Immunopathology and Cancer Biomarkers Unit, IRCCS CRO Centro di Riferimento Oncologico Aviano, 33081, Aviano (PN), Italy; Department of Molecular Sciences and Nanosystems, University of Venice, 30170, Mestre (VE), Italy.
Medical Oncology and Cancer Prevention Unit, IRCCS CRO Centro di Riferimento Oncologico Aviano, 33081, Aviano (PN), Italy.
J Pharm Biomed Anal. 2020 Jun 5;185:113261. doi: 10.1016/j.jpba.2020.113261. Epub 2020 Mar 17.
Few time-consuming bioanalytical methods are currently available for trabectedin quantification in clinical investigations. Here we present a novel, fast and sensitive method for trabectedin determination in human plasma based on hydrophilic interaction liquid chromatography and tandem mass spectrometry (HILIC-MS/MS). Plasma samples are treated with acetonitrile-0.1 % formic acid and the solvent extract is directly injected into an Acquity BEH Amide column (2.1 × 100 mm, 1.7 μm) operating in HILIC mode at 0.2 mL/min with 80:20 acetonitrile-0.1 % formic acid in water. The analyte is separated by an organic solvent gradient and quantified by an Agilent Ultivo triple quadrupole mass spectrometer operating in multiple reaction monitoring (MRM) mode. The quantitative MRM transitions were m/z 762→234 and m/z 765→234 for trabectedin and its d-labeled derivative, respectively. The lower limit of quantification (LLOQ) was 0.01 ng/mL and the assay was linear up to 2.5 ng/mL. The intra- and inter-day relative error ranged from 1.19 % to 8.52 %, while the relative standard deviation was less than 12.35 %. The method was used to determine the pharmacokinetic profiles of trabectedin in 26 patients with soft tissue sarcoma, showing that this new HILIC-MS/MS method is suitable for use in clinical research.
目前，用于临床研究中 trabectedin 定量分析的耗时的生物分析方法很少。在此，我们提出了一种基于亲水相互作用色谱和串联质谱（HILIC-MS/MS）的 trabectedin 测定的新型、快速和灵敏的方法。用乙腈-0.1%甲酸处理血浆样品，然后将溶剂提取物直接注入 Acquity BEH Amide 柱（2.1×100mm，1.7μm）中，在 HILIC 模式下以 0.2mL/min 的流速运行，流动相为 80:20 乙腈-0.1%甲酸水溶液。采用有机溶剂梯度分离分析物，并通过安捷伦 Ultivo 三重四极杆质谱仪以多重反应监测（MRM）模式进行定量分析。定量 MRM 跃迁分别为 m/z 762→234 和 m/z 765→234，用于 trabectedin 和其 d 标记衍生物。定量下限（LLOQ）为 0.01ng/mL，测定线性范围为 2.5ng/mL。日内和日间相对误差范围为 1.19%至 8.52%，而相对标准偏差小于 12.35%。该方法用于 26 例软组织肉瘤患者的 trabectedin 药代动力学研究，表明该新型 HILIC-MS/MS 方法适用于临床研究。
