• Asian-Australasian Journal of Animal Sciences
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• 제15권7호
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• pp.1051-1056
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• 2002

In vitro binding efficacy of esterified glucomannan (E-GM) (0.1%) on aflatoxin B1 (AF) (300 ppb), ochratoxin A (OA) (2 ppm) and T-2 toxin (T-2) (3 ppm), when present alone or in combination, was evaluated in toxin-contaminated feed at pH 4.5 and 6.5. Esterified glucomannan showed significantly (p<0.01) higher binding with AF (81.6%), whereas those recorded with T-2 (27.8%) and OA (25.6%) were moderate. Binding of each toxin decreased as the number of toxins in feed increased. pH of medium showed no effect on mycotoxin binding ability of E-GM. A $2{\times}2{\times}2{\times}2$ factorial experiment of 5 week duration was conducted to study the effects of two dietary levels each of AF (0 and 300 ppb), OA (0 and 2 ppm), T-2 (0 and 3 ppm ) and E-GM (0 and 0.1%) on the immune competence of a total of 960 day-old commercial broilers. Reductions in size of thymus (by AF and T-2) and bursa (by AF) and antibody titers against Newcastle disease and Infectious Bursal disease (by all the toxins) were noted. Additive and antagonistic interactions were seen among the toxins on certain parameters. Esterified glucomannan significantly (p<0.01) improved antibody titers and weights of bursa ofFabricius and thymus indicating its counteracting efficacy against immunosuppression in mycotoxicosis of multiple origin.

• Food Science and Biotechnology
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• 제14권6호
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• pp.866-870
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• 2005

The 14 lactic acid bacteria (LAB) have been evaluated to determine the binding capacity to HT29 cell and Aflatoxin $B_1$ ($AFB_1$). The interaction of LAB to HT29 cells has been further investigated to identify the possibility of competing the binding sites with $AFB_1$. Of 14 LAB strains, Lactobacillus casei KCTC 3260 demonstrated the higher adhesiveness to HT29 and $AFB_1$ with the rate of 19.6% and 46.3%, respectively. In competitive analysis for binding sites, the adhesion of L. casei KCTC 3260 to HT29 cells was reduced with 100 nmol $AFB_1$ by 31.2%. The protoplast of L. casei KCTC 3260 showed no binding capacity to HT29 cells with increment of $AFB_1$ concentration, indicating that cell wall components might serve as a critical factor for the binding. To discriminate the major component influencing on L. casei KCTC 3260 binding to HT29 cells and $AFB_1$, four different pre-treatments (lipase, pronase E, sodium m-periodate, and urea) were employed. Of those, sodium m-periodate treatment caused the lower adhesion of L. casei KCTC 3260 to HT29 cells with the increment of $AFB_1$ concentration. These results indicated that carbohydrate moiety on the cell wall of L. casei KCTC 3260 might be the most critical component in binding to both HT29 cells and $AFB_1$.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 춘계총회 및 학술대회
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• pp.93.3-94
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• 2000

• 한국식품과학회지
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• 제45권2호
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• pp.235-240
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• 2013

본 연구에서는 분유 중 아플라톡신 $M_1$을 면역친화성칼럼을 이용하여 정제하고 HPLC로 분리하였으며 형광검출기로 정량 분석하는 시험법을 확립하였다. 이 시험법을 이용하여 439건의 시료에 대한 분석을 실시하였다. 그 결과, 아플라톡신 $M_1$의 검출농도는 평균 2.6 ng/kg(불검출-14.9 ng/kg)로 나타났으며, 검출률은 64%로 조사되었다. 본 연구의 오염도자료를 이용한 분유 중 아플라톡신 $M_1$의 노출량 및 위해 수준은 연령별로 비교했을 때 신생아에게서 가장 높게 나타났으나 초과간암발생률은 1년에 백만명 중 0.02명 정도로서 위해수준은 매우 낮은 것으로 평가 되었다.

• Food Science and Biotechnology
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• 제17권3호
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• pp.623-630
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• 2008

A non-instrumental immunochromatographic (ICG) strip-test and direct competitive enzyme-linked immunosorbent assay (DC-ELISA) for aflatoxin B1 (AFB1) determination were developed and optimized. The detection limits of ICG strip-test and DC-ELISA were 0.5 and 0.004 ng/mL, respectively, and these methods possessed a cross-reaction to aflatoxins. The results of spiked samples by both methods were coincided with the amount spiked AFB1 and the comparative analyses of 172 real samples by 2 immunoassays and high performance liquid chromatography (HPLC) showed a good agreement. Especially, the ICG strip-test is easier to perform and quicker, but less sensitivity than DC-ELISA. Both methods could analyze a high sample throughput with short time, but the sample throughput of ICG strip-test was better. Therefore, the ICG strip-test can be used as a simple, easy, non-instrumental, and fast screening technique for AFB1 determination.

