Highly Sensitive Spectrofluorometric Determination of Nucleic Acid Based on the Aggregation of two Oppositely Charged Kinds of Water-Soluble Porphyrins
作者:
Junko Nakamura,
Shukuro Igarashi,
期刊:
Analytical Letters
(Taylor Available online 1996)
卷期:
Volume 29,
issue 14
页码: 2453-2461
ISSN:0003-2719
年代: 1996
DOI:10.1080/00032719608002783
出版商: Taylor & Francis Group
关键词: tetrakis(4-N-trimethylaminophenyl)porphine;tetrakis(4-sulfophenyl)-porphine;nucleic acid;aggregation, spectrofluorimetry
数据来源: Taylor
摘要:
When cationic 5, 10, 15, 20-tetrakis(4-N-trimethylaminophenyl)porphine (TTMAPP) and anionic 5, 10, 15, 20-tetrakis(4-sulfophenyl)porphine (TSPP) were mixed in aqueous solution, these compounds were rapidly aggregated due mainly to their electrostatic interaction. The fluorescences of both porphyrins were quenched. However, if small amounts of a bulky molecule such as a polymer electrolyte coexisted in this reaction system, the aggregation of TTMAPP and TSPP was inhibited and their fluorescences reappeared. Based on these findings, a new highly sensitive spectrofluorometric determination of nucleic acids (DNA, RNA) was developed. Each calibration curve was linear in the concentration range of 0.03–0.4 μg/ml (DNA) and 0–05–0.6 μg/ml (RNA). Further, the detection limit (S/N = 3) was 0.016 μg/ml for DNA and for RNA, 0.020 μg/ml. The relative standard deviations were DNA: 1.70% and RNA: 1.83% (5 determinations). When the proposed method was applied to the determination of DNA originating from a bacteriophage, the results were satisfactory.
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