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Detoxification of fumonisins by isothiocyanates
S98
Abstracts / Toxicology Letters 211S (2012) S43–S216
P12-08 Occurrence and risk assessment of emerging myctoxins in nuts and dried fruits Guillermina Font, Josefa Tolosa, Ana Belén Serrano, Emilia Ferrer Valencia University, Spain Mycotoxins are secondary metabolites produced by filamentous fungi in foods and feeds. Emerging Fusarium mycotoxins have acquired importance, since their severe potential toxic effects. Several authors have indicated the presence of some mycotoxins in nuts and dried fruits. In the literature review, no data are available about the incidence of emerging mycotoxins in these matrices. The purpose of this study is to determine the emerging mycotoxins in nuts and dried fruits and approach to the risk assessment. To do this, 71 samples of nuts and dried fruits (peanuts, almonds, pistachios, walnuts, toasted maize, hazelnuts, sunflower seeds, dates, dried apricots, figs, raisins and dried plums) from different markets were analyzed for the determination of five emerging mycotoxins. Ultrasound extraction was performed with acetonitrile followed by C18 purification and determination in reverse phase with C18 column, acetonitrile and methanol with 20 mM ammonium formate phase mobile, by liquid chromatography–triple quadrupole-tandem mass spectrometry (LC–MS–MS QqQ). The results showed that 67% of samples were contaminated. Enniatin A was the most predominant mycotoxin. The concentrations ranged from 0.046 to 1.490 mg/kg for Enniatin A, from 0.007 to 0.030 mg/kg for Enniatin A1, from 0.030 to 0.784 mg/kg for Enniatin B, from 0.022 to 0.784 mg/kg for Beauvericin. For the risk assessment, the estimated total intake (0.028 g/kg bw/day) was the same order as the lowest tolerable daily intake established by JECFA for Fusarium mycotoxins. This work was supported by the projects AGL2010/17024/ALI (Science and Innovation Spanish Ministry) and PLAT/2009-012 (Valencian Government, Spain). doi:10.1016/j.toxlet.2012.03.367
P12-09 Survey of 50 multiclass pesticide residues in bee products ˜ Jordi 2 , Font Houda Berrada 1 , Molto Juan Carlos 2 , Manes 2 Guillermina 1 Valencia University, Spain, 2 Valencia University, Food Science Depart, Spain
An optimized analytical method employing gas chromatography–triple quadruple mass spectrometry was developed for the GC–ESI-MS/MS determination of 50 pesticides within the major groups of pesticides (organohalogen, organophosphorous, pyrethroids and organonitrogen) in representative samples of locally sold honeys, pollens and royal jellies A sample preparation approach based on QuEChERS was implemented and validated for pesticides in bee products. The procedure involved homogenization of a 5 g sample with acetonitrile/water mixture followed by salting out with citrate buffer, magnesium sulphate and sodium chloride. Cleanup was carried out using primary secondary amine (PSA), and graphitized carbon black (GCB). This combination of cleanup steps ensured efficient extract purification. Matrix effects were evaluated by comparing the slopes of matrixmatched and solvent-based calibration curves. The recovery results
found ranged from 73% to 102% and the limits of detection were lower than 0.01 mg/kg in the multiple ion monitoring mode. The occurrence of the selected pesticides in 115 honey samples, 20 pollens and 15 royal jellies was investigated and more than 6% of the investigated samples have a contamination level exceeding the European tolerance limits. The results indicated that bifenthrin was the most prevalent agent detected in all the honey samples even at concentration below 10 g/kg. Pesticides found in Royal jellies and pollen samples belonged to the organohalogen group and lower levels of residues of bifenthrin were also detected. The mean estimated daily intakes of the selected pesticides were compared with the tolerable daily intake. doi:10.1016/j.toxlet.2012.03.368
P12-10 Protection against health hazards of multi-mycotoxins Mosaad Abdel-Wahhab 1 , Mohamed Ibrahim 2 , Ahmed Ibrahim 3 , Aziza El-Nekeety 4 , Nabila Hassan 2 , Amira Mohamed 5 Professor, National Research Center, Egypt, 2 Professor, Egypt, 3 Asst. Professor, Egypt, 4 Researcher, Egypt, 5 MSc student, Egypt
