Metabolic stability tests are one of the fundamental steps at the preclinical stages of new drug development. Microsomes, used as a typical enzymatic model of liver biotransformation, can be a challenging matrix for analytical scientists due to a high concentration of cellular proteins and membrane lipids. In the work, we propose a new procedure integrating biotransformation reaction with SPME-like protocol for sample clean-up. It is beneficial to increase the overall quality of results in contrary to the typical protein precipitation approach. A set of ten arylpiperazine analogs, six of which are considered promising drug candidates (and four are accepted drugs) were used as a probe to assess the goodness of the newly proposed approach. In order to promote an efficient extraction protocol, a new, miniaturized shape of a sorbent, suitable to perform the extraction in 100 µL of the sample has been designed. Termination of the biotransformation process by protein denaturation with hot water was additionally evaluated. A quantitative structure-property relationship (QSPR) study using Orthogonal Partial Least Squares (OPLS) technique to reveal insights to the sorption mechanism was also performed. The obtained results showed the new 3D-printed sorbent can be an attractive basis for the new sample preparation approach for metabolic stability studies and an alternative for commercially available protocols based on solid-phase microextraction (SPME) or solid-phase extraction (SPE) principles.

代谢稳定性测试是新药开发的临床前阶段的基本步骤之一。由于高浓度的细胞蛋白和膜脂质，微粒体被用作肝脏生物转化的典型酶促模型，对于分析科学家而言可能是具有挑战性的基质。在工作中，我们提出了一种将生物转化反应与SPME样规程相结合的新程序，用于样品净化。与典型的蛋白质沉淀方法相反，增加结果的整体质量是有益的。一组十个芳基哌嗪类似物（其中六个被认为是有前途的候选药物（四个是公认的药物））用作评估新提议方法的有效性的探针。为了促进有效的提取方案，一种新的，小型化的吸附剂 设计了适合于以100 µL样品进行萃取的样品。另外评估了通过热水使蛋白质变性而终止的生物转化过程。还进行了使用正交偏最小二乘（OPLS）技术的定量结构-性质关系（QSPR）研究，以揭示对吸附机理的见解。获得的结果表明，新的3D打印吸附剂可以作为新的样品制备方法进行代谢稳定性研究的有吸引力的基础，并且可以作为基于固相微萃取（SPME）或固相萃取（SPE）原理的市售方案的替代方案。还进行了使用正交偏最小二乘（OPLS）技术的定量结构-性质关系（QSPR）研究，以揭示对吸附机理的见解。获得的结果表明，新的3D打印吸附剂可以成为新的样品制备方法（用于代谢稳定性研究）的有吸引力的基础，并且是基于固相微萃取（SPME）或固相萃取（SPE）原理的市售方案的替代方案。还进行了使用正交偏最小二乘（OPLS）技术的定量结构-性质关系（QSPR）研究，以揭示对吸附机理的见解。获得的结果表明，新的3D打印吸附剂可以作为新的样品制备方法进行代谢稳定性研究的有吸引力的基础，并且可以作为基于固相微萃取（SPME）或固相萃取（SPE）原理的市售方案的替代方案。