• Toxicological Research
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• v.31 no.3
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• pp.299-305
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• 2015

An HPLC analytical method was validated for the quantitative determination of biogenic amines in agricultural products. Four agricultural foods, including apple juice, Juk, corn oil and peanut butter, were selected as food matrices based on their water and fat contents (i.e., non-fatty liquid, non-fatty solid, fatty liquid and fatty solid, respectively). The precision, accuracy, recovery, limit of detection (LOD) and quantification (LOQ) were determined to test the validity of an HPLC procedure for the determination of biogenic amines, including tryptamine, ${\beta}$-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, in each matrix. The LODs and LOQs for the biogenic amines were within the range of 0.01~0.10 mg/kg and 0.02~0.31 mg/kg, respectively. The relative standard deviation (RSD) of intraday for biogenic amine concentrations ranged from 1.86 to 5.95%, whereas the RSD of interday ranged from 2.08 to 5.96%. Of the matrices spiked with biogenic amines, corn oil with tyramine and Juk with putrescine exhibited the least accuracy of 84.85% and recovery rate of 89.63%, respectively, at the lowest concentration (10 mg/kg). Therefore, the validation results fulfilled AOAC criteria and recommendations. Subsequently, the method was applied to the analysis of biogenic amines in fermented agricultural products for a total dietary survey in Korea. Although the results revealed that Korean traditional soy sauce and Doenjang contained relatively high levels of histamine, the amounts are of no concern if these fermented agricultural products serve as condiments.

• Journal of Dairy Science and Biotechnology
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• v.34 no.3
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• pp.187-191
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• 2016

Biogenic amines have been used as chemical indicators of fermented foods. So far, several chromatography methods have been developed to detect biogenic amines in foods. However, few methods have identified these compound in domestic cheese. We analyzed the biogenic amines (histamine dihydrochloride, tyramine hydrochloride, ${\beta}$-phenylethylamine hydrochloride, putrescine dihydrochloride, cadaverine, spermidine, tryptamine hydrochloride, ethanolamine hydrochloride and butylamine) in cheese by using HPLC. The calibration curves of the biogenic amines were found to be linear over the concentration range of 10-50 ppm with a correlation coefficient of above 0.99. The limit of detection (LOD) and limit of quantitation (LOQ) of the biogenic amines in the given order were 3.7 and 11.3 ppm, 3.4 and 10.4ppm, 3.4 and 10.3 ppm, 4.0 and 12.2 ppm, 3.4 and 10.4 ppm, 3.4 and 10.5 ppm, 3.5 and 10.7 ppm, 4.1 and 12.5 ppm, and 3.4 and 10.4 ppm, respectively. Recovery rates of the biogenic amines in the given order were 112, 104, 93, 108, 91, 102, 101, and 92%, respectively. The findings of this study suggest that HPLC is a suitable method for the determination of biogenic amines, thereby indicating its potential application in the quality control of aging cheese.

• YAKHAK HOEJI
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• v.35 no.4
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• pp.288-294
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• 1991

A new fluorescent derivatizing reagent was developed to be used in HPLC for the trace determination of primary and secondary amines. This new reagent, 1-(N,N-dimethylamino)pyrene-6-sulfonyl chloride, was synthesized by the chlorination of sodium 1-(N,N-dimethylamino)pyrene-6-sulfonate which was obtained from 1-(N,N-dimethylamino)pyrene after sulfonation. Ephedrine and norephedrine were derivatized quantitatively by this reagent. The optimum conditions for the derivatization such as pH, reagent concentration, reaction time and reaction temperature ware examined. The structures of derivatives were identified by IR, $^{1}$H-NMR and MS methods. The fluorescence properties and the stability of the derivatives were examined. The derivatives were separated on silica column with an isocratic elution using the mixture of n-hexane and ethylacetate and monitored by fluorescene detector. Linear calibration curves were obtained and detection limits in a 10 $\mu$l injection volume were 5 picomole for ephedrine and norephedrine.

