Simultaneous measurement of trace monoadenosine and diadenosine monophosphate in biomimicking prebiotic synthesis using high-performance liquid ... [An article from: Analytica Chimica Acta]

This digital document is a journal article from Analytica Chimica Acta, published by Elsevier in 2006. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.

Description:
The method for simultaneous separation and determination of trace monoadenosine and diadenosine monophosphate (i.e. 2'-AMP, 3'-AMP, 5'-AMP and 3'-5' ApA) in biomimicking prebiotic synthesis was developed using high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection and electrospray ionization mass spectrometry (ESI-MS) identification. The separation was performed on a Supelco C"1"8 column with a gradient elution (solvent A: 10mM NH"4Ac aqueous solution; solvent B: MeOH). The flow rate was set at 1.0ml/min. The quantitative determination was achieved by HPLC with UV detection at 260nm. The linearity ranged from 0.5 to 100@mg/ml for each nucleotide. The limits of detection (LODs) for the four nucleotides were less than 0.30@mg/ml. The recovery ranged from 95.2 to 100.7%. The intra-day relative standard deviations (RSDs) of the retention times were between 0.7 and 1.1%. Both full-scan ESI-MS and -MS^2 for the four nucleotides under both positive and negative polarity were carried out and the possible cleavage pathways of them were depicted. The specific ions, [AMP+H]^+ at m/z 348 and [ApA+H]^+ at m/z 597, were chosen to characterize the four nucleotides in biomimicking prebiotic synthesis between N-(O,O-diisopropyl) phosphoryl amino acid (Dipp-aa) and adenosine. Using the proposed HPLC/UV/ESI-MS method, the concentration of 2'-AMP, 3'-AMP, 5'-AMP and 3'-5' ApA in the biomimicking prebiotic synthesis samples were determined.