Batista Junior Almir Custodio, Rocha Yuri Arrates, Souza Gabriela Guimarães, Chaves Andréa Rodrigues
Federal University of Goiás, Chemistry Institute, Goiânia, Goiás 74690-900, Brazil.
Anal Methods. 2024 May 30;16(21):3318-3330. doi: 10.1039/d4ay00554f.
This study outlines the development and optimization of an analytical method using Disposable Pipette Extraction (DPX) followed by high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis to determine NAs in medicines. HPLC-MS analysis utilized a reversed-phase and positive mode electrospray ion source. DPX parameters were optimized through univariate and multivariate analyses, including extraction phase, desorption solvent, sample pH, equilibrium time, and extraction/desorption cycles. The optimized conditions included a C18 extraction phase, methanol desorption solvent, pH at 7, an equilibrium time of 30 seconds, 2 extraction cycles, and 5 desorption cycles. Considering this method, it was possible to achieve a sample preparation step for the analysis of NAs in medicines using a minimal amount of extraction phase, sample, and desorption solvent. Furthermore, the total extraction procedure enables the extraction of NAs in around 4 minutes with NA recovery up to 98%. Analytical performance demonstrated precision and accuracy below 15% and a quantification limit of 1 ng mL, meeting validation requirements set by regulations worldwide. Thus, the DPX/HPLC-MS technique offers a faster and cost-effective method for analyzing NAs in medicines compared to traditional approaches. Besides, this method reduces solvent consumption and residue generation, enhancing environmental sustainability according to green chemistry principles.
本研究概述了一种分析方法的开发与优化，该方法采用一次性移液器萃取（DPX），随后进行高效液相色谱 - 质谱（HPLC - MS）分析以测定药品中的核苷类似物（NAs）。HPLC - MS分析采用反相和正模式电喷雾离子源。通过单变量和多变量分析对DPX参数进行了优化，包括萃取相、解吸溶剂、样品pH值、平衡时间以及萃取/解吸循环次数。优化条件包括C18萃取相、甲醇解吸溶剂、pH值为7、平衡时间为30秒、2次萃取循环和5次解吸循环。考虑到该方法，使用最少的萃取相、样品和解吸溶剂就可以实现药品中NAs分析的样品制备步骤。此外，整个萃取过程能够在约4分钟内完成NAs的萃取，NAs回收率高达98%。分析性能表明精密度和准确度低于15%，定量限为1 ng/mL，符合全球法规设定的验证要求。因此，与传统方法相比，DPX/HPLC - MS技术为药品中NAs的分析提供了一种更快且更具成本效益的方法。此外，该方法减少了溶剂消耗和残留物产生，根据绿色化学原则提高了环境可持续性。
