Department of Clinical Biochemistry, Faculty of Pharmacy, Nicolaus Copernicus University in Toruń, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland.
Methods Mol Biol. 2021;2198:27-35. doi: 10.1007/978-1-0716-0876-0_3.
Stable-isotope-dilution tandem mass spectrometry is the most advanced technique used for quantitative determination of a wide spectrum of endogenously generated DNA nucleobase modifications. It is regarded as a gold standard for such analyses. Here, we consider the requirements for reliable identification and quantification of DNA adducts/modifications, whether endogenously derived or not, and discuss how their quantification can provide information on the mechanism of action and the biological relevance of individual nucleobase modifications. A clinical application of such measurements will only be possible after a full validation of the assay and once we have gained a better understanding of the exact role that these DNA modifications play in disease pathogenesis. Once these prerequisites are satisfied, DNA modification measurements may be helpful as clinical parameters for treatment monitoring, for risk group identification and for the development of prevention strategies.
稳定同位素稀释串联质谱法是用于定量测定广泛范围内内源性生成的 DNA 碱基修饰物的最先进技术。它被认为是此类分析的金标准。在这里，我们考虑了可靠鉴定和定量分析 DNA 加合物/修饰物（无论内源性还是外源性）的要求，并讨论了它们的定量分析如何提供关于单个碱基修饰物作用机制和生物学相关性的信息。只有在对测定方法进行全面验证后，并且在我们对这些 DNA 修饰在疾病发病机制中的确切作用有了更好的了解后，这种测量的临床应用才成为可能。一旦满足了这些前提条件，DNA 修饰的测量就可能作为治疗监测、风险群体识别和预防策略开发的临床参数。
