HPLC/MS考察大鼠和肝微粒体CYP450活性及硝苯地平代谢研究

温彬宇;王嗣岑

中国药学杂志 ›› 2009, Vol. 44 ›› Issue (07) : 506-509.

中国药学杂志 ›› 2009, Vol. 44 ›› Issue (07) : 506-509.
论著

HPLC/MS考察大鼠和肝微粒体CYP450活性及硝苯地平代谢研究

  • 温彬宇;王嗣岑
作者信息 +

Determination of in Vitro CYP450 Activity with Nifedipine as a Probe Drug by HPLC-MS

  • WEN Bin-yu,WANG Si-cen
Author information +
文章历史 +

摘要

目的建立以硝苯地平为探针药物测定大鼠和犬肝微粒体活性的HPLC/MS。方法采用SHIMADZU VP-ODS(2.0mm×150mm,5μm)色谱柱;柱温为室温;以甲醇-水(55∶45)为流动相,流速:0.2mL·min-1。选择正离子扫描(SIM)m/z 345。以硝苯地平为探针药物与大鼠或Beagle犬的肝微粒体进行体外孵育,以液相色谱串联质谱法测定硝苯地平的代谢产物氧化硝苯地平。结果在所建立的HPLC/MS条件下,硝苯地平的氧化产物保留时间约为7.15min。杂质峰不干扰测定;氧化硝苯地平最低检测限为3μg·L-1(S/N=3),线性范围为0.01~10mg·L-1,回收率及精密度均符合检测要求。结论该HPLC/MS能够准确的检测出在大鼠和Beagle犬肝微粒体中的硝苯地平氧化产物。

Abstract

OBJECTIVE To establish a HPLC/MS method to determine microsomes activity in vitro using nifedipine as a probe drug. METHODS Nifedipine was incubated with rat or Beagle dog liver microsomes in vitro,oxidized nifedipine were determined by HPLC-MS. A SHIMADZU-ODS(2.0 mm×150 mm)column as chromatographic column. The mobile phase consisted of methanol-water (55∶45) at a flow rate of 0.2 mL·min-1. MS detector was set as positive selective ion scan mode at m/z 345. RESULTS The retention time of oxidized nifedipine was 7.15 min. The detection limit in rat or Beagle dog liver microsomes was 3 μg·L-1 for oxidized nifedipine (S/N=3),and the linear range was over the range of 0.01-10 mg·L-1. The recovery and accuracy satisfied the requirement of detection for oxidized nifedipine in rat and Beagle dog liver microsomes. CONCLUSION Oxidized nifedipine can be detected accurately and precisely by using the HPLC/MS method.

关键词

HPLC/MS / 硝苯地平 / 肝微粒体

Key words

HPLC/MS / nifedipine / microsomes

引用本文

导出引用
温彬宇;王嗣岑. HPLC/MS考察大鼠和肝微粒体CYP450活性及硝苯地平代谢研究[J]. 中国药学杂志, 2009, 44(07): 506-509
WEN in-yu;WNG Si-cen. Determination of in Vitro CYP450 Activity with Nifedipine as a Probe Drug by HPLC-MS [J]. Chinese Pharmaceutical Journal, 2009, 44(07): 506-509

参考文献

[1] LENG X F,QIU X H. The Structure Function and Applying Prospect of Cytochrome P450(细胞色素P450酶系的结构、功能与应用前景)[M]. Beijing:Beijing Science Press,2001:56-75. [2] The group of the guard of un-clinic pharmaclkinetic of chemical drug. The Guard of Un-Clinic Pharmaclkinetic of Chemical Drug(化学药物非临床药代动力学研究技术指导原则)[M]. Beijing:SFDA,2005:21. [3] PATKI K C,VON MOLTKE L C,GREENBLATT D J. In vitro metabolism of midazolam,triazolam,nifedipine,and testosterone by human liver microsomes and recombinant cytochomes P450:role of CYP3A4 and CYP3A5 [J]. Drug Metab Dispos,2003,31(7):938-944. [4] DAI C L,LI S B,LIANG Y J,et al. Determination of CYP 3A4 activity in vitro [J]. Chin Pharm J(中国药学杂志),2006,41(18):1420-1423. [5] WILLIAMS J A,RING B J,CANTRELL V E,et al. Comparative metabolic capabilities of CYP3A4,CYP3A5,and CYP3A7[J]. Drug Metab Dispos,2002,30(8):883-891. [6] SHIMADA T,GUENGERICH F P. Evidence for cytochrome P-450NF,the nifedipine oxidase,being the principal enzyme involved in the bioactivation of aflatoxins in human liver [J]. Biochemistry,1989,86(2):462-465. [7] NIWA T,KANEKO H,NARITOMI Y,et al. Species and sex differences of testosterone and nifedipine oxidation in liver microsomes of rat,dog and monkey [J]. Xenobiotica,1995,25(10):1041-1049. [8] ZUBER R,ANZENBACHEROVA E,ANZENBACHER P. Cytochromes P450 and experimental models of drug metabolism [J]. J Cell Mol Med,2002,6(2):189-198. [9] GUENGERICH F,MARTINMV,BEAUNE P H,et al. Characterization of rat and human liver microsomal cytochrome P450 forms involved in nifedipine oxidation,a prototype for genetic polymorphism in oxidative drug metabolism [J]. J Biol Chem,1986,261(11):5051-5055. [10] ZHAO N P,YU L S,YAO T W,et al. Determination of palonosetron in human liver microsomes by high-performance liquid chromatography with UV-detection [J]. Chin Pharm J(中国药学杂志),2006,41(21):1654-1656. [11] ZHOU J Q,TANG Z Q,FENG B,et al. Research on influence of in situ liver perfusion and microsomal incubation on para toluene sulfonamide liver metabolism in rat [J]. Chin Pharm J(中国药学杂志),2006,41(8):615-626. [12] EDLUND P O,BARANCZEWSKI P. Identification of BVT.2938 metabolites by LC/MS and LC/MS/MS after in vitro incubations with liver microsomes and hepatocytes [J]. J Pharm Biomed Anal,2004,34(1):1079-1090. [13] ZENG S. Metabolism of Drug (药物代谢学) [M]. Zhejiang University Press,2004:249-252. [14] JIN K T,SHI S Y ,SHEN J X,et al. Progress in human liver microsomes activity by high performance liquid chromatography[J]. Chin Hosp Pharm J(中国医院药学杂志),2007,27(12):1722-1725. [15] LI Z P,ZHONG M K,SHI X J,et al. A solid phase extracted-HPLC method for determining nifedipine in human plasma [J]. Chin Journal Clin Pharm(中国临床药学杂志),2005,14(2):115-117. [16] ZHOU M J,JIAO J J,DONG W L,et al. Determination of nifedipine in plasma by reversed phase high-performance liquid chromatography [J]. Chin J Clin Pharmacol Ther(中国临床药理学与治疗学),2003,8(2):180-182.

基金

国家自然科学基金重点资助项目(30730110);陕西省自然科学基金资助项目(2006C221)

Accesses

Citation

Detail

段落导航
相关文章

/