Nys Gwenaël, Cobraiville Gaël, Kok Miranda G M, Wéra Odile, Servais Anne-Catherine, Fillet Marianne
Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liège, Quartier hopital, Avenue Hippocrate 15, 4000, Liège, Belgium.
Laboratory of thrombosis and hemostasis, GIGA-Cardiovascular Sciences, University of Liège, Quartier hopital,Avenue Hippocrate 15, 4000, Liège, Belgium.
J Chromatogr A. 2017 Nov 17;1524:160-168. doi: 10.1016/j.chroma.2017.10.006. Epub 2017 Oct 4.
Pharmacokinetic (PK) studies on small animals are challenging as only small volumes of samples are available, in which the analyte is present at low concentration in a complex matrix. In this context, the use of miniaturized analytical techniques may provide undeniable advantages in terms of sensitivity, sample and solvent consumption compared to the reference UHPLC-MS/MS methods In this study, we present the development of a nanofluidic-LC-MS/MS method to analyze two model analytes of therapeutic interest, namely estradiol (E2) and estetrol (E4) after microsampling with volumetric absorptive microsampling (VAMS) devices, an innovative sampling technique to collect small volumes of whole blood. The nanofluidic LC-MS/MS method was developed using an experimental design to find the optimal conditions to analyze both E2 and E4 with the highest sensitivity. Subsequently, the optimized method was validated according to ICH guidelines and compared to a previously developed UHPLC-MS/MS method. A limit of quantitation of 50pg/ml was reached with the LC-chip method, which is 50 times better than UHPLC-MS/MS. Both methods were then critically evaluated from the analytical and operational points of view. Finally, the quantitation of estrogens after whole blood microsampling was compared with the results obtained with the corresponding plasma samples.
对小动物进行药代动力学（PK）研究具有挑战性，因为只能获得少量样本，其中分析物在复杂基质中的浓度较低。在这种情况下，与参考超高效液相色谱-串联质谱（UHPLC-MS/MS）方法相比，使用小型化分析技术在灵敏度、样品和溶剂消耗方面可能具有不可否认的优势。在本研究中，我们展示了一种纳米流体液相色谱-串联质谱方法的开发，该方法用于分析两种具有治疗意义的模型分析物，即雌二醇（E2）和雌三醇（E4），采用体积吸收微采样（VAMS）装置进行微量采样，这是一种收集少量全血的创新采样技术。使用实验设计开发了纳米流体液相色谱-串联质谱方法，以找到分析E2和E4的最佳条件，从而实现最高灵敏度。随后，根据国际人用药品注册技术协调会（ICH）指南对优化后的方法进行了验证，并与先前开发的UHPLC-MS/MS方法进行了比较。液相色谱芯片法的定量限达到50pg/ml，比UHPLC-MS/MS方法高50倍。然后从分析和操作的角度对这两种方法进行了严格评估。最后，将全血微量采样后雌激素的定量结果与相应血浆样本的结果进行了比较。
