메주와 누룩은 한국 전통 발효 식품에 사용되는 스타터로, Aspergillus속 곰팡이나 aflatoxin에 노출되기 쉽다. 본 연구에서는 우리나라에서 시판되는 57개의 메주 시료와 18개의 누룩 시료로부터 Aspergillus 속 곰팡이를 분리하고 동정하였다. 분리주의 aflatoxin 생성 가능성을 평가하기 위하여 multiplex PCR을 통해 aflatoxin 생합성 유전자 (aflO, aflP, aflR)를 확인하고, 이들 분리주에 의해 생성되는 aflatoxin 함량을 HPLC로 조사하였다. 뿐만 아니라 시판 메주와 누룩 시료 중 aflatoxin 함량을 분석하였다. 그 결과, 메주 시료로부터 130개, 누룩 시료로부터 47개 균주가 분리되어 총 177개의 분리주를 확인 및 동정하였다. 각 각 메주와 누룩으로부터 분리된 19.2% (25/130), 10.6% (5/ 47)의 분리주가 3 종류의 aflatoxin 생합성 유전자를 모두 보유하였으며, 그 중 메주로부터 분리된 5개의 분리주가 실제 로 aflatoxin을 생성하였다. 시판 메주와 누룩 시료 중 aflatoxin 함량을 분석한 결과, 88% (51/58)의 메주 시료의 aflatoxin 오염 수준은 모두 검출한계 미만으로 나타났고, 누룩 또한 시료의 39% (7/18)가 검출한계 미만으로 확인되었다. 메주와 누룩에서 분리된 분리주 중 aflatoxin 생합성 유전자를 모두 보유하거나 배지 상에서 aflatoxin 생성을 보여준 aflatoxigenic 균주는 존재하였으나 유통되고 있는 시료에서 aflatoxin 오염 빈도는 낮은 수준임을 확인할 수 있었다.

2016년 6~12월 서울약령시에서 원형 또는 분말 형태로 유통 판매되는 식·약 공용 농산물 총 62건을 구입하여, 후칼럼 유도체화 장치인 광화학 반응장치를 연결한 HPLCFLD를 이용하여 아플라톡신(B1, B2, G1, G2)을 정성 및 정량 분석함으로써 오염도를 조사하였다. 아플라톡신의 검 출한계는 B1 0.086 μg/kg, B2 0.020 μg/kg, G1 0.363 g/kg, G2 0.126 μg/kg이었고, 정량한계는 B1 0.262 μg/kg, B2 0.059 μg/ kg, G1 1.101 g/kg, G2 0.382 μg/kg였다. 회수율은 B1 95.1~ 101.8%, B2 86.9~105.6%, G1 95.2~100.6%, G2 98.6~114.0% 이었으며, 변동계수(Coefficient of Variation, %)는 B1 1.9~ 9.8%, B2 1.2~8.2%, G1 2.1~9.5%, G2 0.9~9.3%였다. 사용부위에 따른 아플라톡신 오염의 특성을 살펴본 결과 종자 (Semen)만 아플라톡신이 검출되었으며, 21건 중 6건(원형 3건, 분말 3건)에서 아플라톡신이 검출되었다. 품목별 아플라톡신 검출 건수를 살펴본 결과, 연자육 중 원형은 7 건을 검사한 결과 2건에서 아플라톡신 B1, B2가 검출되었고, 분말은 6건 중 2건에서 아플라톡신 B1이 검출되었다. 산조인 원형은 5건 중 1건에서 아플라톡신 B1, B2가 검출 되었으며, 분말은 3건 중 1건에서 아플라톡신 B1, B2가 검출되었다. 아플라톡신 검출량을 살펴보면, 연자육 원형은 총아플라톡신(B1, B2, G1 및 G2의 합)이 ND~14.655 μg/kg(B1 ND~11.869 μg/kg) 검출되었고, 분말은 총아플라톡신이 ND~2.223 μg/kg(B1이 ND~2.223 μg/kg) 검출되었다. 산조 인 원형의 아플라톡신 검출량은 총아플라톡신이 ND~9.182μg/kg(B1이 ND~6.612 μg/kg) 검출되었고, 분말은 총아플라 톡신이 ND~21.797 μg/kg(B1이 ND~19.345 μg/kg) 검출되었다. 따라서 이러한 곰팡이독소 검출현황을 토대로 식· 약 공용 농산물에 대한 곰팡이독소 기준을 설정하여 식· 약 공용 농산물의 안전한 유통을 관리할 필요가 있다.

