自制多西紫杉醇脂质体在家兔肝组织中浓度的测定

Determination of docetaxel liposome in rabbit hepatic tissue by HPLC

目的：建立以固相萃取-高效液相色谱法（SPE—HPLC）测定家兔肝组织匀浆中多西紫杉醇的方法，考察自制多西紫杉醇脂质体家兔耳缘静脉给药24h后在肝组织中的浓度。方法：以地西泮为内标，家兔肝组织匀浆样品中多西紫杉醇经固相萃取后，采用大连依利特Hypersil C18（250mm×4.6mm，5μm）色谱分析柱，流动相为乙腈-水（47：53），流速1．0mL·min^-1，检测波长230nm，进样量50μL。家兔耳缘静脉注射给予多西紫杉醇脂质体制剂，剂量为7．5mg·kg^-1，24h后解剖动物，用所建立的HPLC法测定其肝组织中药物浓度。结果-地西泮和多西紫杉醇的保留时间分别是10．9min和13．1min，多西紫杉醇的最低检测质量浓度为5μg·L^-1（S／N=3），肝组织样品中多西紫杉醇在0．025～1．5mg·L^-1范围内与峰面积比之间线性关系良好，回归方程为Y：196．42C-0．6424，r=0．9992（n=7），提取回收率为81．36％～86．72％，方法回收率为91．37％～100．34％，日内RSD为2．72％，3．20％，3．55％，日间RSD为4．68％，4．47％，3．52％，且稳定性良好。实测6例多西紫杉醇脂质体在家兔肝组织中的质量浓度为0．039-0．063mg·L^-1。结论：本试验建立的方法简便灵敏，干扰小，重复性好，可实际应用于多西紫杉醇在家兔肝组织中的测定，并作为评价多西紫杉醇脂质体在靶组织分布的方法学依据。

OBJECTIVE To establish a SPE-HPLC method for determination of docetaxel in rabbit hepatic tissue, and to investigation the docetaxel liposome concentration in rabbit hepatic tissue after 24 h intravenous administration of a single dosage of 7.5 mg. kg^-1. METHODS Diazepam was used as internal standard. The docetaxel was extracted with tert-butyl methyl ether for three times and determined through Hypersil C18 （250 mm × 4. 6 mm, 5 μm） column. The mobile phase consisted of water-acetonitrile （53：47） with the flow rates of 1. 0 mL·min^-1. The detection wavelength was 230 nm. Docetaxel liposome was intravenously administrated to rabbits at a single dosage of 7.5 mg·kg^-1. The rabbits were dissectted after 24 h and docetaxel of hepatic tissue was determined by HPLC. RESULTS In this system, the retention times of diazepam and docetaxel were 10. 9 min and 13. 1 min, respectively. The detection limit for docetaxel was 5μg·L^-1. Docetaxel concentration presented a good linear range of 0. 025 - 1.5 mg. L-1 （r = 0. 999 2）. The extraction recoveries were 81.36% - 86. 72%, and the method recoveries were 91. 37% - 100. 34%, within-day RSDs were 2. 72%, 3.20% and 3.55%, and the between-day RSDs were 4. 68%,4. 47% and 3.52%. The six cases docetaxel concentration in rabbit hepatic tissue were 0. 039 - 0. 063 mg·L^-1. CONCLUSION The method is reliable,convenient and efficient and can be applied to the determination of docetaxel concentration of rabbit hepatic tissue in the tissue distribution research of docetaxel liposome.