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Separation and Purification of Nucleotides, Nucleosides, Purine and Pyrimidine Bases by Ion Exchange

  • P. Senthil KumarEmail author
  • P. R. Yaashikaa
Chapter

Abstract

The examination of a whole arrangement of constituents of biological significance, the nucleic acids, nucleotides, and related substances, has been blocked extensively by the absence of particular strategies for the separation and characterization of their nitrogenous constituents. The strategies accessible for the separation and purification of nucleotides and nucleic acids have been connected to an assortment of materials. Despite the fact that refinement of the nucleotides was endeavored by different systems, for example, precipitation of the nucleotides as insoluble salts and ion-exchange procedures, an appropriate general strategy is still to be developed. The detachment into singular nucleotides by ion-exchange chromatography has been the main dependable method for assessing the nucleotide content. Combination of purines, pyrimidines, amino acids, and different nitrogenous substances was observed to be isolated by ion chromatography on a solitary column of the cation-exchange resin and Dowex 50. The mixture components were eluted with hydrochloric acid and controlled by either their response with ninhydrin or UV adsorption. Ion exchange has turned out to be one of the real techniques for fractionation of organic substances.

Keywords

Ion-exchange chromatography Nucleotides Nucleosides Purines and pyrimidines Purification Applications 

References

  1. 1.
    Yang Y, Hebron HR, Hang J (2008) High performance DNA purification using a novel ion exchange matrix. J Biomol Tech 19(3):205–210PubMedPubMedCentralGoogle Scholar
  2. 2.
    Koubek J, Lin KF, Chen YR, Cheng RP, Huang JJT (2015) Strong anion-exchange fast performance liquid chromatography as a versatile tool for preparation and purification of RNA produced by in vitro transcription. RNA 19(10), Cold Spring Harbor Laboratory PressGoogle Scholar
  3. 3.
    Singh RP, Smesko SA, Abbas NM (1997) Ion chromatographic characterisation of toxic solutions: analysis and ion chemistry of biological liquids. J Chromatogr A 774:21–35CrossRefGoogle Scholar
  4. 4.
    Kasai K (1993) Size-dependent chromatographic separation of nucleic acids. J Chromatogr B Biomed Sci Appl 618(1–2):203–221CrossRefGoogle Scholar
  5. 5.
    Haastert PJMV (1981) High-performance liquid chromatography of nucleobases, nucleosides and nucleotides. J Chromatogr 210:241–254CrossRefGoogle Scholar
  6. 6.
    Ingle J (1963) The extraction and estimation of nucleotides and nucleic acids from plant material. Biochemistry 2:353–370Google Scholar
  7. 7.
    Singhal RP (2006) Separation and analysis of nucleic acids and their constituents by ion-exclusion and ion-exchange column chromatography. Sep Purif Rev 3(2):339–398CrossRefGoogle Scholar
  8. 8.
    Dragull K, Beck JJ (2012) Isolation of natural products by ion exchange methods. Methods Mol Biol 864:189–219CrossRefGoogle Scholar
  9. 9.
    Cramer H, Finn KJ, Herzberg E (2011) Purity analysis and impurities determination by reversed-phase high-performance liquid chromatography. Handbook of analysis of oligonucleotides and related products. CRC Press, Boca Raton, FL, pp 1–46Google Scholar
  10. 10.
    Asteriadis GT, Armbruster MA, Gilham PT (1975) Separation of oligonucleotides, nucleotides and nucleosides on columns of polystyrene anion-exchanges with solvent systems containing ethanol. Anal Biochem 70:64–74CrossRefGoogle Scholar
  11. 11.
    Easton LE, Shibata Y, Lukavsky PJ (2010) Rapid, nondenaturing RNA purification using weak anion-exchange fast performance liquid chromatography. RNA 16:647–653CrossRefGoogle Scholar
  12. 12.
    Singhal RP (1972) Ion-exclusion chromatography: analysis and isolation of nucleic acid components and influence of separation parameters. Arch Biochem Biophys 152:800–810CrossRefGoogle Scholar
  13. 13.
    Singhal RP (1974) Anion-exclusion and anion-exchange chromatography of nucleotides. Eur J Biochem 43:245–252CrossRefGoogle Scholar
  14. 14.
    Zhang Q, Lv H, Wang L, Chen M, Li F, Liang C, Yu Y, Jiang F, Lu A, Zhang G (2016) Recent methods for purification and structure determination of oligonucleotides. Int J Mol Sci 17:2134CrossRefGoogle Scholar
  15. 15.
    Lochmuller CH, Liu Q, Huang L, Li Y (1999) Separation of nucleotide oligomers by unitary anion-exchange. J Chromatogr Sci 37:251CrossRefGoogle Scholar
  16. 16.
    Tomasz J (1979) Separation of nucleic acid bases, nucleosides and nucleotides on strong cation-exchange thin layers. J Chromatogr 169:466–468CrossRefGoogle Scholar
  17. 17.
    Saraswat M, Musante L, Ravida A, Shortt B, Byrne B, Holthofer H (2013) Preparative purification of recombinant proteins: current status and future trends. BioMed Research International, Hindawi Publishing Corporation. http://dx.doi.org/10.1155/2013/312709
  18. 18.
    Wall JS (1953) Simultaneous separation of purines, pyrimidines, amino acids, and other nitrogenous compounds by Ion exchange chromatography. Anal Chem 25(6):950–953CrossRefGoogle Scholar
  19. 19.
    Liu L, Ouyang J, Baeyens RG (2008) Separation of purine and pyrimidine bases by ion chromatography with direct conductivity detection. J Chromatogr 1193:104–108CrossRefGoogle Scholar
  20. 20.
    Westerlund B (2004) Ion-exchange chromatography. In: Simpson RJ (ed) Purifying proteins for proteomics. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of Chemical EngineeringSSN College of EngineeringChennaiIndia

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