돼지고기 중 5종의 곰팡이독소(아플라톡신 B1, B2, G1, G2 및 오크라톡신 A)를 분석하기 위해 0.1% 개미산을 함유한 50% 아세토니트릴용액으로 추출한 후 고체상추출칼럼을 이용하여 정제하고, LC-MS/MS로 동시 정량할 수 있는 시험법을 마련하였다. 분석조건으로 측정한 5종의 곰팡이독소 matrix-matched 표준곡선식에서 모두 상관계수 0.998이상의 상관관계를 나타내었다. 5종의 곰팡이독소 2배의 정량한계에서 10배의 정량한계로 첨가한 시료에서 평균 회수율은 72.1~109.9%로 실험 결과들이 EU 가이드라인에서 제시하는 유효성 확인을 위한 기준을 만족함으로써 시험법의 신뢰성을 확보할 수 있었다. 충청지역 유통되고 있은 돼지고기 20건에 대한 총아플라톡신 및 오크라톡신A에 대한 오염도 조사결과 정량한계 미만으로 조사되었다.

곡류에서 오염되는 곰팡이독소인 아플라톡신과 데옥시 니발레놀을 경기도 내 유통 곡류 및 그 가공품 205개 시료에서 분석하였다. 총아플라톡신은 16개 시료(8%)에서 0.01~27.88 μg/kg 농도로 검출되었고, 데옥시니발레놀은 56 개 시료(27%)에서 2.2~754.4 μg/kg 농도로 검출되었으며, 8개 시료는 아플라톡신과 데옥시니발레놀이 동시에 검출 되었다. 아플라톡신의 검출빈도는 건조옥수수와 부침가루 각각 33%, 튀김가루 25%, 수수 18%, 밀과 기장에서 각각 13%로 나타났고, 데옥시니발레놀의 검출빈도는 율무 85%, 밀 75%, 팝콘용옥수수가공품 60%, 수수 55%, 보리 44%와 건조옥수수 33%로 나타났다. 본 연구 결과를 기초로 한 곡류의 총아플라톡신과 데옥시니발레놀의 성인 1일 인체노출량은 0.80 (×) 10−3 μg/kg b.w./day과 0.18 μg/kg b.w./day 이었으며, 데옥시니발레놀의 성인 1일 인체노출량은 JECFA에서 제정된 PMTDI 1 μg/kg b.w./day 대비 18.5%에 해당하는 농도로 곡류 섭취를 통한 건강상의 위해 가능성은 미약한 것으로 평가되었다. 그러나 총아플라톡신이 가장 높게 검출된 건조옥수수 (27.88 μg/kg)와 데옥시니발레놀이 가장 높게 검출된 율무 (754.4 μg/kg)의 경우 총아플라톡신은 국내·외 규제 최고 농도를 초과한 수준이었으며, 데옥시니발레놀은 일부 국외 규제 최고농도를 초과한 수준이었다. 따라서 지구온난화로 우리나라의 기후조건이 곰팡이가 생육하기 좋은 환경으로 변해가므로 곡류제품이 원산지로 부터 소비에 이르기까지 생산·유통과정 중 어느 시점에서 곰팡이독소가 오염되는지에 대한 연구·관리가 더욱 활발히 수행되어야 할 것으로 판단된다.

