Culture-dependent and -independent analyses of bacterial compositions and its contributions to formation of γ-aminobutyric acid and poly-γ-glutamic acid in Cheonggukjang.

This study was performed to unveil bacterial compositions and their contributions to the formation of γ-aminobutyric acid (GABA) and poly-γ-glutamic acid (γ-PGA) in Cheonggukjang. To predict possible key factors contributing to the content of the bioactive compounds in Cheonggukjang, commercial products were analyzed for various parameters. The content of GABA and γ-PGA showed a negative (R2 = 0.61 - 0.73) and positive correlation (R2 = 0.53 - 0.96) with antioxidative activity. Consistently, GABA content showed a moderate negative correlation with γ-PGA content (R2 = 0.58). Among the physicochemical and microbial parameters, only salinity showed a moderate negative correlation with γ-PGA content (R2 = 0.75), which might be due to the inhibition of bacterial growth. It was also suggested that multiple factors (including bacterial species) were involved in the formation of GABA and γ-PGA in Cheonggukjang. To reveal dominant bacterial species and further presume their contributions to the bioactive compound formation in Cheonggukjang, both culture-independent (metagenomic) and -dependent (culturomic) methods were used. Culture-independent method showed that Bacillus piscis was dominant (23.37 - 94.89 %), followed by B. hisashii (0.00 - 62.45 %) and B. coagulans (0.00 - 13.82 %). Considering the quantitative speciation data on the bioactive compound content in Cheonggukjang (and bacterial production capability) together, it was further elucidated that B. piscis contributed primarily to the bioactive compound formation. Unlike this, culture-dependent analysis revealed that B. licheniformis and B. subtilis were dominant (30.0 - 47.6 and 17.5 - 39.5 %, respectively). Based on the quantitative speciation data on the bacterial production capability of GABA and γ-PGA, B. subtilis was the primarily contributing bacterial species to the bioactive compound formation. Consequently, it was observed that the bacterial compositions and their contributions to the bioactive compound formation determined by the two methods differed considerably, i.e., B. piscis and B. subtilis were identified to be prominent bacterial contributors, respectively, depending on the method used.

Biogenic amine reduction by food additives in Cheonggukjang, a Korean fermented soybean paste, fermented with tyramine-producing heterogeneous bacterial species.

This study was conducted to mitigate the food safety risks related to biogenic amine (BA) by reducing the BA content in Cheonggukjang using applicable food additives. In in-vitro experiments, of the additives tested, tartaric acid (TA), potassium sorbate (PS), and sodium benzoate (SB) considerably inhibited tyramine production of strains of Bacillus spp. and Enterococcus faecium while less affecting their growth. In addition to these three additives, two additives, glycine (GL) and nicotinic acid (NA), reported to have significant inhibitory effects in previous studies, were applied to the Cheonggukjang fermentation with prolific tyramine-producing strains of B. subtilis and E. faecium. The content of tyramine in the Cheonggukjang samples treated with TA, PS, SB, GL, and NA was significantly reduced by 27.5%, 50.7%, 51.4%, 76.1%, and 100.0%, respectively, compared to the control sample. Additionally, the content of polyamines (putrescine, cadaverine, spermidine, and spermine) in the GL-treated sample was reduced by 42.6%-62.4%. The mode of action could be attributed to inhibiting the bacterial decarboxylase activity and/or growth. Consequently, excluding NA that interfered with Cheonggukjang fermentation, GL was the most outstanding additive with an inhibitory effect on tyramine formation in food, followed by SB and PS, all of which showed a more than 50% reduction. Therefore, the use of appropriate additives could be one of the promising strategies to avoid the food safety issues implicated in BAs in Cheonggukjang.

GC-MS/MS Method for Determination of Polycyclic Aromatic Hydrocarbons in Herbal Medicines.

Polycyclic aromatic hydrocarbons (PAHs) are hydrophobic organic contaminants that have a highly carcinogenic and mutagenic nature. This study aimed to develop and validate a sensitive analytical method to determine 8 PAHs in 51 herbal medicines (HMs) using gas chromatography (GC)-tandem mass spectrometry (MS/MS). Liquid--liquid extraction and florisil SPE cartridge purification were basically adopted for pretreatment. For the samples containing essential oil, starch grain, etc., N,N-dimethyl formamide/water mixture (9:1, v/v) was added in the extraction step. The multiple reaction monitoring (MRM) conditions were newly obtained by the infusion of reference solutions of the targeted compounds at a concentration of 100 ng/mL into the GC-MS/MS system used in this study. The 51 items were classified according to whether or not they contained essential oil. Eight PAHs were not detected in 39 (8.3%) of the 459 samples monitored. The total content of 8 PAHs ranged from 0.45 µg/kg in Anemarrhenae Rhizoma to 270.94 µg/kg in Zingiberis Rhizoma. The average content of those ranged from 0.9 µg/kg in Araliae Continentalis Radix to 110.8 µg/kg in Coptidis Rhizoma Preparata cum Vinum. The results of this study prove that the proposed method is useful for determining 8 PAHs in HMs.

