"Detection Of Aromatic Compounds Based On DNA Intercalation Using An Evanescent Wave Biosensor"
Anal. Chem.
1995 Volume 67, Issue 5 Pages 787-792
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P. C. Pandey and H. H. Weetall
Abstract:
A FIA system coupled with an evanescent wave biosensor employing total internal reflection of fluorescence was used to detect compounds that intercalate with DNA. The analyte was detected by competition and displacement of intercalated ethidium bromide (excitation and emission at 499 and 592 nm, respectively). Double-stranded DNA was immobilized over the surface of a silanized optical fiber using acrylamide-methacrylamide-hydrazide prepolymer. The sample solution (20 µL) was introduced into 0.5 M phosphate buffer of pH 7.2 flowing at 24 ml/h and passed to the fluoribiosensor. A constant concentration of ethidium bromide was also pumped at the same rate and, at the steady state, the competitive intercalator was introduced. The response for 9,10-anthraquinone-2,6-disulfonic acid, remazol brilliant blue, decacyclene and 4',6-diamidino-2-phenylindole dihydrochloride at the µg/ml level was significantly higher using the modified fiber than an unmodified one, and increased with increasing concentration of immobilized DNA. A decrease in response to ethidium bromide was obtained on injection of the competing DNA intercalator, which was followed by recovery of the steady state in 6.5-12 min. This decrease in response was a linear function of analyte concentration.
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