Production processes of adenosine include condensation of ribose and adenine by means of chemical or biochemical processes, as well as fermentation. Nowadays, adenosine is commonly produced by alkaline hydrolysis of yeast ribonucleic acid, often in the presence of calcium or lead ions, followed by chromatographic separation of the obtained nucleosides adenosine, cytidine, guanosine and uridine. The current Ph. Eur. monograph for adenosine describes a TLC method for determination of related substances that limits the impurities to 1 per cent. To ensure the quality of adenosine, it is proposed to replace the TLC method by a more sensitive and selective HPLC method, in accordance with the current policy for control of impurities as defined by the Ph. Eur. Commission. An HPLC separation system, originally proposed by the USP, using ion-pair reversed-phase chromatography in combination with tetrabutylammonium hydrogen sulphate as an ion-pair reagent, has been examined for its suitability to limit guanosine, inosine, uridine and adenine. This article describes the experiments as regards the choice of the most suitable commercial column to obtain an appropriate separation, the column temperature, the establishment of a relevant system suitability criterion, and the determination of correction factors for the individual impurities.

• PMID: 17691210
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Reference Type

Evaluation Study | Journal Article

Authors

Kopec S, Almeling S, Holzgrabe U

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