色谱

色谱 ›› 2011, Vol. 29 ›› Issue (02): 157-161.DOI: 10.3724/SP.J.1123.2011.00157

• 研究论文 • 上一篇    下一篇

高效液相色谱-共振瑞利散射光谱法测定大鼠血清中的奈替米星

刘承伟1, 徐勤1, 张丹2, 卢昕2*, 赵书林2   

  1. 1. 桂林医学院基础医学院, 广西 桂林 541004; 2. 药用资源化学与药物分子工程教育部重点实验室, 广西师范大学化学化工学院, 广西 桂林 541004
  • 收稿日期:2010-10-08 修回日期:2010-11-26 出版日期:2011-02-28 发布日期:2007-03-25
  • 通讯作者: 卢昕,教授,研究方向为液相色谱方法及其在生物、医药领域的应用.
  • 基金资助:

    国家自然科学基金项目(No. 20665002)和广西自然科学基金项目(桂科自0991094).

Determination of netilmicin in rat serum using high performance liquid chromatography and resonance Rayleigh scattering

LIU Chengwei1, XU Qin1, ZHANG Dan2, LU Xin2*, ZHAO Shulin2   

  1. 1. College of Basic Medicine, Guilin Medical University, Guilin 541004, China; 2. Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 541004, China
  • Received:2010-10-08 Revised:2010-11-26 Online:2011-02-28 Published:2007-03-25

PDF

459

摘要： 建立了高效液相色谱-共振瑞利散射光谱联用测定大鼠血清中奈替米星的方法。采用的色谱条件为: 以C18柱为分离柱,以甲醇(含0.22%三氟乙酸)-20 mmol/L醋酸钠水溶液(8:92, v/v)为流动相进行等度洗脱。以滂胺天蓝为分子探针,奈替米星在柱后与滂胺天蓝结合生成离子缔合物,产生强烈的共振瑞利散射信号,荧光检测器在激发波长(λex)和发射波长(λem)均为365 nm下进行检测。在优化的检测条件下,奈替米星的检出限(信噪比为3)为0.7 mg/L。以妥布霉素为内标物,以内标法定量,奈替米星与妥布霉素的峰面积比与奈替米星的质量浓度之间的线性响应范围为1.2～30 mg/L。该方法可应用于大鼠血清中奈替米星的药代动力学研究,可为生物样品中氨基糖苷类抗生素的测定提供一种新的选择。

关键词: high performance liquid chromatography (HPLC), netilmicin, pharmacokinetics, pontamine sky blue, rat, resonance Rayleigh scattering, serum

Abstract: A method was developed for the direct determination of netilmicin in rat serum using high performance liquid chromatography and resonance Rayleigh scattering (RRS). The separation was carried out on a C18 column with the isocratic elution of the mixture of 20 mmol/L sodium acetate aqueous solution and methanol containing 0.22% trifluoroacetic acid (92:8, v/v). With the molecular recognition probe of pontamine sky blue, netilmicin formed an ion-association complex and enhanced the intensity of RRS. The RRS signal was detected by a commercial fluorescence detector at 365 nm of both excitation wavelength (λex) and emission wavelength (λem). Under the optimized conditions, the limit of detection (LOD, S/N=3) for the netilmicin was 0.7 mg/L. With the internal standard (IS) of tobramycin, a linear calibration curve ranged from 1.2 mg/L to 30 mg/L was obtained. The presented method can be used for the pharmacokinetics study of netilmicin in rat serum, and can be a new choice for the determination of aminoglycoside antibiotics in biological samples.

Key words: netilmicin, pharmacokinetics, pontamine sky blue, rat serum, resonance Rayleigh scattering, high-performance liquid chromatography