摘要
建立了一种简便、灵敏的氯甲酸芴甲酯(FMOC-Cl)柱前衍生反相高效液相色谱-荧光检测血浆中奈替米星的新方法,同时研究了其药代动力学。对色谱条件进行了优化,采用ZORBAX Eclipse XDB-C8柱(150mm×4.6mm,5μm),流动相为乙腈-水(体积比为85∶15),流速为1.0mL/min,荧光检测激发波长为265nm,发射波长为315nm,得到奈替米星的平均加标回收率为96.62%～100.84%(n=3),对奈替米星检测的线性范围为0.045～8.88mg/L,相关系数为0.999 3,方法的日内与日间精密度分别低于3%与3.5%,最低检出限(S/N=3)与定量限(以3倍检出限计)分别为0.01和0.03mg/L。方法简便、快速、灵敏,样品用量少(30μL奈替米星血浆溶液已能满足该药含量的测定以及药物代谢的研究),为大鼠体内奈替米星的药代动力学研究提供了可靠的分析手段。
A new, simple and sensitive method based on pre-column derivatization by reversed- phase high performance liquid chromatography （ HPLC ） is described for the separation and quantification of netilmicin in plasma, using 9-fluorenylmethyl chloroformate （FMOC-Cl） as the derivatization reagent. Its pharacokinetics is also presented. The derivatization modes and chromatographic conditions were optimized. The separation was performed on an Agilent ZOR-BAX Eclipse XDB-C, column （150 mm×4.6 mm,5 μm） with a mixture of water-acetonitile （15： 85, v/v） as mobile phase and the flow rate was 1.0 mL/min. The excitation wavelength was 265 nm and the emission wavelength was 315 nm. The linear range was 0. 045 -8.88 mg/L and the correlation coefficient （r） was 0. 999 3. The limit of detection （ LOD ） （ S/N= 3 ） was about 0.01 mg/L, and the limit of quantification was 0.03 mg/L （3LOD） for netilmicin. The relative standard deviation was less than 3% for intra-day assay （ n = 5 ） and 3.5% for inter-day assay （ n = 5 ） and the relative recovery was in the range of 96.62% - 100.84% （ n = 3 ）. The plasma volume of 30 μL was sufficient for the determination of netilmicin. The method provides a reliable bioanalytical methodology to carry out netilmicin pharmacokinetics in rat plasma.
出处
《色谱》
CAS
CSCD
北大核心
2009年第6期794-798,共5页
Chinese Journal of Chromatography
基金
国家自然科学基金项目(No.20475045)
重庆市重点实验室资助项目(No.CSTC2006CA8006)
关键词
柱前衍生化
氯甲酸芴甲酯(FMOC-Cl)
高效液相色谱
奈替米星
药代动力学
大鼠血浆
pre-column derivatization
9-fluorenylmethyl chloroformate（FMOC-Cl）
high performance liquid chromatography（HPLC）
netilmicin
pharmacokinetics
rat plasma
