RESUMO
Aflatoxin B1 (AFB1) and ochratoxin A (OTA) are highly toxic mycotoxins present in food and feed, posing serious health risks to humans and animals. This study aimed to validate an efficient and cost-effective analytical method for quantifying AFB1 and OTA in rat urine using immunoaffinity column extraction and liquid chromatography with fluorescence detection (IAC-LC-FD). Additionally, the study evaluated the effect of incorporating fermented whey and pumpkin into the feed on the urinary excretion of these mycotoxins. The limits of detection and quantification were determined to be 0.1 µg/kg and 0.3 µg/kg, respectively, for both mycotoxins in feed, and 0.2 ng/mL and 0.6 ng/mL, respectively, in urine. The method demonstrated robust recovery rates ranging from 74% to 119% for both AFB1 and OTA in both matrices. In feed samples, the levels of AFB1 and OTA ranged from 4.3 to 5.2 µg/g and from 5.4 to 8.8 µg/g, respectively. This validated method was successfully applied to analyze 116 urine samples from rats collected during the fourth week of an in vivo trial. The results indicated that the addition of fermented whey and pumpkin to the feed influenced mycotoxin excretion in urine, with variations observed based on the sex of the rats, type of mycotoxin, and exposure dosage.
Assuntos
Aflatoxina B1 , Ocratoxinas , Animais , Ocratoxinas/urina , Aflatoxina B1/urina , Masculino , Feminino , Ratos , Ração Animal/análise , Cromatografia Líquida , Soro do Leite/química , Ratos Sprague-Dawley , Contaminação de Alimentos/análise , Espectrometria de FluorescênciaRESUMO
Ochratoxin A (OTA) is known to be strongly bound to serum albumin, but it remains unknown how albumin affects its metabolism and kinetics. To close this gap, we used a mouse model, where heterozygous albumin deletion reduces serum albumin to concentrations similar to hypoalbuminemic patients and completely eliminates albumin by a homozygous knockout. OTA and its potential metabolites (OTα, 4-OH-OTA, 7'-OH-OTA, OTHQ, OP-OTA, OTB-GSH, OTB-NAC, OTB) were time-dependently analyzed in plasma, bile, and urine by LC-MS/MS and were compared to previously published hepatotoxicity and nephrotoxicity data. Homozygous albumin deletion strongly accelerated plasma clearance as well as biliary and urinary excretion of the parent compound and its hydroxylation products. Decreasing albumin in mice by the heterozygous and even more by the homozygous knockout leads to an increase in the parent compound in urine which corresponded to increased nephrotoxicity. The role of albumin in OTA-induced hepatotoxicity is more complex, since heterozygous but not homozygous nor wild-type mice showed a strong biliary increase in the toxic open lactone OP-OTA. Correspondingly, OTA-induced hepatotoxicity was higher in heterozygous than in wild-type and homozygous animals. We present evidence that albumin-mediated retention of OTA in hepatocytes is required for formation of the toxic OP-OTA, while complete albumin elimination leads to rapid biliary clearance of OTA from hepatocytes with less formation of OP-OTA. In conclusion, albumin has a strong influence on metabolism and toxicity of OTA. In hypoalbuminemia, the parent OTA is associated with increased nephrotoxicity and the open lactone with increased hepatotoxicity.
Assuntos
Albuminas , Ocratoxinas , Animais , Masculino , Camundongos , Albuminas/metabolismo , Bile/metabolismo , Cromatografia Líquida , Fígado/metabolismo , Fígado/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ocratoxinas/metabolismo , Ocratoxinas/urina , Ocratoxinas/toxicidade , Albumina Sérica/metabolismo , Espectrometria de Massas em TandemRESUMO
Ochratoxin A (OTA) is a widespread food contaminant, with exposure estimated to range from 0.64 to 17.79 ng/kg body weight (bw) for average consumers and from 2.40 to 51.69 ng/kg bw per day for high consumers. Current exposure estimates are, however, associated with considerable uncertainty. While biomarker-based approaches may contribute to improved exposure assessment, there is yet insufficient data on urinary metabolites of OTA and their relation to external dose to allow reliable estimates of daily intake. This study was designed to assess potential species differences in phase II biotransformation in vitro and to establish a correlation between urinary OTA-derived glucuronides and mercapturic acids and external exposure in rats in vivo. In vitro analyses of OTA metabolism using the liver S9 of rats, humans, rabbits and minipigs confirmed formation of an OTA glucuronide but provided no evidence for the formation of OTA-derived mercapturic acids to support their use as biomarkers. Similarly, OTA-derived mercapturic acids were not detected in urine of rats repeatedly dosed with OTA, while indirect analysis using enzymatic hydrolysis of the urine samples prior to LC-MS/MS established a linear relationship between urinary glucuronide excretion and OTA exposure. These results support OTA-derived glucuronides but not mercapturic acids as metabolites suitable for biomonitoring.
