固相萃取-HPLC法测定MCF-7细胞内、外液中多西他赛的含量  

Content Determination of Docetaxel in MCF-7 Cell Extracellular and Intracellular Fluid by SPE-HPLC

作　　者：柯昌毅[1] 车坷科[1] 王丽娟[2] 

机构地区：[1]重庆市第三人民医院,重庆市400014 [2]重庆医药高等专科学校,重庆市400030

出　　处：《中国药房》2011年第17期1569-1571,共3页China Pharmacy

摘　　要：目的:建立测定人乳腺癌细胞MCF-7内、外液中多西他赛(DOC)含量的方法。方法:采用高效液相色谱法。分别制备细胞内液和细胞外液样品,经三氯乙酸除蛋白,固相萃取柱提取,色谱柱为Symmetry-C18,柱温为30℃,流动相为醋酸盐缓冲液(pH5.0)-乙腈=50∶50,检测波长为230nm。结果:DOC在细胞内、外液中检测浓度线性范围均为0.05～5.50μg·mL-1(r=0.9993、r=0.9990),平均提取回收率(n=5)均大于94%,日内、日间RSD(n=5)均小于5%,最低检测限分别为0.05、0.03μg·mL-1;细胞内液中DOC含量远高于细胞外液。结论:本方法简便、灵敏、干扰小,可用于MCF-7细胞内、外液中DOC含量的测定。OBJECTIVE： To establish the method for the content determination of docetaxel （DOC） in extracellular and intracellular fluid of human breast cancer cell MCF-7. METHODS： HPLC method was adopted. The samples of extracellular and intracellular fluid were prepared and the protein was removed using trichloroacetic acid. The DOC was extracted by solid phase extraction column （SPE）. The samples were separated on a symmetry-C18 column with mobile phase consisted of potassium phosphate buffer-acetonitrile （50：50, V/V）. The column temperature was 30 % and detection wavelength was set at 230 nm. RESULTS： The linear range of DOC in extracellular and intracellular fluid was 0.05-5.50 μg. mL^-1 （r= 0.999 3, r=0.999 0） with average recovery more than 94% （n=5）. The RSD of intra-day and inter-day were lower than 5% （n=5）. The lowest detection limits were 0.05 μg. mL^-1 and 0.03 μg. mL^-1. The content of DOC in intracellular fluid was much higher than in extracellular fluid. CONCLUSION： The method is simple, sensitive and with little interference, and could be applied for the content determination of DOC in extracellular and intracellular fluid of MCF-7 cell.

关 键 词：多西他赛 人乳腺癌细胞MCF-7 细胞内液 细胞外液 固相萃取-高效液相色谱法 含量测定 

分 类 号：R979.1[医药卫生—药品]