Hooijerink D, Schilt R, van Bennekom E, Brouwer B
State Institute for Quality Control of Agricultural Products, Department of Food and Environmental Chemistry, Wageningen, The Netherlands.
Analyst. 1994 Dec;119(12):2617-22. doi: 10.1039/an9941902617.
Two different analytical methods are described for the analysis of anabolic steroid esters in oily formulations for veterinary use and animal plasma samples, respectively. For the determination of anabolic steroid esters in oily formulations (at mg kg-1 levels) a reversed-phase high-performance liquid chromatographic method with gradient elution is described. Gradient elution is performed owing to the relatively large variations in polarity of the investigated anabolic steroid esters. For the analysis of anabolic steroid esters in plasma (at ng ml-1 levels) two different strategies are applied. After solid-phase extraction, the plasma samples are introduced into the high-performance liquid chromatography (HPLC) system where the obtained fractions are then analysed by using gas chromatography-mass selective detection (GC-MSD). An alternative method is direct analysis of plasma samples after solid-phase extraction by using GC-MSD without any further clean-up procedure. Prior to GC-MSD the samples are derivatized to corresponding trifluoroacyl (TFA) derivatives. The calibration graph for HPLC is rectilinear over the range 25-150 ng ml-1 plasma and the analytical recoveries for medroxyprogesterone acetate (MPA) and testosterone propionate (TP) are more than 95%. The detection limits for both analytes in GC-MS are 2.5 ng ml-1 plasma for MPA and 0.5 ng ml-1 plasma for TP with an acceptable signal-to-noise ratio (calculated for the derivatized relative molecular mass). In the analysis of plasma obtained from animal experiments concentrations of 6.5 ng ml-1 are found for MPA by using GC-MSD and 5.0 ng ml-1 are found for nortestosterone laurate (NL) by using HPLC.
分别介绍了两种不同的分析方法，用于分析兽用油性制剂和动物血浆样本中的合成代谢类固醇酯。对于油性制剂中合成代谢类固醇酯（毫克/千克水平）的测定，描述了一种采用梯度洗脱的反相高效液相色谱法。由于所研究的合成代谢类固醇酯的极性变化相对较大，因此采用梯度洗脱。对于血浆中合成代谢类固醇酯（纳克/毫升水平）的分析，应用了两种不同的策略。固相萃取后，将血浆样本引入高效液相色谱（HPLC）系统，然后使用气相色谱-质量选择性检测（GC-MSD）对所得馏分进行分析。另一种方法是固相萃取后直接使用GC-MSD分析血浆样本，无需任何进一步的净化程序。在进行GC-MSD分析之前，将样品衍生为相应的三氟乙酰（TFA）衍生物。HPLC的校准曲线在25-150纳克/毫升血浆范围内呈线性，醋酸甲羟孕酮（MPA）和丙酸睾酮（TP）的分析回收率均超过95%。GC-MS中两种分析物的检测限分别为：MPA为2.5纳克/毫升血浆，TP为0.5纳克/毫升血浆，信噪比可接受（针对衍生化相对分子质量计算）。在动物实验获得的血浆分析中，使用GC-MSD测定MPA的浓度为6.5纳克/毫升，使用HPLC测定月桂酸诺龙（NL）的浓度为5.0纳克/毫升。
