在线SPE HPLC-DAD方法测定大鼠血浆环磷酰胺浓度

doi:10.5246/jcps.2012.02.020

在线SPE HPLC-DAD方法测定大鼠血浆环磷酰胺浓度

李晓娜, 于宁, 张建美, 林文斯, 凌笑梅^*, 富戈, 李润涛, 崔景荣

1. 北京大学医学部药物分析教研室, 北京 100191
2. 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

收稿日期:2011-12-15 修回日期:2012-02-10 出版日期:2012-03-15 发布日期:2012-03-15
通讯作者: 凌笑梅*

Quantitative determination of cyclophosphamide in rat plasma using
an on-line SPE HPLC-DAD

Xiao-Na Li, Ning Yu, Jian-Mei Zhang, Wen-Si Lin, Xiao-Mei Ling^*, Ge Fu, Run-Tao Li, Jing-Rong Cui

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

Received:2011-12-15 Revised:2012-02-10 Online:2012-03-15 Published:2012-03-15
Contact: Xiao-Mei Ling*

摘要/Abstract

摘要：

建立快速简便的在线固相萃取液相 (on-line SPE HPLC-DAD) 分析方法, 测定大鼠血浆中环磷酰胺的浓度。该方法为: 将Acclaim^® Polar Advantage II C₁₈保护柱 (PA II柱, 10 mm×4.6 mm, 5 μm) 作为在线SPE柱, 去蛋白的血浆样品进样后, 100% H₂O洗脱1 min; 经PA II C₁₈柱(150 mm×4.6 mm, 5 μm) 分析, 流动相为乙腈-水(40:60, v/v), 流速为1 mL/min, 检测波长为195 nm。经过验证发现该方法的选择性好, 线性、精密度、准确度和灵敏度高, 可用于临床监测环磷酰胺浓度。

关键词: 在线SPE, HPLC-DAD, 环磷酰胺, 大鼠血浆

Abstract:

A rapid and simple liquid chromatography method with on-line solid phase extraction was developed and validated for the quantitative determination of cyclophosphamide in rat plasma. The plasma sample was first extracted on an Acclaim^® Polar Advantage II C₁₈ guard column (PA II C₁₈, 10 mm×4.6 mm, 5 μm), which was also the on-line Extraction Cartridge SPE column, by washing with 100% H₂O for 1 min. The extracted sample was then eluted onto a PA II C₁₈ column (150 mm×4.6 mm, 5 μm) and separated by isocratic elution with acetonitrile–water (40:60, v/v). The mobile phase was run at a flow rate of 1.0 mL/min, and the UV detector was set at 195 nm. Retention time of cyclophosphamide was 4.3 min and the total run-time was 6 min. The linear range of the standard curve was from 1.0 to 200 μg/mL (r² = 0.9999), and the limits of quantification and detection were 1.0 μg/mL (RSD<10%, n = 5) and 0.3 μg/mL(RSD<13%, n = 5), respectively. Both intra- and inter-day variations were less than 5.6%. The developed method can be used for the therapeutic drug monitoring of cyclophosphamide in the clinic.

Key words: On-line SPE, HPLC-DAD, Cyclophosphamide, Rat plasma

中图分类号:

R917.1

Supporting:

Foundation items: National Natural Science Foundation of China (Grant No. 81072612), the Natural Science Foundation of Beijing (Grant No. 7102107), the Open Foundation of State Key Laboratory of Natural and Biomimetic Drugs (Grant No. K20110109) and Specialized Research Fund for the Doctoral Program of Higher Education (Grant No. 20110001110021).
^*Corresponding author. Tel.: 86-10-82801590; Fax: 86-10-82802414

李晓娜, 于宁, 张建美, 林文斯, 凌笑梅^*, 富戈, 李润涛, 崔景荣. 在线SPE HPLC-DAD方法测定大鼠血浆环磷酰胺浓度[J]. , DOI: 10.5246/jcps.2012.02.020.

Xiao-Na Li, Ning Yu, Jian-Mei Zhang, Wen-Si Lin, Xiao-Mei Ling^*, Ge Fu, Run-Tao Li, Jing-Rong Cui. Quantitative determination of cyclophosphamide in rat plasma using
an on-line SPE HPLC-DAD [J]. , DOI: 10.5246/jcps.2012.02.020.

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