当前位置:
X-MOL 学术
›
Nucleic Acids Res.
›
论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Simultaneous determination of ribonucleoside and deoxyribonucleoside triphosphates in biological samples by hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry
Nucleic Acids Research ( IF 14.9 ) Pub Date : 2018-03-15 , DOI: 10.1093/nar/gky203 Ziqing Kong 1 , Shaodong Jia 1 , Anna Lena Chabes 1 , Patrik Appelblad 2, 3 , Richard Lundmark 4, 5 , Thomas Moritz 6 , Andrei Chabes 1, 5
Nucleic Acids Research ( IF 14.9 ) Pub Date : 2018-03-15 , DOI: 10.1093/nar/gky203 Ziqing Kong 1 , Shaodong Jia 1 , Anna Lena Chabes 1 , Patrik Appelblad 2, 3 , Richard Lundmark 4, 5 , Thomas Moritz 6 , Andrei Chabes 1, 5
Affiliation
Information about the intracellular concentration of dNTPs and NTPs is important for studies of the mechanisms of DNA replication and repair, but the low concentration of dNTPs and their chemical similarity to NTPs present a challenge for their measurement. Here, we describe a new rapid and sensitive method utilizing hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry for the simultaneous determination of dNTPs and NTPs in biological samples. The developed method showed linearity (R2 > 0.99) in wide concentration ranges and could accurately quantify dNTPs and NTPs at low pmol levels. The intra-day and inter-day precision were below 13%, and the relative recovery was between 92% and 108%. In comparison with other chromatographic methods, the current method has shorter analysis times and simpler sample pre-treatment steps, and it utilizes an ion-pair-free mobile phase that enhances mass-spectrometric detection. Using this method, we determined dNTP and NTP concentrations in actively dividing and quiescent mouse fibroblasts.
中文翻译:
亲水作用液相色谱-串联质谱法同时测定生物样品中的核糖核苷和脱氧核糖核苷三磷酸
有关dNTPs和NTPs的细胞内浓度的信息对于DNA复制和修复机制的研究很重要,但是dNTPs的低浓度及其与NTPs的化学相似性对其测量提出了挑战。在这里,我们描述了一种新的快速灵敏的方法,该方法利用亲水相互作用液相色谱与串联质谱联用同时测定生物样品中的dNTPs和NTPs。所开发的方法显示出线性(R 2> 0.99)在较宽的浓度范围内,可以准确定量低pmol水平的dNTP和NTP。日内和日间精度低于13%,相对恢复率在92%至108%之间。与其他色谱方法相比,当前方法的分析时间更短且样品的预处理步骤更简单,并且它利用了无离子对流动相来增强质谱检测。使用这种方法,我们确定了活跃分裂和静止的小鼠成纤维细胞中的dNTP和NTP浓度。
更新日期:2018-03-15
中文翻译:
亲水作用液相色谱-串联质谱法同时测定生物样品中的核糖核苷和脱氧核糖核苷三磷酸
有关dNTPs和NTPs的细胞内浓度的信息对于DNA复制和修复机制的研究很重要,但是dNTPs的低浓度及其与NTPs的化学相似性对其测量提出了挑战。在这里,我们描述了一种新的快速灵敏的方法,该方法利用亲水相互作用液相色谱与串联质谱联用同时测定生物样品中的dNTPs和NTPs。所开发的方法显示出线性(R 2> 0.99)在较宽的浓度范围内,可以准确定量低pmol水平的dNTP和NTP。日内和日间精度低于13%,相对恢复率在92%至108%之间。与其他色谱方法相比,当前方法的分析时间更短且样品的预处理步骤更简单,并且它利用了无离子对流动相来增强质谱检测。使用这种方法,我们确定了活跃分裂和静止的小鼠成纤维细胞中的dNTP和NTP浓度。