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1.
Food Chem Toxicol ; 107(Pt A): 108-121, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28645870

RESUMO

Modern agriculture provides the potential for sustainable feeding of the world's increasing population. Up to the present moment, genetically modified (GM) products have enabled increased yields and reduced pesticide usage. Nevertheless, GM products are controversial amongst policy makers, scientists and the consumers, regarding their possible environmental, ecological, and health risks. Scientific-and-political debates can even influence legislation and prospective risk assessment procedure. Currently, the scientifically-assessed direct hazardous impacts of GM food and feed on fauna and flora are conflicting; indeed, a review of literature available data provides some evidence of GM environmental and health risks. Although the consequences of gene flow and risks to biodiversity are debatable. Risks to the environment and ecosystems can exist, such as the evolution of weed herbicide resistance during GM cultivation. A matter of high importance is to provide precise knowledge and adequate current information to regulatory agencies, governments, policy makers, researchers, and commercial GMO-releasing companies to enable them to thoroughly investigate the possible risks.


Assuntos
Ração Animal/análise , Alimentos Geneticamente Modificados/normas , Plantas Geneticamente Modificadas/química , Animais , Qualidade de Produtos para o Consumidor/normas , Ecossistema , Meio Ambiente , Humanos , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento
2.
J Food Sci ; 79(10): M2048-55, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25224778

RESUMO

UNLABELLED: The objective of this study was to develop a 1-step simultaneous lateral flow strip test for the rapid and simple detection of deoxynivalenol (DON) and zearalenone (ZEA) in grains. Two monoclonal antibodies (MAbs) against DON and ZEA were respectively conjugated with gold nanoparticles and used to develop a lateral flow strip test for a single toxin and multiple toxins. First, individual lateral flow strips for a single toxin were optimized, and their conditions were used to develop a simultaneous lateral flow strip for multiple toxins. Limits of detection of both lateral flow strip tests for DON and ZEA were the same (DON: 50 ng/mL, ZEA: 1 ng/mL). Both methods showed cross-reactivity for α-zearalenol and ß-zearalenol, but no cross-reaction to other mycotoxins. The results can be completed obtained within 15 min. The cut-off values of the simultaneous lateral flow strip for the spiked rice and corn were 500 and 10 ng/g for DON and ZEA, respectively. The results demonstrated that the developed simultaneous lateral flow strip test offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site detection of DON and ZEA in food and agricultural commodities. PRACTICAL APPLICATION: Simultaneous lateral strip test is useful for a rapid detection of DON and ZEA at a time in food and grain samples.


Assuntos
Grão Comestível/química , Fitas Reagentes/química , Tricotecenos/química , Zearalenona/química , Anticorpos Monoclonais , Análise de Alimentos , Contaminação de Alimentos , Ouro/química , Nanopartículas Metálicas/química , Sensibilidade e Especificidade , Fatores de Tempo
3.
Anal Bioanal Chem ; 406(3): 859-66, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24337136

RESUMO

Detection of pathogenic bacteria that pose a great risk to human health requires a rapid, convenient, reliable, and sensitive detection method. In this study, we developed a selective filtration method using monoclonal antibody (MAb)-magnetic nanoparticle (MNP) nanocomposites for the rapid and sensitive colorimetric detection of Salmonella typhimurium. The method contains two key steps: the immunomagnetic separation of the bacteria using MAb-MNP nanocomposites and the filtration of the nanocomposite-bound bacteria. Color signals from the nanocomposites remaining on the membrane were measured, which reflected the amount of bacteria in test samples. Immunomagnetic capture efficiencies of 8 to 90 % for various concentrations of the pathogen (2 × 10(4)-2 × 10(1) cells) were obtained. After optimization of the method, 2 × 10(1) cells of S. typhimurium in pure culture solution was detectable as well as in artificially inoculated vegetables (100 cells/g). The method was confirmed to be highly specific to S. typhimurium without cross-reaction to other pathogenic bacteria and could be concluded within 45 min, yielding results in a shorter or similar time period as compared with recently reported antibody immobilized on magnetic-particle-based methods. This study also demonstrated direct application of MAb-MNP nanocomposites without a dissociation step of bacteria from magnetic beads in colorimetric assays in practice.


