反相高效液相色谱法测定人血清中利福平的含量

Determination of rifampin in human serum by RP HPLC

目的 :建立一种反相HPLC法测定人血清中利福平的含量。方法 :以对硝基苯酚为内标 ,样本经有机相提取、纯化后进样 :采用Lichrospher 10 0 (RP 8,4 6μm× 2 5 0mm)为分析柱 ,柱前加YWG C18(Φ4× 4 0mm)保护柱 ,以甲醇 乙腈 0 0 4mol·L-1KH2 PO4(3 1 5∶3 1 5∶3 7,V/V)为流动相 ,流速 1mL·min-1,柱温 4 0℃ ,2 5 4nm处紫外检测。结果 :本法分离良好 ,不受干扰。平均回收率为 99 1% ,RSD =2 2 % (n =6) ,在 0 62 5～ 4 0 0 μg·mL-1范围内 ,峰面积对浓度呈良好的线性关系 (r=0 9998)。结论 :本法简便准确快速 ,灵敏度高 ,重现性好和检测范围广 。

AIM: To establish a RP HPLC method for determination of rifampin in human serum. METHODS: Using p nitrophenol as the internal standard, the serum samples were extracted with methanol.HPLC condition: analytical column: Lichrospher 100 (RP 8, 4 6 μm×250 mm), guard column: YWG C 18 (Φ4×40 mm), the mobile phase: methanol acetonitril 0 04 mol·L -1 KH 2PH 4 (31 5∶31 5∶37, V/V), the flow rate: 1 mL·min -1 , the column temperature: 40 ℃, the wavelength of UV: 254 nm. RESULTS: The chromatography was good and not interfered by the components of the serum. The average recovery were 99 1%, RSD=2 2% ( n=6) , the linear range was 0 625-40 0 μg· mL -1 ,( r = 0 999 8). CONCLUSION: This is a simple, rapid, accurate, sensitive and reliable method to determine the concentration with a wide range. It is stuitable for the clinical monitoring and pharmacokinetic study of rifampin.