高效液相色谱法测定人血浆中茴拉西坦活性代谢产物ABA及其在生物等效性研究中的应用

Determination of active metabolite of aniracetam ABA in human plasma by HPLC and its bioequivalence in healthy volunteers

目的建立人血浆中茴拉西坦代谢产物N-对甲氧基苯甲酰胺基丁酸（ABA）的高效液相色谱法,并研究其在健康人体的生物等效性。方法以氢氯噻嗪为内标,血浆样品用液液-萃取处理。分析柱为SymmetryShield RP18C18（3.9 mm×150 mm,5μm）,柱温：30℃。流动相：20 mmol.L-1甲酸胺-甲醇（83∶17,v/v）,流速：1.0 mL.min-1,进样量：20μL,紫外检测波长250 nm。结果茴拉西坦活性代谢产物ABA的线性范围为53.313 640μg.L-1,最低定量下限为53.3μg.L-1;日内、日间RSD不超过11%;平均回收率为96.9%103.3%。结论该方法专属性强,快速、灵敏,适合血样中茴拉西坦代谢产物ABA的浓度检测。

Objective To establish a method to determine active metabolite of aniracetam ABA concentration in the plasma by HPLC and study its bioequivalence in healthy volunteers.Methods Hydrochlorothiazide was used as the internal standard,and ABA in the plasma sample was determined by HPLC with liquid-liquid extraction,and achieved by the column of SymmetryShield RP18 C18（3.9 mm×150 mm,5 μm）at 30 ℃.The mobile phase consisted of 20 mmol·L-1 ammonium formate-methanol= 83∶17（v/v） at 1.0 mL·min-1.The injection volume was 20 μL.The UV detection wavelength was 250 nm.Results The method was linear within 53.3-13 640 μg·L-1 for ABA,and the lowest detectable concentration was 53.3 μg·L-1.The intra-day and inter-day precision was no more than 11%.The recovery was 96.9%-103.3%.Conclusion The method is accurate and sensitive with no interference,which is suitable for the determination of active metabolite of aniracetam ABA in human plasma.