Citation: ZHU Huiyu, WU Danni, WANG Hailin. Quantification of intracellular adenosine 5'-triphosphate and its metabolites by high performance liquid chromatography analysis[J]. Chinese Journal of Chromatography, ;2017, 35(1): 54-58. doi: 10.3724/SP.J.1123.2016.08031 shu

Quantification of intracellular adenosine 5'-triphosphate and its metabolites by high performance liquid chromatography analysis

State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China

Received Date: 26 August 2016

Fund Project: National Natural Science Foundation of China (No. 21327006).

This study was aimed to provide insight regarding the intracellular metabolites of adenosine 5'-triphosphate (ATP) and whether 2-tert-butyl-1,4-benzoquinone (TBBQ) affects cell metabolites. A rapid high performance liquid chromatography (HPLC) protocol was developed for the separation and quantitation of ATP and its metabolites (adenosine diphosphate (ADP) and adenosine monophosphate (AMP)) in cells. Chromatographic separation was performed using a Shimadzu HPLC system equipped with an Agela Venusil MP C18 column; isocratic elution was adopted. The mobile phase comprised solvent A (50 mmol/L disodium hydrogen phosphate and 15 mmol/L trimethylamine (TEA); pH adjusted to 7.88 using acetic acid (HAc)) and solvent B (methanol). The correlation coefficients of the three analytes were very high (R²≥0.9996), and the contents of the three metabolites in the MRC-5 cells were within the linear ranges (0.1-100 μ mol/L). The limits of detection for the detected three compounds were low. Samples were extracted from cells (after exposure and non-exposure to quinones) using 80% (v/v) methanol aqueous solution. The method developed in this study was successfully applied to detect ATP, ADP and AMP in MRC-5 cells, and the results demonstrated that ATP, ADP, AMP levels in cells were affected by TBBQ, but the relations between the concentration of TBBQ and the level of ATP, ADP and AMP were complex.
- high performance liquid chromatography (HPLC),
- adenosine 5'-triphosphate (ATP),
- quinones,
- metabolites,
- intracellular
1. [1]
2. [2]
3. [3]
4. [4]
5. [5]
6. [6]
7. [7]
8. [8]
9. [9]
10. [10]
11. [11]
12. [12]
13. [13]
14. [14]
15. [15]
16. [16]
17. [17]
18. [18]
19. [19]
20. [20]
21. [21]
22. [22]
23. [23]
24. [24]
25. [25]
26. [26]
1. [1]
  Fan Wu , Wenchang Tian , Jin Liu , Qiuting Zhang , YanHui Zhong , Zian Lin . Core-Shell Structured Covalent Organic Framework-Coated Silica Microspheres as Mixed-Mode Stationary Phase for High Performance Liquid Chromatography. University Chemistry, 2024, 39(11): 319-326. doi: 10.12461/PKU.DXHX202403031
2. [2]
  Siming Bian , Sijie Luo , Junjie Ou . Application of van Deemter Equation in Instrumental Analysis Teaching: A New Type of Core-Shell Stationary Phase. University Chemistry, 2025, 40(3): 381-386. doi: 10.12461/PKU.DXHX202406087
3. [3]
  Yanhui Zhong , Ran Wang , Zian Lin . Analysis of Halogenated Quinone Compounds in Environmental Water by Dispersive Solid-Phase Extraction with Liquid Chromatography-Triple Quadrupole Mass Spectrometry. University Chemistry, 2024, 39(11): 296-303. doi: 10.12461/PKU.DXHX202402017
4. [4]
  Renxiao Liang , Zhe Zhong , Zhangling Jin , Lijuan Shi , Yixia Jia . A Palladium/Chiral Phosphoric Acid Relay Catalysis for the One-Pot Three-Step Synthesis of Chiral Tetrahydroquinoline. University Chemistry, 2024, 39(5): 209-217. doi: 10.3866/PKU.DXHX202311024
5. [5]
  Yuchen Zhou , Huanmin Liu , Hongxing Li , Xinyu Song , Yonghua Tang , Peng Zhou . 设计热力学稳定的贵金属单原子光催化剂用于乙醇的高效非氧化转化形成高纯氢和增值产物乙醛. Acta Physico-Chimica Sinica, 2025, 41(6): 100067-. doi: 10.1016/j.actphy.2025.100067
6. [6]
  Zunxiang Zeng , Yuling Hu , Yufei Hu , Hua Xiao . Analysis of Plant Essential Oils by Supercritical CO₂Extraction with Gas Chromatography-Mass Spectrometry: An Instrumental Analysis Comprehensive Experiment Teaching Reform. University Chemistry, 2024, 39(3): 274-282. doi: 10.3866/PKU.DXHX202309069
