López, B. (Beatriz)
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- Presence of 19 mycotoxins in human plasma in a region of Northern Spain(MDPI, 2020) López, B. (Beatriz); Gonzalez-Peñas, E. (Elena); Irigoyen, A. (Angel); Lizarraga, E. (Elena)This study was conducted to investigate human exposure to19 compounds(mycotoxins and their metabolites) in plasma samples from healthy adults (n = 438, aged 19–68 years) from Navarra, a region of northern Spain. Samples were analyzed by LC-MS/MS, before and after enzymatic hydrolysis for the detection of possible glucuronides and/or sulfates (Phase II metabolites). The most prevalent mycotoxin was ochratoxin A (OTA), with an incidence of 97.3%. Positive samples were in the concentration range of 0.4 ng/mL to 45.7 ng/mL. After enzymatic treatment, OTA levels increased in a percentage of individuals, which may indicate the presence of OTA-conjugates. Regarding ochratoxinB, ithasalso been detected(10% of the samples),and its presence may be related to human metabolism of OTA. Sterigmatocystin was detected with a high incidence (85.8%), but only after enzymatichydrolysis,supporting glucuronidationasa pathway of its metabolism in humans. None of the other studied mycotoxins (aflatoxins B1, B2, G1, G2 and M1; T-2 and HT-2 toxins; deoxynivalenol, deepoxy-deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol; zearalenone; nivalenol; fusarenon-X; neosolaniol; and diacetoxyscirpenol) were detected in any of the samples,neither before nor after enzymatic treatment. To the best of our knowledge, this is the first report carried out in Spain to determine the exposure of the population to mycotoxins and some of their metabolites using plasma, and the obtained results justify the need for human biomonitoring and metabolism studies on mycotoxins.
- Development and validation of a methodology based on Captiva EMR-lipid clean-up and LC-MS/MS analysis for the simultaneous determination of mycotoxins in human plasma(Elsevier, 2020) López, B. (Beatriz); Flores-Flores, M.E. (Myra Evelyn); Gonzalez-Peñas, E. (Elena); Irigoyen, A. (Angel); Lizarraga, E. (Elena)We report the methodology for the quantification of 19 mycotoxins in human plasma using high performance liquid chromatography-mass spectrometry (triple quadrupole). The studied mycotoxins were: deepoxy-deoxynivalenol, aflatoxins (B1, B2, G1, G2 and M1), T-2 and HT-2, ochratoxins A and B, zearalenone, sterigmatocystin, nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, neosolaniol, diacetoxyscirpenol and fusarenon-X. Sample deproteinization and cleanup were performed in one step using Captiva EMR-lipid (3 mL) cartridges and acetonitrile (with 1% formic acid). The extraction step was simple and fast. Validation was based on the evaluation of limits of detection (LOD) and quantification, linearity, precision, recovery, matrix effect, and stability. LOD values ranged from 0.04 ng/mL for aflatoxin B1 to 2.7 ng/mL for HT-2, except for nivalenol, which was 9.1 ng/mL. Recovery was obtained in intermediate precision conditions and at three concentration levels. Mean values ranged from 68.8% for sterigmatocystin to 97.6% for diacetoxyscirpenol (RDS ≤ 15% for all the mycotoxins). Matrix effects (assessed at three concentration levels and in intermediate conditions) were not significant for most of the mycotoxins and were between 75.4% for sterigmatocystin and 109.3% for ochratoxin B (RDS ≤ 15% for all the mycotoxins). This methodology will be useful in human biomonitoring studies of mycotoxins for its reliability.
