains were isolated from the liquid culture media containing the miso starting materials. The Tym-producing strain was a gram-positive coccus, and the Him-producing strain was a gram-positive rod. Thus, the results show that the Tym and Him observed in culture were produced by the two different species. During miso fermentation, Tym and Him are apparently produced within a short period of time when the respective optimum conditions for their growth are present. With the exception of Phm which did not exist in the miso starting materials, other amines including Put, Cad, Spd and Spm were not produced and microorganisms producing them are not believed to be present during miso fermentation.

Non-volatile Amine Contents of Commercial Rice-Bran Pickles of Sardine

Non-volatile Amine Contents of Commercial Rice-Bran Pickles of Sardine

Determination and Contents of Non-Volatile Amines in Soybean Paste and Soy Sauce.

A liquid chromatographic (LC) analysis for the determination of seven amines, tyramine (Tym), histamine (Him), phenethylamine (Phm), putrescine (Put), cadaverine (Cad), spermidine (Spd) and spermine (Spm) in soybean paste (Miso), soy sauce (Shoyu) and its products was studied. The amines in the sample were extracted with 0.1N HCl and applied to a column of Amberlite CG-50 and derivatized with dansyl chloride for the application to the liquid chromatography. The LC separations were performed on a Finepak SIL C18S column with an acetonitrile-water elution gradient. The amines were confirmed by TLC for Him and by GC-MS for others with ethyloxycarbonyl derivatives. In the survey of commercial samples by this method, most of Miso samples and ann of Shoyu samples were found to contain these 7 amines. Tym and Him were determined in the range of nd-194, nd-177 μg/g in every kind of Miso, and 95.8-356, 62.8-232 μg/g in Shoyu and its products as average which were higher than the other amines. Shoyu, in general, had higher Him and Tym contents than Miso samples. However the contents of the other amines in both Shoyu and Miso samples were the same levels. It was considered that Tym, Him and Phm were produced by microorganism in a manufacturing process because these amines were not detected from such raw materials as koji (malted rice) and soy beans of Miso and Shoyu. Put, Cad, Spd and Spm were originated from the raw materials, because the contents of these amines in the samples were nearly the same as those of raw materials.