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.

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.

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.

Reduction of tyramine by addition of Schizandra chinensis baillon in Cheonggukjang.

This study was performed to examine the microorganisms responsible for the high tyramine content of Cheonggukjang, a traditional Korean fermented soy food, and to establish a technology for controlling the growth of these microorganisms. The tyramine content in 13 collected Cheonggukjang samples averaged 604.9 mg/kg. Since the tyramine content measured from most samples was sufficient to cause harm to the human body, it is necessary to control its production in food. Enterococci were confirmed to be the bacterial species producing most of the tyramine through the microbial examination and were present in high numbers from not detected (<10(1)) to 7.0 × 10(10) colony-forming units (CFU)/g. To control the growth of enterococci, various plant extracts with antimicrobial activity, common salts, and variable temperature conditions were tested. It was found that 4 samples among the 159 plant extracts had a strong antimicrobial activity in Cheonggukjang, especially against Enterococcus faecium, showing viable cell counts of <10(1)-10(3) CFU/g after 24 h of ripening, which were significantly lower values compared to the control (10(9)-10(11) CFU/g). The Cheonggukjang with the addition of the four plant extracts showed ∼83%-95% lower concentrations of tyramine compared to the control. Cheonggukjang prepared with the Schizandra chinensis Baillon extract had the lowest tyramine content without sacrificing the sensory quality. Not only was the bacterial species of E. faecium reduced more remarkably, by up to 10(3) CFU/g compared to the 10(9)-10(11) CFU/g shown in the control, but it also decreased the tyramine content by up to 91%.

Effects of minerals on sporulation and heat resistance of Clostridium sporogenes.

In this study, various mineral supplements, such as chloride salts (CaCl2, MgCl2, MnCl2, FeCl2 and KCl) supplying cations and calcium salts (CaCl2, CaCO3, CaSO4, Ca(OH)2 and CaHPO4) supplying anions, were tested if they could stimulate the sporulation of Clostridium sporogenes, a surrogate microorganism for C. botulinum. Of the cations tested, the addition of CaCl2 showed a slightly, but not significantly, greater increase in spore levels within 3 weeks of incubation, compared to that of the other cations. The optimum concentration of CaCl2 was 0.5%, which yielded nearly 10(4) CFU/ml of spores. Of the anions tested, CaCO3 promoted sporulation within one week, which was the most effective compound for promoting rapid sporulation among the minerals tested. CaSO4 produced a pattern of sporulation similar to that of CaCl2. While CaHPO4 resulted in the maximum production of spores after 4 weeks, Ca(OH)2 failed to induce sporulation. With an optimized concentration of 0.5% CaCO3, the spore yield was approximately 10(5) CFU/ml. The spores prepared in sporulation medium with CaCO3 (pH 5.0) had slightly, but not significantly, higher D values than those produced with CaCl2 (pH 5.0) at temperatures ranging from 113 to 121 degrees C. However, no significant differences were observed in Z values (both 10.76 degrees C). In a large scale spore production, D(121 degrees C) values of the spore crops prepared with CaCl2 and CaCO3 and resuspended in phosphate buffer (pH 7.0) were found to be both 0.92 min. In conclusion, our data suggest that CaCO3 is highly effective in reducing sporulation time as well as enhancing heat resistance.

Optimization of production of monacolin K from gamma-irradiated Monascus mutant by use of response surface methodology.

Monascus isolate number 711, which is capable of producing monacolin K as an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, the key enzyme of cholesterol synthesis, was isolated from Ang-kak, the red yeast rice koji. To increase the monacolin K-producing activity of the strain, spore suspensions of the strain were subjected to gamma-irradiation. One thousand mutants were generated via gamma-irradiation and screened using bioassay and high performance liquid chromatography analysis. Several mutants with higher productivities of monacolin K than that of the parent strain were primarily selected. Mutant KU609 was finally selected because of its characteristics of high monacolin K production and non-citrinin-producing activity under our test conditions. Response surface methodology was used to analyze the effect of culture medium on the production of monacolin K in mixed solid-state cultures. The optimal values of nutritional ingredients for the maximal production were soytone, glucose, MgSO4, and barley at concentrations of 0.5 g, 0.48 g, 0.053 g, and 9 g, respectively. The final monacolin K production of Monascus KU609 was increased almost 100-fold compared to that of the parent strain.