Assuntos
Acetilcisteína/urina , Biomarcadores/urina , Monitoramento Ambiental/métodos , Contaminação de Alimentos/análise , Glucuronídeos/urina , Ocratoxinas/metabolismo , Ocratoxinas/urina , Animais , Modelos Animais de Doenças , Técnicas In Vitro , Ratos , Especificidade da Espécie , Suínos , Porco MiniaturaRESUMO
Human are exposed to a wide range of mycotoxins through dietary food intake, including processed food. Even most of the mycotoxin exposure assessment studies are based on analysis of foodstuffs, and evaluation of dietary intake through food consumption patterns and human biomonitoring methods are rising as a reliable alternative to approach the individual exposures, overcoming the limitations of the indirect dietary assessment. In this study, human urine samples were analyzed, seeking the presence of deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZEA), and their metabolites. For this purpose, 40 urine samples from female and male adult residents in the city of Valencia (Spain) were evaluated by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-ESI-qTOF) after salting-out liquid-liquid extraction. Analytical data showed that 72.5% of analyzed samples were contaminated by at least one mycotoxin at variable levels. The most prevalent mycotoxins were de-epoxy DON (DOM-1) (53%), ZEA (40%), and α-zearalenol (αZOL) (43%), while OTA was only detected in one sample. The mean concentrations in positive samples were DON (9.07 ng/mL), DOM-1 (20.28 ng/mL), ZEA (6.70 ng/mL), ZEA-14 glucoside (ZEA-14-Glc) (12.43 ng/mL), αZOL (27.44 ng/mL), αZOL-14 glucoside (αZOL-14-Glc) (12.84 ng/mL), and OTA (11.73 ng/mL). Finally, probable daily intakes (PDIs) were calculated and compared with the established tolerable daily intakes (TDIs) to estimate the potential risk of exposure to the studied mycotoxins. The calculated PDI was below the TDI value established for DON in both female and male adults, reaching a percentage up to 30%; however, this percentage increased up to 92% considering total DON (DON + DOM-1). On the other hand, the PDI obtained for ZEA and its metabolites were higher than the TDI value fixed, but the low urine excretion rate (10%) considered should be highlighted. Finally, the PDI calculated in the detected positive sample for OTA exceeded the TDI value. The findings of the present study confirm the presence of the studied mycotoxins and their metabolites as some of the most prevalent in urine.
Assuntos
Exposição Dietética/análise , Ocratoxinas/urina , Tricotecenos/urina , Zearalenona/urina , Adolescente , Adulto , Idoso , Monitoramento Biológico , Biomarcadores/urina , Cromatografia Líquida , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Medição de Risco , Espectrometria de Massas por Ionização por Electrospray , Adulto JovemRESUMO
In this paper, a novel DNA-based biosensor is proposed, which is based on paramagnetic microbeads carrying an ochratoxin A (OTA) capture aptamer. A sandwich-like detection complex is linked to the capture aptamer and is able to trigger, in presence of OTA, an isothermal rolling circle amplification (RCA) reaction. This latter generated autocatalytic units with a peroxidase activity (DNAzyme) that, in presence of a proper substrate, gave a blue-coloured product visible by the naked eye. The capture aptamer, blocked onto magnetic beads, allowed the specific capture of OTA in liquid samples. The modified detection aptamer, annealed to a circularized probe, was then used to detect the toxin capture event. Indeed, in the presence of OTA and an isothermal enzyme, the circular DNA was amplified, producing a single-stranded and tandem repeated long homologous copy of its sequence. In the DNA strand, a self-catalytic structure was formed with hemin as the catalytic core, inducing the development of blue colour in the presence of ABTS and hydrogen peroxide. The results showed that the biosensor has high sensitivity and selectivity for the detection of OTA, as low as 1.09 × 10-12 ng/mL. Moreover, the proposed biosensor was successfully used for the detection of OTA in naturally contaminated rat urine. Accuracy and repeatability data obtained in recovery experiments were satisfying, being recoveries >95% with relative standard deviations in the range 3.6-15%. For the first time, an aptasensor was successfully applied to detect OTA in biological fluids. It can be used for mycotoxin biomonitoring and assessment of individual exposure.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Ocratoxinas , Animais , DNA , Ocratoxinas/urina , RatosRESUMO
