Matsumoto Kazuko, Tsukahara Yoshie, Uemura Tomonari, Tsunoda Kinichi, Kume Hidehiro, Kawasaki Seiji, Tadano Jutaro, Matsuya Takeshi
Department of Chemistry, Advanced Research Center for Science and Engineering, Waseda University, Shinjuku-ku, Tokyo 169-8555, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Jun 25;773(2):135-42. doi: 10.1016/s1570-0232(02)00137-x.
A new fluorescent europium chelate labeling reagent, 5-(4"-chlorosulfo-1',1"-diphenyl-4'-yl)-1,1,1,2,2-pentafluoro-3,5-pentanedione (CDPP), was synthesized for the time-resolved fluorometric detection of HPLC. The label can be directly bound to amino or phenolic hydroxyl groups of analytes with its chlorosulfonyl group, and the labeled analytes are separated on a HPLC column. After separation, EuCl(3), TOPO (tri-n-octylphosphine oxide), and Triton X-100 were added by post-column introduction to the eluent, and the fluorescence of the europium chelate was measured with the time-resolved fluorometric detector. Estrone (E1), 17beta-estradiol (E2), ethynylestradiol (EE2) and estriol (E3) were measured with the detection limits of 0.65, 0.65, 0.65 and 0.60 ng/ml, respectively. The recovery for river water samples was in the range of 86.0-105.1% with the RSD of 1.9-5.8%. The method was applied to the analysis of a river water sample and estrone (E1) was determined to be 2.1 ng/l. The results and processing have been compared with those of a GC-MS method and a high degree of correlation (r> or =0.98) was observed.
合成了一种新型荧光铕螯合物标记试剂5-(4"-氯磺酰基-1',1"-二苯基-4'-基)-1,1,1,2,2-五氟-3,5-戊二酮(CDPP)，用于高效液相色谱的时间分辨荧光检测。该标记物可通过其氯磺酰基直接与分析物的氨基或酚羟基结合，标记后的分析物在高效液相色谱柱上分离。分离后，通过柱后引入将EuCl(3)、TOPO(三正辛基氧化膦)和Triton X-100加入洗脱液中，并用时间分辨荧光检测器测量铕螯合物的荧光。测定了雌酮(E1)、17β-雌二醇(E2)、乙炔雌二醇(EE2)和雌三醇(E3)，检测限分别为0.65、0.65、0.65和0.60 ng/ml。河水样品的回收率在86.0-105.1%范围内，相对标准偏差为1.9-5.8%。该方法应用于河水样品分析，测定雌酮(E1)为2.1 ng/l。将结果及处理方法与气相色谱-质谱法进行了比较，观察到高度相关性(r≥0.98)。
