LAROVA - Newsletter

Issue November 2011

Nucleotide HPLC Retention Time Standard

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The phosphates of the purine nucleosides adenosine (AMP/ADP/ATP) and guanosine (GMP/GDP/GTP) are pivotal molecules involved in signal transduction pathways where they are enzymatically interconverted (Fig. 1). Their analytical detection is routinely performed by HPLC since characteristic retention times allow convenient detection even in complex biological samples.

Figure 1
Intracellular equilibria of nucleoside monophosphates (NMP), diphosphates (NDP) and its triphosphates (NTP) are maintained by kinase (attachment of one phosphate moiety) and phosphatase (removal of one phosphate moiety) activities

Retention time in HPLC however, is influenced by a number of parameters such as instrumental settings, column, buffer composition, temperature and sample matrix, and it is often desirable to identify analytes by using positive controls as external or internal standards. Therefore, Larova introduces Nucleotide HPLC Retention Time Standards that

contain a nucleoside and its mono, di-, and triphosphate each,
come as buffered aqueous solutions at pre-determined concentrations suitable for direct injection into HPLC,
yield signals of similar intensities well suitable for common detectors,
allow easy evaluation of the HPLC method used (baseline resolution),
enable unambiguous identification and semi-quantitative detection of analytes (Fig. 2).

Figure 2
Adenosine- (a) and Guanosine-Nucleotide (b) HPLC Retention Time Standard (10 µl) was analyzed by C18-Reversed Phase HPLC in 0.1 M TEAA buffer/acetonitrile pH 6.5. The four nucleotides are well separated with NMP eluting first followed by NDP, NTP and finally the most hydrophobic nucleoside.

For more detailed information and for ordering, please visit our website at

www.larova.com/cms/en/1/catalog/317

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Dr. Mathias Grün

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