一种测定MDR肿瘤细胞内外阿霉素浓度的方法

An Approach to Determination of the Intracellular and Extracellular Concentrations of Doxorubicin for MDR Cancer Cells

研究提出K5 62 /A细胞内外阿霉素浓度的反相高效液相色谱 -荧光测定法 .细胞内阿霉素浓度测定采用LichrospherC18色谱柱 ,甲醇 0 0 1%醋酸 ( 5 0∶5 0 ,V/V)流动相 ,流速 1 0mL/min ,柱温 3 5℃ ;细胞外阿霉素浓度测定采用LichrospherC18色谱柱 ,甲醇 0 0 1%醋酸 ( 5 5∶45 ,V/V)流动相 ,流速 0 8mL/min ,柱温 3 5℃ .荧光检测器波长λex=495nm ,λem=5 60nm ,均以盐酸柔红霉素为内标 .研究结果表明 ,该方法简单、准确、线性范围宽、检测限低 ,精确度和回收率良好 ,可用于多药耐药肿瘤细胞内外阿霉素浓度的动态变化规律研究 .

A method using RP-HPLC with fluorescence detector was developed for deter-mining the intracellular and extracellular concentrations of Doxorubicin (Dox) in K562/A cells. To analyze the intracellular concentration of Dox, a Lichrospher C-18 column maintained at 35 degreesC was used, and the mobile phase consisted of methanol-0.01% acetic acid solution (50: 50, V/V) at the flow rate of 1.0 mL/min. A Lichrospher C-18 column maintained at 35 degreesC was used to analyze the extracellular concentration of Dox, and the mobile phase consisted of Methanol-0.01% acetic acid solution (55:45, V/V) at the flow rate of 0.8 mL/min. Daunorubicin was used as the internal standard, and the excitation wavelength and emission wavelength were 495 nm and 560 nm respectively. The proposed method has been verified to be simple, accurate and sensitive. It has low limit of determination, wide linear range, satisfied precision, and good mean recovery. For multi-drug resistance cancer cells, the dynamic status of the intracellular and extracellular concentrations of Dox could be investigated by using this method.