Abstract
OBJECTIVE To develop a method using HPLC with fluorescence detector for determining the Doxorubicin concentrations in K562/DOX cell nuclei.METHODS Nuclei were isolated by modifying a method that employed isotonic conditions and detergent lysis. The mobile phase consisted of methanol-water-acetic acid (50∶50∶0.25). The flow rate of the mobile phase was set at 1.0 mL·min-1, and the eluent was monitored at an excitation wavelength of 497 nm and an emission wavelength of 555 nm. Calibration curves were computed using the area ratio of doxorubicin to daunorubicin (the internal standard).RESULTS The linear calibration curve was observed in the concentration range of 5.0-1 000 μg·L-1. The lower limit of quantification was 5.0 μg·L-1. The spotting recoveries were 98.55%-100.84%. The inter-and intra-day precision (RSD) was below 15%. CONCLUSION The method was proved to be convenient,sensitive and rapid,and suitable for the determination of doxorubicin in nuclei.
Key words
HPLC /
doxorubicin /
nuclei /
K562/DOX cell
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XU Dong-hng;GO Jin-qing;LING Wen-qun;ZHO Yong-xing;.
Determination of Doxorubicin Concentrations in MDR Cancer Cell Nuclei by HPLC [J]. Chinese Pharmaceutical Journal, 2008, 43(16): 1264-1267
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