Ochratoxin A (OTA) and citrinin (CIT) are nephrotoxic mycotoxins, found in various foodstuffs and in animal feed, and may cause adverse effects on animal and human health. Previous biomonitoring data indicate a frequent co-exposure of Bangladeshi adults to these mycotoxins. However, since such data are not yet available for young children, a vulnerable part of the population, we conducted this study to assess their exposure to OTA and CIT and compare it with that of adults in Bangladesh. In total, 154 urine samples were collected from infants and children in Rajshahi (n = 88) and Dhaka (n = 66) district of Bangladesh. OTA, CIT, and their metabolites were analyzed by a sensitive HPLC-FLD or LC-MS/MS method, respectively. Overall, OTA and CIT biomarkers were detectable in 72.7% and 54.9% of urines, respectively. The mean OTA and OTα levels in urines were higher in children (0.13 ng/mL and 0.28 ng/mL, respectively) than in infants (0.08 ng/mL and 0.05 ng/mL, respectively). Regarding region, the mean level of OTA was higher in samples from Rajshahi district (0.13 ng/mL) than from Dhaka district (0.09 ng/mL), while the mean OTα level was 2-fold higher in the Dhaka. The total CIT biomarker concentration was significantly higher in children (2.16 ng/mL) than in infant (0.70 ng/mL) urines (p < 0.05), and the mean concentration of HO-CIT was about 6-fold higher than that of parent compound CIT. A provisional daily intake for CIT was calculated and exceeded a preliminary value set by EFSA (0.2 µg/kg bw) in 23.3% and 11.9% of children and infants, respectively. OTA and CIT biomarker concentrations in the young children cohorts are higher than those found in Bangladeshi adults in summer, but lower than in winter season. The new results indicate frequent co-exposure to nephrotoxic mycotoxins that varies between the cohorts and regions in Bangladesh.
Assuntos
Monitoramento Biológico , Citrinina/urina , Ocratoxinas/urina , Bangladesh , Biomarcadores/urina , Criança , Pré-Escolar , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Citrinina/metabolismo , Estudos de Coortes , Feminino , Geografia , Humanos , Lactente , Recém-Nascido , Masculino , Ocratoxinas/metabolismo , Estações do Ano , Espectrometria de Massas em TandemRESUMO
Cereal foods are commonly contaminated with multiple mycotoxins resulting in frequent human mycotoxin exposure. Children are at risk of high-level exposure because of their high cereal intake relative to body weight. Hence, this study aims to assess multimycotoxin exposure in UK children using urinary biomarkers. Spot urines (n = 21) were analyzed for multimycotoxins (deoxynivalenol, DON; nivalenol, NIV; ochratoxin A, OTA; zearalenone, ZEN; α-zearalenol, α-ZEL; ß-zearalenol, ß-ZEL; T-2 toxin, T-2; HT-2 toxin, HT-2; and aflatoxin B1 and M1, AFB1, AFM1) using liquid chromatography-coupled tandem mass spectrometry. Urine samples frequently contained DON (13.10 ± 12.69 ng/mL), NIV (0.36 ± 0.16 ng/mL), OTA (0.05 ± 0.02 ng/mL), and ZEN (0.09 ± 0.07 ng/mL). Some samples (1-3) contained T-2, HT-2, α-ZEL, and ß-ZEL but not aflatoxins. Dietary mycotoxin estimation showed that children were frequently exposed to levels exceeding the tolerable daily intake (52 and 95% of cases for DON and OTA). This demonstrates that UK children are exposed to multiple mycotoxins through their habitual diet.
Assuntos
Biomarcadores/urina , Micotoxinas/urina , Aflatoxinas/urina , Criança , Pré-Escolar , Dieta/efeitos adversos , Exposição Ambiental/efeitos adversos , Feminino , Contaminação de Alimentos/estatística & dados numéricos , Humanos , Masculino , Ocratoxinas/urina , Inquéritos e Questionários , Toxina T-2/análogos & derivados , Toxina T-2/urina , Tricotecenos/urina , Reino Unido , Zearalenona/urina , Zeranol/análogos & derivados , Zeranol/urinaRESUMO
Ochratoxin A (OTA) is a nephrotoxic mycotoxin with nephrocarcinogenic potential found in a broad spectrum of food commodities. The mode of action of this compound, as well as its metabolism, is still not fully understood. To determine whether the conjugation of OTA with glutathione plays an important role in human OTA metabolism, an ochratoxin-glutathione conjugate (OTB-GSH), as well as the corresponding urinary metabolite ochratoxin-N-acetyl-L-cysteine (OTB-NAC), were synthesized and their structures confirmed by NMR spectroscopy. By means of synthesized stable isotope-labeled d5-OTB-GSH and d5-OTB-NAC references, a sensitive HPLC-MS/MS method has been developed and applied for the screening of human urine samples. OTB-NAC could be detected in 11 of the analyzed 18 urine samples and was quantified in 5 urine samples in the range between 0.023 and 0.176 ng mg-1 creatinine. OTB-GSH has not been detected in the urine samples. In OTB-NAC positive samples, this metabolite contributed to a comparable concentration range to the total OTA excretion as the parent compound. This is the first direct analysis of an OTA phase 2 metabolite in humans.
