Department of Chemistry, Federal Rural University of Pernambuco, Recife, PE, Brazil.
Fundamental Chemistry Department, Federal University of Pernambuco, Recife, PE, Brazil.
Anal Biochem. 2023 Jun 15;671:115135. doi: 10.1016/j.ab.2023.115135. Epub 2023 Apr 3.
Given the importance of identifying the presence of biomarkers of human diseases in DNA samples, the main objective of this work was to investigate, for the first time, the electro-catalytic oxidation of 7-methyl-guanine (7-mGua) and 5-methyl-cytosine (5-mCyt) on a boron doped diamond electrode pre-treated cathodically (red-BDDE), using differential pulse voltammetry (DPV) and cyclic voltammetry (CV). The anodic peak potentials of 7-mGua and 5-mCyt by DPV were at E = 1.04 V and E = 1.37 V at pH = 4.5, indicating excellent peak separation of approximately 330 mV between species. Using DPV, experimental conditions such as supporting electrolyte, pH and influence of interferents were also investigated to develop a sensitive and selective method for individual and simultaneous quantification of these biomarkers. The analytical curves for the simultaneous quantification of 7-mGua and 5-mCyt in the acid medium (pH = 4.5) were: concentration range of 0.50-5.00 μmol L (r = 0.999), detection limit of 0.27 μmol L for 7-mGua; from 3.00 to 25.00 μmol L (r = 0.998), with a detection limit of 1.69 μmol L for 5-mCyt. A new DP voltammetric method for the simultaneous detection and quantification of biomarkers 7-mGua and 5-mCyt using a red-BDDE is proposed.
鉴于鉴定 DNA 样本中人类疾病生物标志物存在的重要性,本工作的主要目的是首次研究经阴极预处理的掺硼金刚石电极(red-BDDE)上电催化氧化 7-甲基鸟嘌呤(7-mGua)和 5-甲基胞嘧啶(5-mCyt),采用差分脉冲伏安法(DPV)和循环伏安法(CV)。在 pH 值为 4.5 时,DPV 法测定 7-mGua 和 5-mCyt 的阳极峰电位分别为 E = 1.04 V 和 E = 1.37 V,表明两种物质之间具有极好的约 330 mV 的峰分离度。采用 DPV 法,还研究了支持电解质、pH 值和干扰物影响等实验条件,以开发一种用于单独和同时定量这些生物标志物的灵敏和选择性方法。在酸性介质(pH 值 = 4.5)中同时定量测定 7-mGua 和 5-mCyt 的分析曲线为:浓度范围为 0.50-5.00 μmol L(r = 0.999),7-mGua 的检测限为 0.27 μmol L;从 3.00 至 25.00 μmol L(r = 0.998),5-mCyt 的检测限为 1.69 μmol L。提出了一种使用 red-BDDE 同时检测和定量生物标志物 7-mGua 和 5-mCyt 的新 DP 伏安法。