高效液相色谱法定量测定美氟尼酮(吡非尼酮的新衍生物)的浓度及其在肝微粒体中的初始药代动力学研究

doi:10.5246/jcps.2018.03.020

中国药学(英文版) ›› 2018, Vol. 27 ›› Issue (3): 193-200.DOI: 10.5246/jcps.2018.03.020

高效液相色谱法定量测定美氟尼酮(吡非尼酮的新衍生物)的浓度及其在肝微粒体中的初始药代动力学研究

陈燕芬^2#, 孙萌^1#, 张世喜³, 程泽能^1*, 胡高云¹, 朱曲波^1*

1. 中南大学药学院, 湖南长沙410013
2. 厦门大学附属中山医院药学部, 福建厦门 361004
3. 广州南鑫药业有限公司, 广东广州 510630

收稿日期:2017-12-10 修回日期:2018-01-15 出版日期:2018-03-31 发布日期:2018-02-27
通讯作者: Tel.: +86-731-82650451, Fax: +86-731-82650446, E-mail: chengzn@csu.edu.cn; biqbz@hotmail.com
基金资助:
The National Natural Science Foundation of China (Grant No. 81302819), the National Natural Science Foundation of China (Grant No. C0709-31201056), the Fundamental Research Funds for the Central Universities of China (Grant No. 7601110179) and the Fundamental Research Funds for the Central south Universities of China (Grant No. 2016zzts494).

Development and validation of an HPLC assay for the quantitation of mefunidone, a novel derivative of pirfenidone, and an initial pharmacokinetics study in liver microsomes

Yanfen Chen^2#, Meng Sun^1#, Shixi Zhang³, Zeneng Cheng^1*, Gaoyun Hu¹, Qubo Zhu^1*

1. The School of Pharmaceutical Sciences in Central South University, Changsha 410013, Hunan, China
2. Department of Pharmacy, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, China
3. The Guangzhou South Xin Pharmaceutical Co, Guangzhou 510630, Guangdong, China

Received:2017-12-10 Revised:2018-01-15 Online:2018-03-31 Published:2018-02-27
Contact: Tel.: +86-731-82650451, Fax: +86-731-82650446, E-mail: chengzn@csu.edu.cn; biqbz@hotmail.com
Supported by:
The National Natural Science Foundation of China (Grant No. 81302819), the National Natural Science Foundation of China (Grant No. C0709-31201056), the Fundamental Research Funds for the Central Universities of China (Grant No. 7601110179) and the Fundamental Research Funds for the Central south Universities of China (Grant No. 2016zzts494).

摘要/Abstract

摘要：

In the present study, a simple and reliable HPLC-UV method was developed for the determination of mefunidone. The bioanalytical specific procedure involved extraction of mefunidone from a 500-µL hepatic microsomal system through protein precipitation by methanol. Chromatographic separation was achieved using an Agilent TC-C₁₈ column (4.6 mm×250 mm, 5 µm) with an isocratic mobile phase consisting of 10 mM ammonium formate (pH 2.9, later adjusted by using 10% formic acid)–acetonitrile (70:30, v/v) at a flow rate of 1.0 mL/min. The UV detection wavelength was set at 245 nm. Mefunidone and pirfenidone (PFD, internal standard, IS) were eluted at 6.0 and 9.7 min, separately, with the total running time of 12 min. According to US Food and Drug Administration bioanalytical guidelines, method validation was performed, and the results met the acceptance criteria in details. The calibration curve of mefunidone in liver microsomes was linear over the concentration range of 0.5–16 µg.mL^–1. Intra- and inter-run precisions of mefunidone were less than 9.0%, and the biases were within ±10.0%. After incubation of mefunidone in liver microsomes, this method was successfully applied to the pharmacokinetic study.

关键词: Mefunidone, HPLC assay, Pharmacokinetics, Liver microsome, Derivative

Abstract:

本研究开发了一种简单可靠的HPLC-UV方法用于美氟尼酮的测定。生物分析步骤包括从500 μL肝微粒体系统中通过甲醇沉淀蛋白质提取美氟尼酮。色谱条件:色谱柱为Agilent TC-C₁₈柱(4.6 mm×250 mm, 5 μm), 流动相为10 mM甲酸铵(用10%的甲酸调PH至2.9)–乙腈(70:30, v/v),流速为1.0 mL/min, UV检测波长设定在245 nm。美氟尼酮和吡非尼酮(内标物)分别在6.0和9.7分钟洗脱,总运行时间为12分钟。根据美国食品和药物管理局生物分析指南,进行了方法验证,结果符合验收标准。美氟尼酮在肝微粒体中的标准曲线在0.5–16 μg/mL范围内呈线性关系。美氟尼酮内和外间精确度低于9.0％,偏差在±10.0％以内。美氟尼酮在肝微粒体中孵育后,该方法成功应用于药代动力学研究。

Key words: 美氟尼酮, HPLC分析, 药代动力学, 肝微粒体, 衍生物

中图分类号:

R969.1

Supporting:

陈燕芬, 孙萌, 张世喜, 程泽能, 胡高云, 朱曲波. 高效液相色谱法定量测定美氟尼酮(吡非尼酮的新衍生物)的浓度及其在肝微粒体中的初始药代动力学研究[J]. 中国药学(英文版), 2018, 27(3): 193-200.

Yanfen Chen, Meng Sun, Shixi Zhang, Zeneng Cheng, Gaoyun Hu, Qubo Zhu. Development and validation of an HPLC assay for the quantitation of mefunidone, a novel derivative of pirfenidone, and an initial pharmacokinetics study in liver microsomes[J]. Journal of Chinese Pharmaceutical Sciences, 2018, 27(3): 193-200.

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高效液相色谱法定量测定美氟尼酮(吡非尼酮的新衍生物)的浓度及其在肝微粒体中的初始药代动力学研究

Development and validation of an HPLC assay for the quantitation of mefunidone, a novel derivative of pirfenidone, and an initial pharmacokinetics study in liver microsomes

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