HPLC测定人血清中阿米替林及去甲阿米替林的浓度

STIMULANEOUS DETERMINATION OF AMITRIPTYLINE AND N-DESMETHYL-AMITRIPTYLINE IN HUMAN SERUM BY HPLC

目的：建立测定血清中阿米替林（AM）及其产物N-去甲阿米替林（N-AM）浓度HPLC法。方法：血清样品碱化后采用乙醚萃取,其有机相吹干检测。以甲醇∶乙腈∶水∶正丁胺∶冰乙酸（55∶90∶45∶2∶1.5）为流动相,检测波长为254nm,VP-ODS柱。结果：血清中阿米替林（AM）及其产物N-去甲阿米替林（N-AM）线性范围均为25-400μg/L,方法回收率分别为92.3%-125.7%及109.9%-121.8%,日内RSD分别为5.52%-7.70%及1.76%-9.46%,日间RSD分别为4.59%-7.70%及1.96%-9.59%。结论：本方法在阿米替林血药浓度监测应用上具有可操作性强,重现性好,临床监测周期短等优点,适合人体药动学研究。

Objective： To develop a method of HPLC for the simultaneous determination of Amitriptyline and its active metabolite N-desmethyl Amitriptyline in serum. Methods： N-desmethyl Amitriptyline and Amitriptyline in serum were extracted by ether and the oganic phase must blow dry to detected. The chromatographic separation was performed on the VP- ODS column using the mixture of methanol： water： butyl-Amine： glacial acetic acid（ 55： 90 ∶2 ∶1. 5） as the mobile phase. The eluate was monit-ored at 229 nm. Results： The linearity was obtained over the range of 31. 25 - 500μg / Lwith the recoveries 92. 3% - 125. 7% and 109. 9% - 121. 8% for N- desmethyl Amitriptyline and Amitriptyline. respectively. The intraday RSD were 5. 52% - 7. 70% and 1. 76% - 9. 46%,and the inter-day RSD were 4. 59% - 7. 70% and 1. 96% - 9. 59%,for N-desmethyl Amitriptyline and Amitriptyline. respectively. Conclusion： The amitriptyline blood concentration has a good operability,reproducibility and short clinical monitoring periodidty,Which suit for human pharmacokinetic.