Document Detail


A novel and specific fluorescence reaction for uracil.
MedLine Citation:
PMID:  20678635     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Facile and specific methods to quantify a nucleobase in biological samples are of great importance for diagnosing disorders in nucleic acid metabolism. In the present study, a novel fluorogenic reaction specific for uracil has been developed. The reaction was carried out in an alkaline medium containing benzamidoxime and K(3)[Fe(CN)(6)] which were heated for 2.0 min. Under the optimum reaction conditions, strong fluorescence was produced only from uracil, not from other many biogenic compounds tested such as cytosine, thymine, adenine, guanine, nucleobases, nucleosides, nucleotides, amino acids, saccharides, creatine, creatinine and urea. The sensitivity of this method was compared with a known fluorogenic reaction using phenacylbromide which does not react with uracil but reacts with cytosine, adenine and their analogues. The proposed uracil-specific reaction showed approximately 400-fold higher sensitivity than the phenacylbromide reaction. The lower detection limit of uracil by the present method was 100 pmol mL(-1), and a good linearity of the calibration curve was obtained up to 100 nmol mL(-1) uracil. Due to its high sensitivity and specificity, the quantitative determination of uracil was possible by the proposed fluorimetric method.
Authors:
Takayuki Shibata; Shin-ya Kawasaki; Jun-ya Fujita; Tsutomu Kabashima; Masaaki Kai
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-06-30
Journal Detail:
Title:  Analytica chimica acta     Volume:  674     ISSN:  1873-4324     ISO Abbreviation:  Anal. Chim. Acta     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-03     Completed Date:  2010-12-01     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370534     Medline TA:  Anal Chim Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  234-8     Citation Subset:  IM    
Copyright Information:
2010 Elsevier B.V. All rights reserved.
Affiliation:
Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-Machi, Nagasaki 852-8521, Japan.
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MeSH Terms
Descriptor/Qualifier:
Acetophenones / metabolism
Benzamidines / metabolism
Cytosine / metabolism
Fluorometry / methods*
Limit of Detection
Uracil / metabolism*
Chemical
Reg. No./Substance:
0/Acetophenones; 0/Benzamidines; 613-92-3/benzamidoxime; 66-22-8/Uracil; 70-11-1/phenacyl bromide; 71-30-7/Cytosine

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