Assuntos
Acetilcisteína/urina , Biomarcadores/urina , Glutationa/urina , Micotoxinas/urina , Ocratoxinas/urina , Cromatografia Líquida de Alta Pressão/métodos , Contaminação de Alimentos/análise , Humanos , Ocratoxinas/análiseRESUMO
Ochratoxin A (OTA) is a mycotoxin detected worldwide. Urine biomonitoring is a more realistic and non-invasive way to assess exposure when compared to traditional methods based on food occurrence and consumption data. Few studies have investigated children's exposure to OTA, although it is a more susceptible population. Our main goal was the OTA biomonitoring in urine of Portuguese children to better characterize the health risk of this population. The validated analytical methodology was based on an immunoaffinity clean-up, followed by LC-FD. First morning samples were collected, between 2018 and 2019, from 85 healthy children, 41 boys and 44 girls, aging between 2 and 13 years old. Overall, from the 85 analysed samples 79 (92.94%) were found positive. The average OTA concentration level was of 0.020â¯ng/mL (29.41â¯ng/g of creatinine), with a maximum value of 0.052â¯ng/mL (114.45â¯ng/g of creatinine). No statistical differences were found between the OTA contamination levels, the anthropometric data and dietary habits considered. Depending on the excretion rate considered for the PDI calculation, the risk assessed for the average OTA contamination varied from 10% to 194%, indicating a potential concern. This is the first study regarding OTA exposure in Portuguese children, showing their clear exposure.
Assuntos
Ocratoxinas/urina , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Limite de Detecção , Masculino , Ocratoxinas/normas , Portugal , Padrões de ReferênciaRESUMO
The Czech Republic occupies the first place in the world in the frequency of renal and other urinary tract tumours, but their aetiology is unknown. To explore whether carcinogenic and nephrotoxic mycotoxins may contribute to kidney diseases in the Czech population, biomarkers of ochratoxin A (OTA) and citrinin (CIT) exposure were determined in biological specimens from a cohort of 50 patients with malignant renal tumours. Biomarker analyses in blood and urine samples used validated targeted methods for measuring OTA and CIT plus dihydrocitrinone (DH-CIT) after enrichment of analytes by specific immunoaffinity clean-up. OTA and CIT plus its metabolite DH-CIT were frequently detected in patient urine samples (OTA 62%; CIT 91%; DH-CIT 100%). The concentration ranges in urine were 1-27.8 ng/L for OTA, 2-87 ng/L for CIT and 2-160 ng/L for DH-CIT. The analyses of blood samples revealed also a frequent co-occurrence of OTA and CIT, in the ranges of 40-870 ng/L serum for OTA and 21-182 ng/L plasma for CIT. This first analysis of biomarkers in blood and urine samples of Czech patients revealed no major differences in comparison with published data for the general healthy Czech and European populations. Nonetheless, a frequent co-occurrence of CIT and OTA biomarkers in patient samples may be of interest with regard to potential interactions with other risk factors for renal disease.
Assuntos
Neoplasias Renais/química , Neoplasias Renais/urina , Micotoxinas/urina , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Biomarcadores/urina , Cromatografia Líquida , Citrinina/sangue , Citrinina/urina , Estudos de Coortes , Tchecoslováquia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Micotoxinas/sangue , Ocratoxinas/sangue , Ocratoxinas/urina , Espectrometria de Massas em TandemRESUMO
SCOPE: The aim of this study is to obtain a deeper knowledge of the kinetics of 2'R-ochratoxin A (2'R-OTA), the thermal degradation product of the mycotoxin ochratoxin A (OTA). To investigate the correlation between the amount of this compound in roasted coffee and human blood samples, a human study is performed. METHODS AND RESULTS: An 18-week human study is carried out. During the first eight weeks, all known 2'R-OTA-containing food sources are excluded from the diet and the reduction of 2'R-OTA in venous blood is analyzed. Afterwards, participants are allowed to consume coffee with known OTA and 2'R-OTA concentrations. On a biweekly scale, 2'R-OTA and OTA blood levels are determined. After eight weeks of fasting on 2'R-OTA-containing foods, the 2'R-OTA blood concentration decreased by about 10%. Based on this, a long biological half-life of over seven months is estimated. In the 24 h urine samples collected before and after the coffee fasting period, only traces of 2'R-OTA are detected. CONCLUSION: Results show that 2'R-OTA has a more than seven-fold higher biological half-life in human blood compared to OTA (approx. 35 days). The reason for the long persistence of 2'R-OTA in human blood is still unclear and further research is needed.
