Ion-pair reversed-phase high-performance liquid chromatography of adenine nucleotides and nucleoside using triethylamine as a counterion

Journal of Chromatography. B, Biomedical Sciences and Applications
T UesugiK Mohri

Abstract

An isocratic HPLC method for the simple and selective determination of adenine nucleoside and nucleotides has been developed. The separation is achieved at room temperature by reversed-phase chromatography (Shiseido, Capcell Pak C18). A mixture of 0.1 M triethylamine (TEA) phosphate buffer and methanol (95:5, v/v) is used as a standard eluent. Influence of pH and concentrations of organic modifiers and TEA ion on capacity factors of adenine compounds has been investigated. It has been also found that the TEA ion in the eluent is adsorbed onto the reversed-phase surface. The results clearly demonstrate that ion-pair formation with TEA ion occurs probably both in the mobile phase and on the stationary phase and governs the retention of adenine and nucleotides in the present system. The HPLC system is applied to the analysis of adenine nucleotides formed as intermediates in the synthesis of 3'-phosphoadenosine 5'-phosphosulphate (PAPS) and to the assays of ATPases and 5'-nucleotidase activities in rat liver plasma membrane. This method is a new type of ion-pair reversed-phase HPLC system and is suitable for the separation of highly polar organic anions, especially for adenine nucleotides.

References

Aug 7, 1992·Journal of Chromatography·Q C MengS Oparil
Dec 1, 1986·Archives of Biochemistry and Biophysics·C FresneauP Volfin
Nov 1, 1981·Journal of Pharmacobio-dynamics·K YamaokaT Uno

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Citations

Jan 5, 2002·Journal of the American Society for Mass Spectrometry·Carmai SetoBerton Gunter
Oct 29, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Rolf W SparidansJos H Beijnen
Dec 31, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Dan HuangXiaoguang Chen
Mar 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·M CichnaP Markl
Dec 1, 2001·Biological & Pharmaceutical Bulletin·K SanoT Uesugi
Jul 24, 2010·Journal of Thoracic Oncology : Official Publication of the International Association for the Study of Lung Cancer·Yoichi NakamuraShigeru Kohno
Nov 14, 2012·Journal of Pharmaceutical and Biomedical Analysis·Mirlinda BibaRebecca A Arvary
Oct 27, 2006·Biochemical and Biophysical Research Communications·Adel BakhtiarovaDaniel M Kemp
Apr 21, 2007·Methods in Cell Biology·Cristofol Vives-BauzaGiovanni Manfredi
Sep 30, 2018·Analytical and Bioanalytical Chemistry·Ludivine FereyKaren Gaudin

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