HPLC-MS/MS测定二至方中特女贞苷和蟛蜞菊内酯含量及其在大鼠体内的药动学研究

Determination of the specnuezhenide and wedelolactone in Erzhi Formula and their pharmacokinetics in rats by HPLC-MS/MS

目的:阐明二至方中特女贞苷和蟛蜞菊内酯的含量及其在大鼠体内的药动学特性。方法:首次建立HPLC-MS/MS法同时测定特女贞苷和蟛蜞菊内酯在二至方及大鼠血清和组织中的含量,再利用DAS 2.0程序分别计算其各自主要的药动学参数,并绘制血药浓度-时间曲线。结果:特女贞苷和蟛蜞菊内酯的回归方程分别为Y=123.408+181.663X(R2=0.9999)、Y=92 127.2+3 822.94X(R2=0.9987),线性范围在25～1 000μg/L,LOD分别为2.04、2.60μg/L,提取回收率分别为101.2%和100.3%;二至方中特女贞苷的含量范围在0.27%～9.79%,蟛蜞菊内酯的含量范围在0.16%～0.61%;特女贞苷Tmax=1.50h,蟛蜞菊内酯Tmax=2.0h,蟛蜞菊内酯的Cmax=(30.24±1.65)μg/L,特女贞苷C_(max)=(6.39±0.05)μg/L,蟛蜞菊内酯的AUC(0-∞)=(123.30±2.68)μg·L^(-1)·h^(-1),特女贞苷AUC(0-∞)=(16.56±0.98)μg·L^(-1)·h^(-1)。结论:该法准确可靠,专属性强,适用于特女贞苷和蟛蜞菊内酯在中药复方、大鼠体内的药代动力学研究。

Objective： To clarify the content and pharmacokinetic properties of specnuezhenide and wedelolactone in Erzhi Formula. Methods： HPLC-MS/MS with ESI source in Negative ion mode and multiple response monitoring system was applied to establish a method for simultaneous determination of specnuezhenide and wedelolactone in Erzhi Formula, serum and tissue distribution. Immediately, the plasma concentration-time curve was drawn out and their respective major pharmacokinetic parameters was calculated by the DAS 2.0 program. Results： The regression equation of specnnezhenide and wedelolactone were Y=123.408＋181.663X （R^2=0.9999）, Y=92 127.2＋3 822.94X （R^2=0.9987）, respectively. Linear range was in 25-1000μg/L, and their LOD were 2.04 and 2.60μg/L, respectively. The recovery rates were 101.2% and 100.3% respectively. In the Erzhi Formula, the content of specnuezhenide ranged from 0.27%-9.79%, and wedelolactone was in the range of 0.16%-0.61%. The Tmax of specnuezhenide and wedelolactone was 1.50h and 2.0h, respectively. The Cmax of specnuezhenide and wedelolactone was （30.24±1.65） and （6.39±0.05）lag/L, respectively. The AUC（0-∞） of specnuezhenide and wedelolactone was （123.30±2.68） and （16.56±0.98） μg/· L^-1· h^-1, respectively. Conclusion： The method is suitable for the determination of the specnuezhenide and wedelolactone in Chinese herbal formula, rat plasma concentration and pharmacokinetics mainly due to its accurate and strong specificity.