HPLC法测定血清中异烟肼、利福平和吡嗪酰胺浓度

Determination of isoniazid, rifampicin and pyrazinamide in serum by HPLC

目的 建立同时测定血清中异烟肼 (INH)、利福平(RFP)、吡嗪酰胺 (PZA)浓度的反相高效液相色谱法。方法 RFP检测条件 :SymmetryC8色谱柱 (5 μm ,15 0mm× 3 9mm) ,流动相为磷酸盐缓冲液 (pH 5 5 )∶甲醇 =30∶70 ,流速0 4 5ml/min ,测定波长 340nm ;INH、PZA检测条件 :Sym metryC18色谱柱 (5 μm ,2 5 0mm× 4 6mm) ,流动相为磷酸盐缓冲液 (pH 6 0 )∶乙腈 =91∶9,流速 0 8ml/min ,测定波长2 5 4nm。结果 以峰高外标法定量 ,线性范围 :利福平为0 5～ 32mg/L ,INH为 0 5～ 2 0mg/L ,PZA为 1 0～ 6 0 0mg/L ,均具有良好的线性关系。 3种药物的日内、日间变异均 <10 %。回收率 >80 %。结论 该方法简便、快速 ,灵敏度和准确度较高 ,无干扰 ,用于上述 3种药物复合制剂卫非特片的药代动力学研究 。

Objective To develop effective HPLC method for the analysis of isoniazid(INH),rifampicin(RFP) and pyrazinamide(PZA) in serum at the same time. Methods Symmetry C8(5 μm,150 mm×3 9 mm)column was used to detect RFP concentration. The mobile phase was consisted of a mixture of methanol phosphate(pH 5 5).The flow rate was 0 45 ml/min.The detecting wave was 340 nm.Symmetry C18(5 μm,250 mm×4 6 mm) column was used to detect INH and PZA concentrations. The mobile phase was consisted of a mixture of acetonitrile phosphate(pH 6 0).The flow rate was 0 8 ml/min.The detecting wave 254 nm. Results A good linearity was obtained for RFP( r = 0 999 5 within 0 5～32 mg/L),INH ( r = 0 999 6 within 0 5～20 mg/L) and PZA( r= 0 999 2 within 1 0～60 0 mg/L).The RSD of intra day and inter day variation of INH,RFP and PZA was less than 10%.The recovery of the two methods was 85%～100%. Conclusion The two methods are simple, accurate and efficient for determination of INH, RFP and PZA in serum and has already been proved in the pharmacokinetics of rifeter.