Yuan J, Matsumoto K
Department of Chemistry, Waseda University, Tokyo, Japan.
J Pharm Biomed Anal. 1997 Jun;15(9-10):1397-403. doi: 10.1016/s0731-7085(96)02036-5.
A new tetradentate beta-diketone chelator, 1,10-bis(8'-chlorosulfo-dibenzothiophene-2'-yl)-4,4,5,5,6,6,7,7, -octafluorodecane-1,3,8,10-tetraone (BCOT), was synthesized. This compound forms a stable chelate with europium (III) and can be used as a fluorescence label for time-resolved fluorescence spectrometry. BCOT can be covalently bound to a protein under relatively mild conditions and, when complexed with europium(III), both in 0.1 M Tris-HCl solution and 1.0 x 10(-5) M tri-n-octyl phosphine oxide (TOPO)-0.05% sodium n-dodecyl sulfate (SDS)-0.1 M NaHCO3 solution, forms a strongly fluorescent chelate having lifetimes of 225 microseconds and 240 microseconds, respectively. As a model reaction, bovine serum albumin (BSA) was labeled with BCOT in a carbonate buffer solution. Fluorescence properties of the labeled BSA solution were studied. A time-resolved fluorescence determination of the BSA(BCOT)40-Eu3+ solution was carried out. Detection limits of 9.3 x 10(-14) M (TOPO-SDS-NaHCO3 solution) and 6.7 x 10(-13) M (Tris-HCl solution) for BSA were obtained by the method.
合成了一种新型四齿β-二酮螯合剂1,10-双(8'-氯磺基-二苯并噻吩-2'-基)-4,4,5,5,6,6,7,7-八氟癸烷-1,3,8,10-四酮(BCOT)。该化合物与铕(III)形成稳定的螯合物，可作为时间分辨荧光光谱法的荧光标记物。BCOT能在相对温和的条件下与蛋白质共价结合，当与铕(III)络合时，在0.1 M Tris-HCl溶液和1.0×10(-5) M三正辛基氧化膦(TOPO)-0.05%十二烷基硫酸钠(SDS)-0.1 M NaHCO3溶液中分别形成寿命为225微秒和240微秒的强荧光螯合物。作为模型反应，在碳酸盐缓冲溶液中用BCOT标记牛血清白蛋白(BSA)。研究了标记的BSA溶液的荧光性质。对BSA(BCOT)40-Eu3+溶液进行了时间分辨荧光测定。该方法对BSA的检测限在TOPO-SDS-NaHCO3溶液中为9.3×10(-14) M，在Tris-HCl溶液中为6.7×10(-13) M。
