HPLC测定人血浆中表柔比星浓度

 目的 建立人血浆中表柔比星的HPLC测定法，为人体药动学研究和药效学研究提供方法学基础。方法 取血浆0.5 mL，以柔红霉素作为内标，采用氯仿-异丙醇（4∶1）两次提取。常温下氮气挥干，残渣用流动相复溶，40 μL进样。色谱条件：YWG-C₁₈柱；流动相：乙腈-磷酸二氢氨溶液-冰醋酸（35∶65∶0.5），流速为1.0 mL·min^-1；荧光检测器的激发波长480 nm, 发射波长560 nm。结果 本方法在5～500 ng·mL^-1的范围内线性良好，相关系数r=0.999 2（n=6）。高、中、低3个浓度的质控样品批间及批内变异RSD都在2.84%～5.22%之间，方法学回收率在99.5%～101.7%之间。结论 本法取样量小，操作简便、快速，适于国内多数实验室和临床研究的需要。

Improved HPLC assay for the determination of epirubicin in human plasma

OBJECTIVE To develop a high-performance liquid chromatographic (HPLC) assay for the determination of epirubicin in human plasma.METHOD Duplicate 0.5 mL plasma samples containing daunorubicin as the internal standard were extracted with chloroform:isopropanol (4∶1) in a two step extraction. The organic layer was evaporated to dryness under nitrogen stream at room temperature. The residue was reconstituted in 60 μL of the mobile phase, of which 40 μL was analyzed through a YWG-C_{18 column. The mobile phase was consisted of acetonitrile:NH4H2PO4 ^{solution:glacial acetic acid (35∶65∶0.5) at a flow rate of 1.0 mL·min-1.The drug was analyzed with a fluorescence detector using an excitation wavelength at 480nm and emission wavelength at 560 nm.RESULTS A linearity was obtained from 5 to 500 ng·mL-1 of epirubicin in plasma with a good correlation coefficient (r=0.9992, n=6).The intraday and interday validations were less than 5.22%, and the mean recoveries were 99.5%, 100.4% and 101.7% for the low, middle and high concentrations of check samples, respectively.CONCLUSION The improved method was sensitive, specific and simple. It is practical and suitable for clinical pharmacokinetic study.}}