Ultrasensitive capillary electrophoresis of sulfated disaccharides in chondroitin dermatan sulfates by laser-induced fluorescence after derivatization with 2-aminoacridone

An ultrasensitive capillary electrophoretic method for separating the vario usly sulfated chondroitin/dermatan sulfate-derived Delta-disaccharides afte r digestion with chondro/dermatolyases and derivatization with the fluoroph ore 2-aminoacridone is described. All known mono-, di- and tri-sulfated Del ta-disaccharides were completely separated using 15 mM orthophosphate buffe r (pH 3.0) at 20 kV without any interference of the excess derivatizing rea gent. They were detected at the anode (reversed polarity) using either an A r-ion laser-induced fluorescence (LIF) detector (excitation wavelength 488 nm) or a UV detector. The sensitivity obtained by LIF (0.51 pmol/l) was at least 100 and 10 times higher as compared to those obtained by UV detection at 232 nm of underivatized Delta-disaccharides and at 254 nm of those deri vatized with aminoacridone, respectively. The method has been easily applie d to the analysis of chondroitin/dermatan sulfates from various tissues at the attogram level, including chondrotin/dermatan sulfates from normal and aneurysmal human abdominal aortas. (C) 1999 Elsevier Science B.V. All right s reserved.