Vrobel Ivo, Janečková Hana, Faber Edgar, Bouchalová Kateřina, Mičová Kateřina, Friedecký David, Adam Tomáš
*Laboratory of Metabolomics, Institute of Molecular and Translational Medicine; †Laboratory of Inherited Metabolic Disorders; ‡Department of Hemato-Oncology; §Laboratory of Experimental Medicine, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University in Olomouc; and ¶Department of Clinical Biochemistry, University Hospital Olomouc, Czech Republic.
Ther Drug Monit. 2016 Aug;38(4):516-24. doi: 10.1097/FTD.0000000000000309.
With an increasing number of cancer patients receiving tyrosine kinase inhibitors (TKIs), therapeutic drug monitoring of these molecules is becoming more widespread today. It is mainly based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) methods with typical run times of several minutes. In an online solid phase extraction-MS/MS (SPE-MS/MS) system, the chromatography column is replaced with a reusable solid phase extraction (SPE) cartridge and the analysis time is shortened to less than half a minute. The aim of this study was to develop such a method and test the performance of this high-throughput system in the analysis of imatinib (IMA), nilotinib (NIL), and lapatinib (LAP) in human plasma.
Samples were prepared by simple protein precipitation with methanol containing deuterated internal standards. After centrifugation, the supernatant was diluted 10 fold with a mixture of methanol and water (1:1). A C4 cartridge was used for SPE and the analytes were eluted by acetonitrile. All the analytes were measured within a wide calibration range (50-5000 ng/mL for nilotinib and imatinib, 100-10,000 ng/mL for lapatinib). The method was compared with the LC-MS/MS method by the analysis of 176 clinical samples.
Intraday and interday inaccuracies within 15% and a coefficient of variation less than 15% were achieved for all the TKIs that were measured. Even though the matrix effects were higher in comparison with LC-MS/MS methods, their effect on the performance of the method was eliminated by the usage of deuterated internal standards. The total run time of the new method was 29 seconds for one analysis and the results were fully comparable with LC-MS/MS.
Routine clinical practice requiring high-throughput methods for therapeutic drug monitoring of TKIs may benefit from the online SPE-MS/MS method that provides fast, low-cost analysis, and results that are comparable with conventional methods.
随着越来越多的癌症患者接受酪氨酸激酶抑制剂（TKIs）治疗，如今对这些分子的治疗药物监测变得越来越普遍。它主要基于液相色谱-串联质谱（LC-MS/MS）方法，典型运行时间为几分钟。在在线固相萃取-质谱/质谱（SPE-MS/MS）系统中，色谱柱被可重复使用的固相萃取（SPE）小柱取代，分析时间缩短至不到半分钟。本研究的目的是开发这样一种方法，并测试该高通量系统在分析人血浆中伊马替尼（IMA）、尼洛替尼（NIL）和拉帕替尼（LAP）方面的性能。
样品通过用含有氘代内标的甲醇进行简单蛋白质沉淀来制备。离心后，上清液用甲醇和水（1:1）的混合物稀释10倍。使用C4小柱进行固相萃取，分析物用乙腈洗脱。所有分析物在较宽的校准范围内进行测定（尼洛替尼和伊马替尼为50 - 5000 ng/mL，拉帕替尼为100 - 10,000 ng/mL）。通过分析176份临床样品将该方法与LC-MS/MS方法进行比较。
所有测定的TKIs的日内和日间误差均在15%以内，变异系数小于15%。尽管与LC-MS/MS方法相比基质效应更高，但通过使用氘代内标消除了其对方法性能的影响。新方法一次分析的总运行时间为29秒，结果与LC-MS/MS完全可比。
对于需要高通量方法进行TKIs治疗药物监测的常规临床实践，在线SPE-MS/MS方法可能有益，该方法提供快速、低成本的分析，且结果与传统方法可比。