Evaluation of Bioactive Compounds and Antioxidative Activity of Fermented Green Tea Produced via One- and Two-Step Fermentation.

This study investigated the influence of one- and two-step fermentation on bioactive compound production in fermented green tea, i.e., one-step fermented green tea (OFG) and two-step fermented green tea (TFG). One-step fermentation entailed acetic acid fermentation, while two-step fermentation consisted of lactic acid fermentation followed by acetic acid fermentation. Acetobacter pasteurianus PCH 325, isolated from an over-ripened peach, was selected for acetic acid fermentation based on its growth and organic acid production characteristics. Acetic acid fermentation conditions were optimized for one- and two-step fermentation: 3% fermentation alcohol for both processes; 8% and 4% sucrose, respectively; and fermentation at 25 °C for both processes. For lactic acid fermentation of TFG, the inoculum and optimized conditions reported previously were used. Under the optimized conditions, the acetic acid content in OFG and TFG increased 21.20- and 29.51-fold, respectively. Furthermore, through two-step fermentation, γ-aminobutyric acid and lactic acid were produced up to 31.49 ± 1.17 mg/L and 243.44 ± 58.15 mg/L, respectively, which together with acetic acid could contribute to the higher DPPH scavenging activity of TFG. This study suggests that two-step fermentation may be a valuable strategy in industry for raising the amount of acetic acid and/or providing additional bioactive compounds.

Biofilm-associated heat resistance of Bacillus cereus spores in vitro and in a food model, Cheonggukjang jjigae.

Bacillus cereus, a foodborne pathogen, is capable of forming spores and biofilms as methods to withstand environmental stresses. These bacterial structures are an issue for food safety as they aid the bacteria survive heat sterilisation processes of foods and food contact surfaces. This study was conducted to investigate the role of the biofilm structure in providing an extra layer of protection to spores against heat treatments. For this, heat resistance of B. cereus spores in intact biofilms was compared to that of planktonic spores in vitro and in a Cheonggukjang jjigae food model. Using methods developed in this study to measure the wet and dry heat resistance of spores in intact biofilms, it was found that B. cereus spores have significantly higher heat resistances when present in biofilms rather than as planktonic spores, and that dry heat is less effective than wet heat at killing spores in biofilms. In further detail, for wet heat treatments, spores in biofilms of the strain isolated from Cheonggukjang (Korean fermented whole soybean), B. cereus CH3, had generally higher wet heat resistances than the reference strain, B. cereus ATCC 10987, both in vitro and in the Cheonggukjang jjigae food model. However, the spores in biofilms of the two strains showed similar heat resistance to dry heat, with some exceptions, when biofilms were formed in vitro or in Cheonggukjang jjigae broth. Meanwhile, B. cereus ATCC 10987 spores in biofilms had higher or similar wet heat resistances in vitro compared to in Cheonggukjang jjigae broth. Wet heat resistances of B. cereus CH3 spores in biofilms were all statistically similar regardless of biofilm formation media (brain heart infusion and Cheonggukjang jjigae broths). For dry heat, spores in biofilms of both B. cereus strains were more heat resistant when biofilms were formed in the Cheonggukjang jjigae food model rather than in vitro. Altogether, heat resistances of spores in biofilms formed in vitro and in the food environment were found to be different depending on the tested B. cereus strain, but higher than planktonic spores in any case. This is the first study examining the heat resistance of B. cereus spores in intact biofilms matrices attached to the surface, both in vitro and in a food model. Therefore, this research is valuable to understand the protective effects of biofilms formed in food environments and to reduce the food safety risks associated with B. cereus.

Reduction in Biogenic Amine Content in Baechu (Napa Cabbage) Kimchi by Biogenic Amine-Degrading Lactic Acid Bacteria.

