RP-HPLC法测定人血浆中磷酸苯丙哌林的浓度

Determination of Benproperine Phosphate in Human Plasma by RP-HPLC

目的:建立反相高效液相色谱法(RP-HPLC)测定人血浆中苯丙哌林浓度的方法。方法:血浆经过两次萃取后,进行HPLC-UV分析,色谱柱为Phenomenex C18柱,流动相为0.3%的磷酸水溶液(三乙胺调pH3.0)-乙腈(62∶38),检测波长为220nm,流速为1.0mL·min-1。结果:苯丙哌林血浆浓度在20～800ng·mL-1范围内线性关系良好(r=0.9992),方法回收率在98%～107%之间,萃取回收率为70%。日内相对标准偏差为3.88%～10.25%,日间标准偏差为4.66%～9.12%。健康志愿者单次口服80mg磷酸苯丙哌林缓释胶囊后,主要药动学参数:Cmax为(146.40±53.68)ng·mL-1,tmax为(4.39±1.75)h,t1/2为(24.70±7.29)h;AUC0-36和AUC0-∞分别为(3144.20±1313.56),(5234.31±2495.85)ng·h·mL-1。结论:本方法操作简单,结果准确可靠,可用于临床常规检测苯丙哌林血药浓度及其药代动力学研究。

Objective： To establish a RP-HPLC method for the determination of benproperine in human plasma. Methods： The blood samples were analyzed by RP-HPLC with UV detector after twice extraction. The separation of benproperine was performed on Phenomenex C18 column with the mobile phase consisting of 0.3% phosphoric acid solution （adjust to pH 3.0 with triethylamine）-acetonitrile （62 ： 38） at a flow rate of 1.0 mL·min^-1 and the detection wavelength was 220 nm. Results： The linear range of benproperine was 20 - 800 ng·mL ^-1 （r = 0.999 2）. The methodology recovery ranged between 98% - 107% and the extraction recovery was about 70%. The intra-day RSD and inter-day RSD were 3.88% - 10.25% and 4.66% - 9.12%, respectively. The main pharmacokinetic parameters after a single dose of 80 mg benproperine phosphate sustained-release capsule were as follows： Cmax was （146.40 ＋ 53.68） ng·mL^-1, tmax was （4.39 ± 1.75） h, t1/2 was （24.70 ± 7.29） h; AUC0-36 was （3 144.20 ± 1 313.56） ng·h·mL^-1 and AUC0-∞ was （5 234.31 ± 2 495.85 ） ng·h·mL^-1, respectively. Conclusion： The method is simple, accurate, reliable and suitable for the determination of henproperine in therapeutic drug monitor and its pharmacokinetics study.