Assuntos
Anticorpos Monoclonais/metabolismo , Técnicas de Tipagem Bacteriana/métodos , Colorimetria/métodos , Nanopartículas Metálicas/química , Nanocompostos/química , Salmonella typhimurium/isolamento & purificação , Filtração , Humanos , Limite de Detecção , Reprodutibilidade dos Testes , Fatores de Tempo
4.
J Food Sci ; 78(10): M1575-M1581, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24024744

RESUMO

A rapid immunochromatographic (ICG) strip based on a conjugate of colloidal gold and monoclonal antibody (mAb) was developed for the rapid, sensitive, and on-site detection of sulfamethazine in meat and egg samples. The detection limit of the ICG strip is 2 ng/mL, and the assay can be completed in 10 min. A cross-reactivity test indicated that the ICG strip was highly specific to sulfamethazine with no cross-reaction with sulfonamide compounds and other antibiotics. The results of the recovery test from meat and egg samples spiked with sulfamethazine were in good agreement with those obtained by the indirect competitive enzyme-linked immunosorbent assay. These results demonstrated that the ICG strip can be used as a rapid and qualitative tool for on-site screening of sulfamethazine in meat and egg samples.


Assuntos
Anti-Infecciosos/isolamento & purificação , Cromatografia de Afinidade/métodos , Ovos/análise , Carne/análise , Sulfametazina/isolamento & purificação , Animais , Anticorpos Monoclonais/metabolismo , Bovinos , Galinhas , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Suínos
5.
J Microbiol Biotechnol ; 23(4): 527-33, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23568207

RESUMO

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was 10(5) CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.


Assuntos
Técnicas Bacteriológicas/métodos , Vibrio parahaemolyticus/isolamento & purificação , Vibrio vulnificus/isolamento & purificação , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/métodos , Cabras , Peroxidase do Rábano Silvestre/metabolismo , Imunoglobulina G/imunologia , Imunoglobulina G/isolamento & purificação , Imunoglobulinas/imunologia , Camundongos , Codorniz/imunologia , Sensibilidade e Especificidade , Coloração e Rotulagem/métodos , Triazinas/metabolismo
6.
J Food Prot ; 75(11): 1991-9, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23127708

RESUMO

The objective of this study was to investigate the occurrence of aflatoxins in herbal medicines distributed in South Korea. A total of 700 herbal medicine samples (10 samples each for 70 types of herbal medicine) were analyzed by an enzyme-linked immunosorbent assay (ELISA) for aflatoxin B(1) (AFB(1)), and levels of total aflatoxins were quantified and confirmed by liquid chromatography combined with tandem mass spectrometry (LC-MS/MS). The levels of recovery of the methods were 84.30 to 102.68% (ELISA for AFB(1)) and 72.17 to 90.92% (LC-MS/MS for total aflatoxins). Fifty-eight (8.29%) of 700 samples were AFB(1) positive by ELISA, and 17 (2.43%) of them were finally confirmed as positive for total aflatoxins by LC-MS/MS. Total aflatoxin levels in the herbal medicines were from 4.51 to 108.42 µg/kg. Among the 17 samples, the AFB(1) content of 6 samples (11.95 to 73.27 µg/kg) and the total aflatoxin content of 10 (12.12 to 108.42 µg/kg) samples exceeded the legal limits set by the Korea Food and Drug Administration for AFB(1) (10 µg/kg) and by the European Commission for total aflatoxins (10 µg/kg), respectively. These results demonstrate the risk to consumers of herbal medicine contamination by aflatoxins and encourage further studies to investigate the transfer rate of mycotoxins to decoction, which is the final product for consumption.


Assuntos
Aflatoxinas/isolamento & purificação , Contaminação de Alimentos/análise , Plantas Medicinais/química , Cromatografia Líquida , Qualidade de Produtos para o Consumidor , Contaminação de Medicamentos , Ensaio de Imunoadsorção Enzimática , Humanos , República da Coreia , Espectrometria de Massas em Tandem
7.
J Microbiol Biotechnol ; 22(10): 1423-31, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23075795