Assuntos
Café/química , Ocratoxinas/sangue , Adulto , Feminino , Meia-Vida , Humanos , Masculino , Ocratoxinas/química , Ocratoxinas/farmacocinética , Ocratoxinas/urina , EstereoisomerismoRESUMO
Objectives: Gene-environment interaction is an emerging hypothesis to expound not only the autism pathogenesis but also the increased incidence of neurodevelopmental disorders (such as autistic spectrum disorder, attention-deficit, hyperactivity disorder). Among xenobiotics, mycotoxins are worldwide contaminants of food that provoke toxicological effects, crucially resembling several symptoms associated with autism such as oxidative stress, intestinal permeability, and inflammation. Here, we focused on a group of mycotoxins to test their role in the manifestation of autism, try to explain their mechanism of action, and discuss possible preventive and therapeutic interventions. Methods: Autistic children (n = 52) and healthy children [n = 58 (31 siblings and 27 unrelated subjects)] were recruited and body fluids and clinical data collected. The diagnosis of autism was made according to DSM V criteria, then with GMDS 0-2, WPPSI, and ADOS. Ochratoxin A (OTA), gliotoxin, zearalenone, and sphingosine/sphinganine ratio were determined by LC analysis in sera and urines. Statistical analysis was performed by the Wilcoxon Rank Sum (Mann-Whitney) test and Spearman test. Results: By comparing the results of autistic patients with those of unrelated controls, a significant association was found for OTA levels in urines (P = 0.0002) and sera (P = 0.0017), and also comparing patients with siblings and unrelated controls together (P = 0.0081). Discussion: Our results are the first describing a possible role of OTA in the pathobiology of autism. Recalling the male prevalence of ASD (male/female = 4-5/1), it is noted that, in animal models, OTA exerts its neurotoxicity especially in males. Moreover, in vitro, OTA increases microRNA-132 that is dysregulated in autistic patients and involved in reciprocal regulation of the autism-related genes MeCP2 and PTEN. A personalized diet coupled with probiotic administration, especially OTA adsorbing Lactobacillus, could ameliorate autistic symptoms in OTA-positive patients.
Assuntos
Transtorno do Espectro Autista , Micotoxinas/sangue , Micotoxinas/urina , Transtorno do Espectro Autista/sangue , Transtorno do Espectro Autista/etiologia , Transtorno do Espectro Autista/urina , Feminino , Humanos , Masculino , Ocratoxinas/sangue , Ocratoxinas/urinaRESUMO
Ochratoxin A (OTA), a mycotoxin known for its nephrotoxic and carcinogenic properties, is a worldwide occurring contaminant in a variety of food commodities. Biomonitoring (i.e. analysis in biological fluids) can serve to assess human internal exposure from all consumed foods and beverages. We now determined the concentration of OTA and its metabolite ochratoxin alpha (OTα) in plasma and in urine of two male volunteers with different food habits, in order to assess intra-individual temporal fluctuations and inter-individual differences in their biomarker levels. Moreover, the urinary levels of both OTA and OTα were analyzed in a cohort of German adults (23 males, 27 females) on their regular diet. All samples were subjected to an enzymatic hydrolysis of biomarker conjugates prior to clean-up by liquid-liquid extraction and HPLC-FD analysis. The profile in the first individual showed small fluctuations over time: mean levels in plasma were 0.42 and 0.45ng/mL for OTA and OTα, respectively, and in urine means of 0.06ng/mL for both analytes. The other individual had mean levels of 1.64 and 0.20ng/mL for OTA and OTα in plasma, and 0.24 and 2.22ng/mL for these analytes in urine. It is concluded that inter-individual differences in biomarker levels reflect dissimilar dietary exposure and/or disposition of ingested mycotoxin, with an apparently more efficient detoxification of OTA to OTα in the second individual. In the German cohort (n=50), analytes were detected in 100% (OTA: range 0.02-1.82ng/mL mean level 0.21±0.31ng/mL) and 78% (OTα: range 0.01-14.25ng/mL, mean level 1.33±2.63ng/mL) of all urines. Parameters such as gender, age and body mass index did not show a significant association with urine biomarker levels. This study indicates frequent exposure to OTA among German adults. The new results are discussed in the context of biomarker data from other countries and some methodological issues.
