Guddat Sven, Thevis Mario, Thomas Andreas, Schänzer Wilhelm
Institute of Biochemistry, German Sport University, Cologne, Germany.
Biomed Chromatogr. 2008 Jul;22(7):695-701. doi: 10.1002/bmc.986.
The increasing number of samples and target substances in doping control requires continuously improved screening methods, combining high-throughput analysis, simplified sample preparation, robustness and reliability. Hence, a rapid screening procedure based on liquid chromatography-electrospray ionization-tandem mass spectrometry with in-source collision-induced dissociation was developed. The detection of the polysaccharide-based plasma volume expanders dextran and hydroxyethyl starch (HES) in human urine was established without further sample preparation. The in-source fragmentation strategy of the approach represented a valuable tool in the analysis of the polysaccharide-based compounds, allowing the use of tandem mass spectrometry. After direct injection of urine specimens, analytes were chromatographically separated on a monolithic reverse-phase column and detected via multiple reaction monitoring of diagnostic ions at detection limits of 10 microg/mL for HES and 30 microg/mL for dextran. Validation was performed regarding the parameters specificity, linearity, precision (8-18%) and accuracy (77-105%) and the method was applied to the investigation of approximately 400 doping control samples and seven dextran and two hydroxyethyl starch post-administration samples. The approach demonstrated its capability as a rapid screening tool for the detection of dextran and hydroxyethyl starch and represents an alternative to existing screening procedures since time consuming hydrolysis or derivatization steps were omitted.
兴奋剂检测中样本和目标物质数量的不断增加,要求筛选方法持续改进,要兼具高通量分析、简化样本制备、稳健性和可靠性。因此,开发了一种基于液相色谱 - 电喷雾电离 - 串联质谱并结合源内碰撞诱导解离的快速筛选程序。无需进一步样本制备即可实现人尿中基于多糖的血浆容量扩充剂右旋糖酐和羟乙基淀粉(HES)的检测。该方法的源内碎裂策略是分析基于多糖化合物的宝贵工具,可用于串联质谱分析。直接注入尿样后,分析物在整体反相柱上进行色谱分离,并通过对诊断离子的多反应监测进行检测,HES的检测限为10μg/mL,右旋糖酐的检测限为30μg/mL。针对特异性、线性、精密度(8 - 18%)和准确度(77 - 105%)等参数进行了验证,并将该方法应用于约400份兴奋剂检测样本以及7份右旋糖酐和2份羟乙基淀粉给药后样本的调查。该方法展示了其作为检测右旋糖酐和羟乙基淀粉的快速筛选工具的能力,并且由于省去了耗时的水解或衍生步骤,代表了现有筛选程序的一种替代方法。