• Analytical Science and Technology
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• v.20 no.2
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• pp.115-123
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• 2007

The heterocyclic amines (HAs) are a family of mutagenic/carcinogenic chemicals formed from the cooking of muscle meats such as beef, meat, fowl, and fish. A major draw back in the analysis of HAs from foods is their very low level of concentration (ng/g) and a number of matrix interferences in samples. Solid-phase extraction (SPE) is one of the procedures widely used for the extraction and purification of HAs in food samples. In this study, several SPE procedures of HAs determination were performed. Recoveries of the HAs were obtained from comparing a matrix such as a standard methanolic solution and pre-cooked meat extracts. Recovery values were ranging between 25.3 and 93.0 % at a concentration of 25.0 ng/g. HAs were determined with high sensitivity by micro-HPLC (${\mu}$-HPLC) analysis with electrospray ionization mass spectrometry (ESI-MS). Another developed method, which is freezing filtration method, shows better extraction recoveries and good precisions. The established method will be applicable to monitoring of heterocyclic amines from the cooked meat.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.491-496
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• 2001

A modification of decarboxylating agar medium as described by niven was performed to improve the detection method of biogenic amine-producing bacteria and to eliminate the false-positive. A total of 120 bacterial strains isolated from kimchi were used to evaluate different dicarboxylating agar media and for screening biogenic amines. Potential false-positives ranged from approximately 66 to 79% of the strains tested in the already well-known media. In our improved medium, none of the 120 strains showed the potential false-positives. There was a good agreement (81.7%-87.5%) between the results obtained by the improved medium and by HPLC analysis. Consequently, this medium was greatly improved in screening biogenic amine-producing bacteria and discarding false-positives. Of the 120 kimchi isolates, 14.2, 18.3, 37.5, and 0.8% were found by HPLC to be the producers of histamine, tyramine, putrescine (as a form of spermine), and cadaverine, respectively. The proportion of biogenic amine producer during kimchi fermentation increased to a maximum at an immature period and decreased thereafter.

• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.692-699
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• 2003

The changes of physicochemical and microbiological states of Myeolchi-jeot, a Korean salted and fermented anchovy, were investigated during 20 days of storage at $4^{\circ}C,\;15^{\circ}C,\;and\;30^{\circ}C$. A total of 314 bacterial strains isolated from Myeolchi-jeot samples at different time intervals were identified, and their abilities to produce biogenic amines were determined by both decarboxylating agar media and HPLC analysis. The salinity and water activity of Myeolchi-jeot changed little, while the pH increased slightly over 20 days at the tested temperatures. A significant increase of total plate count was observed in the sample stored at $30^{\circ}C$. Staphylococcus spp. were dominant in Myeolchi-jeot, and its amine productivity was very weak. Bacillus spp. appeared increasingly with the progress of storage at all temperatures tested, and the increase was considerably dependent on the increase of storage temperature. Also, 58-73%, 83-90%, 81-90%, and 83-93% of Bacillus strains had the ability to produce histamine, tyramine, putrescine, and cadaverine, respectively. Therefore, the main amine producer in Myeolchi-jeot stored for a long period seems to be the genus Bacillus, especially B. licheniformis.

• Korean Journal of Food Science and Technology
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• v.38 no.6
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• pp.730-737
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• 2006

High performance liquid chromatographic methods for the simultaneous determination of 12 biogenic amines were developed and contents of biogenic amines in 23 varieties of Korean commercial fermented food were analyzed. Dansyl derivatives of biogenic amine were very stable and had good peak resolution. Except agmatine, the recovery of biogenic amines from soybean paste with extraction of 0.1 N HC1 added biogenic amines to soybean paste was greater 85%. The calibration curve showed good linearity over a concentration range up to 50 ${\mu}g/mL$. In the determination of biogenic amine level in Korean commercial fermented foods, doenjang and chungkukjang (fermented soybean pastes), soy sauce, anchovy and pacific sand lance liquid jeotkal (fermented fish sauce products), and cabbage kimchi had high level biogenic amines. Especially, traditional doenjang had a histamine (HIS) level of 952.0 mg/kg, a tyramine (TYR) level of 1,430.7 mg/kg. Most cheese had low level of biogenic amines, but one Gouda cheese had a tyramine (TYR) level of 97.5 mg/kg. A low level of biogenic amines was detected in wines, beer, yoghurt, and sausage. Putrescine (PUT), cadaverine (CAD), tryptamine (TRY), histamine (HIS), tyramine (TYR), and 2-phenylethylamine (PHE) were mainly formed in fermented foods by the action of microorganism, so their levels were high with a range. On the other hand, spermidine (SPD), spermine(SPM), serotonin (SER), noradrenaline (NOR), and dopamine (DOP) were formed originally via biosynthesis with consequent low level.