Assuntos
Monitoramento Ambiental/métodos , Poluentes Ambientais/urina , Ocratoxinas/urina , Adulto , Biomarcadores/sangue , Biomarcadores/metabolismo , Biomarcadores/urina , Estudos de Coortes , Poluentes Ambientais/sangue , Poluentes Ambientais/metabolismo , Feminino , Contaminação de Alimentos/análise , Alemanha , Humanos , Limite de Detecção , Masculino , Ocratoxinas/sangue , Ocratoxinas/metabolismo , Adulto JovemRESUMO
Ochratoxin A (OTA), a mycotoxin with nephrotoxic and carcinogenic properties, is an important contaminant of food and feed. Analysis of OTA in human biological fluids (blood, urine, or breast milk) has documented frequent exposure to this mycotoxin, yet at quite variable levels in different population groups across the world. Urine is the preferred matrix in biomonitoring since sample collection is non-invasive and better accepted by study participants. As only a small fraction of the ingested OTA is excreted in urine, determination of urinary OTA requires sensitive analytical techniques, and phase-II-metabolites should be also considered as biomarkers of exposure. Yet, data published so far on the presence of OTA-glucuronide/sulfate in human urine have been contradictory. In this study, urines (n = 38) from two groups of breastfed infants (German and Turkish) and from German adults were now analysed for the presence of OTA glucuronides or sulfates by an indirect method, i.e. by comparing the levels of OTA (aglycone) in urines without and after enzymatic hydrolysis with ß-glucuronidase/arylsulfatase. Additionally, ochratoxin A-8-ß-glucuronide and open lactone ochratoxin A-8-ß-glucuronide were synthesized to serve as reference materials for metabolite analysis. Attempts for definitive confirmation of glucuronides of OTA via direct identification in LC-MS/MS analysis were hampered by the lower ionizability of the conjugates compared to the parent compound. Considerable increases in OTA levels were found after enzymatic hydrolysis in several (not all) urine samples and provide clear evidence for the excretion of OTA-conjugates. The latter observation is of importance, since OTA phase-II-metabolites may escape detection when direct methods are applied for urinary biomarker analysis. In conclusion, enzymatic hydrolysis of urine samples is highly advisable in order to avoid an underestimation of the OTA-exposure.
Assuntos
Desintoxicação Metabólica Fase II , Ocratoxinas/metabolismo , Ocratoxinas/urina , Urinálise/métodos , Adulto , Alemanha , Glucuronídeos/metabolismo , Glucuronídeos/urina , Humanos , Lactente , Sulfatos/metabolismo , Sulfatos/urina , TurquiaRESUMO
Ochratoxin A (OTA) and citrinin (CIT) are both nephrotoxic and teratogenic in animals, and the occurrence of these mycotoxins in food may cause adverse health effects in humans. Data on the combined exposure to these food contaminants are still scarce, especially in pregnancy. Therefore, a biomonitoring study was conducted to determine the presence of urinary biomarkers of exposure to OTA and CIT in pregnant women in Bangladesh. In total, 54 spot urine samples were collected from residents of a rural and a suburban area of the Savar region in Dhaka district for analysis of OTA and CIT urinary biomarkers by previously validated HPLC-FD and LC-MS/MS methods. Most urines were positive for OTA and CIT biomarkers, with OTA being detected in 93 % (range 0.01-0.84 ng/mL) and CIT biomarkers in 87 % (range 0.02-6.93 ng/mL) of all samples. The mean levels of OTA were different between the rural (0.06 ± 0.07 ng/mL) and suburban (0.15 ± 0.19 ng/mL) study participants. CIT and its metabolite dihydrocitrinone (HO-CIT) were more than twofold higher in the rural (0.42 ± 1.20 and 0.55 ± 1.04 ng/mL, respectively) than the suburban (CIT 0.15 ± 0.13 ng/mL; HO-CIT 0.23 ± 0.18 ng/mL) participants. When a provisional daily intake for CIT was calculated, it exceeded the preliminary tolerable value set by European Food Safety Authority (0.2 µg/kg/day) in 9 % of the rural participants but in none of the urban participants. Urinary biomarker levels for OTA and CIT did not show significant association with intake of certain types of food consumed by the pregnant women, although total CIT biomarker levels were considerably higher among participants who consumed more rice in a day. Overall, this study indicates a frequent co-exposure to OTA and CIT among pregnant women in Bangladesh, at levels similar to those determined recently in the general population of this country.
