Quantitation of the dexrazoxane hydrolysis product ADR-925 by fluorescence detection of its terbium(III) complex after high-performance liquid chromatographic separation.
An HPLC fluorescence detection method was developed to quantitate the complexing agent ADR-925 (II). Compound II is the metal-ion-binding rings-opened hydrolysis product of the doxorubicin cardioprotective drug dexrazoxane (I). II formed a strong complex with the fluorescent metal-ion terbium(III) and this complex could be chromatographed by HPLC and detected by its fluorescence, with excitation and emission wavelengths of 200 and 544 nm, respectively. The terbium(III)-II complex was separated isocratically on a C18 reversed-phase column with an eluent consisting of 50% methanol and 50% 4 mM aqueous solution of the ion-pairing reagent 1-heptanesulfonate. The lower limit of detection of II, quantitated as its fluorescent terbium(III) complex, was estimated to be 25 pmol, which was some twenty times lower than with UV-Vis absorbance detection. The fluorescent detection method was used to follow the hydrolysis of I to II in buffer and in blood plasma.
Hasinoff
B B
BB
Faculty of Pharmacy, University of Manitoba, Winnipeg, Canada.
D002614:Chelating Agents / Q000032:analysis* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D005029:Ethylenediamines / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D005998:Glycine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000302:isolation & purification; D006801:Humans; D011929:Razoxane / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013725:Terbium / Q000737:chemistry*
D002614:Chelating Agents; D005029:Ethylenediamines; D013725:Terbium; D011929:Razoxane; C059866:ICRF 198; D005998:Glycine