This study was performed to mine biogenic amine (BA)-degrading lactic acid bacteria (LAB) from kimchi and to investigate the effects of the LAB strains on BA reduction in Baechu kimchi fermentation. Among 1448 LAB strains isolated from various kimchi varieties, five strains capable of considerably degrading histamine and/or tyramine were selected through in vitro tests and identified as Levilactobacillus brevis PK08, Lactiplantibacillus pentosus PK05, Leuconostoc mesenteroides YM20, L. plantarum KD15, and Latilactobacillus sakei YM21. The selected strains were used to ferment five groups of Baechu kimchi, respectively. The LB group inoculated with L. brevis PK08 showed the highest reduction in tyramine content, 66.65% and 81.89%, compared to the control group and the positive control group, respectively. Other BA content was also considerably reduced, by 3.76-89.26% (five BAs) and 7.87-23.27% (four BAs), compared to the two control groups, respectively. The other inoculated groups showed similar or less BA reduction than the LB group. Meanwhile, a multicopper oxidase gene was detected in L. brevis PK08 when pursuing the BA degradation mechanism. Consequently, L. brevis PK08 could be applied to kimchi fermentation as a starter or protective culture to improve the BA-related safety of kimchi where prolific tyramine-producing LAB strains are present.

Effect of Calcium and Manganese Supplementation on Heat Resistance of Spores of Bacillus Species Associated With Food Poisoning, Spoilage, and Fermentation.

Bacterial spores often survive thermal processing used in the food industry, while heat treatment leads not only to a decrease in the nutritional and organoleptic properties of foods, but also to a delay in fermentation of fermented foods. Selective reduction of undesirable spores without such impediments is an ongoing challenge for food scientists. Thus, increased knowledge of the spore-forming bacteria is required to control them. In this study, the heat resistance results (D 100 ° C) of the spores of four Bacillus species were determined and compared to previous literature, and found that B. cereus has significantly lower heat resistance than the other Bacillus species, B. coagulans, B. subtilis, and B. licheniformis. Using the spores of these strains, this study also evaluated the effects of single and combined supplementation of calcium (0.00-2.00 mM) and manganese (0.00-0.50 mM) on heat resistance (D 100 ° C). The results revealed that the spores of B. licheniformis and B. cereus displayed the smallest heat resistance when sporulated on media rich in calcium. Conversely, B. coagulans spores and B. subtilis spores exhibited the greatest heat resistance when sporulated under calcium-rich conditions. The opposite results (stronger heat resistance for B. licheniformis spores and B. cereus spores, and smaller heat resistance for B. coagulans spores and B. subtilis spores) were obtained when the spores were formed on media poor in the minerals (particularly calcium). Based on the results, the Bacillus species were divided into two groups: B. licheniformis and B. cereus; and B. coagulans and B. subtilis. The study provides valuable insight to selectively reduce spores of undesirable Bacillus species in the food industry.

Development of a colorimetric enzymatic assay method for aromatic biogenic monoamine-producing decarboxylases.

Biogenic amines (BAs) produced by the action of bacterial amino acid decarboxylases in fermented foods cause various health problems in human. Despite the importance, detailed characterizations of the BA-producing decarboxylases are relatively less progressed than the studies on BA-producing bacteria, due to the time-consuming chromatography-based assay method. In this study, a simple and general colorimetric assay for aromatic amino acid decarboxylases coupled with an amine oxidase from Arthrobacter aurescens (AMAO) and horseradish peroxidase was developed using a tyrosine decarboxylase from Enterococcus faecium DSM20477 (EfmTDC) as a model enzyme. The activity profiles over pH and temperature and the kinetic analysis for EfmTDC revealed that the results by the colorimetric assay are compatible with those by the chromatographic assay. In addition, due to the broad substrate specificity of AMAO for histamine and 2-phenylethylamine, the colorimetric assay would be applicable to the characterization of other aromatic amino acid decarboxylases including histidine decarboxylases. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-00938-4.

Tyrosinase Inhibitory Activity of Soybeans Fermented with Bacillus subtilis Capable of Producing a Phenolic Glycoside, Arbutin.