Assuntos
Citrinina/urina , Monitoramento Ambiental , Ocratoxinas/urina , Gestantes , Bangladesh , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Fluorometria , Humanos , População Rural , População Suburbana , Espectrometria de Massas em Tandem , UrináliseRESUMO
Biomonitoring studies can provide valuable insights into human mycotoxin exposure, especially when food contaminant data are scarce or unavailable as in Bangladesh. First biomonitoring data in Bangladeshi adults indicated exposure to the nephrotoxic mycotoxins ochratoxin A (OTA) and citrinin (CIT). This led us to conduct a follow-up study with analysis of urinary biomarkers for both CIT and OTA to investigate regional and seasonal influences on mycotoxin exposure in two Bangladeshi cohorts. In total, 164 urines were collected (n = 69 in summer, n = 95 in winter) from residents of a rural and an urban area, among which there were 62 participants enrolled in both sampling periods. Most urines had detectable biomarker levels (OTA, CIT and its metabolite dihydrocitrinone, HO-CIT), with more or less pronounced differences with regard to season and region. In both cohorts, OTA was found at a mean level of 0.06 ± 0.10 ng/mL urine (range 0.01-0.55 ng/mL) in summer and a mean of 0.19 ± 0.38 ng/mL (range 0.01-1.75 ng/mL) in winter season. A season difference was significant in the rural cohort, but not in the urban cohort, and slightly higher mean OTA levels in the rural compared to the urban cohort were only observed in winter urines. CIT biomarkers showed more pronounced variations, with a CIT mean of 0.10 ± 0.17 ng/mL (range 0.02-1.22 ng/mL) and HO-CIT mean of 0.42 ± 0.98 ng/mL (range 0.02-5.39 ng/mL) in summer, and CIT mean of 0.59 ± 0.98 ng/mL (range 0.05-5.03 ng/mL) and HO-CIT mean of 3.18 ± 8.49 ng/mL (range 0.02-46.44 ng/mL) in winter urines of both cohorts. In both seasons, total CIT biomarker concentrations were significantly higher in the rural cohort than in the urban cohort. A provisional daily intake for CIT was calculated and exceeded a preliminary value set by EFSA (0.2 µg/kg/d) in 10 and 24 % of participants in summer and winter, respectively. No significant correlations were found between urinary biomarker levels and intake of certain types of food, except for a positive trend for higher rice consumption. Our results in the Bangladeshi population indicate frequent co-exposure to nephrotoxic mycotoxin food contaminants that vary by season and region.
Assuntos
Carcinógenos Ambientais/toxicidade , Citrinina/toxicidade , Exposição Ambiental/efeitos adversos , Ocratoxinas/toxicidade , Oryza , Saúde da População Rural , Saúde da População Urbana , Adulto , Bangladesh , Biomarcadores/urina , Carcinógenos Ambientais/análise , Carcinógenos Ambientais/metabolismo , Citrinina/análogos & derivados , Citrinina/metabolismo , Citrinina/urina , Estudos de Coortes , Países em Desenvolvimento , Dieta/efeitos adversos , Dieta/etnologia , Monitoramento Ambiental , Feminino , Seguimentos , Contaminação de Alimentos , Humanos , Masculino , Ocratoxinas/metabolismo , Ocratoxinas/urina , Oryza/efeitos adversos , Oryza/química , Saúde da População Rural/etnologia , Estações do Ano , Sementes/efeitos adversos , Sementes/química , Toxicocinética , Saúde da População Urbana/etnologiaRESUMO
The purposes of this review are to study potential biomarkers of exposure for ochratoxin A (OTA) in biological fluids (blood, urine and breast milk) for the period 2005-14, calculate the estimated daily intake (EDI) of OTA by using database consumption for the Spanish population, and, finally, to correlate OTA levels detected in blood and EDI values calculated from food products. The values of OTA detected in potential biomarkers of exposure for blood, breast milk and urine ranged from 0.15 to 18.0, from 0.002 to 13.1, and from 0.013 to 0.2 ng ml(-1), respectively. The calculated EDI for OTA in plasma ranged from 0.15 to 26 ng kg(-1) bw day(-1), higher than that obtained in urine (0.017-0.4 ng kg(-1) bw day(-1)). All these values are correlated with the range of EDI for OTA calculated from food products: 0.0001-25.2 ng kg(-1) bw day(-1).
Assuntos
Exposição Ambiental/análise , Contaminação de Alimentos/análise , Leite Humano/química , Ocratoxinas/sangue , Ocratoxinas/urina , Biomarcadores/análise , Biomarcadores/sangue , Biomarcadores/urina , Humanos , Estrutura Molecular , Ocratoxinas/administração & dosagem , Ocratoxinas/químicaRESUMO
Mycotoxin producing moulds may contaminate numerous agricultural commodities either before harvest or during storage. A varied diet consisting of different foods may therefore be contaminated with a range of mycotoxins. The aim of the present study was to study concurrent exposure to mycotoxins through urinary multi-biomarker analysis, as well as its possible associations with the diet. Urinary samples from 252 adults, participating in the Swedish national dietary survey Riksmaten 2010-11, were collected together with a 4-day diet record. Concurrent mycotoxin exposure was studied using a multi-biomarker LC-MS/MS method. The results revealed that exposure to mycotoxins is common and concurrent exposure to more than one toxin was found in 69% of the study population. However, when comparing the number of toxins detected with the reported consumption data it was difficult to distinguish food patterns which would indicate an increased risk of exposure to many mycotoxins simultaneously. This is the first study to investigate concurrent mycotoxin exposure and urinary levels of fumonisin B1 (FB1), fumonisin B2 (FB2), nivalenol (NIV), ochratoxin A (OTA), zearalenone (ZEA), α-zearalenol (α-ZOL), ß-zearalenol (ß-ZOL) and de-epoxydeoxynivalenol (DOM-1) among adults in Sweden.
