School of Food Science and Engineering, Wuhan Polytechnic University, Wuhan 430023, China.
NHC Key Laboratory of Food Safety Risk Assessment, Food Safety Research Unit (2019RU014) of Chinese Academy of Medical Science, China National Center for Food Safety Risk Assessment, Beijing 100021, China.
Toxins (Basel). 2020 Mar 18;12(3):193. doi: 10.3390/toxins12030193.
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, and emerging mycotoxins (fumonisin B, B, and B, hydrolyzed fumonisin B and B, ochratoxin A, B, and alpha, alternariol, alternariol monomethyl ether, altenuene, tentoxin, tenuazonic acid, beauvericin, enniatin A, A, B, and B) 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 R 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.
各种来源的真菌毒素经常污染农产品，对动物和人类构成潜在的毒理学关注。真菌毒素暴露一直是世界各国政府关注的焦点。迄今为止，生物标志物用于监测真菌毒素暴露，并促进对其在慢性疾病中作用的新认识。本研究的目的是开发和验证一种使用同位素标记内标物的灵敏 UHPLC-MS/MS 方法，该方法适用于准确测定 18 种真菌毒素生物标志物，包括伏马菌素、黄曲霉毒素和新兴真菌毒素（伏马菌素 B、B 和 B、水解伏马菌素 B 和 B、黄曲霉毒素 A、B 和 α、交链孢酚、交链孢酚单甲基醚、Alternuene、 tentoxin、 tenuazonic acid、 beauvericin、 enniatin A、A、B 和 B）在人尿中的含量。经过β-葡萄糖醛酸酶酶解后，人尿样品用 HLB 固相萃取小柱进行净化，然后进行仪器分析。多真菌毒素和分析物特异性方法在内部进行了验证，结果令人满意。该方法在测试浓度范围内（对于不同的分析物，从 LOQ 到 25-500ng/mL）提供了良好的线性，R 值从 0.997 到 0.999。所有分析物在尿液中的定量限从 0.0002 到 0.5ng/mL 不等。加标样品的回收率在 74.0%到 133%之间，日内精密度为 0.5%-8.7%，日间精密度为 2.4%-13.4%。该方法应用于北京采集的 60 名健康志愿者的尿液样本，发现了 10 种生物标志物。除了一个样本外，所有样本中都发现了至少一种生物标志物。该方法具有高灵敏度和准确性，非常适合人体生物监测和真菌毒素暴露评估。