The production of arbutin, an effective tyrosinase inhibitor as well as an outstanding antioxidant, by 691 Bacillus strains isolated from soybean-based foods was tested to enhance the tyrosinase inhibitory activity of soybeans via fermentation with the strains. Among the strains tested, the 5 strains capable of significantly producing arbutin were identified as B. subtilis via 16S rRNA sequencing. When soybeans were fermented with each of the selected strains, the arbutin content was highest on day 1 of fermentation and decreased thereafter. However, the tyrosinase inhibitory activity of the fermented soybeans continuously increased as fermentation progressed, whereas the activity of non-inoculated soybeans was consistently low. The results indicate that arbutin enhances the tyrosinase inhibitory activity of soybeans in the early period of fermentation, while other substances besides arbutin contribute to the activity in the later period. Consequently, soybeans fermented with arbutin-producing B. subtilis strains could be considered as a natural source of cosmeceuticals and nutricosmetics used in skin lightening and may be of interest in the food industry because they contain well-known and powerful antioxidants such as arbutin and other substances.

The Role of Enterococcus faecium as a Key Producer and Fermentation Condition as an Influencing Factor in Tyramine Accumulation in Cheonggukjang.

The study evaluated the role of Enterococcus faecium in tyramine production and its response to fermentation temperature in a traditional Korean fermented soybean paste, Cheonggukjang. Tyramine content was detected in retail Cheonggukjang products at high concentrations exceeding the recommended limit up to a factor of 14. All retail Cheonggukjang products contained Enterococcus spp. at concentrations of at least 6 Log CFU/g. Upon isolation of Enterococcus strains, approximately 93% (157 strains) produced tyramine at over 100 µg/mL. The strains that produced the highest concentrations of tyramine (301.14-315.29 µg/mL) were identified as E. faecium through 16S rRNA sequencing. The results indicate that E. faecium is one of the major contributing factors to high tyramine content in Cheonggukjang. During fermentation, tyramine content in Cheonggukjang groups co-inoculated with E. faecium strains was highest at 45 °C, followed by 37 °C and 25 °C. The tyramine content of most Cheonggukjang groups continually increased as fermentation progressed, except groups fermented at 25 °C. At 45 °C, the tyramine content occasionally exceeded the recommended limit within 3 days of fermentation. The results suggest that lowering fermentation temperature and shortening duration may reduce the tyramine content of Cheonggukjang, thereby reducing the safety risks that may arise when consuming food with high tyramine concentrations.

Occurrence and Reduction of Biogenic Amines in Kimchi and Korean Fermented Seafood Products.

Biogenic amines produced during fermentation may be harmful when ingested in high concentrations. As current regulations remain insufficient to ensure the safety of fermented vegetable products, the current study determined the risks associated with the consumption of kimchi by evaluating the biogenic amine concentrations reported by various studies. Upon evaluation, some kimchi products were found to contain histamine and tyramine at potentially hazardous concentrations exceeding the recommended limit of 100 mg/kg for both histamine and tyramine. The biogenic amines may have originated primarily from metabolic activity by microorganisms during fermentation, as well as from Jeotgal (Korean fermented seafood) and Aekjeot (Korean fermented fish sauce) products commonly used as ingredients for kimchi production. Many studies have suggested that Jeotgal and Aekjeot may contribute to the histamine and tyramine content in kimchi. Microorganisms isolated from kimchi and Jeotgal have been reported to produce both histamine and tyramine. Despite the potential toxicological risks, limited research has been conducted on reducing the biogenic amine content of kimchi and Jeotgal products. The regulation and active monitoring of biogenic amine content during kimchi production appear to be necessary to ensure the safety of the fermented vegetable products.

Effects of Calcium and Manganese on Sporulation of Bacillus Species Involved in Food Poisoning and Spoilage.

Spores are resistant against many extreme conditions including the disinfection and sterilization methods used in the food industry. Selective prevention of sporulation of Bacillus species is an ongoing challenge for food scientists and fermentation technologists. This study was conducted to evaluate the effects of single and combined supplementation of calcium and manganese on sporulation of common pathogenic and food spoilage Bacillus species: B. cereus, B. licheniformis, B. subtilis and B. coagulans. Sporulation of Bacillus vegetative cells was induced on sporulation media supplemented with diverse concentrations of the minerals. Under the various mineral supplementation conditions, the degree of sporulation was quantified with colonies formed by the Bacillus spores. The results revealed that B. licheniformis and B. cereus displayed the weakest sporulation capabilities on media with minimal supplementation levels of calcium and manganese. The lowest sporulation of B. subtilis and B. coagulans was observed on media supplemented with the highest level of calcium and low levels of manganese. Depending on effect of supplementation on sporulation, the Bacillus species were divided into two distinct groups: B. licheniformis and B. cereus; and B. subtilis and B. coagulans. The information provides valuable insight to selectively reduce sporulation of Bacillus species undesirable in the food industry.

