PRECOLUMN DERIVATIZATION OF NUCLEOTIDES BASED ON FLUORESCENT CARBAMATE FORMATION ON THE SUGAR MOIETIES IN HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

The fluorescence derivatization of nucleotides with 2-(5-chlorocarbonyl-2-oxazolyl)-5,6-methylenedioxybenzofuran in the presence of sodium azide and the separation of the derivatives by high-performance liquid chromatography are described. The reagent reacts with 5'-terminal hydroxyl groups of...

Full description

Saved in:
Bibliographic Details
Published inChemical & pharmaceutical bulletin Vol. 40; no. 9; pp. 2559 - 2561
Main Authors NAGAOKA, H, NOHTA, H, SAITO, M, OHKURA, Y
Format Journal Article
LanguageEnglish
Published TOKYO Pharmaceutical Soc Japan 1992
Maruzen
Subjects
Analytical chemistry
Base Sequence
Carbamates
Chemistry
Chemistry, Medicinal
Chemistry, Multidisciplinary
Chromatographic methods and physical methods associated with chromatography
Chromatography, High Pressure Liquid
Exact sciences and technology
Fluorescent Dyes
Life Sciences & Biomedicine
Molecular Sequence Data
Oligonucleotides - analysis
Other chromatographic methods
Pharmacology & Pharmacy
Physical Sciences
Science & Technology
NUCLEOTIDE
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
SODIUM AZIDE
PRECOLUMN FLUORESCENCE DERIVATIZATION
2-(5-CHLOROCARBONYL-2-OXAZOLYL)-5,6-METHYLENEDIOXYBENZOFURAN
ADENINE
ALKALINE PHOSPHATASE
5'-TERMINAL HYDROXYL GROUP
Precolumn
Detection limit
HPLC chromatography
Nucleotide
Derivatization
Organic carbamate
Fluorescence spectrometry
Signal to noise ratio
Online AccessGet full text

Cover

More Information
Summary:The fluorescence derivatization of nucleotides with 2-(5-chlorocarbonyl-2-oxazolyl)-5,6-methylenedioxybenzofuran in the presence of sodium azide and the separation of the derivatives by high-performance liquid chromatography are described. The reagent reacts with 5'-terminal hydroxyl groups of nucleotides to produce the corresponding fluorescent carbamates. The derivatives of mono- and oligonucleotides are separated by chromatography on a reversed phase column (TSKgel ODS-80T(M)) and the derivatives of octa- and deca-nucleotides on a size exclusion column (TSKgel G3000SWXL). The detection limits (signal-to-noise ratio = 3) are 0.8-6.0 pmol on column. 5'-Phosphorylated nucleotide also gives a fluorescent derivative after alkaline phosphatase-mediated dephosphorylation.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ObjectType-Article-1
ObjectType-Feature-2
ISSN:0009-2363
1347-5223
DOI:10.1248/cpb.40.2559