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[采用非凝胶筛分毛细管电泳法对恒河猴血浆中反义寡脱氧核苷酸坎地德进行定量测定]

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

作者信息

Wang Xiuzhong, Wang Qingqing, Wang Shihong, Li Weiping, Song Haifeng, Lu Dandan, Wang Shengqi

机构信息

Department of Pharmacology, Anhui Medical University, Hefei 230032, China.

出版信息

Se Pu. 2010 Jun;28(6):561-5. doi: 10.3724/sp.j.1123.2010.00561.

DOI:10.3724/sp.j.1123.2010.00561
PMID:20873576
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 degrees C for 24 h, room temperature.for 4 h, -80 degrees C for 30 days and freeze-thaw for 2 cycles. This method was finally successfully applied to pharmacokinetic study of Cantide in rhesus monkeys.

摘要

建立了一种双固相萃取(SPE)预处理结合非凝胶筛分毛细管电泳(NGCE)分析方法,用于定量测定恒河猴血浆中的反义寡脱氧核苷酸坎地德。对SPE条件和NGCE分析条件进行了优化。在优化条件下(SPE条件:上样缓冲液pH为9.0;阴离子交换柱的上样体积和洗脱液体积分别为5 mL和3 mL。NGCE分析条件:上样凝胶时间为30 min,分离电压为24 kV),坎地德在恒河猴血浆中的线性动态范围为1.95 - 250 mg/L,相关系数(r)大于0.998。定量限为1.95 mg/L。批内准确度范围为93.38%至100.71%,批内相对标准偏差(RSD)小于11%。批间准确度为89.46%至103.46%,批间RSD小于9%。稳定性实验表明,坎地德血浆样品在4℃储存24 h、室温储存4 h、-80℃储存30天以及冻融2个循环时均稳定。该方法最终成功应用于坎地德在恒河猴体内的药代动力学研究。

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