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目的建立反相高效液相色谱法测定人尿中右美沙芬及其代谢产物去甲右美沙芬浓度的方法。方法以非那西丁为内标,尿样经水解,碱化后用正己烷-正丁醇(9:1)萃取,采用DiamonsilTMC18柱(250mm×4.6mm,5μm)分析。色谱条件:流动相为乙腈(A)-1%三乙胺(磷酸调节pH=2.2,B),梯度洗脱,0～15min,20%～35%A,流速为1.0ml/min,检测波长为280nm,柱温40℃。结果右美沙芬在0.05～2.0μg/ml范围内线性良好(r=0.9999,n=5),检测限为0.04μg/ml;去甲右美沙芬尿样浓度在0.5～20.0μg/ml范围内线性良好(r=0.9999,n=5),检测限为0.4μg/ml。两者日内、日间精密度RSD均<10%,低、中、高浓度的提取回收率在94%～108%之间。结论此方法简便准确、重复性好,适用于CYP2D6表型分析以及右美沙芬与其代谢产物的人体药代动力学研究。
Objective To establish a method for the determination of dextromethorphan and its metabolite dextromethorphan in human urine by reversed-phase high performance liquid chromatography. Methods Phenacetin was used as internal standard. The urine samples were hydrolyzed and alkalized and extracted with n-butanol (9: 1) and analyzed by Diamonsil TMC18 column (250 mm × 4.6 mm, 5 μm). Chromatographic conditions: mobile phase acetonitrile (A) -1% triethylamine (phosphoric acid to adjust the pH = 2.2, B), gradient elution, 0 ~ 15min, 20% ~ 35% A, flow rate of 1.0ml / min, 280nm, column temperature 40 ℃. Results Dextromethorphan was linear in the range of 0.05-2.0μg / ml (r = 0.9999, n = 5) with the detection limit of 0.04μg / ml. The dextromethorphan urine concentration was in the range of 0.5-20.0μg / ml The linearity was good (r = 0.9999, n = 5) with a detection limit of 0.4 μg / ml. The RSD of intra-day and inter-day precision were all less than 10%, and the extraction recoveries of low, medium and high concentrations were between 94% and 108%. Conclusion This method is simple, accurate and reproducible. It is suitable for CYP2D6 phenotype analysis and human pharmacokinetics study of dextromethorphan and its metabolites.