The increase in the consumption of herb medicines have made their use a public health problem due to the potential fungal contamination and the risk of the presence of mycotoxins. 360 samples of herb medicines were evaluated for the aflatoxin contamination. The natural occurrence of aflatoxins in these samples were determined using immunoaffinity column clean up and high performance liquid chromatography (HPLC) with post-column derivatization. For samples analyzed, mean levels (incidence) of AFB1, AFB2, AFG1 and AFG2 in positive samples were 1.4 μg/kg (46.4%), 0.4 μg/kg (25.4%), 1.1 μg/kg (37.8%) and 0.9 μg/kg (24.3%), respectively. Recoveries of the full analytical procedure were 71.7~99.7% for AFB1, 88.1~99.2% for AFB2, 82.8~95.5% for AFG1 and 77.9~90.0%for AFG2. The excess cancer risk estimated using the cancer potency of aflatoxin B1 (7 (mg/kg/day)−1 for HBsAg− and 230 (mg/kg/day)−1 for HBsAg+) were 1.30 × 10^(−5) ~ 1.22 × 10^(−7) for hepatits B surface antigen negative (HBsAg−) and 3.31 × 10^(−4)~ 3.12 × 10^(−6) for hepatits B surface antigen positive (HBsAg+) respectively. In conclusion, although the contamination levels of samples used in the study were low, further actions are also required to undertake a program of herbal surveys in order to access mycotoxin contamination overall so that the safety of public will be protected.

This study was performed to investigate contamination levels of aflatoxins, the secondary metabolites produced by fungi Aspergillus flavus and A. parasiticus, in herbal medicine. Herbs is susceptible to these fungi infections through its growth harvest, transport and storage. This study determine the aflatoxin B₁, B₂, G₁ and G₂levels by HPLC-florescence detector coupled with photochemical enhancement in 558 samples herbal medicine distributed in Korea and China. Also, We checked a transfer ratio of aflatoxins from raw herbal medicines to herbal medicine extract. Hot water extraction of herbal medicines was prepared by air pressure and high pressure condition. The analytical method for aflatoxins was validated in this method. In results recoveries of the analytical method were ranged from 67.4% to 96.2% and, limits of detection and quantitation for aflatoxins were 0.015~0.138 μg/kg and 0.046~0.418 μg/kg, respectively. According to the results of monitoring on aflatoxins in herbal medicine, aflatoxins 1.7 ug/kg B1 and 0.9 ug/kg G₁were detected in only one sample of Strychni Ignatii Semen, and 0.8 ug/kg G₁ in Strychni Semen. About 13.6~51.3% of aflatoxins were transferred to hot water extract. Although the detected levels are under the permitted levels for aflatoxins in herbal medicine, these amounts should be considered in regard to overall daily exposure to mycotoxins.

Aflatoxins produced by Aspergillus spp. are recognized as a major concern in animal and human health. In pigs, ingestion of aflatoxin-contaminated feeds causes immunosuppression, hepatotoxicosis and poor feed efficiency. In this study, we screened the contaminated level of aflatoxins in 449 pig feeds from 12 swine farms in Korea. For rapid and efficient screening of aflatoxins in pig feeds we evaluated the feasibility of three commercial ELISA kits for the screening of aflatoxins in pig feeds. Twenty-nine pig feed samples were examined for total aflatoxins using three ELISA kits, simultaneously. From three repetitions of each assay, the average intra-assay precisions and the average inter-assay precisions expressed as coefficient of variation (CV, %) for VeratoxⓇ Quantitative Aflatoxin test were 6.90 and 12.29, respectively. A statistical comparison of the results between HPLC and ELISAs showed that the correlation coefficient values for VeratoxⓇ was 0.96. The results demonstrated that we can apply the VeratoxⓇ Quantitative Aflatoxin test for the detection of aflatoxins in pig feed from the field. The screening of field samples with this ELISA kit showed that 11 out of 265 pig feeds for growers and weaners were contaminated with total aflatoxin levels exceeding 10 ppb, maximum tolerable limits for their compound feeds and the aflatoxins levels of remaining 184 pig feeds for other age groups of pigs were confirmed as below 20 ppb. The results from the screening indicated overall low levels of aflatoxins contamination in pig feeds.

