液相色谱-串联质谱法测定健康人体内阿德福韦浓度及其药动学研究(英文)

目的建立人血浆和尿样中阿德福韦浓度的液相色谱-串联质谱(LC-MS/MS)测定方法。方法10名健康受试者单剂量口服阿德福韦10mg。于服药前(0h)和服药后抽取静脉血及留取尿样。采用BDS-Phenyl column(250mm×4.6mm,5μm)色谱柱,流动相为甲醇-0.35mol·L-1冰醋酸梯度洗脱,流速为0.3mL.min-1;电喷雾离子化正离子选择性反应检测;检测离子为m/z274.1→162.1(阿德福韦),m/z254.1→135.0(喷昔洛韦,内标)。结果LC-MS/MS测定阿德福韦血浆样品线性范围为0.50～200μg.L-1,尿样线性范围为50～20000μg·L-1,线性关系良好。血浆样品分析的回收率、精密度和准确度均良好,定量限为0.50μg·L-1。主要药动学参数为:ρmax(24±s4)μg·L-1,tmax(1.0±0.6)h,AUC0～t(278±45)h.μg·L-1,AUC0～∞(285±45)h.μg·L-1,t1/2(8.8±1.6)h,CL(F)(36±6)L.h-1,Vd(F)(456±136)L,0～48h的尿累计排泄量为(48±12)%。结论建立的LC-MS/MS测定法专属性强、灵敏度适宜,可以用于阿德福韦的药动学研究。

Determination of adefovir concentrations in healthy volunteers by LC-MS/MS and its pharmacokinetics

AIM To develop a sensitive and selective LC-MS/MS method for the determination of adefovir in human plasma and urine. METHODS Ten healthy Chinese volunteers participated in the single dose pharmacokinetics study. Venous blood samples and urine samples were collected pre-treatment and after the oral administration of adefovir 10 mg. The analyte was eluted on a BDS-Phenyl column (250 mm × 4.6 mm, 5 μm) with methanol-0.35 mol·L-1 acetic acid at a flow rate of 0.3 mL.min-1 with gradient elution and detected with a triple quad LC-MS / MS using ESI with positive ionization. Ions monitored in the multiple selected reaction monitoring mode were m / z 274.1 →162.1 for adefovir, and m / z 254.1 →135.0 for penciclovir ( internal standard), respectively. RESULTS A good linear correlation was found between the HPLC peak areas and concentrations of adefovir in the range from 0.50 to 200 μg·L-1 for plasma and 50 to 20 000 μg·L-1 for urine. The recovery, precision, and accuracy were validated perfect for plasma sample analysis with a lower limit of quantitation of 0.50 μg·L-1. The main pharmacokinetic parameters found for adefovir were as follows: ρmax (24 ± s 4) μg·L-1, tmax (1.0 ± 0.6) h, AUC0-t (278 ± 45) h.μg·L-1, AUC0-∞ (285 ± 45) h.μg·L-1, t1/2 (8.8 ± 1.6) h, CL (F) (36 ± 6) L.h-1, Vd (F) (456 ± 136) L. The accumulate excrete quantity of urine was (48 ± 12) % within 0 -48 h. CONCLUSION The established LC-MS/MS method was proved to be accurate and sensitive, and can be applied to study the pharmacokinetics of adefovir in healthy Chinese volunteers.