高效液相离子对色谱法测定糖尿病大鼠血浆中二甲双胍浓度及其药代动力学研究

doi:10.5246/jcps.2012.03.027

高效液相离子对色谱法测定糖尿病大鼠血浆中二甲双胍浓度及其药代动力学研究

陈烨, 李汉青, 许娇娇, 酒向飞, 邓晨辉, 李新刚, 李良, 徐小晴, 周田彦^*, 卢炜^*

1. 北京大学医学部天然药物及仿生药物国家重点实验室, 北京 100191
2. 北京大学医学部药学院药剂学系, 北京 100191

收稿日期:2011-11-17 修回日期:2012-02-20 出版日期:2012-05-15 发布日期:2012-05-15
通讯作者: 周田彦*, 卢炜*

Determination of metformin in diabetic rat plasma by an improved ion-pair high-performance liquid chromatography: application to a pharmacokinetic study

Ye Chen, Han-Qing Li, Jiao-Jiao Xu, Xiang-Fei Jiu, Chen-Hui Deng, Xin-Gang Li, Liang Li, Xiao-Qing Xu, Tian-Yan Zhou^*, Wei Lu^*

1. State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center, Beijing 100191, China
2. Department of Pharmaceutics, School of Pharmaceutical Science, Peking University Health Science Center, Beijing 100191, China

Received:2011-11-17 Revised:2012-02-20 Online:2012-05-15 Published:2012-05-15
Contact: Tian-Yan Zhou*, Wei Lu*

摘要/Abstract

摘要：

本研究建立并验证了测定糖尿病大鼠血浆中二甲双胍浓度的高效液相色谱方法, 以阿替洛尔为内标, 加入10%高氯酸沉淀蛋白来制备血浆样品。色谱条件: AQ-C18极限色谱柱 (250 mm×4.6 mm, 5 μm), 流动相 (pH 5.05) 为乙腈-水 (31:69, v/v, 水相中含有0.002 M 十二烷基磺酸钠, 0.0125 M 磷酸二氢钾, 0.015 M 三乙胺), 流速1.0 mL/min。二甲双胍在7.5-4000 ng/mL范围内具有良好的线性关系 (r>0.994), 最低检测限 (LLOQ)为7.5 ng/mL, 描述精密度的相对标准偏差 (RSD) 范围为1.87%-15.70%, 准确度为93.98%-106.89%。二甲双胍和内标的回收率分别为95.40%和95.31%。不同实验条件下质控样品的稳定性相对偏差范围在±9.00%之内。该方法成功地运用到糖尿病大鼠单剂量灌胃10 mg/kg二甲双胍后的药代动力学研究中, 结果显示本研究采用AQ-C18极限色谱柱建立的离子对色谱法对强极性化合物如二甲双胍具有良好的分离效果。

关键词: 二甲双胍, 离子对高效液相色谱, 糖尿病大鼠血浆, 药代动力学, AQ-C18极限色谱柱

Abstract: An efficient and sensitive ion-pair HPLC-UV method using atenolol as internal standard (IS) was developed and validated for the determination of metformin in the plasma of diabetic rats. Plasma samples were deproteinated with 10% (v/v) perchloric acid. Separation was achieved on a Ultimate^TM AQ-C18 column (250 mm×4.6 mm, 5 μm) with a mobile phase (pH 5.05) composed of acetonitrile-water (31:69, v/v, containing 0.002 M sodium dodecyl sulfate, 0.0125 M potassium dihydrogen phosphate, 0.015 M triethylamine) at a flow rate of 1.0 mL/min. The calibration curve was linear (r>0.994) between 7.5 and 4000 ng/mL. The lower limit of quantification (LLOQ) was 7.5 ng/mL. The precision was validated and the relative standard deviation was in the range of 1.87% to 15.70%; the accuracy was between 93.98%-106.89%. The mean recoveries were 95.40% and 95.31% for metformin and IS, respectively. The relative error (RE) of stability at different storage conditions was within ±9.00%. This method was used to determine the concentration-time profile of metformin in diabetic rat plasma following an oral administration of metformin at the dose of 10 mg/kg. Our results indicated that ion-pair HPLC-UV method using Ultimate^TM AQ-C18 column was effective for the pharmacokinetic studies of high polarity compounds like metformin.

Key words: Metformin, Ion-pair HPLC, Diabetic rat plasma, Pharmacokinetics, Ultimate TM AQ-C18

中图分类号:

R969.1

Supporting:

Foundation items: National Integrity Innovational Technology Platform of New Drug and Research and Development (Grant No. 2009ZX09201-010) and Innovation Team of Ministry of Education (Grant No. BMU20110263).
^*Corresponding author. Tel./Fax: 86-10-82805763; 86-10-82801717

陈烨, 李汉青, 许娇娇, 酒向飞, 邓晨辉, 李新刚, 李良, 徐小晴, 周田彦^*, 卢炜^*. 高效液相离子对色谱法测定糖尿病大鼠血浆中二甲双胍浓度及其药代动力学研究[J]. , DOI: 10.5246/jcps.2012.03.027.

Ye Chen, Han-Qing Li, Jiao-Jiao Xu, Xiang-Fei Jiu, Chen-Hui Deng, Xin-Gang Li, Liang Li, Xiao-Qing Xu, Tian-Yan Zhou^*, Wei Lu^*. Determination of metformin in diabetic rat plasma by an improved ion-pair high-performance liquid chromatography: application to a pharmacokinetic study[J]. , DOI: 10.5246/jcps.2012.03.027.

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