• Title/Summary/Keyword: immunoaffinity column

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Effect of Various Culture Conditions on the Production of Mycotoxin by Aspergillus sp. (배양 조건이 Aspergillus sp.의 독소 생산에 미치는 영향)

  • Lee, Yu Na;Kim, Nam Yeun;Lee, Seung Eun;Ji, Geun Eog
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.36-41
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    • 2016
  • Ochratoxin A and aflatoxin may be detected from naturally fermented foods due to the contamination of the mycotoxin-producing molds or un-prudential use of the mycotoxin producing starter strains during the fermentation. This study was carried out to analyze the production of ochratoxin A and aflatoxin under the various environmental conditions. For the experiment, the effects of different temperature, culture media, and fermentation time on the production of ochratoxin A by Aspergillus usamii KFRI 999 and A. awamori KFRI 983 were analyzed. Additionally, the production of aflatoxin was assessed under the various temperature, initial pH, fermentation time and culture media during fermentation by A. flavus KACC 41403 and A. oryzae KACC 46471. The levels of ochratoxin A and aflatoxin were analyzed by HPLC. The result showed that the production of mycotoxin was greatly affected by the fermentation temperature. A. oryzae KACC 46471 did not produce aflatoxin. All of the mycotoxin producing strains showed the highest level of mycotoxin at $30^{\circ}C$. A. awamori KFRI 983 showed the lowest level of ochratoxin A in PDA media among the experimental medium. The results of the present study may be useful for the reduction of ochratoxin A and aflatoxin in various foods.

Analysis of Total Aflatoxins in Spices and Dried Fruits (향신료 및 건조과실류 중 총 아플라톡신의 분석)

  • Kang, Young-Woon;Cho, Tae-Yong;Park, Hee-Ra;Oh, Keum-Soon;Kim, Dong-Sul
    • Journal of Food Hygiene and Safety
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    • v.25 no.1
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    • pp.65-72
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    • 2010
  • 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 $0.1\;{\mu}g/kg$. 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 27 dried fruits (19.7%) and in 87 spices (48.6%).

Contents of Mineral Elements and Cytokinins in Xylem Sap of Two Oriental Melon Cultivars Affected by Rootstocks (참외품종과 대목종류에 따른 목부액 내의 무기성분 및 시토키닌 함량)

  • Kim, Sung-Eun;Jung, Ki-Hwan;Lee, Jung-Myung
    • Horticultural Science & Technology
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    • v.17 no.6
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    • pp.742-746
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    • 1999
  • Contents of mineral element and cytokinin in the xylem sap of 'Keumdongee' and 'Tongilhwang' oriental melons were compared with those in oriental melons grafted onto 8 rootstocks. The effect of grafting on the fruit quality of oriental melon was also investigated. Flesh firmness varied with rootstocks. Soluble solids contents in the placenta tissue of grafted 'Tongilhwang' were higher than that in the 'Keumdongee'. Electric conductivity of the xylem sap in own-rooted plants was higher in 'Keumdongee' than in 'Tongilhwang', but it increased in 'Tongilhwang' once they were grafted. The sap volume per plant was greater in 'Keumdongee' than in 'Tongilhwang'. The mineral concentrations varied considerably depending on the rootstock used. Xylem sap of grafted oriental melons contained a higher amount of mineral ions, especially $NO_3{^-}$ and $PO_4{^-}$, than did the sap in own-rooted plants. The increase in the mineral levels in sap due to grafting was most apparent in 'Tongilhwang'. Xylem sap from both 'Keumdongee' and 'Tongilhwang' contained trans-zeatin (t-Z), trans-zeatin riboside (t-ZR), and dihydrozeatin riboside (DHZR). Small amounts of isopentenyl adenine (IPA) and isopentenyl adenine riboside (IPAR) were also detected. Trans-zeatin riboside was the most abundant, followed by t-Z. Cytokinin concentration in 'Keumdongee' was not significantly influenced by rootstock type used, although the highest concentration of cytokinins in 'Keumdongee' was obtained with 'Chamtozwa' rootstock. However, the cytokinin concentration in 'Tongilhwang' increased with grafting irrespective of rootstock type used.

