Detection of synthetic testosterone use by novel comprehensive two-dimensional gas chromatography combustion-isotope ratio mass spectrometry.
Detection of synthetic testosterone use by novel comprehensive two-dimensional gas chromatography combustion-isotope ratio mass spectrometry.
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DOI：
10.1021/ac2015849
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发表时间：
2011-09-15
影响因子：
7.4
通讯作者：
Brenna, J. Thomas
中科院分区：
文献类型：
--
作者：
Tobias, Herbert J.;Zhang, Ying;Auchus, Richard J.;Brenna, J. Thomas
我们报告了首次演示综合二维气相色谱燃烧同位素比质谱法 (GC×GCC-IRMS)，用于分析尿类固醇，以检测非法合成睾酮的使用，这在运动兴奋剂中很重要。 GC 与 IRMS 联用 (GCC-IRMS) 目前用于测量反兴奋剂工作中尿类固醇的碳同位素比（CIR、δ13C）；然而，在分析之前需要对尿液提取物进行大量净化，以实现目标类固醇的基线分离。凭借其更强的分离能力，GC×GC 有可能降低样品制备要求，并能够对经过最低限度处理的尿液提取物进行 CIR 分析。解决的挑战包括最小化尺寸的在线反应器以保留窄峰形状、在某些情况下峰的基线分离以及从切片类固醇色谱峰重建同位素信息。仍然存在的困难包括在线反应器的长期稳定性和尿液基质效应，这些效应妨碍了低浓度和痕量成分的基线分离和同位素分析。在这项工作中，使用精制的自制 GC×GCC-IRMS 系统对类固醇进行提取、乙酰化和分析。由于信号强度令人满意，11-羟基雄酮 (11OHA) 和 11-酮乙胆酮 (11KE) 被选为内源性参考化合物 (ERC)，并将其 CIR 与目标化合物 (TC) 雄酮 (A) 和乙胆酮 (E) 进行比较。另外，GC×GC-qMS 系统用于测量睾酮 (T)/EpiT 浓度比。对职业运动员的尿液提取物以及一名接受睾酮凝胶（T-gel）的个体和一名接受睾酮注射（T-shot）的个体的尿液进行了分析。 δ13C 和 Δδ13C 测量的平均精度为 SD(δ13C) 约± 1‰ (n=11)。 T-shot 样本的 T 使用结果呈阳性，T/EpiT 比率 > 9，CIR 测量值 Δδ13C > 5‰，均满足世界反兴奋剂机构的标准。这些数据首次表明，合成类固醇的使用可以通过 GC×GCC-IRMS 进行检测，而无需进行大量的尿液净化。
We report the first demonstration of Comprehensive Two-dimensional Gas Chromatography Combustion Isotope Ratio Mass Spectrometry (GC×GCC-IRMS) for the analysis of urinary steroids to detect illicit synthetic testosterone use, of interest in sport doping. GC coupled to IRMS (GCC-IRMS) is currently used to measure the carbon isotope ratios (CIR, δ13C) of urinary steroids in anti-doping efforts; however, extensive cleanup of urine extracts is required prior to analysis to enable baseline separation of target steroids. With its greater separation capabilities, GC×GC has the potential to reduce sample preparation requirements and enable CIR analysis of minimally processed urine extracts. Challenges addressed include on-line reactors with minimized dimensions to retain narrow peaks shapes, baseline separation of peaks in some cases, and reconstruction of isotopic information from sliced steroid chromatographic peaks. Difficulties remaining include long-term robustness of on-line reactors and urine matrix effects that preclude baseline separation and isotopic analysis of low concentration and trace components. In this work, steroids were extracted, acetylated, and analyzed using a refined, home-built GC×GCC-IRMS system. 11-hydroxy-androsterone (11OHA) and 11-ketoetiocolanolone (11KE) were chosen as endogenous reference compounds (ERC) because of their satisfactory signal intensity, and their CIR was compared to target compounds (TC) androsterone (A) and etiocholanolone (E). Separately, a GC×GC-qMS system was used to measure testosterone (T)/EpiT concentration ratios. Urinary extracts of urine pooled from professional athletes, and urine from one individual that received testosterone gel (T-gel) and one individual that received testosterone injections (T-shot) were analyzed. The average precisions of δ13C and Δδ13C measurements were SD(δ13C) approximately ± 1‰ (n=11). The T-shot sample resulted in a positive for T use with a T/EpiT ratio of > 9 and CIR measurements of Δδ13C > 5‰, both fulfilling World Anti-Doping Agency criteria. These data show for the first time that synthetic steroid use is detectable by GC×GCC-IRMS without need for extensive urine cleanup.
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DOI：
10.1073/pnas.94.4.1049
发表时间：
1997-02-18
影响因子：
11.1
作者：
Corso, TN;Brenna, JT
通讯作者：
Brenna, JT
DOI：
10.1016/s0378-4347(99)00066-3
发表时间：
1999-04-30
期刊：
JOURNAL OF CHROMATOGRAPHY B
影响因子：
--
作者：
Aguilera, R;Catlin, DH;Hatton, CK
通讯作者：
Hatton, CK
DOI：
10.1007/978-3-642-13145-5_9
发表时间：
2011-01-01
期刊：
PROGRESS IN BOTANY 72
影响因子：
--
作者：
Gessler, Arthur
通讯作者：
Gessler, Arthur
影响因子：
2
作者：
Catlin, DH;Sekera, MH;Hatton, CK
通讯作者：
Hatton, CK
影响因子：
7.4
作者：
Burgoyne, TW;Hayes, JM
通讯作者：
Hayes, JM