Destrez Blandine, Bichon Emmanuelle, Rambaud Lauriane, Courant Frédérique, Monteau Fabrice, Pinel Gaud, Antignac Jean-Philippe, Le Bizec Bruno
Laboratoire d'Etude des Résidus et Contaminants dans les Aliments (LABERCA), Ecole Nationale Vétérinaire de Nantes (ENVN), BP 50707, Route de Gachet, 44307 Nantes Cedex 3, France.
Steroids. 2009 Oct;74(10-11):803-8. doi: 10.1016/j.steroids.2009.04.010. Epub 2009 May 4.
Boldenone is banned in the European Union (Directive 96/22/EC) as growth promoter for meat producing animals. Boldione (ADD), boldenone and boldenone esters (mainly the undecylenate form) are commercially available as anabolic preparations, either to the destination of human, horse or cattle. Since the late 90s, the natural occurrence of boldenone metabolites has been reported in cattle. According to EU regulation, the unambiguous demonstration of boldenone administration in bovine urine should be provided on the basis of boldenone identification in the corresponding conjugate fraction. An analytical method has been developed and validated according to current standards with main concern to the measurement of intact 17beta-boldenone-sulphate. The analytical procedure included direct extraction-purification of target analyte on octadecylsilyl cartridges and direct detection of phase II metabolite by liquid chromatography (negative electrospray), tandem mass spectrometry (QqQ) or high resolution mass spectrometry (Orbitrap). Decision limit (CCalpha) and detection capability (CCbeta) were respectively 0.2 microg L(-1) and 0.4 microg L(-1) on triple quadrupole and 0.1 microg L(-1) and 0.2 microg L(-1) on hybrid system. The method was successfully applied to the analysis of incurred samples collected in different experiments. 17beta-Boldenone-sulphate was measurable up to 36h after oral administration of boldione, and 30 days after 17beta-boldenone undecylenate intra-muscular injection. This conjugate form was never detected in non-treated animals, confirming its status of definitive candidate marker for boldenone administration in calf.
在欧盟（指令96/22/EC）中，去甲雄三烯醇酮被禁止用作产肉动物的生长促进剂。ADD、去甲雄三烯醇酮及其酯类（主要是十一烯酸酯形式）作为合成代谢制剂在市场上销售，用于人类、马匹或牛。自20世纪90年代末以来，已报道在牛体内自然存在去甲雄三烯醇酮代谢物。根据欧盟规定，应基于相应共轭物组分中去甲雄三烯醇酮的鉴定，明确证明在牛尿液中施用了去甲雄三烯醇酮。已根据现行标准开发并验证了一种分析方法，主要关注完整的17β - 去甲雄三烯醇酮硫酸盐的测量。分析程序包括在十八烷基硅烷柱上直接萃取 - 纯化目标分析物，并通过液相色谱（负电喷雾）、串联质谱（QqQ）或高分辨率质谱（Orbitrap）直接检测II相代谢物。在三重四极杆上，决策限（CCα）和检测能力（CCβ）分别为0.2 μg L⁻¹和0.4 μg L⁻¹，在混合系统上分别为0.1 μg L⁻¹和0.2 μg L⁻¹。该方法已成功应用于不同实验中收集的实际样品分析。口服ADD后36小时内以及肌肉注射17β - 去甲雄三烯醇酮十一烯酸酯后30天内均可检测到17β - 去甲雄三烯醇酮硫酸盐。在未处理的动物中从未检测到这种共轭物形式，证实了其作为犊牛体内去甲雄三烯醇酮施用的确定性候选标志物的地位。
