色谱

色谱

• 研究论文 • 上一篇    下一篇

无胶筛分毛细管电泳法测定猕猴血浆中的反义寡核苷酸药物癌泰得

王秀中1,2, 王清清2, 王诗鸿2, 李卫平1, 宋海峰2*, 鲁丹丹2, 王升启2   

  1. 1. 安徽医科大学药理学教研室, 安徽 合肥 230032; 2. 军事医学科学院放射与辐射医学研究所, 北京 100850
  • 收稿日期:2009-12-16 修回日期:2010-02-16 出版日期:2010-06-28 发布日期:1980-06-25
  • 通讯作者: 宋海峰

Quantitative determination of Cantide, an antisense oligodeoxynucleotide in rhesus monkey plasma using non-gel sieving capillary electrophoresis method

WANG Xiuzhong1,2, WANG Qingqing2, WANG Shihong2, LI Weiping1, SONG Haifeng2*, LU Dandan2, WANG Shengqi2   

  1. 1. Department of Pharmacology, Anhui Medical University, Hefei 230032, China; 2. Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China
  • Received:2009-12-16 Revised:2010-02-16 Online:2010-06-28 Published:1980-06-25
  • Contact: SONG Haifeng

PDF

499

被引次数

18

摘要： 采用两步固相萃取(SPE)法结合无胶筛分毛细管电泳(NGCE)技术建立了猕猴血浆中的反义寡核苷酸药物癌泰得的定量分析方法。优化并确定了SPE的相关条件(阴离子交换柱,上样缓冲液pH值为9.0,上样体积及洗脱体积分别为5 mL和3 mL)和NGCE的分析条件(灌胶时间为30 min,分离电压为24 kV)。在优化的条件下,猕猴血浆中癌泰得在1.95～250 mg/L范围内呈良好的线性关系,定量限(LOQ)为1.95 mg/L。批内准确度为93.38%～100.71%,批内相对标准偏差＜11%;批间准确度为89.46%～103.46%,批间相对标准偏差＜9%。在不同条件(室温下存放4 h; 4 ℃下存放24 h；反复冻融（-80 ℃至室温）2次；-80 ℃下保存1个月)下癌泰得在猕猴血浆中的稳定性良好。已将该方法成功地应用于癌泰得的猕猴药代动力学研究。

关键词: 癌泰得, 反义寡核苷酸, 固相萃取, 猕猴 , 无胶筛分毛细管电泳

Abstract: A dual solid phase extraction (SPE) pretreatment coupling with non-gel sieving capillary electrophoresis (NGCE) analysis method was established for the quantitative determination of an antisense oligodeoxynucleotide, Cantide, in rhesus monkey plasma. The conditions of SPE and the NGCE analysis were optimized. Under the optimized conditions (the SPE conditions: the pH of loading buffer was 9.0; the volumes of loading and the elution solution for the anion-exchange column were 5 mL and 3 mL, respectively. The NGCE analysis conditions: loading gel time was 30 min and the separation voltage was 24 kV), the linear dynamic range of Cantide in rhesus monkeys plasma was 1.95-250 mg/L, and the correlation coefficient (r) was more than 0.998. The limit of quantitation was 1.95 mg/L. The intra-batch accuracies ranged from 93.38% to 100.71% with the intra-batch relative standard deviation (RSD) less than 11%. The inter-batch accuracies were from 89.46% to 103.46% with the inter-batch RSD less than 9%. The stability experiment showed that the Cantide plasma sample was stable when stored at 4 ℃ for 24 h, room temperature for 4 h, -80 ℃ for 30 days and freeze-thaw for 2 cycles. This method was finally successfully applied to pharmacokinetic study of Cantide in rhesus monkeys.

Key words: antisense oligodeoxynucleotide, Cantide, non-gel sieving capillary electrophoresis (NGCE), rhesus monkey , solid phase extraction (SPE)