反相高效液相色谱法测定兔房水中莫索尼定的含量  

Detection of moxonidine levels in rabbit aqueous humor by reversed phase high performance liquid chromatography

作　　者：张倩[1] 孙乃学[2] 王嗣岑[3] 

机构地区：[1]西安市第四医院眼科,陕西省西安市710004 [2]西安交通大学第二医院眼科,陕西省西安市710004 [3]西安交通大学药学院,陕西省西安市710068

出　　处：《眼科新进展》2010年第4期301-303,共3页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30170995)~~

摘　　要：目的应用反相高效液相色谱法（reversed phase high performance liquid chroma-tography,RP-HPLC）建立兔房水中莫索尼定（moxonidine,MOX）的定量分析方法。方法兔眼滴入2g.L-1MOX滴眼液50μL后,于预定时间点行耳缘静脉空气栓塞处死兔,迅速摘除眼球,抽取房水样品0.2mL经固相萃取后,用RP-HPLC检测MOX的标准曲线、MOX的回收率、MOX的日内和日间精密度。色谱条件：色谱柱为Kromasil,C18（5μm,4.6mm×250.0mm）柱,检测波长254nm,柱温为室温,流速0.3mL·min-1,流动相：甲醇-水-三乙胺（50.050.00.5）。结果MOX保留时间为12.023min,兔房水的MOX药物峰与杂质峰分离良好,无杂质峰干扰,以含量C（μg）对峰面积AUC回归得标准曲线方程：A=293612C-127.37,r=0.9988,R2=0.9978,在0.002~0.040μg范围内含量与测定面积间具有良好的线性关系,最低定量限为0.002μg;在0.1mg·mL-1、0.4mg·mL-1和1.0mg·mL-13种浓度的平均回收率分别为77.50%、75.63%、84.25%,日内和日间变异系数分别为2.93%、2.89%,均在10%以下。结论建立了简便、敏感的RP-HPLC检测法,可用于MOX的眼内药代动力学研究。Objective To establish the method for quantitatively analyzing the moxonidine（MOX）levels in rabbit aqueous humor by reversed phase high performance liquid chromatography（RP-HPLC）.Methods After dropping 2 g·L-1 MOX eyedrops for 50 μL into eyeballs,rabbits were sacrificed at preset times by air embolism of vein at ear limbus.The eyeballs were quickly taken out.About 0.2 mL aqueous humor was collected and treated with solid phase extraction.RP-HPLC was used to detect the stan-dard curve,recovery rate,intra-day and inter-day precision of MOX.About chromatographic conditions,chromatographic column was a Kromasil,C18 column（5 μm,4.6×250 mm）;The wave length was 254 nm;The temperature of column was room temperature;The flow-rate was 0.3 mL·min-1;The mobile phase consisted of metnanol-water-triethylamine（50.050.00.5）.Results The remaining time of MOX was 12.023 min.The drug peak was separated well from foreign matter peak.The standard curve equation was get between the concentration（μg）and peak area AUC（A=293 612C-127.37,r=0.998 8,R2=0.997 8）.There was linear correlation between peak area and concentration within the range of 0.002 μg to 0.040 μg,and the minimal detectable limit was 0.002 μg.The average recovery rates were 77.50%,77.63% and 84.25% at the concentration of 0.1 μg·mL-1,0.4 μg·mL-1 and 1.0 μg·mL-1,respectively.The coefficient of variability for the inter-day and intra-day were all less than 10%.Conclusion RP-HPLC is a simple and sensitive assay,which can be used in ocular pharmacokinetic studies of MOX.

关 键 词：莫索尼定 反相高效液相色谱法 兔房水 定量检测 

分 类 号：R777.2[医药卫生—眼科]