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Direct quantification of deoxynivalenol glucuronide in human urine as biomarker of exposure to the Fusarium mycotoxin deoxynivalenol

机译:直接定量人类尿液中的脱氧雪腐酚葡萄糖醛酸作为暴露于镰刀菌真菌毒素脱氧雪腐酚的生物标志物

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摘要

The direct quantification of deoxynivalenol glucuronide (DON-GlcA) by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application as a biomarker of exposure to the Fusarium mycotoxin deoxynivalenol (DON) is reported. Usually, DON exposure is estimated from dietary average intakes or by measurement of the native toxin in urine after enzymatic hydrolysis with β-glucuronidase. These methods are time-consuming, expensive, and fail to determine the ratio of DON to DON-GlcA in a simple one-step procedure. One of the main reasons for the use of indirect methods is the unavailability of DON-GlcA standards. Consequently, DON-3-O-glucuronide (D3GlcA) was synthesized and used to develop a method allowing quantification of both DON and D3GlcA by a simple "dilute and shoot" approach without the need for any cleanup. Limit of detection and apparent recovery of D3GlcA was 3 μg l~(-1) and 88%, respectively. The identity of D3GlcA in human urine was confirmed by comparison with LC-MS/MS measurements of the synthetically produced D3GlcA standard which was also used for external calibration. The applicability of the method was demonstrated through the analysis of urine samples obtained from a volunteer during regular and cereal-restricted diet, respectively. In regular-diet urine samples, D3GlcA was quantified in concentrations >30 μg l~(-1) by this approach. [Figure not available: see fulltext.]
机译:据报道,通过液相色谱-串联质谱法(LC-MS / MS)对脱氧雪腐酚葡萄糖醛酸(DON-GlcA)进行直接定量,并将其用作暴露于镰刀菌真菌毒素脱氧雪腐酚(DON)的生物标志物。通常,从膳食平均摄入量或通过使用β-葡萄糖醛酸苷酶酶水解后尿液中天然毒素的含量来估算DON暴露量。这些方法耗时,昂贵并且无法通过简单的一步步骤确定DON与DON-GlcA的比例。使用间接方法的主要原因之一是DON-GlcA标准的不可用。因此,合成了DON-3-O-葡糖醛酸(D3GlcA),并用于开发一种方法,该方法可通过简单的“稀释并射击”方法定量定量DON和D3GlcA,而无需进行任何纯化。 D3GlcA的检出限和表观回收率分别为3μgl〜(-1)和88%。通过与合成生产的D3GlcA标准品的LC-MS / MS测量结果进行比较,证实了人尿中D3GlcA的身份,该标准品也用于外部校准。该方法的适用性通过分析分别从常规饮食和谷物限制饮食中从志愿者那里获得的尿液样本来证明。在常规饮食的尿液样本中,通过这种方法将D3GlcA的浓度定为> 30μgl〜(-1)。 [图不可用:请参见全文。]

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