RESUMO

The inhibitory effect of IgY against Vibrio parahaemolyticus and Vibrio vulnificus responsible for seafood-borne diseases was investigated in this study. Water-soluble fractions (WSF) of protein containing IgYs were isolated from the egg yolk of hens initially immunized with formalin inactivated V. parahaemolyticus or V. vulnificus. Protein, total and specific IgY contents of the WSF were determined. The inhibitory and protective effects of IgYs on the growth of V. parahaemolyticus and V. vulnificus were assayed in liquid medium and in mice. IgYs showed high affinity to their corresponding antigens with high titer from day 28 onwards. Protein contents and total IgY concentrations remained stable throughout the immunization period, whereas specific IgY concentrations increased steadily and reached a plateau at day 49. Specific IgY powder (150 mg/ml) significantly inhibited further multiplication of both V. parahaemolyticus and V. vulnificus in liquid medium as compared with the control IgY. The bacteria count in mice feces was lower in mice pretreated with specific IgYs than in those pretreated with PBS or control IgY. Higher survival of mice was observed in the experimental groups pretreated with either anti-V. parahaemolyticus (75% survival) or anti-V. vulnificus (87% survival) IgYs, compared with those in the control groups pretreated with PBS or nonspecific IgY. All mice in the control groups died within three days after bacteria inoculation; hence, the protective effect of specific IgYs against infection caused by V. parahaemolyticus and V. vulnificus was demonstrated.


Assuntos
Anticorpos Antibacterianos/imunologia , Imunoglobulinas/imunologia , Vibrio parahaemolyticus/imunologia , Vibrio vulnificus/imunologia , Animais , Antígenos de Bactérias/imunologia , Carga Bacteriana , Galinhas/imunologia , Reações Cruzadas , Meios de Cultura/metabolismo , Avaliação Pré-Clínica de Medicamentos , Gema de Ovo/imunologia , Gema de Ovo/microbiologia , Fezes/microbiologia , Feminino , Microbiologia de Alimentos , Formaldeído/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Alimentos Marinhos/microbiologia , Vacinação/métodos , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrioses/terapia , Vibrio parahaemolyticus/crescimento & desenvolvimento , Vibrio vulnificus/crescimento & desenvolvimento
8.
J Food Prot ; 75(7): 1213-8, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22980003

RESUMO

Sample preparation methods (pummeling, pulsifying, sonication, and shaking by hand) were compared for achieving maximum recovery of foodborne pathogens from iceberg lettuce, perilla leaves, cucumber, green pepper, and cherry tomato. Antimicrobial and dehydration effects also were examined to investigate causes of poor recovery of pathogens. Each produce type was inoculated with Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus at 6.0 log CFU/cm(2), and samples were prepared using the four methods. Bacterial populations recovered from the five types of produce were significantly different (P < 0.05) according to sample preparation methods and produce type. The bacterial populations recovered from pummeled and pulsified samples were higher (P < 0.05) than those recovered from sonicated and hand-shaken samples, except for cherry tomato. The number of bacteria recovered from produce was reduced (P < 0.05) from that of the inoculum by 0.16 to 2.69 log CFU/cm(2). Although extracts of iceberg lettuce, perilla leaves, cucumber, and green pepper had no antimicrobial activity, the populations of E. coli O157:H7, Salmonella Typhimurium, B. cereus, and L. monocytogenes in cherry tomato extract were slightly reduced after these treatments (P < 0.05). The pathogen populations on perilla leaves and cherry tomatoes decreased by >2 log CFU/cm(2) after exposure to 40% relative humidity for 1 h. No reduction was observed when the five pathogens were exposed to 90% relative humidity. These data suggest that pummeling and pulsifying are optimal sample preparation methods for detection of microorganisms. Acidic produce such as cherry tomato should be treated with a method that does not cause sample breakdown so that acid stress on the bacteria can be minimized. Dehydration stress also affects recovery of pathogens from produce.


Assuntos
Contaminação de Alimentos/análise , Manipulação de Alimentos/métodos , Microbiologia de Alimentos , Verduras/microbiologia , Bacillus cereus/isolamento & purificação , Capsicum/microbiologia , Contagem de Colônia Microbiana , Cucumis sativus/microbiologia , Escherichia coli O157/isolamento & purificação , Contaminação de Alimentos/prevenção & controle , Humanos , Lactuca/microbiologia , Listeria monocytogenes/isolamento & purificação , Solanum lycopersicum/microbiologia , Salmonella typhimurium/isolamento & purificação , Staphylococcus aureus/isolamento & purificação
9.
J Chromatogr A ; 1218(42): 7627-33, 2011 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-21831391

