Tagliaro F, Moffa M, Gentile M M, Clavenna G, Valentini R, Ghielmi S, Marigo M
Institute of Forensic Medicine, University of Verona, Italy.
J Chromatogr B Biomed Appl. 1994 Jun 3;656(1):107-13. doi: 10.1016/0378-4347(94)00084-0.
The aim of the present work was the development of a simple capillary electrophoretic strategy, complementary to high-performance liquid chromatography, for the separation of different calcitonins (CTs) and calcitonin tryptic digests. Capillary electrophoresis was carried out with a manual capillary electropherograph with "on column" UV absorbance detection at 200 nm. The separation was accomplished in a 70 cm x 50 microns I.D. bare silica capillary. About 6 nl was loaded into the capillary by means of a split-flow system. Except in particular cases, electric fields of 300 V/cm were used at constant voltage. Separations were carried out in 0.05 M citrate buffer pH 2.5 or, alternatively, in 0.05 M borate buffer pH 9.5. A complete resolution of salmon, ASU1,7-eel, and human calcitonins was obtained in citrate and borate buffers. Other CT analogues could be separated only in one of the two buffers. Capillary electrophoresis in citrate buffer was also successful in the separation of the four final trypsin cleavage fragments of salmon calcitonin and, at least tentatively, of the nine intermediate cleavage products.
本研究的目的是开发一种简单的毛细管电泳方法，作为高效液相色谱的补充，用于分离不同的降钙素（CTs）和降钙素胰蛋白酶消化产物。使用手动毛细管电泳仪进行毛细管电泳，在200 nm处进行“柱上”紫外吸光度检测。分离在一根内径为50微米、长度为70厘米的未涂层石英毛细管中完成。通过分流系统将约6纳升样品注入毛细管。除特殊情况外，在恒定电压下使用300 V/cm的电场。分离在pH 2.5的0.05 M柠檬酸盐缓冲液或pH 9.5的0.05 M硼酸盐缓冲液中进行。在柠檬酸盐和硼酸盐缓冲液中，鲑鱼降钙素、ASU1,7-鳗鱼降钙素和人降钙素得到了完全分离。其他CT类似物只能在两种缓冲液中的一种中分离。在柠檬酸盐缓冲液中进行的毛细管电泳也成功分离了鲑鱼降钙素的四个最终胰蛋白酶裂解片段，以及至少九个中间裂解产物（初步结果）。