• Korean Journal of Food Science and Technology
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• v.44 no.2
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• pp.196-201
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• 2012

Microorganisms, having the lower decarboxylase activity, among the isolated strains from $cheonggukjang$ and rice-straw in this study were selected by using biogenic amine (BA) media. The selected strains were identified as $Bacillus$ $subtilis$ HH12, $B.$ $subtilis$ HR254, and $Paenibacillus$ $barcinonensis$ KR97, by using 16S rRNA analysis. PCR analysis showed that the histidine decarboxylase ($hdc$) gene was absent in the HH12, HR254, and KR97 strains. However, PCR analysis showed that the tyrosine decarboxylase ($tdc$) gene was present in the HH12, HR254, and KR97 strains. Quantitative analysis of the selected strains by using high-performance liquid chromatography showed that histamine was absent in the HH12, HR254, and KR97 strains. However, these 3 strains showed tyramine concentrations of 6.09, 3.68, and 6.30 mg/L, respectively. These strains produced lower concentrations of amines (approximately 7.9, 0, and 9.3% amines in the HH12, HR254, and KR97 strains, respectively) than the $B.$ $subtilis$ MC138 strain, which showed the higher protease activity.

• Journal of Korean Society on Water Environment
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• v.25 no.2
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• pp.227-234
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• 2009

In this research, a retrofitting process, which consists of a pretreatment system (coagulation) for dye wastewater combined with a biological nutrient system (MLE process using media), for a sewage treatment plant that has to treat dye wastewater efficiently with domestic wastewater were developed and a pilot plant was operated for verifying adoptability and performance of the developed advanced process for dye wastewater. From the results of the pilot plant operation, BOD 52.9%, $COD_{Cr}$ 55.9%, and color 71.3% were removed in pretreatment of coagulation process and the biodegradability of dye wastewater was improved to $0.32{\sim}0.59BOD/COD_{Cr}$ of the coagulated wastewater from $0.29{\sim}0.43BOD/COD_{Cr}$ of the raw dye wastewater. The final effluent concentrations were BOD of 8 mg/L, $COD_{Cr}$ of 43 mg/L, $COD_{Mn}$ of 18 mg/L, T-N of 8 mg/L, and T-P of 1.3 mg/L, respectively. Color was removed from 1655 to 468 unit by coagulation and then to 123 unit by MLE process. The HPLC analysis of aromatic amines in wastewater showed that decolorization was achieved by cometabolism while aromatic amines were produced by cleavage of azo bonds under anaerobic conditions and these products were removed in an aerobic tank subsequently. Nitrification rates of attached and suspended microorganisms were evaluated comparatively and the acclimating conditions of bacteria on media were validated by the scanning electron microscope.

• Food Science of Animal Resources
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• v.29 no.6
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• pp.719-725
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• 2009

Heterocyclic aromatic amines (HCAs) are potent mutagens and possible human carcinogens that are formed during the heating of protein-rich foods. The effects of preheating treatment of beef patties using a microwave prior to frying at $220^{\circ}C$ for 10 min on each side on the reduction of HCAs (amino-carbolines and amino-imidazo-azaarenes) were evaluated. The amount of HCAs was then evaluated by solid-phase extraction and high pressure liquid chromatography (HPLC) analysis. The beef patties were treated by microwaving for various times (0, 1, 1.5, 2, or 3 min) before pan-frying. The results revealed the presence of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), 3-amino-1,4-dimethyl-5H-pyrido-[4,3-b] indole (Trp-P-1), 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1), 2-aminodipyrido[1,2-a:3',2'-d]imidazole (Glu-P-2), 9H-pyrido[3,4-b]indole (Norharman), 1-methyl-9H-pyrido[3,4-b]indole (Harman), 2-amino-9H-pyrido[2,3-b] indole ($A{\alpha}C$), 2-amino-3,8 dimethylimidazo[4,5-f]-quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP) in all samples. However, microwave pretreatment for 1 min inhibited the formation of these HCAs by up to 90% when compared to the control.