• 한국동물위생학회지
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• 제36권4호
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• pp.303-309
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• 2013

This study was conducted to determine the content of 7 mycotoxins (aflatoxin $B_1$, $B_2$, $G_1$, $G_2$, $M_1$, ochratoxin A and zearalenone) using LC-MS/MS in pork available on the Korean markets. The analysis was carried out using following conditions; C18 column ($2.1{\times}100mm$, $1.7{\mu}m$), mobile phase composed of $H_2O$ (0.1 mM $NH_4Ac$ 0.01% HCOOH) : Methanol (0.1 mM $NH_4Ac$ 0.01% HCOOH), binary pump at a flow rate of 0.5 mL/min and $2{\mu}L$ of injection volume, MS/MS detector with ESI positive and negative mode. The quantication of mycotoxins was based on matrix-matched calibration curves with a correlation coefficient in excess of 0.99 for the 7 mycotoxins. The dectection limits were ranged 0.74~2.13 ng/g, with mean recoveries between 73.10~97.46% except aflatoxin $B_1$ (61.31%). We also monitored mycotoxin residues in 208 pork samples. The test results, mycotoxins were not found except one sample. Ochratoxin A in one sample of the test samples was detected below the quantification limit.

• Toxicological Research
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• 제27권2호
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• pp.125-131
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• 2011

Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with ${\lambda}$-type light chains. The $IC_{50}s$ of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml ($R^2$ > 0.99) and from 1 to 100 ng/ml ($R^2$ > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.

• 한국식품과학회지
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• 제42권1호
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• pp.27-32
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• 2010

고시된 아플라톡신 시험법의 정량한계, 검출한계를 개선한 분석법을 유효화하였으며, 개선한 분석법으로 강활, 개자 등의 생약 400개의 시료를 분석하였다. 사군자와 제니 각 1건에서 아플라톡신 $B_1$이 각 $2.3\;{\mu}g$/kg 검출되었으며, 사군자와 대풍자 1건에서 정량한계 미만의 아플라톡신이 검출되었다. 탕제로의 이행률은 아플라톡신 $B_1$의 경우 약 20% 정도의 이행률을 나타냈다. 아플라톡신 $B_1$ 기준치보다 낮게 검출되어 현재까지는 아플라톡신으로부터 안전한 것으로 판단되었다.

• Food Science and Biotechnology
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• 제18권3호
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• pp.641-648
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• 2009

A one-step simultaneous immunchromatographic (OS-ICG) assay using colloidal gold-monoclonal antibody (gold-MAb) conjugates was developed for the rapid multianalysis of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feed samples. Visual detection limits for AFB1 and OTA were 0.5 and 2.5 ng/mL, respectively, and the results were obtained within 15 min. Matrix interference from the feed extracts was efficiently reduced by appropriate dilution with buffer. Cut-off values of the OS-ICG assay for the feed spiked with AFB1/OTA mixtures (5/5, 10/10, 25/25, 50/55, 100/100 ${\mu}g/kg$) were 10 and 50 ${\mu}g/kg$ for AFB1 and OTA. The comparative analyses of 65 feed samples by OS-ICG, enzyme-linked immunosorbent assay (ELISA), and high performance liquid chromatography (HPLC) showed good agreement. In this study, we confirmed that simultaneous analysis based on immunoassay is possible and it can be used as an on-site multianalysis of AFB1 and OTA in feed, food, and agricultural products.

• Toxicological Research
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• 제3권1호
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• pp.1-8
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• 1987

In vitro induction of cytochrome 450 associated monooxygenase activities by phenobarbital (PB) and 3-methylcholanthrene (MC) was investigated in primary cultures of adult rat hepatocytes. PB and MC were added to the culture 24 hr after the initial plating of hepatocytes. A signiftcant increase of the activities of 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase were observed in MC and PB treated culture. MC caused about 500% induction of the initial oxidation rates of both enzymes in 48 hr. However the PB maintained both enzyme activities close to the level of freshly isolated hepatocytes. Biphenyl 4-hydroxylase and aminopyrine N-demethylase activities were also induced by MC and PB. But the level of induction was less than that occuring with 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase. When aflatoxin $B_1$ was added to the hepatocyte cultures which have been treated with MC or PB, it caused a significant increase of the unscheduled DNA synthesis at higher dose of aflatoxin $B_1$ as compared to those of untreated control hepatocyte cultures. The results suggest that microsomal enzyme activities can be selectively controlled preferably in hepatocyte cultures by the in vitro induction method. This principle may be useful for studying the metabolism and other toxicological studies.