高效液相色谱法测定大鼠血浆及组织中多柔比星含量

目的：建立高效液相色谱法测定大鼠血浆及组织样品中多柔比星的含量,并研究其在大鼠体内的分布特征。方法：生物样品经过液-液萃取后,采用HPLC-FLD法,色谱柱为Zorbax SB-C1（8150mm×4.6mm,5μm）,流动相为乙腈和0.01mol·L-1磷酸二氢钾缓冲液（pH2.3）,梯度洗脱,流速为1mL·min-1,柱温为35℃;荧光检测器,激发波长为237nm,发射波长为555nm。结果：该法测定血浆中多柔比星的线性范围为2.5-500ng·mL-1,r=0.9991。其他组织的分析方法也均满足生物样品的测定要求。静脉给予5mg·kg-1多柔比星,结果显示其在大鼠体内分布广泛且持久,肾脏和脾脏及毒性靶器官心脏中浓度较高。结论：该方法准确度、灵敏度高,可以测定72h内多柔比星在主要组织中的分布,可用于研究心脏毒性与多柔比星＂量＂之间相关性。

Determination of doxorubicin in rat plasma and tissues by HPLC

Objective： To establish an HPLC method for determination of doxorubicin in rat plasma and tissues, and study the distribution of doxorubicin in rat tissues. Methods： The biological samples were prepared based on a simple liquid-liquid extraction and analyzed by HPLC-FLD. The Zorbax SB-C18 （150 mm×4.6 mm, 5 μm） was used as analytical column. The mobile phase consisted of acetonitrile and 0.01 mol·L-1 monopotassium phosphate buffer （pH 2.3） at a flow rate of 1 mL·min-1 with gradient elution. The column temperature was 35 ℃. The analytes were measured by fluorescence detector with excitation wavelength of 237 nm and emission wavelength of 555 nm. Results： A linear calibration curve of doxorubicin in plasma was obtained in the concentration range of 2.5 ~ 500 ng·mL-1 （ r = 0.999 1）. The analysis methods of other tissues also satisfied the demand of bio- analysis. After iv injection of 5 mg·kg-1 of doxorubicin, the drug distributed extensively and persistently in rat. The concentrations in kidney and spleen were relatively high, and the concentration was at a high level in heart which was the toxicity target tissue. Conclusion： The method is accurate and sensitive, which can be applied to determine the distribution of doxorubicin for 72 hours in main tissues of rat and research the correlation between cardiotoxicity and quantity of doxorubicin.