毛细管凝胶电泳法测定血浆中的癌泰得  

Quantitative Analysis of Cantide in Plasma by Capillary Gel Electrophoresis

作　　者：张嵬[1] 杨秉呼[1] 梁乾德[1] 鲁丹丹[1] 林汝仙[1] 王升启[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850

出　　处：《色谱》2005年第4期374-377,共4页Chinese Journal of Chromatography

基　　金：国家"863"项目(No.2001AA215261;2001AA234041);国家重大科技专项(No.2002AA2Z337);国家自然科学基金资助项目(No.39870879).

摘　　要：为了进行硫代反义寡核苷酸药物癌泰得的药代动力学研究,建立了毛细管凝胶电泳测定血浆中癌泰得含量的方法.样品经强阴离子交换柱除去血浆中的蛋白和油脂,通过反相C18柱脱盐,再通过渗滤膜除去残留盐分后,以长度为24个碱基的寡核苷酸作为内标,采用毛细管凝胶电泳测定血浆中癌泰得的含量.结果表明,毛细管凝胶电泳测定血浆中癌泰得含量的线性范围为12.5～400 mg/L(r=0.999 8),日内、日间的相对标准偏差分别为0.398% ～2.46% 、2.75% ～6.07% ,回收率为99.53% ～102.1% .毛细管凝胶电泳法用于血浆中反义硫代寡核苷酸的含量测定具有良好的准确性、稳定性和重现性.Cantide is a 20-mer antisense phosphorothioate oligonucleotide that inhibits telomerase catalytic subunit hTERT, pharmacologic results showed that it had promising antitumor activity. In order to study the pharmacokinetic properties of Cantide, a capillary gel electrophoretic （CGE） method with internal standard was used for the determination of Cantide in rat plasma. Cantide and the internal standard had approximately equal percentage of base composition. Extraction of the phosphorothioate oligonucleotides from plasma was accomplished using two solid-phase extraction columns, a strong anion-exchange column to remove plasma proteins and lipids, followed by a reversed-phase column to remove plasma salts. A second desalting step, achieved by dialysis utilizing a membrane, was required to remove residual ionic material from the extracted sample. The size of the capillary column was 31 cm × 100 μm i. d.with an effective length of 20 cm. The running buffer was a mixture of Tris-boric acid-urea （ pH8.5 ）. The calibration curve was linear in the range of 12.5 -400 mg/L, with correlation coefficient （r） of 0. 999 8. Intra-day and inter-day relative standard deviations （RSDs） for the extracted samples were 0. 398% - 2.45% and 2.75% - 5.07%, respectively. The range of recoveries was 99.53% - 102. 1%. The results demonstrate the high accuracy, stability and reproducibility of the procedure.

关 键 词：毛细管凝胶电泳 反义寡核苷酸 癌泰得 药代动力学 

分 类 号：O658[理学—分析化学]