Fabresse Nicolas, Grassin-Delyle Stanislas, Etting Isabelle, Alvarez Jean-Claude
MassSpecLab, Plateforme de Spectrométrie de Masse, UFR des Sciences de la Santé Simone Veil, Université Versailles Saint-Quentin, 2 Avenue de la Source de la Bièvre, 78180, Montigny le Bretonneux, France.
Laboratoire de Pharmacologie-Toxicologie, Centre Hospitalier Universitaire Raymond Poincaré, AP-HP, 104 Boulevard R. Poincaré, 92380, Garches, France.
Int J Legal Med. 2017 Jul;131(4):989-999. doi: 10.1007/s00414-017-1552-3. Epub 2017 Feb 24.
We developed and validated a method to detect and quantify 12 anabolic steroids in blood (androstenedione, dihydrotestosterone, boldenone, epitestosterone, mesterolone, methandienone, nandrolone, stanozolol, norandrostenedione, tamoxifene, testosterone, trenbolone) and eight more in hair samples (nandrolone phenylpropionate, nandrolone decanoate, testosterone propionate, testosterone benzoate, testosterone cypionate, testosterone decanoate, testosterone phenylpropionate, testosterone undecanoate) using liquid chromatography coupled to high-resolution mass spectrometry. This method used a benchtop Orbitrap mass spectrometer operating with an APCI probe under positive ionization mode. Analysis was realized in full scan experiment with a nominal resolving power of 140,000. After addition of the internal standard (testosterone-D3) and incubation in phosphate buffer pH = 5 for hair, 200 μL of blood and 30 mg of hair samples were extracted with heptane. LOQ and LOD were determined at 5 and 1 ng mL in whole blood and 10 to 100 pg mg and 2 to 20 pg mg in hair according to the compounds, respectively. The method was linear in the 5-1000 ng mL range in whole blood and between 10 or 100 pg mg and 1000 pg mg in hair with correlation coefficients >0.99, and intra- and inter-day accuracy and precision were <14.8% for all compounds except for some esters in hairs (<19.9%) probably due to an important matrix effect for these compounds. This sensitive and specific method to detect anabolic steroids has been successfully applied to two real cases, for which various anabolic steroids in whole blood, urine, and hair were identified and quantified.
我们开发并验证了一种方法，可使用液相色谱-高分辨率质谱联用技术检测和定量血液中的12种合成代谢类固醇（雄烯二酮、双氢睾酮、苯丙酸诺龙、表睾酮、美睾酮、甲睾酮、诺龙、司坦唑醇、去甲雄烯二酮、他莫昔芬、睾酮、群勃龙）以及毛发样本中的另外8种（苯丙酸诺龙、癸酸诺龙、丙酸睾酮、苯甲酸睾酮、环戊丙酸睾酮、癸酸睾酮、苯丙酸睾酮、十一酸睾酮）。该方法使用一台台式Orbitrap质谱仪，在正离子模式下通过APCI探头进行操作。分析在全扫描实验中进行，标称分辨率为140,000。在加入内标（睾酮-D3）并在pH = 5的磷酸盐缓冲液中对毛发进行孵育后，用庚烷提取200 μL血液和30 mg毛发样本。根据化合物不同，全血中的定量限（LOQ）和检测限（LOD）分别为5和1 ng/mL，毛发中的为10至100 pg/mg和2至20 pg/mg。该方法在全血中5 - 1000 ng/mL范围内以及毛发中10或100 pg/mg至1000 pg/mg之间呈线性，相关系数>0.99，除毛发中的某些酯类化合物（<19.9%）外，所有化合物的日内和日间准确度及精密度均<14.8%，这可能是由于这些化合物存在重要的基质效应。这种检测合成代谢类固醇的灵敏且特异的方法已成功应用于两个实际案例，对全血、尿液和毛发中的各种合成代谢类固醇进行了鉴定和定量。
