摘要
目的 建立灵敏、简便、适合临床监测阿米替林及其代谢物去甲替林总浓度与游离浓度的反相高效液相色谱法(RP HPLC)。方法 血样在碱性条件下 ,经一步液 液提取法后 ,在C 18柱上进行分离。流动相为乙腈∶水 =30∶70(V/V) ,其中含三乙胺 0 5 %和磷酸 0 3%。血浆样品药物总浓度测定为RP HPLC ,游离药物测定为超滤离心 /RP HPLC。结果 阿米替林和去甲替林的标准曲线在 4～ 2 0 0μg·L-1(总浓度 )和 4～ 6 4μg·L-1(游离浓度 )范围内呈线性。两药的平均回收率分别为 10 2 0 %± 3 77%与 99 3 %± 7 13 % ;日内RSD分别为 2 40 %～ 4 39% ,3 0 2 %～4 2 8% ;日间RSD分别为 4 92 %～ 6 15 % ,6 35 %～7 48%。测定了 7例健康志愿者单剂量口服盐酸阿米替林片 5 0mg后 6h的血药浓度。血浆阿米替林总浓度为 18 0～ 2 7 2 μg·L-1,游离浓度为 1 4～ 2 5 μg·L-1。血浆去甲替林总浓度为 (2 0± 0 4) μg·L-1(1 5～ 2 5 μg·L-1)。阿米替林的血浆蛋白结合率在 89 8%～ 92 6 %之间。结论 本方法简便、快速 ,灵敏度与选择性高 ;成本低 。
AIM To develop a sensitive, specific and reliable reversed phase high performance liquid chromatographic method(RP\|HPLC) to determine the total and unbound(free) concentrations in human plasma of amitriptyline and its major metabolite, nortriptyline. METHODS\ The assay involved a simple extraction procedure. The mobile phase consisted of acetonitrile and distilled water(30∶70, V/V ), containing triethylamine(0 5%) and orthophosphoric acid(0 3%), pH 3 1. Separation was achieved on the C18 ODS column and the effluent was measured for UV absorption at 240 nm. RESULTS The calibration curves were linear in the range of 4～400 μg·L -1 for total concentration, and in the range of 4～64 μg·L -1 for free concentration for both amitriptyline and nortriptyline. The lowest limits of detection were 4 μg·L -1 for both compounds. The absolute recovery rates were 102 0%±3 77% for amitriptyline and 99 3%±7 13% for nortriptyline. The precision values(RSD) of intra day and inter day for both amitriptyline and for nortriptyline were determined to be <5%, and <8%, respectively. The method was applied to determine the total and free concentrations in plasma of the healthy volunteers after a single oral dose of 50 mg amitriptyline. CONCLUSION The assay was simple, repid, highly selective and sensitive. It is suitable for the routine analysis of total and free drug concentrations in plasma using readily available instruments with lower cost.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2001年第1期104-107,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金!(No 396 70 843)
国家人事部科研基金!(No1997-2 0 1)资助项目&&
