HPLC Separation of Inositol Polyphosphates
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High performance liquid chromatography (HPLC) is an essential analytical tool in the study of the large number of inositol phosphate isomers. This chapter focuses on the separation of inositol polyphosphates from [3H]myo-inositol labeled tissues and cells. We review the different HPLC columns that have been used to separate inositol phosphates and their advantages and disadvantages. We describe important elements of sample preparation for effective separations and give examples of how changing factors, such as pH, can considerably improve the resolving ability of the HPLC chromatogram.
Key words:High performance liquid chromatography Inositol phosphates Acid extraction HPLC columns HPLC gradients
The authors would like to thank Prof. R.H. Michell and Dr. C.J. Kirk for permission to use data obtained while C.J.B was a research fellow in their laboratory. The authors own work was supported by grants from Karolinska Institutet, Novo Nordisk Foundation, the Swedish Research Council, the Swedish Diabetes Association, EFSD, The Family Erling-Persson Foundation, Berth von Kantzow’s Foundation, and EuroDia (LSHM-CT-2006-518153).
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