Laboratory of Pharmaceutical Analysis, Department of Pharmaceutical Technology, School of Pharmacy, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece.
J Chromatogr A. 2022 Sep 13;1680:463432. doi: 10.1016/j.chroma.2022.463432. Epub 2022 Aug 19.
In this study, an "all-in-one" microextraction device was designed and fabricated for the extraction of doxorubicin and its two metabolites from rat plasma prior to their determination by high performance liquid chromatography coupled to fluorescence detector. A sol-gel-based sorbent was synthesized in situ and incorporated within two conjoined porous polypropylene tubes together with a cylindrical magnetic bar in order to avoid the need of an external stirring bar. Among other sorbents investigated, the moderately polar sol-gel poly(tetrahydrofuran) was found to be advantageous due to its high affinity toward the target analytes. Systematic investigation of the critical parameters affecting the adsorption and the desorption step was carried out. Due to the "built-in" filtration mechanism of the porous microextraction capsules, the isolation of the analytes was performed directly in the plasma matrix without any previous sample pretreatment (i.e., protein precipitation, centrifugation, etc.). The proposed method was validated in terms of linearity, accuracy, precision, specificity, sensitivity, and stability according to the FDA guidelines. The limits of detection ranged between 1 - 2 ng mL while the lower limits of quantitation of the analytes were calculated as 10 ng mL. The accuracy (% relative error) was found within -9.7 - 15.3% under both intra- and inter-day conditions. The precision was better than 13.4% in all cases. ComplexGAPI index was employed to present the green attributes of the developed protocol from the preparation of the microextraction device to the final determination of the analytes. Finally, the applicability of the fabricated stand-alone extraction device was demonstrated in the analysis of the target analytes in rat plasma after intravenous administration of doxorubicin in order to assess its pharmacokinetic profile.
在这项研究中，设计并制造了一种“一体化”微萃取装置，用于在高效液相色谱-荧光检测法测定大鼠血浆中的阿霉素及其两种代谢物之前，从大鼠血浆中萃取阿霉素及其两种代谢物。采用溶胶-凝胶法原位合成了一种中等极性的溶胶-凝胶聚（四氢呋喃）作为吸附剂，并将其与两个相连的多孔聚丙烯管和一个圆柱形磁棒一起装入，以避免使用外部搅拌棒。在所研究的其他吸附剂中，中等极性的溶胶-凝胶聚（四氢呋喃）由于对目标分析物具有高亲和力而具有优势。系统研究了影响吸附和解吸步骤的关键参数。由于多孔微萃取胶囊的“内置”过滤机制，无需对样品进行预处理（即蛋白沉淀、离心等），即可直接在血浆基质中对分析物进行分离。根据 FDA 指南，该方法在线性、准确性、精密度、特异性、灵敏度和稳定性方面进行了验证。检测限范围为 1-2ng/mL，分析物的定量下限计算为 10ng/mL。在日内和日间条件下，准确度（%相对误差）均在-9.7-15.3%之间。在所有情况下，精密度均优于 13.4%。采用复杂 GAPI 指数从微萃取装置的制备到分析物的最终测定，展示了所开发方案的绿色属性。最后，该自主式萃取装置在静脉注射阿霉素后分析大鼠血浆中目标分析物中的应用，评估其药代动力学特征。
