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Study on Determination of Purine Nucleotides by Ultra Performance Liquid Chromatography-Mass Spectrometry |
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DOI:10.3969/j.issn.1004-4957.年份.月份 |
KeyWord:UPLC-MS cellular ATP ADP AMP NAD+ NADP+ |
Author | Institution |
姜丹丹,李伟,周怀彬,张婷,许芳,武佳,吕建新 |
温州医学院检验医学院 生命科学学院检验医学教育部重点实验室 |
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Abstract: |
An ultra performance liquid chromatography-mass spectrometric(UPLC-MS)method was developed for the simultaneous determination of purine nucleotides(e.g.ATP,ADP,AMP,NAD+ and NADP+) in mammalian cell.The HPLC separation was carried out on a KinetexTM HILIC column(50 mm×4.6 mm,2.1 μm).The experimental parameters,including-extraction method,compositions of mobile phase and MS parameters were optimized.The results demonstrated that the standard curves were linear in the range of 1.0-100 μmol/L,with correlation coefficients above 0.99.The average recoveries were in the range of 95.7%-101.1%with RSDs of 1.2%-6.1%.The method is sensitive and repeatable,and could be applied in the analysis of intracellular energy metabolism. |
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