HPLC检测大鼠血浆中姜黄素的浓度及其药动学研究

目的建立大鼠血浆姜黄素检测的高效液相色谱方法,研究姜黄素的药动学。方法血浆经乙酸乙酯萃取,以ZORBAX SB-C18（4.6 mm×150 mm,5μm）为色谱柱;流动相为乙腈-水-0.1%TFA（50∶30∶20）,流速为1.0 mL.min-1;检测波长为302（0～5.5 min）、426 nm（5.5～7.5 min）。6只SD大鼠,♂,单剂量静脉给予10 mg.kg-1姜黄素,分别在给药后多点尾静脉采血。用该方法检测血浆中姜黄素的浓度;用DAS计算药动学参数。结果姜黄素浓度在0.05～6.00 mg.L-1内线性关系良好（r=0.999 8）;定量下限为0.05 mg.L-1;低（0.10 mg.L-1）、中（1.00 mg.L-1）、高（4.00 mg.L-1）3个浓度的回收率分别为（99.3±5.4）%,（104.2±4.7）%和（99.8±2.0）%;日内RSD分别为4.49%,3.90%和1.72%,日间RSD分别为4.61%,4.27%和2.00%。大鼠姜黄素静脉注射后,姜黄素在大鼠体内符合二室模型;α相和β相消除半衰期分别为0.08 h和0.75 h。结论本方法准确可靠、简便快速,适用于大鼠血浆姜黄素浓度的测定及其药代动力学研究。

Determination of Curcumin in Rat Plasma by HPLC and Study of its Pharmacokinetics

OBJECTIVE To develop a high performance liquid chromatography method for the determination of curcumin in rat plasma and study its pharmacokinetics.METHODS Plasma was extracted by ethyl acetate.The analytical column was packed with ZORBAX SB-C18(4.6 mm×150 mm,5 μm).The mobile phase was acetonitrile-water-0.1% trifluoroacetic acid(50:30:20) and the flow rate was 1.0 mL.min-1.The UV detection wavelength was 302 nm(0-5.5 min) and 426 nm(5.5-7.5 min).Six SD rats,♂,were given a single dose of 10 mg.kg-1 curcumin with intravenously injection into the sublingual vein.Blood samples were collected from the tail vein at different time points after injection.The concentration of curcumin in plasma were detected by the established HPLC method.The pharmacokinetics parameters were analyzed by DAS program.RESULTS Excellent liner relationship was obtained in the range of 0.05-6.00 mg.L-1(r=0.999 8),the limit determination of curcumin was 0.05 mg.L-1.The recoveries were(99.3±5.4)%,(104.2±4.7)%and(99.8±2.0)% respectively at three concentrations(0.10,1.00,4.00 mg.L-1),the intra-day RSD were 4.49%,3.90% and 1.72% and inter-day RSD were 4.61%,4.27% and 2.00%,respectively.After intravenously injection of curcinum,curcinum was fitted the two-compartment model,the half-life of α phase and β phase was 0.08 h and 0.75 h,respectively.CONCLUSION The method is accurate,simple,rapid and could be used to determine the curcinum concentration in rat plasma and study its pharmacokinetics.