1
Propose: The current study was conducted to evaluate the protective effects of Panax ginseng extract (PGE) against the toxicity and oxidative stress in rats fed aflatoxin (AFs) and/or fumonisin (FB)-contaminated diet. Methods: Eighty female Sprague-Dawley rats were divided into eight experimental groups included the control group, the group treated orally with PGE (0.5 mg/kg b.w.) and the groups fed AFs (1.4 mg/kg diet) and/or FB (20 mg/kg b.w.) contaminated diet alone or plus PGE for 11 weeks. Blood, liver and kidney tissue samples were collected at the end of treatment period for biochemical and histological studies. Results and conclusion of the study: The results indicated that PGE increased SOD level in liver however; the other parameters were comparable to controls. Animals fed AFs and/or FB-contaminated diet showed a significant increase in serum biochemical parameters and oxidative stress markers accompanied with a significant decrease in antioxidant parameters levels and a severe histological changes in the liver tissue. These changes were pronounced in the group fed AFs plus FB. PGE succeeded to induce a significant improvement in all biochemical parameters and the histological picture towards the control although it did not normalize them. It could be concluded that PGE is a promise candidate against the exposure to multi-mycotoxins in food doi:10.1016/j.toxlet.2012.03.369
P12-11 Detoxification of fumonisins by isothiocyanates Guillermina Font, Ines Azaiez, Meca Giuseppe, Fernández-Franzón Mónica Faculty of Pharmacy, Univ. of Valencia, Spain Fumonisins (FBs) are a family of mycotoxins produced mainly by the maize pathogens Fusarium verticillioides and Fusarium moniliforme. FB1, the most abundant and toxic metabolite, is known to cause a range of species-specific toxic responses. Fumonisin B1 has been declared by IARC as a class 2B carcinogen, which means a possible human carcinogen. The isothiocyanates (ITCs) are natu-
Abstracts / Toxicology Letters 211S (2012) S43–S216
ral compounds produced by enzymatic conversion of metabolites called glucosinolates through the enzyme myrosinase. The aim of this study was to reduce the occurrence of the FB1 and FB2, employing a chemical reduction based on the reaction between the mycotoxins (1 mg/L) and the ITCs: Allyl (AITC), Phenil (PITC) and Benzyl isothiocyanates (BITC) (1 mg/L). The antifungal activity of the ITCs on Fusarium mycotoxigenic strains has also been tested. The ITCs employed in this study were active against several Fusarium mycotoxigenic strains such as Fusarium verticillioides in a range variable from 0.005 to 500 g. The reduction of the FB1 and FB2 after their reaction with PITC ranged from 2.32% to 100%, from 3.9% to 99% with AITC, and from 2.0% to 81.3% with BITC. In the food system composed by the corn grain naturally contaminated with F. verticillioides, the reduction of the mycotoxins object of this study was similar to the previous results. The addition of ITCs to food contaminated by FBs might be a possible way of detoxification. Acknowledgement: This work was supported by Science and Innovation Spanish Ministry (AGL2010-17024/ALI). doi:10.1016/j.toxlet.2012.03.370
P12-12 Analysis of trichothecenes in infant cereals from Spanish markets Guillermina Font, Hui Lu, Maria José Ruiz, Mónica Fernández-Franzón Faculty of Pharmacy, Univ of Valencia, Spain Fifty-seven samples of infant cereals from Spanish supermarkets, and organic food shops were collected during 2010 and analysed by liquid chromatography (LC) and electrospray ionization tandem mass spectrometry with a triple quadrupole (QqQ) analyser (LC–MS/MS) for the simultaneous determination of seven trichothecenes: T-2, HT-2, diacetoxyscirpenol (DAS), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15acetyldeoxynivalenol (15-ADON), and fusarenon X (FUS-X). The QuEChERS (quick, easy, cheap, effective, rugged and safe) method was performed to extract and purified the selected mycotoxins. Recoveries ranged from 72% to 83% with relative standard deviation (RSD) from 4% to 12%. The method was also validated with respect