高效液相色谱法测定大鼠血浆中的西他列汀浓度及其药动学研究

doi:10.5246/jcps.2011.01.009

高效液相色谱法测定大鼠血浆中的西他列汀浓度及其药动学研究

酒向飞, 尚德为, 陈烨, 李新刚, 万小蒙, 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:2010-07-21 修回日期:2010-10-15 出版日期:2011-01-15 发布日期:2011-01-15
通讯作者: 周田彦*, 卢炜*

A high performance liquid chromatography method for the quantitative determination assay of sitagliptin in rat plasma and its application in pharmacokinetics study

Xiang-Fei Jiu, De-Wei Shang, Ye Chen, Xin-Gang Li, Xiao-Meng Wan, 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 Sciences, Peking University Health Science Center, Beijing 100191, China

Received:2010-07-21 Revised:2010-10-15 Online:2011-01-15 Published:2011-01-15
Contact: Tian-Yan Zhou*, Wei Lu*

摘要/Abstract

摘要：

本研究建立并验证了测定大鼠血浆中磷酸西他列汀浓度的高效液相色谱法, 并在大鼠体内进行药动学研究。血浆中的药物和内标(氢化可的松)由乙酸乙酯萃取。所用色谱柱为Agilent Zorbax Extend-C18柱 (250 mm×4.6 mm, 4 µm), 进样量为20 µL, 检测波长为267 nm, 柱温为30 ºC。流动相为甲醇-水( 60:40, 内含10 mM Tris和10 mM三乙胺, 并用1 M盐酸调至pH 9.0)。流速为1.0 mL/min。色谱峰具有良好的特异性。在0.75-100.0 µg/mL范围内具有良好的线性关系(r>0.9957)。定量下限为0.75 µg/mL。日间, 日内精密度均小于10%。在0.75, 10.0, 100.0 µg/mL的相对回收率为105.3%, 99.8%, 99.0%, 提取回收率分别为81.5%, 82.4%, 84.5%。样品的稳定性良好。运用该方法测定了大鼠口服磷酸西他列汀水溶液后的血药浓度, 说明本方法可用于药物动力学的研究。

关键词: 西他列汀, 高效液相色谱, 大鼠血浆, 药物动力学

Abstract:

A new high-performance liquid chromatography (HPLC) method for the quantitative determination of sitagliptin in rat plasma was developed and validated for pharmacokinetics study. The plasma was spiked with the internal standard (hydrocortisone, IS), treated with sodium hydroxide, and extracted with ethyl acetate. The extracted analyte was injected into an Agilent Zorbax Extend-C18 column (250 mm×4.6 mm, 4 µm) maintained at 30 ºC and monitored at 267 nm. The mobile phase consisting of methanol-water (60:40, v/v, containing 10 mM Tris and 10 mM triethylamine) was titrated to pH 9.0 using 1 mol/L hydrochloric acid. The flow rate was 1.0 mL/min. The method showed high specificity. Calibration curves of the peak area ratio of each analyte/IS versus sitagliptin concentration were linear in the range of 0.75-100.0 µg/mL (r>0.9957). The lower limit of quantification (LLOQ) was 0.75 µg/mL. The intra-day and inter-day coefficient of variation was lower than 10%. The accuracy (relative recovery) at three levels was 105.3% (0.75 µg/mL), 99.8% (10.0 µg/mL) and 99.0% (100.0 µg/mL). The extraction recovery was 81.5%, 82.4% and 84.5% at the concentrations of 0.75, 10.0 and 100.0 µg/mL, respectively. The short-term, long-term, freeze-thaw storage stability of plasma samples and the stability of standard solutions were satisfactory. This HPLC method is suitable for determining the concentration of sitagliptin in rat plasma and it was applied to determine the concentration-time profiles of sitagliptin in rat plasma following oral administration of sitagliptin.

Key words: Sitagliptin, HPLC, Rat plasma, Pharmacokinetics

中图分类号:

R917

Supporting:

Foundation items: National Integrity Innovational Technology Platform of New Drug and Research and Development (Grant No. 2009ZX09301-010) and National Fund for Talent Training in Basic Science.

^*Corresponding author. Tel./fax: 86-10-82801717

酒向飞, 尚德为, 陈烨, 李新刚, 万小蒙, 周田彦^*, 卢炜^*. 高效液相色谱法测定大鼠血浆中的西他列汀浓度及其药动学研究[J]. , DOI: 10.5246/jcps.2011.01.009.

Xiang-Fei Jiu, De-Wei Shang, Ye Chen, Xin-Gang Li, Xiao-Meng Wan, Tian-Yan Zhou^*, Wei Lu^*. A high performance liquid chromatography method for the quantitative determination assay of sitagliptin in rat plasma and its application in pharmacokinetics study[J]. , DOI: 10.5246/jcps.2011.01.009.

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