Chair of the Environmental Chemistry & Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, 7 Gagarin St., PL-87 100 Toruń, Poland.
Chair of the Environmental Chemistry & Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, 7 Gagarin St., PL-87 100 Toruń, Poland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Jul 15;1090:90-100. doi: 10.1016/j.jchromb.2018.05.025. Epub 2018 May 18.
Antisense oligonucleotides have been successfully investigated for the treatment of different types of diseases. Detection and determination of antisense oligonucleotides and their metabolites are necessary for drug development and evaluation. This review focuses mainly on the first step of the analysis of oligonucleotides i.e. the sample preparation stage, and in particular on the techniques used for liquid chromatography and liquid chromatography coupled with mass spectrometry. Exceptional sample preparation techniques are required as antisense oligonucleotides need to be determined in complex biological matrices. The text discusses general issues in oligonucleotide sample preparation and approaches to their solution. The most popular techniques i.e. protein precipitation, protein enzyme digestion and liquid-liquid extraction are reviewed. Solid phase extraction methods are discussed and the issues connected with the application of each method are highlighted. Other newly reported promising techniques are also described. Finally, there is a summary of actually used techniques and the indication of the direction of future research.
反义寡核苷酸已成功应用于多种疾病的治疗研究。为了药物研发和评估，需要对反义寡核苷酸及其代谢物进行检测和定量分析。本篇综述主要聚焦于寡核苷酸分析的第一步，即样品制备阶段，特别是与液相色谱和液相色谱-质谱联用相关的技术。由于反义寡核苷酸需要在复杂的生物基质中进行检测，因此需要使用特殊的样品制备技术。本文讨论了寡核苷酸样品制备的常见问题及解决方案。综述了最常用的技术，如蛋白质沉淀、蛋白酶消化和液-液萃取。还讨论了固相萃取方法，并强调了每种方法应用中存在的问题。此外，还描述了一些新报道的有前景的技术。最后，对实际应用的技术进行了总结，并指出了未来研究的方向。