to parameters of linearity, precision and limit of quantification. The most frequently detected mycotoxin was 3-ADON; 6 (10.5%) samples were contaminated at levels ranging from 0.095 to 0.165 mg kg−1 . FUS-X was found in 2 samples (3.5%) in the range of 0.66–1.66 mg kg−1 . Only one sample was contaminated with T-2 at a level of 0.039 mg kg−1 and other sample with 15-ADON at a level of 0.119 mg kg−1 . One sample contained two mycotoxins, 15ADON and 3-ADON. The amounts found was relatively low, only one sample contained a quite high amount of FUS-X (1.66 mg kg−1 ), although this mycotoxin is not regulated, it should be carefully controlled. The results obtained suggest the low probability of health risks from these trichothecenes through the consumption of infant cereals. Acknowledgement: This work was supported by Science and Innovation Spanish Ministry (AGL2010-17024/ALI). doi:10.1016/j.toxlet.2012.03.371
S99
P12-13 Comparison lithium concentration in water of Ahvaz city, Iran Abdolazim Behfar 1 , Zahra Nazari Khorasgani 1 , Ahmadzeza Barzan 1 , Gholamreza Reissi 2 , Maysam Sheikhzadeh 1 1
Ahvaz Jundishapur University of Medical, Iran, 2 Ahvaz Water & Wastewater Company, Iran Introduction: Due to the misconception that bottled water is safer or healthier than tap water, bottled water is now one of the largest industries in the world. However, studies show that most of these bottled water also come from municipal sources. There are no drinking water standards for lithium set by any government. There is extremely limited data on Li concentration of bottled waters and tap water in Ahvaz city – Iran, so this research was done. Methods and materials: In order to quantification of Li in produced bottled water of Dezoosh and Anahita factories and also Tap water from outlet of Treated Water Plant No. 2 in Ahvaz, 14times, each time, one composite sample which comprising 20 grab samples was taken from each of them. Flame atomic absorption spectrometry with direct nebulization was applied for determination of Li concentration in collected water samples using wavelength of 671 nm. Calibration curve was made by 1, 0.5, 0.125 and 0.0625 ppm of Li standard solutions. The recovery by standard addition method ranged from 94.3% to 96.9%. Results and discussion: The least and highest concentrations of Li were seen in Deznoosh bottled water and tap water at concentrations 0.070 and 0.750 ppm, respectively. The highest and lowest concentrations of Li belonged to tap water and Deznoosh water at concentration 0.561 and 0.1300 ppm, respectively. The use of highly purified water may have deprived us of a trace amount of lithium found in tap water which appears to be an essential element needed for good mental health. doi:10.1016/j.toxlet.2012.03.372
P12-14 Assessment of heavy metals concentrations in imported raw sugar at Ahvaz markets, Iran Rezvan Ebrahimi, Zahra Nazari Khorasgani, Abdolazim Behfar, Azam Hossieni Ahvaz Jundishapur University of Medical, Iran Purpose: The control of heavy metals in sugar is important because of the existence of sugar in all food we consume. Look at the statistics shows that Iran currently one of 10 large sugar importers in the world is known. It is estimated per capita sugar consumption in Iran is 30 kg per year, thus it should be carefully monitored and therefore, the levels of lead and cadmium in imported raw sugar was investigated. Methods: For this purpose, 10 samples from imported sugar in existence markets in Ahvaz city were purchased. An adequate amount of each milled sample dried at 110–120 ◦ C in an oven. The vessel was placed in cold furnace and temperature was set at 550 ◦ C and holds at this temperature overnight until white Carbon free ash was obtained, then removed and cooled. Residue was dissolved in HNO3 –H2 O (1 + 99), quantitatively transferred to 5 ml volumetric flask and diluted with water. Deionized water was used for blank. Then the concentration of Cd and Pb in raw sugar samples was measured by flame atomic absorption. Results: All samples were found to be contaminated of with Pb and Cd at levels ranging from 28 to 233 and 1.6 to 60 ppb, respectively. Pb levels in 1 sample (10%) were above the Maximum tolerable limit of Pb (200 ppb) and Cd (100 ppb)
