Turković Lu, Mutavdžić Pavlović Dragana, Mlinarić Zvonimir, Skenderović Anamarija, Silovski Tajana, Sertić Miranda
Department of Pharmaceutical Analysis, Faculty of Pharmacy and Biochemistry, University of Zagreb, Ante Kovacica 1, 10000 Zagreb, Croatia.
Department of Analytical Chemistry, Faculty of Chemical Engineering and Technology, University of Zagreb, Marulicev trg 20, 10000 Zagreb, Croatia.
Pharmaceuticals (Basel). 2023 Oct 12;16(10):1445. doi: 10.3390/ph16101445.
In the development of bioanalytical LC-MS methods for the determination of drugs in plasma samples in a clinical setting, adequate sample preparation is of utmost importance. The main goals are to achieve the selective extraction of the analytes of interest and attain thorough matrix removal while retaining acceptable ecological properties, cost-effectiveness, and high throughput. Solid-phase extraction (SPE) offers a versatile range of options, from the selection of an appropriate sorbent to the optimisation of the washing and elution conditions. In this work, the first SPE method for the simultaneous extraction of six anticancer drugs used in novel therapeutic combinations for advanced breast cancer treatment-palbociclib, ribociclib, abemaciclib, anastrozole, letrozole, and fulvestrant-was developed. The following sorbent chemistries were tested: octylsilyl (C8), octadecylsilyl (C18), hydrophilic-lipophilic balance (HLB), mixed-mode cation-exchange (MCX and X-C), and mixed-mode weak cation-exchange (WCX), with different corresponding elution solvents. The samples were analysed using LC-MS/MS, with a phenyl column (150 × 4.6 mm, 2.5 μm). The best extraction recoveries (≥92.3%) of all analytes were obtained with the C8 phase, using methanol as the elution solvent. The optimised method was validated in the clinically relevant ranges, showing adequate precision (inter-day RSD ≤ 14.3%) and accuracy (inter-day bias -12.7-13.5%). Finally, its applicability was successfully proven by the analysis of samples from breast cancer patients.
在临床环境中开发用于测定血浆样本中药物的生物分析液相色谱-质谱（LC-MS）方法时，充分的样品前处理至关重要。主要目标是实现对目标分析物的选择性提取，并在保留可接受的生态特性、成本效益和高通量的同时，彻底去除基质。固相萃取（SPE）提供了多种选择，从合适吸附剂的选择到洗涤和洗脱条件的优化。在这项工作中，开发了第一种用于同时提取六种用于晚期乳腺癌治疗的新型治疗组合中的抗癌药物的SPE方法，这些药物为哌柏西利、瑞博西利、阿贝西利、阿那曲唑、来曲唑和氟维司群。测试了以下吸附剂化学类型：辛基硅烷基（C8）、十八烷基硅烷基（C18）、亲水亲脂平衡（HLB）、混合模式阳离子交换（MCX和X-C）以及混合模式弱阳离子交换（WCX），并使用了不同的相应洗脱溶剂。使用苯基柱（150×4.6 mm，2.5μm）通过LC-MS/MS对样品进行分析。使用C8固定相，以甲醇作为洗脱溶剂时，所有分析物均获得了最佳提取回收率（≥92.3%）。该优化方法在临床相关范围内进行了验证，显示出足够的精密度（日内相对标准偏差≤14.3%）和准确度（日内偏差-12.7-13.5%）。最后，通过对乳腺癌患者样本的分析成功证明了其适用性。
