Lee H S, Lee J S, Lee H, Jung Y S, DeLuca P P, Lee K C
Screening and Toxicology Center, Korea Research Institute of Chemical Technology, Yoosung, Taejeon, South Korea.
J Chromatogr B Biomed Appl. 1995 Nov 3;673(1):136-41. doi: 10.1016/0378-4347(95)00254-g.
For the determination of salmon calcitonin and its degradation products in biological samples, a reversed-phase HPLC method with column switching and flow-through radioisotope detection has been developed using high specific activity [125I]salmon calcitonin. Effects of the precolumn packing material and washing solvent were examined in terms of [125I]salmon calcitonin recovery. Spiked samples of [125I]salmon calcitonin in plasma and kidney homogenate were injected onto a LiChroprep RP-8 precolumn after dilution with 0.1% trifluoroacetic acid. After washing the polar interfering compounds with 0.1% trifluoroacetic acid, the concentrated [125I]salmon calcitonin and its degradation products were eluted and separated on a W-Porex C18 column with a gradient of 0.1% trifluoroacetic acid in acetonitrile-water. Detection and calibration of [125I]salmon calcitonin were possible down to picogram levels. Reproducible kinetic data for the degradation of intact [125I]salmon calcitonin were possible down to picogram levels. Reproducible kinetic data for the degradation of intact [125I]salmon calcitonin by rat kidney homogenate could be traced.
为了测定生物样品中的鲑鱼降钙素及其降解产物，已开发出一种采用柱切换和流通式放射性同位素检测的反相高效液相色谱法，该方法使用高比活度的[125I]鲑鱼降钙素。从[125I]鲑鱼降钙素回收率的角度考察了预柱填充材料和洗涤溶剂的影响。将用0.1%三氟乙酸稀释后的血浆和肾脏匀浆中的[125I]鲑鱼降钙素加标样品注入LiChroprep RP - 8预柱。用0.1%三氟乙酸洗涤极性干扰化合物后，浓缩的[125I]鲑鱼降钙素及其降解产物在W - Porex C18柱上以乙腈 - 水体系中含0.1%三氟乙酸的梯度进行洗脱和分离。[125I]鲑鱼降钙素的检测和校准可低至皮克水平。完整的[125I]鲑鱼降钙素降解的可重复动力学数据可低至皮克水平。可以追踪大鼠肾脏匀浆对完整的[125I]鲑鱼降钙素降解的可重复动力学数据。