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Survey on Fusarium Mycotoxin Contamination in Oat, Sorghum, Adlay, and Proso Millet during the Harvest Season in Korea (귀리, 수수, 율무, 기장의 수확기에 발생하는 Fusarium 곰팡이독소 오염도 조사)

  • Lee, Mi Jeong;Wee, Chi-Do;Ham, Hyenheui;Choi, Jung-Hye;Baek, Ji Sun;Lim, Soo Bin;Lee, Theresa;Kim, Jeom-Soon;Jang, Ja Yeong
    • Journal of Food Hygiene and Safety
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    • v.35 no.1
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    • pp.13-22
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    • 2020
  • A total of 244 cereal samples (oat, sorghum, adlay, and proso millet) were collected from fields to examine the contamination of Fusarium mycotoxins in cereals during harvest season in 2017 and 2018. The contamination levels of deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEA) were analyzed individually by using the immunoaffinity column clean-up method with ultra performance liquid chromatography, and fumonisins (FUM) were analyzed by using the QuEChERS method with liquid chromatography-mass spectrometry. Highest level of NIV contamination (120.0-3277.0 mg/kg) was observed in oat samples among the analyzed cereals. In the adlay samples, DON contamination was the highest (maximum level 730.0 ㎍/kg). The proso millet samples had a high frequency of detection of NIV and ZEA (61.5% and 57.9%, respectively), but the levels were low (average detection level of NIV, 75.6 ㎍/kg, for ZEA, 21.5 ㎍/kg). Among the cereal samples, sorghum had the highest contamination frequency of DON, ZEA, and FUM, and the co-occurrence of Fusarium mycotoxin was 70.0%, which was higher than the average of 29.9%. In order to safely manage Fusarium mycotoxin levels in cereals, continuous research on the development of contamination prevention technologies together with monitoring of mycotoxin contamination is needed.

A study on Aflatoxins Analysis in The Herb Medicines (한약재 중 아플라톡신 분석에 관한 연구)

  • Lee, Sung-Deuk;Kim, Yeon-Sun;Kim, Nam-Hoon;Jung, Hee-Jung;Jung, Sam-Joo;Kim, Hwa-Soon;Kim, Kyung-Sik;Han, Ki-Young
    • Journal of Food Hygiene and Safety
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    • v.26 no.4
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    • pp.424-434
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    • 2011
  • 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 my cot ox ins. 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 $1.4\;{\mu}g/kg$(46.4%), $0.4\;{\mu}g/kg$(25.4%), $1.1\;{\mu}g/kg$(37.8%) and $0.9\;{\mu}g/kg$(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 $1.30{\times}10^{-5}{\sim}1.22{\times}10^{-7}$ for hepatits B surface antigen negative ($HBsAg^-$) and $3.31{\times}10^{-4}{\sim}3.12{\times}10^{-6}$ 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.

A Monitoring of Aflatoxins in Commercial Herbs for Food and Medicine (식·약공용 농산물의 아플라톡신 오염 실태 조사)

  • Kim, Sung-dan;Kim, Ae-kyung;Lee, Hyun-kyung;Lee, Sae-ram;Lee, Hee-jin;Ryu, Hoe-jin;Lee, Jung-mi;Yu, In-sil;Jung, Kweon
    • Journal of Food Hygiene and Safety
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    • v.32 no.4
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    • pp.267-274
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    • 2017
  • This paper deals with the natural occurrence of total aflatoxins ($B_1$, $B_2$, $G_1$, and $G_2$) in commercial herbs for food and medicine. To monitor aflatoxins in commercial herbs for food and medicine not included in the specifications of Food Code, a total of 62 samples of 6 different herbs (Bombycis Corpus, Glycyrrhizae Radix et Rhizoma, Menthae Herba, Nelumbinis Semen, Polygalae Radix, Zizyphi Semen) were collected from Yangnyeong market in Seoul, Korea. The samples were treated by the immunoaffinity column clean-up method and quantified by high performance liquid chromatography (HPLC) with on-line post column photochemical derivatization (PHRED) and fluorescence detection (FLD). The analytical method for aflatoxins was validated by accuracy, precision and detection limits. The method showed recovery values in the 86.9~114.0% range and the values of percent coefficient of variaton (CV%) in the 0.9~9.8% range. The limits of detection (LOD) and quantitation (LOQ) in herb were ranged from 0.020 to $0.363{\mu}g/kg$ and from 0.059 to $1.101{\mu}g/kg$, respectively. Of 62 samples analyzed, 6 semens (the original form of 2 Nelumbinis Semen and 2 Zizyphi Semen, the powder of 1 Nelumbinis Semen and 1 Zizyphi Semen) were aflatoxin positive. Aflatoxins $B_1$ or $B_2$ were detected in all positive samples, and the presence of aflatoxins $G_1$ and $G_2$ were not detected. The amount of total aflatoxins ($B_1$, $B_2$, $G_1$, and $G_2$) in the powder and original form of Nelumbinis Semen and Zizyphi Semen were observed around $ND{\sim}21.8{\mu}g/kg$, which is not regulated presently in Korea. The 56 samples presented levels below the limits of detection and quantitation.