RESUMO

This paper describes the application of sol-gel immunoaffinity columns for clean up of ochratoxin A contaminated cereal crops. Monoclonal antibodies selective for OTA have been entrapped into the pores of a sol-gel matrix in order to prepare immunoaffinity columns. Different parameters such as amount of entrapped antibodies and loading conditions were optimized to obtain highest possible recoveries of OTA. The method has been found to be a suitable tool in sample preparation prior to HPLC-FLD determination and as selective as conventional commercially available immunoaffinity columns. In the clean up of different cereals mean recoveries of 82±5%, 90±6% and 91±3%, were obtained for wheat, barley and rye, respectively, with sol-gel columns containing 1mg of anti-OTA antibodies. The detection limit (signal-to-noise ratio, 3) was 0.5 µg/kg and the limit of quantification (signal-to-noise ratio, 10) determined to be 1 µg/kg. Sol-gel columns can be reused 7 times without significant loss of recovery. After 10 applications the recovery decreased to approx. 50%.


Assuntos
Cromatografia de Afinidade/métodos , Grão Comestível/química , Técnicas de Imunoadsorção , Ocratoxinas/isolamento & purificação , Adsorção , Anticorpos Monoclonais/metabolismo , Cromatografia Líquida de Alta Pressão , Grão Comestível/normas , Reutilização de Equipamento , Hordeum , Ocratoxinas/química , Ocratoxinas/metabolismo , Reprodutibilidade dos Testes , Secale , Sensibilidade e Especificidade
10.
Anal Bioanal Chem ; 400(8): 2615-22, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21461614

RESUMO

The paper presents a new sample clean-up method based on immuno-ultrafiltration for the analysis of ochratoxin A in cereals. In contrast to immunoaffinity chromatography, in immuno-ultrafiltration, the antibodies are used in non-immobilised form. Ochratoxin A was extracted with ACN/water (60/40, v/v), and the extract was loaded onto the ultrafiltration device. After a washing step with phosphate-buffered saline, containing 0.05% Tween 20, ochratoxin A was eluted with MeOH/acetic acid (99/1, v/v). The detection of ochratoxin A was carried out with high-performance liquid chromatography and a fluorescence detector coupled to an electrochemical cell (Coring cell). The electrochemical cell was used to eliminate matrix interferences by oxidising matrix compounds. The method was validated by repeatedly analysing spiked barley and rye samples as well as a certified wheat reference material. Recoveries and standard deviations (1 SD) were found to be 71 ± 9%, 77 ± 12% and 77 ± 8% in wheat, barley and rye, respectively. The limit of detection (S/N = 3) and limit of quantitation (S/N = 10) were determined to be 0.4 µg kg(-1) and 1 µg kg(-1). The analysis of the certified reference material resulted in ochratoxin A concentrations which were in the range assigned by the producer. Additionally, the effect of the electrochemical cell on other widely used clean-up techniques, namely the immunoaffinity clean-up and multifunctional columns (Mycosep #229), was evaluated. In all clean-up methods, an improvement of the chromatogram quality was registered.


Assuntos
Grão Comestível/química , Ocratoxinas/análise , Cromatografia Líquida de Alta Pressão , Eletroquímica , Fluorescência , Ultrafiltração
11.
Foodborne Pathog Dis ; 7(12): 1511-9, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21121865

RESUMO

This study conducted microbiological assessment in tunnel style strawberry greenhouses and packaging centers and suggested recommendations to establish a good agricultural practice for strawberry production. The samples from irrigation water, workers' gloves, harvest bins, soil, strawberry leaves and strawberries in greenhouses, packers' gloves, conveyor belts, packaging tables, and door knobs of entrances in packaging centers were collected. Bacterial cell counts of aerobic plate counts, coliforms, Escherichia coli, E. coli O157:H7, Salmonella, Staphylococcus aureus, and Bacillus cereus were then enumerated on appropriate selective media. In general, bacterial populations were similar (p ≥ 0.05) among strawberry greenhouses but not among packaging houses. E. coli and E. coli O157:H7 were negative in all samples, and low levels of Salmonella and B. cereus were detected. However, high bacterial cell counts of aerobic plate counts, coliforms, and S. aureus were found in most samples. These results suggest that food safety practice in strawberry greenhouses and packaging centers should be improved, and the results may be useful in the establishment of a good agricultural practice system for strawberry production.


