https://scholars.lib.ntu.edu.tw/handle/123456789/631324
標題: | Using an In-Sample Addition of Medronic Acid for the Analysis of Purine- and Pyrimidine-Related Derivatives and Its Application in the Study of Lung Adenocarcinoma A549 Cell Lines by LC-MS/MS | 作者: | Lin, Ya-Ting Chan, Shan-An Chen, Yi-Jung KUEI-PIN CHUNG CHING-HUA KUO |
關鍵字: | lung adenocarcinoma A549 cell; mass spectrometry; medronic acid; mitochondrial dysfunction; purine- and pyrimidine-related derivatives | 公開日期: | 5-五月-2023 | 出版社: | AMER CHEMICAL SOC | 卷: | 22 | 期: | 5 | 起(迄)頁: | 1434 | 來源出版物: | Journal of proteome research | 摘要: | Intracellular purine- and pyrimidine-related derivatives are vital molecules for preserving genetic information and are essential for cellular bioenergetics and signal transduction. This study developed a practical liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying intracellular purine- and pyrimidine-related derivatives. To solve the distorted peak shape related to di- and triphosphate nucleotides, in-sample addition of medronic acid and ammonium phosphate was performed. Using the BEH-amide column, the results showed that adding 0.5 mM medronic acid to the sample significantly improved the peak shape without causing an obvious ion suppressive effect. Method validation confirmed that the coefficients of determination (R2) values for linearity evaluation were above 0.94 for all analytes. The intraday and interday accuracies ranged from 85.1 to 128.4%, with the precision below 16.6%. The validated method was successfully applied in characterizing the alterations of purine- and pyrimidine-related derivatives in the A549 cell line with perturbed mitochondrial fission or blockade of the electron transport chain. Collectively, this study demonstrates that the strategy of in-sample medronic acid addition is effective in improving the quantification of intracellular purine- and pyrimidine-related derivatives. We believe that our proposed platform can facilitate the development of novel drugs targeting purine and pyrimidine metabolism in the future. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/631324 | ISSN: | 1535-3893 | DOI: | 10.1021/acs.jproteome.2c00736 |
顯示於: | 醫學院附設醫院 (臺大醫院) |
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