Assuntos
Dieta/efeitos adversos , Contaminação de Alimentos , Micotoxinas/toxicidade , Adulto , Idoso , Doenças Assintomáticas/epidemiologia , Biomarcadores/urina , Registros de Dieta , Monitoramento Ambiental , Feminino , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/etiologia , Doenças Transmitidas por Alimentos/urina , Humanos , Internet , Masculino , Pessoa de Meia-Idade , Micotoxinas/metabolismo , Micotoxinas/urina , Inquéritos Nutricionais , Ocratoxinas/metabolismo , Ocratoxinas/toxicidade , Ocratoxinas/urina , Prevalência , Suécia/epidemiologia , Tricotecenos/metabolismo , Tricotecenos/toxicidade , Tricotecenos/urina , Zearalenona/metabolismo , Zearalenona/toxicidade , Zearalenona/urinaRESUMO
A direct, fast and sensitive LC-MS/MS method was developed to measure biomarkers for mycotoxin exposure in human urine. In total, 32 biomarkers were quantitatively or semi-quantitatively measured in 32 urine samples of Belgian volunteers using two injections. All urine samples contained deoxynivalenol-15-glucuronide, the major detoxification metabolite of deoxynivalenol, in the ng/mL range. Also deoxynivalenol-3-glucuronide and de-epoxy-deoxynivalenol-glucuronide were present in, respectively, 90 and 25% of the samples, while deoxynivalenol was detected in 60% of the samples, in lower concentrations. Deoxynivalenol glucuronides were the major biomarkers for deoxynivalenol exposure. Ochratoxin A was detected in 70% of the samples in pg/mL. Citrinin and/or dihydrocitrinone were detected in 90% of the samples, also in concentrations of pg/mL. The presence of ochratoxin A and citrinin was confirmed by a second method using sample cleanup by immunoaffinity columns, followed by LC-MS/MS. Our data show that humans are much more exposed to citrinin than realized before and suggest further work on citrinin exposure in relation with ochratoxin A exposure, as both mycotoxins are nephrotoxic.
Assuntos
Cromatografia Líquida/métodos , Micotoxinas/urina , Espectrometria de Massas em Tandem/métodos , Tricotecenos/urina , Adulto , Bélgica , Biomarcadores/urina , Citrinina/análogos & derivados , Citrinina/urina , Feminino , Glucuronídeos/urina , Humanos , Masculino , Ocratoxinas/urinaRESUMO
SCOPE: In this study, the exposure of a German population (n = 101) to mycotoxins was assessed using an LC-MS/MS urinary multibiomarker approach. Food consumption of the participants was documented with a food frequency questionnaire to correlate mycotoxin exposure with individual nutritional habits. METHODS AND RESULTS: The presence of 23 urinary biomarkers including trichothecenes (deoxynivalenol (DON), DON-3-glucuronide (DON-3-GlcA), T-2 toxin, HT-2 toxin (HT-2, HT-2-toxin-4-glucuronide (HT-2-GlcA), fumonisins (fumonisin B1, fumonisin B2), aflatoxins (aflatoxin B1, aflatoxin G2, aflatoxin B2, aflatoxin M1), zearalenone and derivatives (zearalanone, α-zearalenol, ß-zearalenol, zearalenone-14-O-glucuronide, zearalanone-14-O-glucuronide, α-zearalenol-14-O-glucuronide/ß-zearalenol-14-O-glucuronide), ochratoxin A, ochratoxin alpha, enniatin B and dihydrocitrinone was evaluated using a validated, sensitive "dilute and shoot"-LC-MS/MS method applying Scheduled MRM(TM) technology. Six mycotoxins and urinary metabolites were detected (DON, DON-3-GlcA, zearalenone-14-O-glucuronide, T-2 toxin, enniatin B, and dihydrocitrinone) in 87% of the samples in single- or co-occurence. Only DON and DON-3-GlcA were detectable in quantifiable amounts. A provisional mean daily intake of 0.52 µg DON/kg body weight was calculated. No statistical evidence for the correlation of staple food intake and urinary biomarker concentration could be determined. CONCLUSION: The results of this study suggest a low everyday exposure of the investigated German population to mycotoxins, but reveal peak exposures above the widely accepted tolerable daily intake to DON in parts of the population.