Assuntos
Contaminação de Alimentos , Manipulação de Alimentos/normas , Microbiologia de Alimentos , Inocuidade dos Alimentos , Fragaria/microbiologia , Irrigação Agrícola/normas , Agricultura/normas , Bacillus cereus/isolamento & purificação , Contagem de Colônia Microbiana/métodos , Qualidade de Produtos para o Consumidor/normas , Escherichia coli O157/isolamento & purificação , Salmonella/isolamento & purificação , Staphylococcus aureus/isolamento & purificação
12.
J Agric Food Chem ; 58(12): 7135-40, 2010 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-20507064

RESUMO

Rapid detection of enterotoxigenic Clostridium perfringens in meat samples was accomplished with an immunomagnetic separation polymerase chain reaction (IMS-PCR). First, a monoclonal antibody (mAb) specific to C. perfringens was generated. The antibody showed strong binding to C. perfringens and no binding to non- Clostridia bacteria, except a weak cross-reaction to Staphylococcus aureus based on the enzyme-linked immunosorbent assay (ELISA). Then, magnetic beads were coated with the mAb, and the IMS-PCR system was developed. With the optimized conditions, the IMS-PCR assay was capable of detecting as few as 10 colony forming units (CFU)/g of C. perfringens cells in the meat sample within 10 h. Of the 116 collected samples (26 chicken samples, 20 beef samples, 30 pork samples, 20 fish samples, and 20 processed meat samples) examined with IMS-PCR, 36 (31%) were C. perfringens -positive samples and 2 (1.7%) were enterotoxigenic C. perfringens -positive samples. The IMS-PCR results gave a good agreement with the results obtained by conventional culture methods. In comparison to conventional culture methods, the IMS-PCR is a rapid and specific method and has potential use as a screening tool for enterotoxigenic C. perfringens in food samples.


Assuntos
Clostridium perfringens/isolamento & purificação , Separação Imunomagnética/métodos , Carne/análise , Reação em Cadeia da Polimerase/métodos , Animais , Bovinos , Galinhas , Clostridium perfringens/genética , Suínos
13.
Mycotoxin Res ; 26(4): 279-86, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23605491

RESUMO

A large amount-260,000 tons-of red paprika is consumed annually in Korea, where the people prefer hot and pungent to sweet foods. Concern has recently grown among consumers over contamination of paprika powder by mycotoxins; contamination can occur at any stage from pre-harvest to drying, storage, grinding, and eventually transport to the retail market. This study had dual aims: to investigate the current level of contamination of hot peppers by ochratoxin A and to identify the critical control points in the food chain. We measured the ochratoxin A (OTA) content of 200 samples from various sources including supermarkets, an online shopping mall, small stakeholder mills, Hazard Analysis and Critical Control Points (HACCP)-implemented factories, and an import company. Mycotoxin was determined using immunoaffinity column cleanup/HPLC quantification as well as strong anion exchange cleanup/stable isotope dilution assay. Monitoring revealed that approximately 30% of red paprika samples were OTA-positive, indicating a need to establish a maximum level for regulation. We selected two model factories that had adopted HACCP in different ways, and compared data in order to develop guidelines for alleviation of mitigation of the mycotoxin contamination.

14.
Biosci Biotechnol Biochem ; 73(6): 1412-5, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19502760

RESUMO

Sequence comparison of individual genes in the prfA virulence gene cluster (pVGC), a central virulence gene cluster, from different Listeria species indicated that priming sites within the genes appeared to be specific for Listeria monocytogenes exclusively. Therefore, the pVGC was targeted for polymerase chain reaction (PCR) assays to detect and specifically identify L. monocytogenes. Each gene of the pVGC was specifically amplified in L. monocytogenes but not in other Listeria species.


Assuntos
Listeria monocytogenes/patogenicidade , Família Multigênica , Reação em Cadeia da Polimerase/métodos , Virulência/genética , Sequência de Bases , Primers do DNA , Genes Bacterianos , Listeria monocytogenes/genética , Especificidade da Espécie
15.
J Sep Sci ; 32(10): 1729-39, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19472274

RESUMO

The present paper describes the development of a new clean-up strategy for the analysis of aflatoxins (AFs) in food. The sample preparation method is based on immuno-ultrafiltration (IUF) which, in contrast to immunoaffinity chromatography, makes use of antibodies in free form. After selecting an appropriate ultrafiltration (UF) device and optimizing different operation conditions the IUF method was applied to the clean-up of maize and rice. Quantification of AFs was carried out by HPLC and fluorescence detection, after postcolumn derivatization in a Kobracell. The IUF method was shown to be as selective as sample clean-up using commercial immunoaffinity columns. Recovery rates and RSD for the AFs G(2), G(1), B(2) and B(1) in spiked rice were found to be 76 +/- 3, 76 +/- 2, 83 +/- 5 and 99 +/- 14%, respectively. The analysis of a FAPAS (food analysis performance assessment scheme) maize material resulted in AFs concentrations which were in the range assigned by the producer of the reference material.