Abstracts / Toxicology Letters 211S (2012) S43–S216
P12-08 Occurrence and risk assessment of emerging myctoxins in nuts and dried fruits Guillermina Font, Josefa Tolosa, Ana Belén Serrano, Emilia Ferrer Valencia University, Spain Mycotoxins are secondary metabolites produced by filamentous fungi in foods and feeds. Emerging Fusarium mycotoxins have acquired importance, since their severe potential toxic effects. Several authors have indicated the presence of some mycotoxins in nuts and dried fruits. In the literature review, no data are available about the incidence of emerging mycotoxins in these matrices. The purpose of this study is to determine the emerging mycotoxins in nuts and dried fruits and approach to the risk assessment. To do this, 71 samples of nuts and dried fruits (peanuts, almonds, pistachios, walnuts, toasted maize, hazelnuts, sunflower seeds, dates, dried apricots, figs, raisins and dried plums) from different markets were analyzed for the determination of five emerging mycotoxins. Ultrasound extraction was performed with acetonitrile followed by C18 purification and determination in reverse phase with C18 column, acetonitrile and methanol with 20 mM ammonium formate phase mobile, by liquid chromatography–triple quadrupole-tandem mass spectrometry (LC–MS–MS QqQ). The results showed that 67% of samples were contaminated. Enniatin A was the most predominant mycotoxin. The concentrations ranged from 0.046 to 1.490 mg/kg for Enniatin A, from 0.007 to 0.030 mg/kg for Enniatin A1, from 0.030 to 0.784 mg/kg for Enniatin B, from 0.022 to 0.784 mg/kg for Beauvericin. For the risk assessment, the estimated total intake (0.028 g/kg bw/day) was the same order as the lowest tolerable daily intake established by JECFA for Fusarium mycotoxins. This work was supported by the projects AGL2010/17024/ALI (Science and Innovation Spanish Ministry) and PLAT/2009-012 (Valencian Government, Spain). doi:10.1016/j.toxlet.2012.03.367
P12-09 Survey of 50 multiclass pesticide residues in bee products ˜ Jordi 2 , Font Houda Berrada 1 , Molto Juan Carlos 2 , Manes 2 Guillermina 1 Valencia University, Spain, 2 Valencia University, Food Science Depart, Spain
An optimized analytical method employing gas chromatography–triple quadruple mass spectrometry was developed for the GC–ESI-MS/MS determination of 50 pesticides within the major groups of pesticides (organohalogen, organophosphorous, pyrethroids and organonitrogen) in representative samples of locally sold honeys, pollens and royal jellies A sample preparation approach based on QuEChERS was implemented and validated for pesticides in bee products. The procedure involved homogenization of a 5 g sample with acetonitrile/water mixture followed by salting out with citrate buffer, magnesium sulphate and sodium chloride. Cleanup was carried out using primary secondary amine (PSA), and graphitized carbon black (GCB). This combination of cleanup steps ensured efficient extract purification. Matrix effects were evaluated by comparing the slopes of matrixmatched and solvent-based calibration curves. The recovery results
found ranged from 73% to 102% and the limits of detection were lower than 0.01 mg/kg in the multiple ion monitoring mode. The occurrence of the selected pesticides in 115 honey samples, 20 pollens and 15 royal jellies was investigated and more than 6% of the investigated samples have a contamination level exceeding the European tolerance limits. The results indicated that bifenthrin was the most prevalent agent detected in all the honey samples even at concentration below 10 g/kg. Pesticides found in Royal jellies and pollen samples belonged to the organohalogen group and lower levels of residues of bifenthrin were also detected. The mean estimated daily intakes of the selected pesticides were compared with the tolerable daily intake. doi:10.1016/j.toxlet.2012.03.368
P12-10 Protection against health hazards of multi-mycotoxins Mosaad Abdel-Wahhab 1 , Mohamed Ibrahim 2 , Ahmed Ibrahim 3 , Aziza El-Nekeety 4 , Nabila Hassan 2 , Amira Mohamed 5 Professor, National Research Center, Egypt, 2 Professor, Egypt, 3 Asst. Professor, Egypt, 4 Researcher, Egypt, 5 MSc student, Egypt
