Luminarin 3 as a derivatization reagent for the liquid chromatographic determination of cytidine, adenosine and related nucleotides with fluorimetric detection
✍ Scribed by F. Traoré; C. Fente; P. Prognon; G. Mahuzier
- Publisher
- Elsevier Science
- Year
- 1994
- Tongue
- English
- Weight
- 739 KB
- Volume
- 290
- Category
- Article
- ISSN
- 0003-2670
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✦ Synopsis
Luminarin 3 is a quinolizinocoumarine labelling reagent for carbonyl compounds, reacting via its hydrazide function. It was found to be a highly sensitive fluorescence reactive probe for nucleosides and nucleotides in liquid chromatography. Luminarin 3 reacts with the dialdehyde induced by preliminary periodate oxidation of the ribose moiety of nucleosides and nucleotides in 100 mM acetate buffer (at pH 3.0 or 4.0, depending on the derivative), to give highly fluorescent morpholino derivatives. The luminarin 3 derivatives of cytidine (Cyd), adenosine (Ado) and corresponding nucleotides were separated by chromatography on a Spherisorb cyano 5-pm column (300 X 4.6 mm id.), with a mixture of methanol and 100 mM phosphate buffer pH 6.0 (30/70, v/v), followed by fluorescence detection. In such a way the separation can easily be optimized for the eight nucleotides by variation of the methanol content of the mobile phase. Typical detection limits (in amount injected with signal/noise ratio of 3) were 40 fmol for Cyd; 52 fmol for CMP; 71 fmol for CDP; 36 fmol for CTP; 667 fin01 for Ado, 70 fmol for AMP, 28 fmol for ADP and 39 fmol for ATP.