Assuntos
Aflatoxinas/isolamento & purificação , Análise de Alimentos/métodos , Ultrafiltração/métodos , Zea mays/química , Cromatografia Líquida de Alta Pressão , Reprodutibilidade dos Testes
16.
J Food Prot ; 72(2): 436-40, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19350995

RESUMO

To survey fumonisins B1 (FB1) and B2 (FB2) in agricultural products consumed in South Korea and provide an exposure assessment, ground samples were extracted (80% MeOH), filtered (0.2 microm), and cleaned up. After evaporation, dry residues were reconstituted in 50% MeOH, and a 50-micro1 aliquot of this sample was mixed with 200 micro1 of o-phthaldialdehyde for derivatization. The derivatives were analyzed with a high-performance liquid chromatography system equipped with a fluorescence detector. For validation of the detection procedure, linearity, accuracy, precision, detection limit, and quantification limit were determined. The validated detection method was then used to survey fumonisins in white rice, brown rice, barley, barley tea, beer, wheat flour, millet, dried corn, corn flour, corn tea, canned corn, popcorn, and breakfast cereal. Retention times for FB1 and FB2 standards were 7 and 18 min, respectively. Linearity (R2 = 0.99995 to 0.99998), accuracy (81.47 to 108.83%), precision (2.35 to 5.77), detection limit (25 ng/g or ng/ml), and quantification limit (37 ng/g or ng/ml) indicated that this procedure is capable of quantifying fumonisins in agricultural products. Only FB1-positive samples (5.12%, three dried corn samples and five corn flour samples) were found at 90.89 to 439.67 ng/g. According the survey results, an estimated daily intake of FB1 and FB2 in Korea was 0.087 ng/kg of body weight per day. These results indicate that continuous monitoring of these mycotoxins is necessary to establish appropriate risk assessment, and the maximum tolerable daily intake of fumonisins in Korea is lower than the 2 microg/kg set by the Joint Food and Agriculture Organization-World Health Organization Expert Committee.


Assuntos
Contaminação de Alimentos/análise , Fumonisinas/análise , Medição de Risco , Cerveja/análise , Carcinógenos Ambientais/análise , Cromatografia Líquida de Alta Pressão/métodos , Qualidade de Produtos para o Consumidor , Exposição Ambiental , Hordeum/química , Humanos , Coreia (Geográfico) , Sensibilidade e Especificidade , Zea mays/química
17.
J Agric Food Chem ; 57(10): 4035-41, 2009 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-19348422

RESUMO

A monoclonal antibody (mAb)-based gold nanoparticle immunochromatographic assay (ICG) for zearalenone detection was developed, optimized, and validated. The detection limits of ICG optimized with appropriate amounts of zearalenone-bovine serum albumin and gold nanoparticle-mAb to zearalenone were 2.5 ng/mL and 30 µg/kg for the standard solution and spike sample, respectively, and a weak cross-reaction for α-zearalenol and ß-zearalenol was observed. The assay required only 15 min to obtain results and one step to perform the assay. In validation, the results obtained from spiked corn (10, 20, 30, 50, and 100 µg/kg) and naturally contaminated corn samples by the ICG were in good agreement with those obtained by direct competitive enzyme-linked immunosorbent assay (DC-ELISA) and high-performance liquid chromatography (HPLC). Therefore, the results obtained in this study could be used as basic research for the development of zearalenone-ICG, and the ICG developed could be a useful on-site screening tool for the rapid detection of zearalenone in corn without special instrumentation.


Assuntos
Cromatografia/métodos , Estrogênios não Esteroides/análise , Imunoensaio/métodos , Zearalenona/análise , Anticorpos Monoclonais , Contaminação de Alimentos/análise , Ouro , Nanopartículas , Reprodutibilidade dos Testes , Zea mays/química
18.
J Microbiol Biotechnol ; 19(1): 83-92, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19190413

RESUMO

Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, resepectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and 20/30 microg/kg). The cut-off values of the OS-ICG for the spiked corn were 5 and 10 microg/kg for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS-ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle.