Formation of Biogenic Amines in Pa (Green Onion) Kimchi and Gat (Mustard Leaf) Kimchi.

In this study, biogenic amine content in Pa (green onion) kimchi and Gat (mustard leaf) kimchi, Korean specialty kimchi types, was determined by high-performance liquid chromatography (HPLC). Many kimchi samples contained low levels of biogenic amines, but some samples had histamine and tyramine content over the safe levels. Based on the comparative analysis between the ingredient information on food labels and biogenic amine content of kimchi samples, Myeolchi-aekjeot appeared to be an important source of biogenic amines in both kimchi. Besides, through the 16s rRNA sequence analysis, Lactobacillus brevis appeared to be responsible for the formation of biogenic amines (tyramine, ß-phenylethylamine, putrescine, and cadaverine) in both kimchi, in a strain-dependent manner. During fermentation, a higher accumulation of tyramine, ß-phenylethylamine, and putrescine was observed in both or one (for putrescine) of kimchi types when L. brevis strains served as inocula. The addition of Myeolchi-aekjeot affected the initial concentrations of most biogenic amines (except for spermidine in Gat kimchi) in both kimchi. Therefore, this study suggests that using appropriately salted and fermented seafood products for kimchi preparation and using biogenic amine-negative and/or biogenic amine-degrading starter cultures would be effective in reducing biogenic amine content in Pa kimchi and Gat kimchi.

Bacterial Production and Control of Biogenic Amines in Asian Fermented Soybean Foods.

Fermented soybean foods possess significant health-promoting effects and are consumed worldwide, especially within Asia, but less attention has been paid to the safety of the foods. Since fermented soybean foods contain abundant amino acids and biogenic amine-producing microorganisms, it is necessary to understand the presence of biogenic amines in the foods. The amounts of biogenic amines in most products have been reported to be within safe levels. Conversely, certain products contain vasoactive biogenic amines greater than toxic levels. Nonetheless, government legislation regulating biogenic amines in fermented soybean foods is not found throughout the world. Therefore, it is necessary to provide strategies to reduce biogenic amine formation in the foods. Alongside numerous existing intervention methods, the use of Bacillus starter cultures capable of degrading and/or incapable of producing biogenic amines has been proposed as a guaranteed way to reduce biogenic amines in fermented soybean foods, considering that Bacillus species have been known as fermenting microorganisms responsible for biogenic amine formation in the foods. Molecular genetic studies of Bacillus genes involved in the formation and degradation of biogenic amines would be helpful in selecting starter cultures. This review summarizes the presence and control strategies of biogenic amines in fermented soybean foods.

High Prevalence of Cefotaxime Resistant Bacteria in Grazing Beef Cattle: A Cross Sectional Study.

Although the over-use of antibiotics during food animal production is a potential driver of antimicrobial resistant microorganisms (ARMs), a high prevalence of cefotaxime resistant bacteria (CRB) has been observed in grazing animals raised without antibiotic supplementation. In this cross-sectional study, the prevalence and concentration of CRB in beef cattle on grazing farms were investigated. Fecal samples from the recto-anal junction of cattle (n = 840) and environmental samples (n = 258) were collected from 17 farms in North and Central Florida in the United States, and a survey of farm characteristics, animal husbandry practices, and antibiotic usage was conducted. CRB were detected in fecal samples from 47.4% of all cattle, with the prevalence ranging from 21.1 to 87.5% on farms, and significantly higher (P < 0.001) in calves compared to adult cows (54.1 vs. 41.8%). Environmental samples had a higher prevalence than fecal samples (P < 0.001), with CRB detected in 88.6% of water, 98.7% of soil, and 95.7% of forage samples. Compared to the concentration (log CFU/g) of CRB in fecal samples (2.95, 95% CI: 2.89, 3.02), the concentration of CRB was higher (P < 0.001) in soil and forage samples (5.37, 95% CI: 5.16, 5.57) and lower (P < 0.001) in water samples (1.08, 95% CI: 0.82, 1.36). Soil microbiota from farms with high prevalence of CRB clustered closer together and the proportion of Phylum Proteobacteria was higher on farms with high prevalence of CRB resistance. Large farming operations were associated with a 58% higher likelihood of CRB detection in fecal samples. Regular cleaning of drinking troughs and the addition of ionophores to feed were associated with CRB reduction in fecal samples. Taken together, the widespread of CRB into both cattle seldom treated with cephalosporin antibiotics and the surrounding environment suggests the environment is a natural source of antimicrobial resistance in beef cattle.

