固相萃取-高效液相色谱法测定大鼠血浆中大黄素甲醚及其药动学研究

Determination of physcion in plasma by SPE-HPLC and its pharmacokinetic study in rats

目的:建立血浆中大黄素甲醚的血药浓度测定方法,研究大黄素甲醚在大鼠体内的药代动力学.方法:大鼠分高(105.6 mg/kg)、中(52.8 mg/kg)、低(26.4 mg/kg)3个质量分数灌胃给予大黄素甲醚后,于0.083~24.000 h内12个不同时间点采集大鼠血浆,采用固相萃取法处理血浆样品,高效液相色谱-荧光检测法(HPLCFLD)内标法测定血药浓度.采用Venusil XBP C18(L)(4.6 mm×250 mm,5μm)色谱柱,流动相为V_(甲醇)∶V_(0.1%磷酸水)=70∶30,柱温30℃,激发波长435 nm,发射波长515 nm,以DAS2.0软件拟合计算药动学参数.结果:大黄素甲醚的血药质量浓度在0.024 2~5.920 0μg/m L范围内线性关系良好(r=0.992 7),检测限为0.008 1μg/m L,定量限为0.024 2μg/m L,提取回收率均大于90%,日内、日间精密度RSD均小于6%,高(105.6 mg/kg)、中(52.8 mg/kg)、低(26.4 mg/kg)3个质量分数大黄素甲醚药代动力学参数t_(1/2)分别为(10.97±6.6)、(14.23±11)、(13.25±5.6)min;ρ_(max)质量浓度分别为(0.49±0.17)、(0.41±0.15)、(0.29±0.12)μg/m L,AUC_(0-t)每小时质量浓度分别为(3.28±1.3)、(1.98±1.4)、(1.91±1.5)μg/m L.结论:本实验所建立的SPE-HPLC适用于大黄素甲醚在大鼠体内的血药浓度测定及其药代动力学研究,操作简便、快速、灵敏.

Aim： To establish a method for determination of physcion in rats plasma by solid phase extraction-high performance liquid chromatography,and study the pharmacokinetic. Methods： Rats were i. g administered with physcion at doses of 105. 6、52. 8 and 26. 4 mg/kg,blood samples were collected at time intervals. The plasma concentrations of sample in rats were assayed by SPE. The plasmaconcentrations of physcion were determined by HPLC-FLD method and internal standard method.Physcion was eluted on a Venusil XBP C18（L）（4. 6 mm × 250 mm,5 μm） column,The mobile phase was composed of 0. 1% phosphoric acid water water and methanol with gradient elution. The flow rate was1. 0 m L/min and the column temperature was 30 ℃; The excitation wavelength and emission wavelength for fluorescence detector were 435 nm and 515 nm. The pharmacokinetic parameters were calculated by the software of DAS 2. 0. Results： In rat plasma,physcion was linear in the range of 0. 024 2 - 5. 920 μg/m L（r = 0. 992 7）. Detection limit（LOD） and quantification limit（LOQ） were 0. 008 1 and 0. 024 2 μg/m L respectively. The extraction recoveries were higher than 90%,and RSDs of intra-day and inter-day were both lower than 6%. The main pharmacokinetic parameters of three test groups were as follows： t_（1/2）were（10. 97 ± 6. 6） 、（14. 23 ± 11） and（13. 25 ± 5. 6） min,respectively; ρ_（max）were（0. 49 ± 0. 17） 、（0. 41± 0. 15） and（0. 29 ± 0. 12） μg/m L,respectively; AUC_（0-t）were（3. 28 ± 1. 3） 、（1. 98 ± 1. 4） and（1. 91 ±1. 5） μg/m L per hour,respectively. Conclusion： The established method is accurate,precise and specific,and can be applied to determine the pharmacokinetic of physcion.