1
Propose: The current study was conducted to evaluate the protective effects of Panax ginseng extract (PGE) against the toxicity and oxidative stress in rats fed aflatoxin (AFs) and/or fumonisin (FB)-contaminated diet. Methods: Eighty female Sprague-Dawley rats were divided into eight experimental groups included the control group, the group treated orally with PGE (0.5 mg/kg b.w.) and the groups fed AFs (1.4 mg/kg diet) and/or FB (20 mg/kg b.w.) contaminated diet alone or plus PGE for 11 weeks. Blood, liver and kidney tissue samples were collected at the end of treatment period for biochemical and histological studies. Results and conclusion of the study: The results indicated that PGE increased SOD level in liver however; the other parameters were comparable to controls. Animals fed AFs and/or FB-contaminated diet showed a significant increase in serum biochemical parameters and oxidative stress markers accompanied with a significant decrease in antioxidant parameters levels and a severe histological changes in the liver tissue. These changes were pronounced in the group fed AFs plus FB. PGE succeeded to induce a significant improvement in all biochemical parameters and the histological picture towards the control although it did not normalize them. It could be concluded that PGE is a promise candidate against the exposure to multi-mycotoxins in food doi:10.1016/j.toxlet.2012.03.369
P12-11 Detoxification of fumonisins by isothiocyanates Guillermina Font, Ines Azaiez, Meca Giuseppe, Fernández-Franzón Mónica Faculty of Pharmacy, Univ. of Valencia, Spain Fumonisins (FBs) are a family of mycotoxins produced mainly by the maize pathogens Fusarium verticillioides and Fusarium moniliforme. FB1, the most abundant and toxic metabolite, is known to cause a range of species-specific toxic responses. Fumonisin B1 has been declared by IARC as a class 2B carcinogen, which means a possible human carcinogen. The isothiocyanates (ITCs) are natu-
Abstracts / Toxicology Letters 211S (2012) S43–S216
ral compounds produced by enzymatic conversion of metabolites called glucosinolates through the enzyme myrosinase. The aim of this study was to reduce the occurrence of the FB1 and FB2, employing a chemical reduction based on the reaction between the mycotoxins (1 mg/L) and the ITCs: Allyl (AITC), Phenil (PITC) and Benzyl isothiocyanates (BITC) (1 mg/L). The antifungal activity of the ITCs on Fusarium mycotoxigenic strains has also been tested. The ITCs employed in this study were active against several Fusarium mycotoxigenic strains such as Fusarium verticillioides in a range variable from 0.005 to 500 g. The reduction of the FB1 and FB2 after their reaction with PITC ranged from 2.32% to 100%, from 3.9% to 99% with AITC, and from 2.0% to 81.3% with BITC. In the food system composed by the corn grain naturally contaminated with F. verticillioides, the reduction of the mycotoxins object of this study was similar to the previous results. The addition of ITCs to food contaminated by FBs might be a possible way of detoxification. Acknowledgement: This work was supported by Science and Innovation Spanish Ministry (AGL2010-17024/ALI). doi:10.1016/j.toxlet.2012.03.370
P12-12 Analysis of trichothecenes in infant cereals from Spanish markets Guillermina Font, Hui Lu, Maria José Ruiz, Mónica Fernández-Franzón Faculty of Pharmacy, Univ of Valencia, Spain Fifty-seven samples of infant cereals from Spanish supermarkets, and organic food shops were collected during 2010 and analysed by liquid chromatography (LC) and electrospray ionization tandem mass spectrometry with a triple quadrupole (QqQ) analyser (LC–MS/MS) for the simultaneous determination of seven trichothecenes: T-2, HT-2, diacetoxyscirpenol (DAS), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15acetyldeoxynivalenol (15-ADON), and fusarenon X (FUS-X). The QuEChERS (quick, easy, cheap, effective, rugged and safe) method was performed to extract and purified the selected mycotoxins. Recoveries ranged from 72% to 83% with relative standard deviation (RSD) from 4% to 12%. The method was also validated with respect to parameters of linearity, precision and limit of