Abstract
A variety of mycotoxins from different sources frequently contaminate farm products, presenting a potential toxicological concern for animals and human. Mycotoxin exposure has been the focus of attention for governments around the world. To date, biomarkers are used to monitor mycotoxin exposure and promote new understanding of their role in chronic diseases. The goal of this research was to develop and validate a sensitive UHPLC-MS/MS method using isotopically-labeled internal standards suitable for accurate determination of 18 mycotoxin biomarkers, including fumonisins, ochratoxins, Alternaria and emerging Fusarium mycotoxins (fumonisin B1, B2, and B3, hydrolyzed fumonisin B1 and B2, ochratoxin A, B, and alpha, alternariol, alternariol monomethyl ether, altenuene, tentoxin, tenuazonic acid, beauvericin, enniatin A, A1, B, and B1) in human urine. After enzymatic digestion with β-glucuronidase, human urine samples were cleaned up using HLB solid phase extraction cartridges prior to instrument analysis. The multi-mycotoxin and analyte-specific method was validated in-house, providing satisfactory results. The method provided good linearity in the tested concentration range (from LOQ up to 25–500 ng/mL for different analytes), with R2 from 0.997 to 0.999. The limits of quantitation varied from 0.0002 to 0.5 ng/mL for all analytes in urine. The recoveries for spiked samples were between 74.0% and 133%, with intra-day precision of 0.5%–8.7% and inter-day precision of 2.4%–13.4%. This method was applied to 60 urine samples collected from healthy volunteers in Beijing, and 10 biomarkers were found. At least one biomarker was found in all but one of the samples. The high sensitivity and accuracy of this method make it practical for human biomonitoring and mycotoxin exposure assessment.
Highlights
To expand the research scope, acquire more extensive occurrence data about multiple mycotoxins and characterize their potential risks, we developed a method for simultaneous quantitation of 18 mycotoxin biomarkers, including common toxins and emerging toxins in urine
Mycosep and Multisep cartridges are multi-functional columns containing a combination of adsorbents that was designed for mycotoxin analysis
This study reports on the development of an accurate and sensitive UHPLC-MS/MS method for the determination of 18 mycotoxins in human urine
Summary
Mycotoxins are toxic secondary metabolites produced under favorable conditions by different species of fungi that can grow in a wide variety of cereals and foods from production to storage [1,2].Their occurrence and concentrations vary considerably among foods due to factors such as crop susceptibility, climate change, storage and transportation conditions, as well as sanitary standards [3,4].Mycotoxins are hepatotoxic, nephrotoxic, teratogenic, carcinogenic, cytotoxic, immunosuppressive, inflammatory, neurotoxic, and estrogenic, posing diverse health hazards to humans and animals [5,6,7,8].In particular, fumonisins (FBs) and ochratoxin a (OTA) have been classified as possible human carcinogens (Group 2B). Mycotoxins are toxic secondary metabolites produced under favorable conditions by different species of fungi that can grow in a wide variety of cereals and foods from production to storage [1,2]. Their occurrence and concentrations vary considerably among foods due to factors such as crop susceptibility, climate change, storage and transportation conditions, as well as sanitary standards [3,4]. Fumonisins (FBs) and ochratoxin a (OTA) have been classified as possible human carcinogens (Group 2B). Alternaria toxins, including alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), tentoxin (TEN), and tenuazonic acid (TeA), are produced by Alternaria species and have strong evidence of acute and chronic toxicity [11]
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