Experimental and Clinical Pharmacology, CRO- National Cancer Institute, Aviano, Pordenone, Italy.
Oncology Unit B, CRO- National Cancer Institute, Aviano, Pordenone, Italy.
PLoS One. 2018 Feb 23;13(2):e0193500. doi: 10.1371/journal.pone.0193500. eCollection 2018.
Paclitaxel belongs to the taxanes family and it is used, alone or in multidrug regimens, for the therapy of several solid tumours, such as breast-, lung-, head and neck-, and ovarian cancer. Standard dosing of chemotherapy does not take into account the many inter-patient differences that make drug exposure highly variable, thus leading to the insurgence of severe toxicity. This is particularly true for paclitaxel considering that a relationship between haematological toxicity and plasma exposure was found. Therefore, in order to treat patients with the correct dose of paclitaxel, improving the overall benefit-risk ratio, Therapeutic Drug Monitoring is necessary. In order to quantify paclitaxel and its main metabolite, 6α-hydroxy-paclitaxel, in patients' plasma, we developed a new, sensitive and specific HPLC-MS/MS method applicable to all paclitaxel dosages used in clinical routine. The developed method used a small volume of plasma sample and is based on quick protein precipitation. The chromatographic separation of the analytes was achieved with a SunFire™ C18 column (3.5 μM, 92 Å, 2,1 x 150 mm); the mobile phases were 0.1% formic acid/bidistilled water and 0.1% formic acid/acetonitrile. The electrospray ionization source worked in positive ion mode and the mass spectrometer operated in selected reaction monitoring mode. Our bioanalytical method was successfully validated according to the FDA-EMA guidelines on bioanalytical method validation. The calibration curves resulted linear (R2 ≥0.9948) over the concentration ranges (1-10000 ng/mL for paclitaxel and 1-1000 ng/mL for 6α-hydroxy-paclitaxel) and were characterized by a good accuracy and precision. The intra- and inter-day precision and accuracy were determined on three quality control concentrations for paclitaxel and 6α-hydroxy-paclitaxel and resulted respectively <9.9% and within 91.1-114.8%. In addition, to further verify the assay reproducibility, we tested this method by re-analysing the incurred samples. This bioanalytical method was employed with success to a genotype-guided phase Ib study of weekly paclitaxel in ovarian cancer patients treated with a wide range of drug's dosages.
紫杉醇属于紫杉烷类，单独或联合多种药物方案用于治疗多种实体瘤，如乳腺癌、肺癌、头颈部癌和卵巢癌。标准化疗剂量并未考虑到许多个体间差异，这些差异导致药物暴露高度可变，从而导致严重毒性的出现。对于紫杉醇来说尤其如此，因为已经发现血液学毒性与血浆暴露之间存在关系。因此，为了用正确剂量的紫杉醇治疗患者，提高总体获益风险比，有必要进行治疗药物监测。为了定量检测患者血浆中的紫杉醇及其主要代谢物 6α-羟基紫杉醇，我们开发了一种新的、灵敏且特异的 HPLC-MS/MS 方法，适用于临床常规中使用的所有紫杉醇剂量。所开发的方法使用小体积的血浆样本，并基于快速蛋白沉淀。分析物的色谱分离采用 SunFire™ C18 柱（3.5μm，92Å，2.1×150mm）；流动相为 0.1%甲酸/双蒸水和 0.1%甲酸/乙腈。电喷雾离子源在正离子模式下工作，质谱仪在选择反应监测模式下工作。我们的生物分析方法根据 FDA-EMA 关于生物分析方法验证的指南成功验证。校准曲线呈线性（R2≥0.9948），浓度范围为（1-10000ng/mL 用于紫杉醇，1-1000ng/mL 用于 6α-羟基紫杉醇），具有良好的准确性和精密度。紫杉醇和 6α-羟基紫杉醇的三个质控浓度的日内和日间精密度和准确度分别为<9.9%和 91.1-114.8%。此外，为了进一步验证该方法的重现性，我们通过重新分析处理过的样品来测试该方法。该生物分析方法成功用于卵巢癌患者每周紫杉醇治疗的基因型指导的 Ib 期研究，药物剂量范围广泛。
