Korecka Magdalena, Solari Sandra G, Shaw Leslie M
Department Pathology and Laboratory Medicine, University of Pennsylvania, School of Medicine, Philadelphia, PA 19104, USA.
Ther Drug Monit. 2006 Aug;28(4):484-90. doi: 10.1097/00007691-200608000-00002.
A new HPLC-MS/MS method for everolimus measurement was developed that includes the following features: small sample volume, short run time, fast, simple and cost-efficient sample preparation, assessment of performance of two internal standards (IS), SDZ RAD 223-756 and ascomycin and comparison of the method with an HPLC-MS/MS reference method. The authors established a multiple reaction monitoring positive ion HPLC-MS/MS method with on-line extraction and sample cleanup. This procedure includes: an API 2000 triple quadrupole mass spectrometer with turbo-ion spray, built-in Valco switching valve, an HPLC system; guard column; a Nova-Pak C18 analytical column; washing solution, methanol:30 mM ammonium acetate pH 5.1 (80:20); eluting solution, methanol:30 mM ammonium acetate pH 5.1 (97:3); flow rate 0.8 mL/min; and a run time of 2.8 minutes. The first and third quadrupoles were set to detect the ammonium adduct ion and a high mass fragment of everolimus (m/z 975.5-->908.5), and two ISs: SDZ RAD 223-756 (m/z 989.8-->922.8) and ascomycin (m/z 809.5-->756.5). The LLOQ was 1.0 microg/L for everolimus using either IS. Between day precision ranged from 3.1% to 5.7% for SDZ RAD 223-756 and 6.0% to 8.6% for ascomycin using spiked blood with everolimus concentrations 2.0 to 25.0 microg/L. Absolute recoveries using spiked samples over the range of 2.5 to 25 mug/L averaged 77.3% (SDZ RAD 223-756) and 76.8% (ascomycin). No matrix effect on everolimus was demonstrated based on the mean observed signal detection of 98.6% (SDZ RAD 223-756) and 105% (ascomycin). Comparison of everolimus concentrations obtained using this method with two internal standards with a reference laboratory demonstrated that the mean everolimus concentration obtained with ascomycin was statistically different (lower) than results with the reference method and the method that used SDZ RAD 223-756 as the internal standard gave equivalent results compared with the reference method.
开发了一种用于测定依维莫司的新型高效液相色谱-串联质谱（HPLC-MS/MS）方法，该方法具有以下特点：样本体积小、运行时间短、快速、简单且成本效益高的样本制备、两种内标（IS）即SDZ RAD 223-756和子囊霉素性能的评估以及该方法与HPLC-MS/MS参考方法的比较。作者建立了一种带有在线萃取和样本净化的多反应监测正离子HPLC-MS/MS方法。该程序包括：一台配备涡轮离子喷雾的API 2000三重四极杆质谱仪、内置的瓦尔科切换阀、一个HPLC系统；保护柱；一根Nova-Pak C18分析柱；洗涤溶液，甲醇:30 mM醋酸铵pH 5.1（80:20）；洗脱溶液，甲醇:30 mM醋酸铵pH 5.1（97:3）；流速0.8 mL/min；运行时间2.8分钟。第一和第三个四极杆设置为检测依维莫司的铵加合物离子和高质量碎片（m/z 975.5-->908.5），以及两种内标：SDZ RAD 223-756（m/z 989.8-->922.8）和子囊霉素（m/z 809.5-->756.5）。使用任一种内标时，依维莫司的定量下限（LLOQ）为1.0 μg/L。使用添加了依维莫司浓度为2.0至25.0 μg/L的血液样本时，SDZ RAD 223-756的日间精密度范围为3.1%至5.7%，子囊霉素的日间精密度范围为6.0%至8.6%。使用添加样本在2.5至25 μg/L范围内时，绝对回收率平均为77.3%（SDZ RAD 223-756）和76.8%（子囊霉素）。基于98.6%（SDZ RAD 223-756）和105%（子囊霉素）的平均观察信号检测结果，未证明对依维莫司有基质效应。将使用该方法和两种内标获得的依维莫司浓度与一家参考实验室进行比较，结果表明，使用子囊霉素获得的依维莫司平均浓度在统计学上与参考方法的结果不同（更低），而使用SDZ RAD 223-756作为内标的方法与参考方法相比给出了等效结果。
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