Assuntos
Cromatografia/métodos , Imunoensaio/métodos , Ocratoxinas/análise , Fitas Reagentes , Zearalenona/análise , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Carcinógenos/análise , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Estrogênios não Esteroides/análise , Análise de Alimentos/métodos , Coloide de Ouro , Ocratoxinas/imunologia , Controle de Qualidade , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Zea mays/química , Zearalenona/imunologia
19.
J Agric Food Chem ; 57(3): 791-6, 2009 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-19140717

RESUMO

A fluorescence polarization immunoassay (FPIA) for the quantitative determination of 6-chloronicotinic acid (6-CNA) using polyclonal antibody was developed. The 6-CNA-protein (bovine serum albumin and soybean trypsin inhibitor) conjugates and fluorescein-labeled 6-CNA derivative (tracer) were prepared and used as the immunogens and tracer, respectively. The synthesized tracer was purified by thin layer chromatography (TLC) and showed a good binding to antiserum (73/5) which was obtained from the immunized rabbit (No. 73) with 6-CNA-BSA conjugate. The detection limit (10% inhibition) of FPIA was 4 microg/mL, and IC(50) value was 32 microg/mL. The FPIA showed a cross-reaction for 5-amino-2-chloropyridine (60%), but no cross-reaction for other pesticides was observed. Recoveries for spiked apple, urine, soil, and water samples (5, 50, and 500 ppm) averaging between 78.6 +/- 8.8 and 114 +/- 18% were reasonable and in good agreement with the amounts spiked. Although the developed FPIA possesses low sensitivity, this assay is more simple and quick than other analytical methods, such as high performance liquid chromatography and gas chromatography. Thus, the developed FPIA method could be a useful tool for express screening 6-CNA in agricultural, environmental, and biological samples.


Assuntos
Imunoensaio de Fluorescência por Polarização/métodos , Inseticidas/metabolismo , Ácidos Nicotínicos/análise , Agroquímicos/química , Agroquímicos/metabolismo , Cromatografia Líquida de Alta Pressão , Frutas/química , Imidazóis/metabolismo , Inseticidas/química , Malus/química , Neonicotinoides , Ácidos Nicotínicos/urina , Nitrocompostos/metabolismo , Sensibilidade e Especificidade , Solo/análise , Água/química
20.
J Food Prot ; 71(4): 781-9, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18468033

RESUMO

An enzyme-linked immunosorbent assay (ELISA), immunochromatography (ICG) strip test, and immunomagnetic bead separation (IMBS) system based on a monoclonal antibody were individually developed for the detection and isolation of Listeria monocytogenes in meat samples. The three methods showed a strong reaction with Listeria species and a weak reaction with Staphylococcus aureus. To increase the rapidity of L. monocytogenes detection, combinations of the ELISA and ICG strip test with the IMBS system (ELISA-IMBS and ICG-IMBS) were investigated. In comparative analyses of artificially inoculated meat and samples of processed meat, the ELISA and ICG strip test required 24 h of enrichment time to detect the inoculated meat samples with > or =1 X 10(2) CFU/10 g, whereas the ELISA-IMBS and ICG-IMBS required only 14 h of enrichment. Analyses of naturally contaminated meat samples (30 pork samples, 20 beef samples, 26 chicken samples, 20 fish samples, and 20 processed meat samples) performed by ELISA-IMBS, ICG-IMBS, and API kit produced similar results. The ELISA-IMBS and ICG-IMBS provide a more rapid assay than the individual ELISA and the ICG strip test and are appropriate for rapid and qualitative detection of L. monocytogenes (or Listeria species) in meat samples. With the ICG-IMBS, L. monocytogenes could be detected in meat samples within 15 h and the method has potential as a rapid, cost-effective on-site screening tool for the detection of L. monocytogenes in food samples and agricultural products at a minimum detection level of approximately 100 CFU/10 g.


Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Contaminação de Alimentos/análise , Imunoensaio/métodos , Separação Imunomagnética/métodos , Listeria monocytogenes , Carne/microbiologia , Anticorpos Monoclonais/biossíntese , Contagem de Colônia Microbiana , Listeria monocytogenes/imunologia , Listeria monocytogenes/isolamento & purificação , Produtos da Carne/microbiologia , Fitas Reagentes , Sensibilidade e Especificidade , Especificidade da Espécie , Staphylococcus aureus/imunologia , Staphylococcus aureus/isolamento & purificação , Fatores de Tempo
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