Optimization of Analytical Methods for Ochratoxin A and Zearalenone by UHPLC in Rice Straw Silage and Winter Forage Crops (UHPLC를 이용한 볏짚 사일리지와 동계사료작물의 오크라톡신과 제랄레논 분석법 최적화)

  • Ham, Hyeonheui;Mun, Hye Yeon;Lee, Kyung Ah;Lee, Soohyung;Hong, Sung Kee;Lee, Theresa;Ryu, Jae-Gee
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.4
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    • pp.333-339
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    • 2016
  • The objective of this study was to optimize analytical methods for ochratoxin A (OTA) and zearalenone (ZEA) in rice straw silage and winter forage crops using ultra-high performance liquid chromatography (UHPLC). Samples free of mycotoxins were spiked with $50{\mu}g/kg$, $250{\mu}g/kg$, or $500{\mu}g/kg$ of OTA and $300{\mu}g/kg$, $1500{\mu}g/kg$, or $3000{\mu}g/kg$ of ZEA. OTA and ZEA were extracted by acetonitrile and cleaned-up using an immunoaffinity column. They were then subjected to analysis with UHPLC equipped with a fluorescence detector. The correlation coefficients of calibration curves showed high linearity ($R^2{\geq_-}0.9999$ for OTA and $R^2{\geq_-}0.9995$ for ZEA). The limit of detection and quantification were $0.1{\mu}g/kg$ and $0.3{\mu}g/kg$, respectively, for OTA and $5{\mu}g/kg$ and $16.7{\mu}g/kg$, respectively, for ZEA. The recovery and relative standard deviation (RSD) of OTA were as follows: rice straw = 84.23~95.33%, 2.59~4.77%; Italian ryegrass = 79.02~95%, 0.86~5.83%; barley = 74.93~97%, 0.85~9.19%; rye = 77.99~96.67%, 0.33~6.26%. The recovery and RSD of ZEA were: rice straw = 109.6~114.22%, 0.67~7.15%; Italian ryegrass = 98.01~109.44%, 1.65~4.81%; barley = 98~113.53%, 0.25~5.85%; rye = 90.44~108.56%, 2.5~4.66%. They both satisfied the standards of European Commission criteria (EC 401-2006) for quantitative analysis. These results showed that the optimized methods could be used for mycotoxin analysis of forages.

Preliminary Monitoring of Mycotoxins for Safety Management of Medicinal Herbs (한약재 안전관리를 위한 곰팡이독소 선제적 모니터링 연구)

  • Lee, Hyun-Kyung;Kim, Ae-Kyeong;Kim, Ouk-Hee;Kim, Sung-Dan;Lee, Young-Ju;Lee, Sea-Ram;Kim, Il-Young;Lee, Jung-Mi;Yu, In-Sil;Jung, Kweon
    • Journal of Food Hygiene and Safety
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    • v.32 no.3
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    • pp.187-192
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    • 2017
  • The consumption of herbal medicines has been increasing with growing interest in health. However, due to recent climate change and the complex distribution process of herbal medicines with high import dependence the likelihood of contamination with mycotoxin has been increased. Mycotoxins are emerging as key indicators for ensuring safety of herbal medicines. A total of 498 herbal medicine samples were screened for mycotoxin contamination in this study. Aflatoxin in the herbal medicine samples was extracted by using immunoaffinity column, then the extracted aflatoxin was quantified via high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD) method. The extraction method was verified by linearity, recovery, LOD and LOQ. Aflatoxins were detected in 39/498 samples in an average of $7.670{\mu}g/kg$ ($0.610-77.452{\mu}g/kg$ range). Although safety standards for Corydalis Tuber is not currently available in korea, five of the 39 samples had high concentration of aflatoxins (average of $14.9{\pm}4.1{\mu}g/kg$). In conclusion, it is urgent to establish safety criteria of aflatoxin in Corydalis Tuber. The results of the current study suggest that continuous monitoring is necessary for proactive management of herbal medicine safety.

