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Analytical investigations about the presence of prednisolone in cow urine

机译：牛尿中泼尼松龙存在的分析研究

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Since 2008, the analyses carried out in the Lombardia region as part of National Residue Control Plans have evidenced unexpected frequent detection of the corticosteroid prednisolone (PRED) in cow urine samples taken to the slaughterhouse. Considering the scarce plausibility of these high frequent findings, analytical investigations were started to ascertain the real presence of this corticosteroid. The applied confirmatory method involved liquid-chromatography low-resolution tandem mass spectrometry (triple quadrupole) as instrumental technique, and it was validated in compliance with the requirements of the Commission Decision 2002/657/EC. However, recently some criticism regarding Commission Decision 2002/657/EC identification criteria has been pointed out, experimentally demonstrating false positive results (wrong identification) although these criteria have been strictly observed. Therefore, considering the serious implications (i.e. the possibility that PRED could be considered endogenous in particular animal conditions), studies were carried out to investigate the reliability of PRED identification through the change of the chromatographic conditions (mobile phases, gradient and analytical column) of the confirmatory procedure routinely applied. Further confirmation came from the application of high-resolution mass spectrometry technique (MS ~2 and MS ~3 experiments) to analyze incurred cow urines samples. All the obtained results confirmed definitively the real presence of this corticosteroid excluding false-positive findings in routine analysis. In addition, other experiments demonstrated that high-resolution mass spectrometers (Time of Flight and Orbitrap technologies) could be successfully applied to routine determination of steroid residues in biological fluids at very low concentrations (< 1 μg L ~(-1)).
机译：自2008年以来，作为国家残留控制计划一部分在伦巴第大区进行的分析表明，出入屠宰场的牛尿液样本中频繁检测到皮质类固醇泼尼松龙（PRED）出乎意料。考虑到这些频繁发现的稀缺合理性，开始进行分析研究以确定这种皮质类固醇的真正存在。应用的确认方法涉及液相色谱法低分辨率串联质谱（三重四极杆）作为仪器技术，并已按照委员会决定2002/657 / EC的要求进行了验证。但是，最近有人指出，对委员会决定2002/657 / EC的识别标准提出了一些批评，尽管严格遵守了这些标准，但实验证明了假阳性结果（错误识别）。因此，考虑到严重的影响（即在特定的动物条件下PRED可能被认为是内源性的），通过改变色谱条件（流动相，梯度和分析柱）来研究PRED鉴定的可靠性。常规应用的确认程序。高分辨率质谱技术（MS〜2和MS〜3实验）在分析已发生的牛尿样中的应用进一步证实了这一点。所有获得的结果均明确证实了该皮质类固醇的真正存在，不包括常规分析中的假阳性结果。此外，其他实验表明，高分辨率质谱仪（飞行时间和Orbitrap技术）可以成功地用于常规测定非常低浓度（<1μgL〜（-1））的生物液体中的类固醇残留。

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