姜黄素及姜黄素衍生物A在大鼠血浆中含量的测定

Determination of curcumin and curcumin derivative A in rat plasma

目的:建立大鼠血浆中姜黄素及姜黄素衍生物A的高效液相色谱(HPLC)测定方法,研究其在大鼠体内的药物动力学,为姜黄素的临床应用提供参考。方法:将姜黄素及姜黄素衍生物A分别灌胃给药,采用HPLC测定其在大鼠血浆中的血药浓度,色谱柱为Hypersil ODS2C18(5μm×4.6mm×200mm);柱温:30℃;姜黄素流动相,乙腈-水-乙酸(体积比45∶55∶1),姜黄素检测波长为420nm;姜黄素衍生物A流动相,乙腈-水-乙酸(体积比70∶30∶1);体积流量为1.0mL.min-1;姜黄素衍生物A检测波长为361nm。结果:姜黄素及姜黄素衍生物A在0.25～100.00 mg.L-1范围内与峰面积呈良好的线性关系,姜黄素回归方程为Y=105.90X-22.70,r=0.999 6;姜黄素衍生物A回归方程为Y=71.20X+24.62,r=0.999 4。两者日内、日间精密度RSD均小于4%,平均回收率均大于96%;大鼠灌胃给药后的药动学行为符合单室模型,姜黄素衍生物A的清除率比姜黄素显著降低,约是姜黄素清除率的3.57倍,曲线下面积是姜黄素的4.09倍,半衰期t1/2约为姜黄素的2.79倍。结论:HPLC测定姜黄素及姜黄素衍生物A在大鼠体内血药浓度的方法准确、简单可行、重复性好,为提高姜黄素在体内的稳定性,延长其在体内半衰期提供了依据。

Objective To develop the method of high performance liquid chromatography（HPLC） for determination of curcumin and curcumin derivative A in rat plasma and study their pharmacokinetics in rats,and to provide basis for clinical application of curcumin.Methods The curcumin and the curcumin derivative A were lavaged to the rats,and the concentrations of curcumin and curcumin derivative A in rat blood at different time points were determined by HPLC.The curcumin and curcumin derivative A were separated well on a Hypersil ODS2 C18（5 μm×4.6 mm ×200 mm）under 30℃.The mobile phase of curcumin was a mixture of acetonitrile-water containing 1% acetic acid（45∶55） at a flow rate of 1 mL·min-1.The detective wavelength was 420 nm;the mobile phase of curcumin derivative A was a mixture of acetonitrile-water containing 1% acetic acid（70∶30） at a flow rate of 1 mL·min-1.The detective wavelength was 361 nm.Results The calibration curve was liner at the range of 0.25-100.00 mg·mL-1.The regression equation of curcumin was Y=105.90X-22.70,r=0.999 6;the regression equation of curcumin derivative A was Y=71.20X＋24.62,r=0.999 4.The RSD of intra-day precision and inter-day precision was less than 4%.The average recovery was more than 96%.The plasma concentration-time curves of curcumin and curcumin derivative A in rats fit to one compartment model respectively.The clearance rate,the area under temperature and t1/2 of curcumin derivative A were 3.57,4.09 and 2.79 times respectively than those of curcumin.Conclusion The method of HPLC for the determination of curcumin and curcumin derivative A in plasma is accurate,simple and reliable,which provides the basis for the improvement of stablility and the extending of the half-time of curcumin.