Department of Clinical Pharmacology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Mar 1;887-888:25-34. doi: 10.1016/j.jchromb.2012.01.004. Epub 2012 Jan 20.
To support clinical pharmacokinetic studies with the anticancer agent E7080 (lenvatinib), liquid chromatography tandem mass spectrometry (LC-MS/MS) methods were developed for the quantification of E7080 and four of its metabolites in human plasma, urine and faeces and of E7080 in whole blood. Cross-analyte interferences between metabolites and parent compound were expected and therefore accounted for early in the method development. Plasma, urine and faeces samples were extracted with acetonitrile. Chromatographic separation was achieved on a 50 mm × 2.1 mm I.D. XTerra MS C18 column, with a 0.2 mL/min flow and gradient elution starting with 100% formic acid in water, followed by an increasing percentage of acetonitrile. Whole blood samples were extracted with diethyl ether and extracts were injected on a 150 mm × 2.1mm I.D. Symmetry Shield RP8 column. Detection was performed using an API3000 triple quadrupole mass spectrometer, with a turbo ion spray interface, operating in positive ion mode. Using 250 μL of plasma, E7080 and its metabolites could be quantified between 0.25 and 50.0ng/mL. The quantifiable ranges of E7080 in whole blood, urine and faeces were 0.25-500 ng/mL, 1.00-500 ng/mL and 0.1-25μg/g, using sample volumes of 250 μL, 200 μL and 250 mg, respectively. Calibration curves in all matrices were linear with a correlation coefficient (r(2)) of 0.994 or better. At the lower limit of quantification, accuracies were within ±20% of the nominal concentration with CV values less than 20%. At the other concentrations the accuracies were within ±15% of the nominal concentration with CV values below 15%. The developed methods have successfully been applied in a mass balance study of E7080.
为了支持抗癌药物 E7080（仑伐替尼）的临床药代动力学研究，开发了液相色谱串联质谱法（LC-MS/MS）来定量测定人血浆、尿液和粪便中的 E7080 及其四种代谢物，以及全血中的 E7080。预计代谢物与母体化合物之间存在交叉分析物干扰，因此在方法开发早期就对此进行了考虑。血浆、尿液和粪便样品用乙腈提取。色谱分离在 50mm×2.1mm ID XTerra MS C18 柱上进行，流速为 0.2mL/min，梯度洗脱从水相中的 100%甲酸开始，然后逐渐增加乙腈的百分比。全血样品用二乙醚提取，提取物在 150mm×2.1mm ID Symmetry Shield RP8 柱上进样。检测使用 API3000 三重四极杆质谱仪，带有涡轮离子喷雾接口，以正离子模式运行。使用 250μL 血浆，可定量测定 E7080 及其代谢物的浓度范围为 0.25-50.0ng/mL。使用 250μL、200μL 和 250mg 的样品量，E7080 在全血、尿液和粪便中的定量范围分别为 0.25-500ng/mL、1.00-500ng/mL 和 0.1-25μg/g。所有基质中的校准曲线均具有线性，相关系数（r(2)）大于或等于 0.994。在定量下限处，准确度在名义浓度的±20%以内，CV 值小于 20%。在其他浓度处，准确度在名义浓度的±15%以内，CV 值低于 15%。所开发的方法已成功应用于 E7080 的质量平衡研究。