7. [7]
  Ruiqing LIU , Wenxiu LIU , Kun XIE , Yiran LIU , Hui CHENG , Xiaoyu WANG , Chenxu TIAN , Xiujing LIN , Xiaomiao FENG . Three-dimensional porous titanium nitride as a highly efficient sulfur host. Chinese Journal of Inorganic Chemistry, 2024, 40(5): 867-876. doi: 10.11862/CJIC.20230441
8. [8]
  Mingyang Men , Jinghua Wu , Gaozhan Liu , Jing Zhang , Nini Zhang , Xiayin Yao . 液相法制备硫化物固体电解质及其在全固态锂电池中的应用. Acta Physico-Chimica Sinica, 2025, 41(1): 2309019-. doi: 10.3866/PKU.WHXB202309019
9. [9]
  Zhaoxin LI , Ruibo WEI , Min ZHANG , Zefeng WANG , Jing ZHENG , Jianbo LIU . Advancements in the construction of inorganic protocells and their cell mimic and bio-catalytical applications. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2286-2302. doi: 10.11862/CJIC.20240235
10. [10]
  Peng XU , Shasha WANG , Nannan CHEN , Ao WANG , Dongmei YU . Preparation of three-layer magnetic composite Fe₃O₄@polyacrylic acid@ZiF-8 for efficient removal of malachite green in water. Chinese Journal of Inorganic Chemistry, 2024, 40(3): 544-554. doi: 10.11862/CJIC.20230239
11. [11]
  Bing Yuan , Fengli Yu , Congxia Xie . Teaching Cases Design of Catalysis Courses for Emerging Engineering Education. University Chemistry, 2024, 39(3): 191-198. doi: 10.3866/PKU.DXHX202309032
12. [12]
  Qian Shao , Jiajing Tan , Yongmei Chen , Jiyue Jing , Zhuo Wang . Exploration and Practice on the Management of Extracurricular Innovation Laboratories in Chemistry. University Chemistry, 2024, 39(4): 19-25. doi: 10.3866/PKU.DXHX202310119
13. [13]
  Zhibei Qu , Changxin Wang , Lei Li , Jiaze Li , Jun Zhang . Organoid-on-a-Chip for Drug Screening and the Inherent Biochemistry Principles. University Chemistry, 2024, 39(7): 278-286. doi: 10.3866/PKU.DXHX202311039
14. [14]
  Xue Dong , Xiaofu Sun , Shuaiqiang Jia , Shitao Han , Dawei Zhou , Ting Yao , Min Wang , Minghui Fang , Haihong Wu , Buxing Han . 碳修饰的铜催化剂实现安培级电流电化学还原CO₂制C₂₊产物. Acta Physico-Chimica Sinica, 2025, 41(3): 2404012-. doi: 10.3866/PKU.WHXB202404012
15. [15]
  Shuyu Liu , Xiaomin Sun , Bohan Song , Gaofeng Zeng , Bingbing Du , Chongshen Guo , Cong Wang , Lei Wang . Design and Fabrication of Phospholipid-Vesicle-based Artificial Cells towards Biomedical Applications. University Chemistry, 2024, 39(11): 182-188. doi: 10.12461/PKU.DXHX202404113
16. [16]
  Chunmei GUO , Weihan YIN , Jingyi SHI , Jianhang ZHAO , Ying CHEN , Quli FAN . Facile construction and peroxidase-like activity of single-atom platinum nanozyme. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1633-1639. doi: 10.11862/CJIC.20240162
17. [17]
  Sunting Xuan , Hang Shen , Xin Wang . Discussion on the Current Situation and Strategies for Academic Master’s Education in Chemistry. University Chemistry, 2024, 39(6): 37-41. doi: 10.3866/PKU.DXHX202401013
18. [18]
  Jinglun Wang , Hu Zhou , Baishu Zheng , Guobin Li , Ming Yue , Zhihua Zhou . Exploration and Practice of “Four Cooperations and Four Integrations” to Cultivate Innovative Talents in Chemical Materials in Local Colleges. University Chemistry, 2024, 39(7): 93-98. doi: 10.12461/PKU.DXHX202405013
19. [19]
  Qiaowen CHANG , Ke ZHANG , Guangying HUANG , Nuonan LI , Weiping LIU , Fuquan BAI , Caixian YAN , Yangyang FENG , Chuan ZUO . Syntheses, structures, and photo-physical properties of iridium phosphorescent complexes with phenylpyridine derivatives bearing different substituting groups. Chinese Journal of Inorganic Chemistry, 2025, 41(2): 235-244. doi: 10.11862/CJIC.20240311
20. [20]
  Fengmiao Yu , Yang Sheng , Chanyue Li , Bao Li . The Three Lives of Aspirin. University Chemistry, 2024, 39(9): 115-121. doi: 10.12461/PKU.DXHX202402033

Get Citation

PDF

XML

Metrics

PDF Downloads(4)
Abstract views(343)
HTML views(59)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142