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Fluorescence Quenching Investigation for Janus Green B and used as Probe in Determination of Nucleic Acids
Author(s) -
LiHua Chen,
LiuZhan Liu,
HanXi Shen
Publication year - 2005
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.200590291
Subject(s) - chemistry , nucleic acid , quenching (fluorescence) , fluorescence , nucleic acid quantitation , detection limit , sodium dodecyl sulfate , dna , micelle , molecular beacon , photochemistry , chromatography , aqueous solution , biochemistry , organic chemistry , oligonucleotide , physics , quantum mechanics
Fluorescence quenching of janus green B (JGB) in sodium dodecyl sulfate (SDS) micelle by nucleic acids (DNA) was studied using UV‐vis absorption, steady state fluorescence emission methods and lifetime measurements. In the SDS micelle, weak fluorescence of JGB was enhanced, and the maximum emission shifted from 425 to 410 nm. In the presence of DNA, the fluorescence of JGB was quenched. Linear relationships between the fluorescence quenching ( F 0 /F ) and concentrations of DNA were observed in the range of 2.4×10 −8 to 1.08×10 −7 mol·L −1 for calf thymus nucleic acids (ct DNA) and 1.9×10 −8 to 3.8×10 −8 mol·L −1 for fish sperm nucleic acids (fs DNA) when 2.5×10 −5 mol·L −1 JGB was employed. The limit detection were 1.3×10 −8 mol·L −1 for ct DNA and 6.4×10 −9 mol·L −1 for fs DNA. At high DNA concentration, there was a systematic deviation from the Stern‐Volmer equation due to the static and dynamic quenching occurring simultaneously. The proposed method was applied to the determination of the nucleic acids in chicken blood extraction and the analytical results were in good agreement with the UV‐method.