- Biomonitoring of Mycotoxins in Plasma of Patients with Alzheimer's and Parkinson's Disease(2021) López, B. (Beatriz); De-Santis, B. (Barbara); Alvarez-Erviti, L. (Lydia); Lopez-de-Cerain, A. (Adela); Marzo-Sola, M.E. (María Eugenia); Vettorazzi, A. (Ariane); Debegnach, F. (Francesca); Gonzalez-Peñas, E. (Elena); Izco, M. (María); López-Calvo, S. (Silvia); Lizarraga, E. (Elena)Exposure to environmental contaminants might play an important role in neurodegenerative disease pathogenesis, such as Parkinson ' s disease (PD) and Alzheimer's disease (AD). For the first time in Spain, the plasmatic levels of 19 mycotoxins from patients diagnosed with a neurodegenerative disease (44 PD and 24 AD) and from their healthy companions (25) from La Rioja region were analyzed. The studied mycotoxins were aflatoxins B1, B2, G1, G2 and M1, T-2 and HT-2, ochratoxins A (OTA) and B (OTB), zearalenone, sterigmatocystin (STER), nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deepoxy-deoxynivalenol, neosolaniol, diacetoxyscirpenol and fusarenon-X. Samples were analyzed by LC-MS/MS before and after treatment with beta-glucuronidase/arylsulfatase in order to detect potential metabolites. Only OTA, OTB and STER were detected in the samples. OTA was present before (77% of the samples) and after (89%) the enzymatic treatment, while OTB was only detectable before (13%). Statistically significant differences in OTA between healthy companions and patients were observed but the observed differences might seem more related to gender (OTA levels higher in men, p-value = 0.0014) than the disease itself. STER appeared only after enzymatic treatment (88%). Statistical analysis on STER, showed distributions always different between healthy controls and patients (patients' group > controls, p-value < 0.0001). Surprisingly, STER levels weakly correlated positively with age in women (rho = 0.3384), while OTA correlation showed a decrease of levels with age especially in the men with PD (rho = -0.4643).
- Estudio de la presencia de 19 micotoxinas en plasma humano: niveles en donantes sanos y enfermos en Navarra y la Rioja(Universidad de Navarra, 2021-07-01) López, B. (Beatriz); Gonzalez-Peñas, E. (Elena); Lizarraga, E. (Elena)Mycotoxins pose a risk to food safety and, consequently, to human health. Exposure assessment of these toxins by human biomonitoring studies (HBM) based on the analysis of these compounds (mycotoxins and their metabolites) in biological samples appears as a promising alternative to the current employed approach: the study of mycotoxin levels in different food matrices. In the present study, the state-of-the-art of HBM has been first reviewed and some challenges to be faced have been identified. Subsequently, an analytical LC-MS/MS (QqQ) method has been developed and validated for the determination of 19 mycotoxins and their metabolites (aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2), M1 (AFM1), ochratoxins A (OTA) and B (OTB), zearalenone (ZEA), sterigmatocystin (STER), deoxynivalenol (DON), deepoxynivalenol (DOM-1), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV), toxins T-2 (T-2) and HT-2 (HT-2), neosolaniol (NEO), diacetoxyscirpenol (DAS) and fusarenon-X (FUS- X)) in human plasma samples. Captiva EMR-Lipid® cartridges were used for the sample clean-up process. Good values of selectivity, linearity, accuracy, precision, stability, recovery and matrix effect were obtained. With this methodology, a total of 610 samples were analysed, distributed as follows: 438 samples from healthy adults (19-68 years), 79 from healthy children and with digestive or behavioural pathologies (attention deficit hyperactivity (ADHD) and autism spectrum disorders (ASD)) (2-16 years) and 93 from healthy adults and with neurodegenerative pathologies (Parkinson¿s and Alzheimer¿s diseases) (44-85 years) were analysed before and after an enzymatic treatment with β-glucuronidase/arylsulfatase (to detect possible conjugates of the mycotoxins studied). OTA proved to be the most prevalent mycotoxin in all the studied samples. Moreover, there is a percentage of individuals in all groups with higher levels of this compound after enzymatic hydrolysis, which would indicate the presence of conjugates of this mycotoxin. The different behaviour after enzymatic hydrolysis could indicate differences in the metabolism of OTA between samples from healthy and sick donors. The occurrence of OTB in a percentage of samples, and always associated with OTA, suggests that it may be a metabolite of OTA. STER, a less studied mycotoxin and whose metabolism route is unknown, appears only after enzymatic hydrolysis with a high incidence in all groups. This supports glucuronidation as a pathway of metabolism of this mycotoxin in humans. Despite the regulations applied in our country on the maximum levels of some mycotoxins in food, exposure to OTA and STER has been observed. The levels found could pose a danger to individuals, especially children, with a longer life expectancy and a less varied diet. In addition, differences in incidence and levels have been found in healthy and sick individuals, both in adults and children. The reason for these differences still remains unknown, but it would be of interest to elucidate whether or not exposure to these environmental toxicants has an influence on the development of these diseases, or if the differences are due to the existence of the disease.