We used fluorescence detector to analyse total aflatoxins (G1, G2, B1, B2) with TFA (Trifluoroacetic acid) derivation method and PHRED (Photochemical reactor enhanced detection) method. PHRED method was superior in reproduction and convenience, but TFA derivation method was superior in selectivity and sensitivity. The recovery rate of aflatoxin B1, B2, G1 were more than 80%, and G2 was more than 70%. The detection limit of B1,B2, G1 and G2 were respectively 0.05, 0.05, 0.2 and 0.1 μg/kg. Confirmed method was used to analyse total aflatoxins in total 316 items as 9 kinds 137 dried fruits and 10 kinds 179 spices. By the result, Aflatoxins were detected in 27dried fruits (19.7%) and in 87 spices (48.6%).

A survey of total aflatoxin levels was conducted on 145 samples(carthamiflos, thujae semen, giycyrrhizae radix et rhizoma) collected in Yakyeang markets in Seoul. Aflatoxin levels were quantified by the immunoaffinity column clean-up method followed by performance liguid chromatography(HPLC)-fluorescence detector(FLD). Aflatoxins were found in 10(6.9%)samples including 5 Arecae semen, 4 Thujae semen, 1 Zizyphi semen with a range of 0.45~79.15 μg/kg. Generally These results show that the contamination level of aflatoxins in Herb Medicines consumed in Korea is high compared with the standard in Korea Herb Medicine Code(10 μg/kg as aflatoxin B1). It is considered that aflatoxin concentration was increased in herb medicines during a storage and drying in herb medicines examined

경기지역 시중마트와 국내 온라인상에서 유통되고 있는 견과류 및 그 가공식품 79건, 장류 29건, 곡류가공품 등 가공식품 50건에 대해 immunoaffinity column 정제방법을 이용하여 HPLC-FLD로 아플라톡신 오염실태를 조사한 결과 총 158건 중 45건(28.5%)에서 아플라톡신 오염이 확인되었고, 오염수준은 아플라톡신 B1으로서 0.02~3.13 μg/kg, 총 아플라톡신으로서 0.02~3.96 μg/kg 범위로 국내 기준치인 10 μg/kg을 초과하는 것은 없었다. 아플라톡신이 검출된 식품은 견과류 및 그 가공식품에서 34건(43.0%) 장류에서 8건(27.6%),그 외 가공식품에서 3건(6.0%)으로 견과류 및 그 가공식품에서 검출빈도가 가장 높았다. 식품을 개봉하여 1개월, 6개월 보관 후의 아플라톡신B1 함량은 저장기간이 길어질수록 땅콩, 옥수수스낵에서는 증가하는 경향을 보였으나 된장의 경우에는 다소 감소하였다.

This study was performed to investigate the possible effect of sunlight on the reduction or degradation of mycelia and aflatoxins. The mycelia and aflatoxins were produced by Aspergillus parasiticus ATCC 15517 in a yeast-extract sucrose broth (YES) and potato-dextrose agar (PDA) and then exposed to sunlight. The weight of mycelia was decreased to 76.8% in 8 hours and to 66.7% in 168 hours(p$lt;0.05). The total aflatoxin level was significantly decreased to below 50% (46.3% in the YES broth and 49.6% in the PDA) in 8 hours (p$lt;0.05). After 168 hours, a 90.4% degradation of aflatoxin in the YES broth and a 77.2% degradation of aflatoxin in the PDA was observed, respectively (p$lt;0.01). The results showed that the degradation ratios of total aflatoxin level increased with increased exposure time to sunlight. These results indicate that sunlight could be an effective factor in aflatoxin degradation although its effect on mycelia was less pronounced.

