National Dope Testing Laboratory, Ministry of Youth Affairs and Sports, J. N. Stadium, New Delhi - 100 03, India.
Indian J Pharmacol. 2009 Dec;41(6):278-83. doi: 10.4103/0253-7613.59928.
Adrafinil and modafinil have received wide publicity and have become controversial in the sporting world when several athletes were discovered allegedly using these drugs as doping agents. By acknowledging the facts, the World Anti-Doping Agency (WADA) banned these drugs in sports since 2004. The present study explores the possibility of differentiating adrafinil and modafinil and their major metabolites under electron impact ionization in gas chromatograph-mass spectrometer (GC-MSD) and electrospray ionization in liquid chromatograph-mass spectrometer (LC-MS/MS) by studying the fragmentation pattern of these drugs.
Adrafinil, modafinil and their major metabolite, modafinilic acid were analyzed on EI-GC-MSD and ESI-LC-MS/MS using various individual parameters on both the instruments. The analytical technique and equipment used in the analysis were an Agilent 6890N GC with 5973 mass selective detector for the GC-MSD analysis and an Agilent 1100 HPLC with API-3200 Triple quadrupole mass spectrometer for the LC-MS/MS analysis. Validation of both methods was performed using six replicates at different concentrations.
The results show that adrafinil, modafinil and their major metabolite modafinilic acid could be detected as a single artifact without differentiation under EI-GC-MSD analysis. However, all drugs could be detected and differentiated under ESI-LCMS/MS analysis without any artifaction. The GC-MSD analysis gives a single artifact for both the drugs without differentiation and thus can be used as a marker for screening purposes. Further, the Multiple Reaction Monitoring (MRM) method developed under LC-MS/MS is fit for the purpose for confirmation of suspicious samples in routine sports testing and in forensic and clinical analysis.
阿德拉非尼和莫达非尼在运动界受到广泛关注，并成为争议话题，因为有几名运动员被发现涉嫌使用这些药物作为兴奋剂。世界反兴奋剂机构（WADA）承认这一事实后，自 2004 年起将这些药物列入运动禁药名单。本研究通过研究这些药物的碎裂模式，探索在电子轰击电离气相色谱-质谱（GC-MSD）和电喷雾电离液相色谱-质谱（LC-MS/MS）中区分阿德拉非尼和莫达非尼及其主要代谢物的可能性。
采用各种仪器参数，在 EI-GC-MSD 和 ESI-LC-MS/MS 上对阿德拉非尼、莫达非尼及其主要代谢产物莫达非尼酸进行分析。GC-MSD 分析采用安捷伦 6890N 气相色谱仪和 5973 质量选择检测器，LC-MS/MS 分析采用安捷伦 1100 HPLC 与 API-3200 三重四极杆质谱联用仪。两种方法均采用不同浓度的 6 个重复进行验证。
结果表明，阿德拉非尼、莫达非尼及其主要代谢产物莫达非尼酸在 EI-GC-MSD 分析中无法区分，只能作为单一的假说来检测。然而，所有药物都可以在 ESI-LCMS/MS 分析中被检测和区分，而没有任何假说来干扰。GC-MSD 分析对这两种药物都给出了单一的假说来进行筛选，因此可作为筛查目的的标记物。此外，在 LC-MS/MS 下开发的多重反应监测（MRM）方法适用于常规运动测试、法医和临床分析中可疑样本的确证。