The Occurrence of Biogenic Amines and Determination of Biogenic Amine-Producing Lactic Acid Bacteria in Kkakdugi and Chonggak Kimchi.

In this study, biogenic amine content in two types of fermented radish kimchi (Kkakdugi and Chonggak kimchi) was determined by high performance liquid chromatography (HPLC). While most samples had low levels of biogenic amines, some samples contained histamine content over the toxicity limit. Additionally, significant amounts of total biogenic amines were detected in certain samples due to high levels of putrefactive amines. As one of the significant factors influencing biogenic amine content in both radish kimchi, Myeolchi-aekjoet appeared to be important source of histamine. Besides, tyramine-producing strains of lactic acid bacteria existed in both radish kimchi. Through 16s rRNA sequencing analysis, the dominant species of tyramine-producing strains was identified as Lactobacillus brevis, which suggests that the species is responsible for tyramine formation in both radish kimchi. During fermentation, a higher tyramine accumulation was observed in both radish kimchi when L. brevis strains were used as inocula. The addition of Myeolchi-aekjeot affected the initial concentrations of histamine and cadaverine in both radish kimchi. Therefore, this study suggests that reducing the ratio of Myeolchi-aekjeot to other ingredients (and/or using Myeolchi-aekjeot with low biogenic amine content) and using starter cultures with ability to degrade and/or inability to produce biogenic amines would be effective in reducing biogenic amine content in Kkakdugi and Chonggak kimchi.

Occurrence and reduction of biogenic amines in traditional Asian fermented soybean foods: A review.

Biogenic amines are harmful substances generated during the fermentation process. Regulations on biogenic amine content in fermented foods are currently insufficient in comparison to the popularity of fermented food consumption in Asian countries. The current review evaluated the biogenic amine content of fermented soybean-based Asian foods to determine whether the food products are safe for consumption. Though the reported ranges of biogenic amine content in fermented soybean foods varied widely, most products contained biogenic amine concentrations at potentially hazardous levels. To ensure the safety of fermented soybean food products, further efforts are required in the improvement of the food manufacturing process, as well as the establishment of regulations on managing biogenic amine content.

Tyramine reduction by tyrosine decarboxylase inhibitor in Enterococcus faecium for tyramine controlled cheonggukjang.

This study was carried out to find a method to control tyrosine decarboxylase activity (TDC) of a strain of Enterococcus faecium capable of producing high levels of tyramine. To select a TDC inhibitor, enzyme assay was first performed using purified TDC enzyme and 0.1% of TDC inhibiting chemicals. When 0.23% of nicotinic acid was added, tyramine content (363 ug/mL) was lower than that of the control group (873 ug/mL). At the same time, bacterial growth was decreased 1 log cycle from 8.62 to 7.56 log CFU/mL. TDC expression level in E. faecium was measured by using RT-qPCR. Lower expression level (below 0.7) was observed after the addition of 0.23% nicotinic acid (in vitro). When cheonggukjang was manufactured with addition of nicotinic acid, tyramine contents were decreased from 698.67 to 117.27 mg/kg when the concentration of nicotinic acid added was increased from 0.10 to 0.30%. These results suggest that nicotinic acid could be used as an agent (TDC inhibitor) to reduce tyramine content in cheonggukjang.

Simultaneous and rapid analysis of chemical preservatives in processed animal products by ultra-performance liquid chromatography.

An ultra-performance liquid chromatography-tunable ultraviolet method was optimized and validated for the simultaneous analysis of nine chemical preservatives in processed animal products. The limits of detection and quantification for the preservatives were within the ranges of 0.02-0.23 and 0.07-0.76 µg/mL, respectively. The relative standard deviations for intraday analyses of retention time and peak area were 0.00-0.23 and 0.03-2.93%, respectively, whereas, those for interday analyses were 0.67-2.30 and 2.12-5.37%, respectively. Of the nine preservatives spiked into six different animal products, dehydroacetic acid spiked into soft cheese exhibited the lowest recovery rate of 72.1 ± 0.36% at the lowest concentration (0.25 g/kg). Comparing data between UPLC and high-performance liquid chromatography with a 5% significance level, the t-statistic was 1.42. Moreover, sorbic acid was detected in 16 animal products (0.11-2.49 g/kg) when 278 products were analyzed for preservatives.