Single Laboratory Validation and Uncertainty Estimation of a HPLC Analysis Method for Deoxynivalenol in Noodles (면류에서 HPLC를 이용한 데옥시니발레놀 분석법의 검증과 불확도 산정)

  • Ee, Ok-Hyun;Chang, Hyun-Joo;Kang, Young-Woon;Kim, Mee-Hye;Chun, Hyang-Sook
    • Journal of Food Hygiene and Safety
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    • v.26 no.2
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    • pp.142-149
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    • 2011
  • An isocratic high performance liquid chromatography (HPLC) method for routine analysis of deoxynivalenol in noodles was validated and estimated the measurement uncertainty. Noodles (dried noodle and ramyeon) were analyzed by HPLC-ultraviolet detection using immunoaffinity column for clean-up. The limits of detection (LOD) and quantification (LOQ) were 7.5 ${\mu}g$/kg and 18.8 ${\mu}g$/kg, respectively. The calibration curve showed a good linearity, with correlation coefficients $r^2$ of 0.9999 in the concentration range from 20 to 500 ${\mu}g$/kg. Recoveries and Repeatabilities expressed as coefficients of variation (CV) spiked with 200 and 500 ${\mu}g$/kg were $82{\pm}2.7%$ and $87{\pm}1.3%$% in dried noodle, and $97{\pm}1.6%$ and $91{\pm}12.0%$ in ramyeon, respectively. The uncertainty sources in measurement process were identified as sample weight, final volume, and sample concentration in extraction volume as well as components such as standard stock solution, working standard solution, 5 standard solutions, calibration curve, matrix, and instrument. Deoxynivalenol concentration and expanded uncertainty in two matrixes spiked with 200 ${\mu}g$/kg and 500 ${\mu}g$/kg were estimated to be $163.8{\pm}52.1$ and $435.2{\pm}91.6\;{\mu}g$/kg for dried noodle, and $194.3{\pm}33.0$ and $453.2{\pm}91.1\;{\mu}g$/kg for ramyeon using a coverage factor of two which gives a level of statistical confidence with approximately 95%. The most influential component among uncertainty sources was the recovery of matrix, followed by calibration curve.

Monitoring of Aflatoxins in Medicinal Herbs (유통 생약재의 아플라톡신 모니터링)

  • Kim, Yong-Hoon;Kang, Han-Saem;Oh, Sun-Woo;Lee, Hwa-Jung;Kim, Mi-Gyeong;Chung, So-Young;Choi, Seon-Hee;Bang, Su-Jin;Han, Kyung-Jin;Lee, Ji-Won;Kim, Young-Seon;Kim, Hee-Yun
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.27-32
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    • 2010
  • This study was conducted to monitor aflatoxins in various medicinal herbs, providing available data for the safety of those products. To monitor aflatoxins in medicinal herbs, a total of 400 samples of 40 different herbs were collected in commercial retailers in Seoul, Daejeon, Gwangju, Daegu, and Busan from March to August, 2008. The samples that passed the sensory evaluation were tested for aflatoxins. Aflatoxins in samples were analyzed by HPLC-florescence coupled with photochemical enhancement. Samples were extracted with 70% methanol and then diluted to the appropriate concentration. A refining process was performed using an immunoaffinity column. The analytical method used in this study was validated. The $R^2$ value for aflatoxin $B_1$ was 0.99946, and the detection range was from 0.25 to 10.0 ng/mL. The accuracy of the analysis was ranged from 83.2% to 101.8%. The relative standard deviation (RSD) in the aflatoxin $B_1$ analysis was 3.4%, demonstrating the precision of this method. In addition, the detection limit and quantitative analysis limit of aflatoxin $B_1$ was $0.53\;{\mu}g/kg$ and $1.76\;{\mu}g/kg$, respectively. These results indicated that the analytical method used in this study was appropriate. The results of HPLC showed that 1% (4 samples) of the samples may contain aflatoxins. The concentration of quantified aflatoxin was $2.3\;{\mu}g/kg$ for both Quisqualis fructus and Remotiflori radix samples. The other samples were below the limit of quantification. Moreover, the concentration of aflatoxin $B_1$ which is made by specific fungi were below the level of regulation. Only 20% of aflatoxin $B_1$ were transferred to hot water. Therefore, the levels of aflatoxins in medicinal herbs were considered to be safe especially considering the aflatoxin transfer ratio.