Comparison between capillary electrophoresis and high performance liquid chromatography for the study of the occurrence of patulin in apple juice intended for infants

Apple juice samples intended for infants purchased in Navarra (Spain) have been analyzed for PAT occurrence. Two capillary electrophoresis methods, based on a MEKC and a CEC system, and an HPLC method were evaluated for the aforementioned study. The CEC system gave less satisfying separations and several practical problems, so samples have been analyzed by MEKC and HPLC. Both methods have been comparable in terms of recovery, precision, limits of detection, volume of organic solvents used and adequate selectivity with regard to PAT and HMF. The analysis time in HPLC has been slightly lower than in the MEKC methodology. The PAT levels obtained in apple juice by both validated methods showed a strong correlation (p<0.001). Therefore, both methodologies are useful for the accurate quantification of patulin in this matrix. The PAT levels obtained in the 20 infant apple juices samples were in a range between <LOD and 29.6 microg L(-1), with a mean concentration of 8.0 microg L(-1) which implies a dietary intake estimation of 104 ng kg(-1)b.w.day(-1) considering a body weight of 10 kg and an apple juice consumption of 130 mL day(-1), 26% of the PMTDI recommended by JECFA

Validation of a UHPLC-FLD method for the simultaneous quantification of aflatoxins, ochratoxin A and zearalenone in barley

A fast and simple UHPLC-FLD method has been developed for the simultaneous determination in barley of aflatoxins (B1, G1, B2 and G2), ochratoxin A (OTA) and zearalenone (ZEA), some of the most important mycotoxins due to their toxicity and occurrence. The procedure is based on the extraction of the six mycotoxins with a mixture of acetonitrile and water, and the purification of the extract with immunoaffinity columns before analysis. Detection of AFB1 and AFG1 is improved using a photochemical reaction. The method has been validated with satisfactory results. Limits of detection were 340 ng kg-1 for ZEA, 13 ng kg-1 for OTA and varied from 0.5 to 15 ng kg-1 for aflatoxins. Recovery percentages were between 78.2 and 109.2%. After being validated, the method has been successfully applied to 20 barley samples cultivated in a region of northern Spain (Navarra)

Validation of an antiviral assay method for quantifying IFN-α5 activity in macaque and human serum

Background: IFN-alpha 5 has been demonstrated to induce stronger signaling and higher expression of antiviral genes than IFN-alpha 2, which is the current treatment in chronic viral hepatitis. However, there is no specific and validated quantification method in order to conduct kinetic studies as part of the preclinical and clinical evaluation for regulatory purposes. Results: A novel integration of an antiviral assay against the cytopathic effect of the encephalomyocarditis virus in HeLa cells with a very sensitive method for assay processing - the Vialight (R) Plus assay - is presented for IFN-alpha 5 activity quantification. The bioassay has been validated in macaque and human serum and it has been demonstrated to be selective, precise and accurate. Conclusion: The validated bioassay meets suitable acceptance criteria for these types of biological assays

Comparison between capillary electrophoresis and high performance liquid chromatography for the study of the occurrence of patulin in apple juice intended for infants

Apple juice samples intended for infants purchased in Navarra (Spain) have been analyzed for PAT occurrence. Two capillary electrophoresis methods, based on a MEKC and a CEC system, and an HPLC method were evaluated for the aforementioned study. The CEC system gave less satisfying separations and several practical problems, so samples have been analyzed by MEKC and HPLC. Both methods have been comparable in terms of recovery, precision, limits of detection, volume of organic solvents used and adequate selectivity with regard to PAT and HMF. The analysis time in HPLC has been slightly lower than in the MEKC methodology. The PAT levels obtained in apple juice by both validated methods showed a strong correlation (p<0.001). Therefore, both methodologies are useful for the accurate quantification of patulin in this matrix. The PAT levels obtained in the 20 infant apple juices samples were in a range between <LOD and 29.6 microg L(-1), with a mean concentration of 8.0 microg L(-1) which implies a dietary intake estimation of 104 ng kg(-1)b.w.day(-1) considering a body weight of 10 kg and an apple juice consumption of 130 mL day(-1), 26% of the PMTDI recommended by JECFA

Occurrence of Ochratoxin A in southern Spanish generous wines under the denomination of origin "Jerez-Xérès-Sherry and 'Manzanilla' Sanlúcar de Barrameda".

The mycotoxin ochratoxin A (OTA) has toxic effects in animals; the most relevant of them is nephrotoxicity. OTA has also been classified as a possible carcinogen for humans (group 2B) by the International Agency for Research on Cancer (IARC). Therefore, exposure to OTA through contaminated food can represent health impairment to humans. The maximum permitted level for this mycotoxin in wine is 2.0 μg/L. The presence of OTA in Spanish wines produced using the traditional methods under the Denomination of Origin "Jerez-Xérès-Sherry andmanzanilla Sanlúcar de Barrameda" was evaluated by a High performance Liquid Chromatography method with fluorescence detection and immunoaffinity column purification. A recovery of 95.4% and a limit of detection and quantification of 0.009 μg/L and 0.02 μg/L respectively, were achieved. In manzanilla, fino, amontillado and oloroso wine, the mean OTA values were 0.042, 0.044, 0.144, and 0.319 μg/L, respectively. These levels are not different from other data given in the reference literature on white wines, although fino and manzanilla wines have very low OTA levels

OTA-producing fungi in foodstuffs: A review

Ochratoxin A (OTA) is a secondary metabolite produced by filamentous fungi of the genera Aspergillus and Penicillium present in a wide variety of foodstuffs. The most relevant OTA-producing species are Penicillium verrucosum (P. verrucosum), Aspergillus ochraceus (A. ochraceus), Aspergillus niger and Aspergillus carbonarius due to their prevalence in foodstuffs (cereals, grapes, coffee, etc.) and the number of strains able to produce OTA. To target pre- and post-harvest control programs, studies concerning the toxigenic fungi in each foodstuff are essential. This paper summarizes the state-of-the-art and the requirements in OTA control

OTA-producing fungi in foodstuffs: A review

Ochratoxin A (OTA) is a secondary metabolite produced by filamentous fungi of the genera Aspergillus and Penicillium present in a wide variety of foodstuffs. The most relevant OTA-producing species are Penicillium verrucosum (P. verrucosum), Aspergillus ochraceus (A. ochraceus), Aspergillus niger and Aspergillus carbonarius due to their prevalence in foodstuffs (cereals, grapes, coffee, etc.) and the number of strains able to produce OTA. To target pre- and post-harvest control programs, studies concerning the toxigenic fungi in each foodstuff are essential. This paper summarizes the state-of-the-art and the requirements in OTA control