HPLC法同时测定人尿液中三苯双脒代谢物氨脒和乙酰氨脒的浓度  

Simultaneous Determination of Tribendimidine Metabolite dADT and Acetylizad dADT in Human Urine by HPLC

作　　者：汪晓静 袁桂艳[2] 张蕊[2] 魏春敏[2] 曲婷婷 郭瑞臣[2] 

机构地区：[1]山东医学高等专科学校,济南250002 [2]山东大学齐鲁医院临床药理研究所,济南250012 [3]德州市食品药品监督管理局德城区分局,山东德州253012

出　　处：《中国药房》2015年第2期213-216,共4页China Pharmacy

摘　　要：目的：建立同时测定人尿液中三苯双脒代谢物氨脒和乙酰氨脒浓度的方法,并进行其排泄动力学研究。方法：8名健康男性受试者空腹口服三苯双脒肠溶片400 mg,收集服药前及服药后0~2、2~4、4~6、6~8、8~12、12~24、24~36、36~48 h的尿液。尿样经沉淀蛋白并稀释后,以比卡鲁胺为内标,采用高效液相色谱法进样测定,色谱柱为Agilent Zorbax Extend C18（250 mm×4.6 mm,5μm）流动相为乙腈-水-三乙胺（60∶40∶0.2,V/V/V）,紫外检测波长为265 nm,流速为0.7 ml/min,进样量为10μl。结果：氨脒和乙酰氨脒尿药浓度分别在0.5~500μg/ml（r=0.996 2）和0.1~200μg/ml（r=0.995 8）范围内线性关系良好,最低定量限分别为0.5、0.1μg/ml,绝对回收率分别为94.78%~95.23%和91.26%~96.99%,相对回收率为98.37%~101.44%和99.26%~102.06%,日内、日间RSD〈7%;-20℃冷冻24 h及7 d和反复冻融均稳定,RSD〈7%。受试者口服400 mg三苯双脒肠溶片后,氨脒和乙酰氨脒48 h尿累积排泄率分别为（55.84±13.60）%和（14.98±7.65）%,采用DAS 2.0程序计算氨脒和乙酰氨脒半衰期分别为（3.46±0.73）h和（4.21±0.43）h,尿药排泄速率常数分别为（0.21±0.07）h^-1和（0.17±0.02）h^-1。结论：该方法简单、快速、灵敏、重复性好,可用于健康受试者口服三苯双脒肠溶片后氨脒和乙酰氨脒尿药浓度的测定。OBJECTIVE ： To establish a method for simultaneous determination of teibendimidine metabolites dADT and acetyl- izad dADT in human urine, and to study its excretion kinetics. METHODS： 8 healthy male subjects were selected and given Triben- dimidine enteric-coated tablets 400 mg. Urine samples were collected before and 0-2 h, 2-4 h, 4-6 h, 6-8 h, 8-12 h, 12-24 h, 24-36 h and 36-48 h after administration. Urine samples were diluted and determined by HPLC method using bicalutamide as inter- nal standard. The determination was performed on Agilent Zorbax Extend C18 （250 mm × 4.6 mm, 5 μ m） column with mobile phase consisted of acetonitrile-water-triethylamine （60：40：0.2,V/V/V） at a flow rate of 0.7 ml/min. The sample size was 10 pl. RESULTS： The linear ranges of dADT and acetylizad dADT were 0.5-500 μg/ml （r=0.996 2） and 0.1-200μg/ml （r=0.995 8）, respectively. The lower limit of quantitation （LLOQ） was 0.5 ~tg/ml and 0.1 μg/ml. The absolute recoveries were 94.78%-95.23% and 91.26%-96.99%. The relative recoveries were 98.37%-101.44% and 99.26%-102.06%. Intra-day and inter-day RSD were both less than 7%. dADT and acetylizad dADT in human urine were stable when frozen at --20℃ for 24 h and 7 days or after frozen repeat- edly （RSD〈 7 % ）. After oral administration of Tribendimidine enteric-coated tablets 400 mg, the excretion rates of dADT and acetyl- izad dADT were （55.84 ± 13.60）% and （14.98 ± 7.65）%, respectively. The half-life period （tl/2） and excretion rate constant （Ke） of dADT and acetylizad dADT calculated by DAS 2.0 program were as follows： t1/2（3.46 ± 0.73）h and （4.21 ± 0.43）h; Ke （0.21 ± 0.07） h^-1 and （0.17 ± 0.02）h^-1. CONCLUSIONS： The developed method is simple, quick, sensitive and reproducible. It can be used for the determination of dADT and acetylizad dADT in urine of healthy volunteers after oral administration of Tribendimidine enter- ic-coated tablets.

关 键 词：三苯双脒 代谢物 氨脒 乙酰氨脒 尿液 高效液相色谱法 

分 类 号：R969.1[医药卫生—药理学] R978[医药卫生—药学]