Karatt Tajudheen K, Sathiq M Anwar, Laya Saraswathy, Ajeebsanu M P Muhammed, Karakka Kal Abdul Khader, Subhahar Michael Benedict, Perwad Zubair
Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Post Graduate and Research Department of Chemistry, Jamal Mohamed College (Affiliated to Bharathidasan University), Tiruchirappalli, India.
Rapid Commun Mass Spectrom. 2023 Sep 30;37(18):e9611. doi: 10.1002/rcm.9611.
Recently, there has been a report suggesting that ecdysteroids can enhance sports performance, making them relevant substances in doping control. Hence, it is imperative to examine the analytical characteristics of ecdysteroids in biological samples to identify their misuse in competitive sports.
To assess the doping of ecdysteroids such as ecdysone, ecdysterone, ponasterone A, turkesterone, and ajugasterone C, a fast and sensitive extraction and detection method was developed, optimized, and validated using equine urine and plasma. Different extraction techniques, namely, solid-phase extraction, liquid-liquid extraction, and dilute-and-inject, were explored to detect ecdysteroids from equine urine and plasma.
The most suitable method of detection was solid-phase extraction using ABS Elut-NEXUS, while liquid-liquid extraction and dilute-and-inject methods encountered difficulties due to the high polarity of ecdysteroids and the presence of significant matrix interferences. Mass spectrometric parameters are optimized on both the Q Exactive high-resolution mass spectrometer and the TSQ Altis triple quadrupole mass spectrometer. However, the study indicated that the triple quadrupole mass spectrometer exhibited improved limit of detection when analyzing samples. To achieve optimal separation of the analytes under investigation from the matrix interferences, various liquid chromatography columns were compared. The Selectra PFPP LC column with a mobile phase consisting of 0.2% formic acid in water (mobile phase A) and acetonitrile (mobile phase B) at a flow rate of 0.5 mL/min demonstrated superior performance.
The findings of this study will significantly contribute to the accurate identification of ecdysteroids, facilitating the investigation of their illicit use in horse racing.
最近，有报告表明蜕皮甾类可以提高运动成绩，使其成为反兴奋剂控制中的相关物质。因此，必须检查生物样品中蜕皮甾类的分析特性，以确定其在竞技体育中的滥用情况。
为了评估蜕皮甾酮、蜕皮甾醇、波那甾酮A、土克甾酮和筋骨草甾酮C等蜕皮甾类的兴奋剂使用情况，开发、优化并验证了一种使用马尿和血浆的快速灵敏的提取和检测方法。探索了不同的提取技术，即固相萃取、液液萃取和稀释进样，以检测马尿和血浆中的蜕皮甾类。
最合适的检测方法是使用ABS Elut-NEXUS进行固相萃取，而液液萃取和稀释进样方法由于蜕皮甾类的高极性和显著的基质干扰而遇到困难。在Q Exactive高分辨率质谱仪和TSQ Altis三重四极杆质谱仪上都对质谱参数进行了优化。然而，研究表明，三重四极杆质谱仪在分析样品时检测限有所提高。为了实现所研究的分析物与基质干扰的最佳分离，比较了各种液相色谱柱。以0.2%甲酸水溶液（流动相A）和乙腈（流动相B）为流动相、流速为0.5 mL/min的Selectra PFPP液相色谱柱表现出优异的性能。
本研究结果将对准确鉴定蜕皮甾类有显著贡献，有助于调查其在赛马中的非法使用情况。