quantification. The most frequently detected mycotoxin was 3-ADON; 6 (10.5%) samples were contaminated at levels ranging from 0.095 to 0.165 mg kg−1 . FUS-X was found in 2 samples (3.5%) in the range of 0.66–1.66 mg kg−1 . Only one sample was contaminated with T-2 at a level of 0.039 mg kg−1 and other sample with 15-ADON at a level of 0.119 mg kg−1 . One sample contained two mycotoxins, 15ADON and 3-ADON. The amounts found was relatively low, only one sample contained a quite high amount of FUS-X (1.66 mg kg−1 ), although this mycotoxin is not regulated, it should be carefully controlled. The results obtained suggest the low probability of health risks from these trichothecenes through the consumption of infant cereals. Acknowledgement: This work was supported by Science and Innovation Spanish Ministry (AGL2010-17024/ALI). doi:10.1016/j.toxlet.2012.03.371
S99
P12-13 Comparison lithium concentration in water of Ahvaz city, Iran Abdolazim Behfar 1 , Zahra Nazari Khorasgani 1 , Ahmadzeza Barzan 1 , Gholamreza Reissi 2 , Maysam Sheikhzadeh 1 1
Ahvaz Jundishapur University of Medical, Iran, 2 Ahvaz Water & Wastewater Company, Iran Introduction: Due to the misconception that bottled water is safer or healthier than tap water, bottled water is now one of the largest industries in the world. However, studies show that most of these bottled water also come from municipal sources. There are no drinking water standards for lithium set by any government. There is extremely limited data on Li concentration of bottled waters and tap water in Ahvaz city – Iran, so this research was done. Methods and materials: In order to quantification of Li in produced bottled water of Dezoosh and Anahita factories and also Tap water from outlet of Treated Water Plant No. 2 in Ahvaz, 14times, each time, one composite sample which comprising 20 grab samples was taken from each of them. Flame atomic absorption spectrometry with direct nebulization was applied for determination of Li concentration in collected water samples using wavelength of 671 nm. Calibration curve was made by 1, 0.5, 0.125 and 0.0625 ppm of Li standard solutions. The recovery by standard addition method ranged from 94.3% to 96.9%. Results and discussion: The least and highest concentrations of Li were seen in Deznoosh bottled water and tap water at concentrations 0.070 and 0.750 ppm, respectively. The highest and lowest concentrations of Li belonged to tap water and Deznoosh water at concentration 0.561 and 0.1300 ppm, respectively. The use of highly purified water may have deprived us of a trace amount of lithium found in tap water which appears to be an essential element needed for good mental health. doi:10.1016/j.toxlet.2012.03.372
P12-14 Assessment of heavy metals concentrations in imported raw sugar at Ahvaz markets, Iran Rezvan Ebrahimi, Zahra Nazari Khorasgani, Abdolazim Behfar, Azam Hossieni Ahvaz Jundishapur University of Medical, Iran Purpose: The control of heavy metals in sugar is important because of the existence of sugar in all food we consume. Look at the statistics shows that Iran currently one of 10 large sugar importers in the world is known. It is estimated per capita sugar consumption in Iran is 30 kg per year, thus it should be carefully monitored and therefore, the levels of lead and cadmium in imported raw sugar was investigated. Methods: For this purpose, 10 samples from imported sugar in existence markets in Ahvaz city were purchased. An adequate amount of each milled sample dried at 110–120 ◦ C in an oven. The vessel was placed in cold furnace and temperature was set at 550 ◦ C and holds at this temperature overnight until white Carbon free ash was obtained, then removed and cooled. Residue was dissolved in HNO3 –H2 O (1 + 99), quantitatively transferred to 5 ml volumetric flask and diluted with water. Deionized water was used for blank. Then the concentration of Cd and Pb in raw sugar samples was measured by flame atomic absorption. Results: All samples were found to be contaminated of with Pb and Cd at levels ranging from 28 to 233 and 1.6 to 60 ppb, respectively. Pb levels in 1 sample (10%) were above the Maximum tolerable limit of Pb (200 ppb) and Cd (100 ppb)