Sensitive determination of DNA based on the interaction between norfloxacin-Tb3+ complex and DNA.
The fluorescence intensity of the norfloxacin (NFX)-Tb3+ complex enhanced by DNA was studied. Therefore, a sensitive fluorescence method for the determination of DNA was developed. The optimal conditions of the method were as follows: the hexamethylenamine (HMA)-HCl buffer was adopted for adjusting the pH to 6.5 +/- 0.1, the concentrations of NFX and Tb3+ were both fixed in 1.0 x 10(-6) mol L(-1), and the excitation and emission wavelengths were selected at 290 and 545 nm, respectively. Under the optimal conditions, the enhanced fluorescence intensity was in proportion to the concentration of DNA in the same range of 5.0 x 10(-9) - 1.0 x 10(-6) g mL(-1) for hsDNA and thermally denatured ctDNA. The detection limits (S/N = 3) were 0.9 and 0.6 ng mL(-1), respectively. In addition, the interaction between NFX-Tb3+ and DNA was discussed in detail. The experimental results from UV absorption spectra, fluorescence spectra, and the salt effect study indicated that the interaction between norfloxacin-Tb3+ complex and DNA had at least two different binding modes: the electrostatic binding and the intercalation binding. The mechanism of the fluorescence enhancement effect was also discussed.
Tong
Changlun
C
Institute of Environmental Science, Zhejiang University, Hangzhou 310029, China. cltong@zju.edu.cn
Hu
Zhou
Z
Liu
Weiping
W
D000818:Animals; D002417:Cattle; D004247:DNA / Q000032:analysis*; D005399:Fishes; D005456:Fluorescent Dyes; D008297:Male; D009643:Norfloxacin; D011868:Radioisotopes; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013094:Spermatozoa / Q000737:chemistry; D013725:Terbium; D013950:Thymus Gland / Q000737:chemistry
D005456:Fluorescent Dyes; D011868:Radioisotopes; D013725:Terbium; D004247:DNA; D009643:Norfloxacin