Study of DNA-ellipticine interaction by capillary electrophoresis with laser-induced fluorescence detection.
Ellipticine (5,11-dimethyl-6H-pyrido[4,3-b]carbazole), an alkaloid isolated from Apocynaceae plants, exhibits an antitumor activity, which is exceptionally high against several specific types of tumors. Ellipticine is also interesting as an anticancer drug as it has limited side effects and lacks of hematological toxicity. Various methods to study intercalating activity of this drug have been developed. However, to our best knowledge, capillary electrophoresis (CE) as a technique combining high separation resolution with various detection options has never been used for these purposes. In this study, a novel separation method based on CE with laser-induced fluorescence (CE-LIF) detection has been developed for the determination of ellipticine and for the monitoring of ellipticine-DNA interaction. Sodium acetate (50 mM, pH 4.5) was used as a background electrolyte and LIF detection at λ(ex) = 488 nm. The limit of detection for ellipticine was determined to be 5 × 10⁻⁸ M. A total of 20% dimethyl sulfoxide was found optimal as sample solvent. Additionally, intercalation of ellipticine into the double-stranded DNA was investigated. Signal corresponding to ellipticine was decreasing and a new peak appeared and was growing. It can be concluded that CE-LIF is a method applicable to in vitro studies of ellipticine-DNA complexes.
Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Brno, Czech Republic.
Marketa
M
Ryvolova
Vojtech
V
Adam
Tomas
T
Eckschlager
Marie
M
Stiborova
Rene
R
Kizek
D000818:Animals; D002645:Chickens; D004247:DNA / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D004611:Ellipticines / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D004912:Erythrocytes / Q000737:chemistry; D057230:Limit of Detection; D016014:Linear Models; D013050:Spectrometry, Fluorescence / Q000379:methods*
D004611:Ellipticines; C034192:ellipticine; D004247:DNA; D004121:Dimethyl Sulfoxide