This study was performed to investigate the possible effect of Bacillus subtilis which is the predominant species of bacteria in Korean soy sauce, soy paste, and Meju (soybean cake) on the growth and aflatoxin production of Aspergillus parasiticus ATCC 15517. The microorganisms were grown in a modified APT broth and incubated at 30℃ for 12 days. Aflatoxins were determined using high performance liquid chromatography (HPLC). A remarkable inhibition of the growth of Aspergillus parasiticus was observed during the incubation period when in the presence of B. subtilis (mixed culture). Dry mycelial weight in the mixed culture was significantly reduced by 85.3% in comparison to the control at the end of the incubation period (p$lt;0.01). Lower levels of aflatoxins were found in the mixed culture than in the monoculture. At the end of the incubation period aflatoxin production was significantly inhibited by more than 50% (p$lt;0.05). These results indicate that B. subtilis mainly inhibites the growth and aflatoxin production of toxigenic Aspergillus in Meju, soy sauce and soy paste. Although its effect on aflatoxin production was less pronounced, we . could expect more inhibition by another bacteria related with fermentation in Meju.

HPLC에 의한 주요 aflatoxins(afatoxin B₁, B₂, G₁ 및 G₂)의 동시 분석에서 postcolumn 유도체화법을 시도하였다. Electrochemical cell(Kobra-cell)을 사용한 postcolumn 유도체화법은 기존의 precolumn 유도체화법보다 분석시간을 단축하였으며(약 1/2 단축), 더 안전하고, 향상된 분석능을 보였다. Aflatoxin B₁과 G₁의 경우 10~100 ppb에서, 그리고 B₂와 G₂의 경우 3~30 ppb에서 직선성을 나타내었다. Aflatoxin B₁과 G₁은 각각 88.9% 및 100.5%로 양호환 회수육을 보였다. Aflatoxin B₂와 G₂의 경우 분리도는 우수하였으나 회수율에 있어서 변이가 크게 나타났다.

A procedure for the determination of Aflatoxins in food and grains which utilizes reversed phased liquid chromatographic (LC) analysis with postcolumn derivatization by an electrochemical cell and determination with a fluorescence detector has been evaluated. The LC mobile phase was water-acetonitrile-methanol (6+2+2) with 1mM KBr and 1 mM HNO₃ which gave baseline separation for the four Aflatoxins (AfB₁, AfB₂, AfG₁, AfG₂). The electrochemical cell set at 7V, generated bromine and derivatized aflatoxins B₁ and G₁, The derivatives were detected by the fluorescence detector. The aflatoxins in naturally contaminated corn samples were isolated by three different cleanup procedures: the AOAC method I column (CB method), a rapid filtrate column (Romer's column), and an immunoaffinity column. The final extract were quantitated with fluorodensitometric TLC and the LC postcolumn derivatization techniques. The results were quite similar, however the LC technique showed less interferences and could be automated. Samples of corn, raw peanuts, peanut butter and dried dates were also analyzed successfully with this procedure.

Our paper shows the results of 302 samples of herb medicines about fungal contamination at Yakyeang markets in Seoul. The sample medicines were treated VICAM pretreatment and analysed by post column derivatisation procedure(PHRED-HPLC) with a fluorescence detector. Aflatoxin B1 was founded from 50.3% of samples, aflatoxin B2 was 39.7%, aflatoxin G1 was 21.2% and aflatoxin G2 was 23.5%. The detected ranges of aflatoxin B1, B2, G1 and G2 were from 0.1 to 57.2 μg/kg, 0.1 to 42.6 μg/kg, 0.1 to 23.5 μg/kg and 0.1 to 9.5 μg/kg respectively. Among total samples, 26 samples contained aflatoxin B1 violated the regulation (less than 10 ug/kg) for aflatoxin B1 of KFDA. From the result, we could presumed that more than a half of samples were contaminated by aflatoxins. Therefore, it seems to be necessary that the new safety giudeline will be established aflatoxin B2, G1 and G2 from herb medicines as aflatoxin B1.