Division of Mass Spectrometry and Chromatography, Institute of Medical and Chemical Laboratory Diagnostics (ZIMCL), University Hospital Innsbruck, Innsbruck, Austria.
Anal Bioanal Chem. 2012 May;403(4):961-72. doi: 10.1007/s00216-011-5612-0. Epub 2011 Dec 10.
Co-administration of synthetic progestin containing hormonal contraceptives (HCs) and antiepileptic drugs (AEDs) is a common clinical situation which needs specific considerations due to drug interactions. Several studies have demonstrated that lamotrigine plasma levels are significantly decreased during co-medication with HCs, and that this interaction is associated with increased seizure frequency in most of the cases. Additionally, an increase in contraceptive failure and unintended pregnancy could be observed during co-medication. Hence, monitoring of progestin plasma levels in patients with AED co-medication is of interest. A rapid and reliable online solid-phase extraction-high performance liquid chromatography-tandem mass spectrometry (online SPE-LC-MS/MS) method using gradient elution in the LC domain was established and validated for the simultaneous quantitative determination of gestodene, dienogest, drospirenone, etonogestrel, cyproterone acetate, and levonorgestrel in human plasma. The online SPE-LC-MS/MS method covered a quantification concentration range of 5-100 ng/ml for dienogest, 1-100 ng/ml for etonogestrel and 2-100 ng/ml for all other analytes. Stable isotope-labeled internal standards were used for analyte quantification based on selected reaction monitoring experiments. Inter- and intra-assay precision and accuracy were determined from quality control (QC) samples at the lower limits of quantification and at low, medium, and high concentration levels within the calibration range. Inter-assay reproducibility at the QC levels was better than 10% (relative standard deviation, RSD), accuracy at these levels ranged from -3.7% to 11.3%. Total extraction efficiency, tested at three concentrations, ranged from 92.5% to 106.4%. Matrix interferences were excluded by post-column infusion experiments. To prove the applicability of the assay in clinical cohorts, a sample set (n = 298) stemming from study patients under AED/oral HC co-medication was screened for progestin plasma levels. This method has to be considered a research-use-only assay and must not be used for diagnostic or therapeutic purposes, since it did not undergo formal performance evaluation in the sense of the IVD directive (98/79/EG) of the European Community.
同时使用合成孕激素避孕药（HCs）和抗癫痫药物（AEDs）是一种常见的临床情况，由于药物相互作用，需要特别考虑。一些研究表明，拉莫三嗪的血浆水平在与 HCs 联合用药时显著降低，并且这种相互作用与大多数情况下的癫痫发作频率增加有关。此外，还可以观察到联合用药期间避孕失败和意外怀孕的增加。因此，监测 AED 联合用药患者的孕激素血浆水平是很有意义的。本文建立并验证了一种快速可靠的在线固相萃取-高效液相色谱-串联质谱（在线 SPE-LC-MS/MS）方法，该方法在 LC 域中采用梯度洗脱，用于同时定量测定人血浆中的孕二烯酮、地诺孕素、屈螺酮、依托孕烯、醋酸环丙孕酮和左炔诺孕酮。在线 SPE-LC-MS/MS 方法涵盖了地诺孕素的定量浓度范围为 5-100ng/ml，依托孕烯为 1-100ng/ml，以及所有其他分析物为 2-100ng/ml。基于选择反应监测实验，使用稳定同位素标记的内标进行分析物定量。通过在定量下限和校准范围内的低、中、高浓度水平上的质控（QC）样品来确定批内和批间精密度和准确度。QC 水平的批间重现性优于 10%（相对标准偏差，RSD），这些水平的准确度范围为-3.7%至 11.3%。总提取效率在三个浓度下测试，范围为 92.5%至 106.4%。通过柱后注入实验排除了基质干扰。为了证明该方法在临床队列中的适用性，对来自接受 AED/口服 HC 联合治疗的研究患者的样本集（n=298）进行了孕激素血浆水平筛查。该方法被认为是一种仅供研究使用的检测方法，不能用于诊断或治疗目的，因为它没有按照欧洲共同体 98/79/EG 指令的要求进行正式的性能评估。
