LC-MS/MS法测定家兔血浆中多奈哌齐的浓度

Determination of donepezil in rabbit plasma by LCMS/MS

目的建立测定家兔血浆中多奈哌齐浓度的LC-MS/MS法,并用该法研究多奈哌齐在家兔体内的药动学特征。方法色谱柱为Diamonsil C_(18)柱(150 mm×4.6 mm,5μm),流动相为甲醇-5 mmol·L^(-1)醋酸铵溶液-甲酸(体积比为50.0:50.0:0.1),采用沉淀蛋白法处理血浆样品,以多反应监测(multiple reaction monitoring,MRM)扫描方式检测,测定家兔给予多奈哌齐贴剂后血浆中的药物浓度。结果血浆中多奈哌齐质量浓度在1.0～500.0μg·L^(-1)内线性关系良好(r=0.996 5);日内精密度RSD≤7.9%,日间精密度RSD≤6.4%;平均提取回收率为(95.1±7.1)%(n=6),基质效应为(109.9±2.8)%(n=6)。多奈哌齐贴剂在家兔血浆中主要药物动力学参数如下:t_(1/2)为(28.2±11.4)h,ρ_(max)为(0.353±0.101)mg·L^(-1),AUC_(0-168)和AUC_(0-∞)分别为(15.6±6.4)和(15.7±6.4)mg·h·L^(-1)。结论该方法适用于多奈哌齐在家兔体内药物动力学的研究。

Objective To develop a simple, rapid and sensitive liquid chromatography-tandem mass spectrom- etry（LC-MS/MS） method for the determination of donepezil in rabbit plasma, and apply the method to study the pharmaceutical characteristic of donepezil in rabbit plasma. Methods The chromatographic separa- tion was accomplished on Diamonsil C18 analytical column （150 mm x 4. 6 mm,5 μm） at a flow rate of 1.0 mL· min - 1 with the mobile phase consisting of methanol ：5 mmol· L - 1 ammonium acetate aqueous solu- tion：formic acid （ V： V： V --50. 0：50. 0：0. 1 ）. Protein precipitation was used for the sample pretreatment with a plasma volume of 50 μL. The plasma concentration was detected by multiple reaction monitoring after the rabbit was given donepezil hydrochloride patch. Results The method was linear over the range of 1.0- 500. 0 μg·L-1 ;the intra- and inter-day precision were 7.9% and 6.4% ,respectively. The average recover- ies were between （95. 1±7. 1 ） %. The matrix effects were between （ 109.9 ±2. 8） %. The main pharma- cokinetic parameters were as follows： t1/2 （ 28.2 ± 11.4 ） h, Pmax （ 0. 353 ± 0. 101 ） mg · L-1 , AUCo-168 （ 15.6 ± 6. 4） mg- h. L - 1, AUCo-∞ （ 15.7 ± 6.4 ） mg. h. L - 1, respectively. Conclusions This simple and sensitive LC-MS/MS method was successfully applied to pharmacokinetic studies of donepezil in rabbit plas- ma.