毛细管区带电泳法测定板蓝根注射液中四种核苷的含量

Determination of Four Nucleosides in Banlangen Injections Using Capillary Zone Electrophoresis

采用毛细管区带电泳法测定了板蓝根注射液中胞苷、腺苷、鸟苷和尿苷的含量。电泳条件:采用未涂层石英毛细管(32 5cm×50μmi d ,有效长度23 5cm),以60mmol/L硼砂溶液 10%(体积分数)异丙醇 20%(体积分数)乙腈为运行缓冲液,在25℃下以20kV恒压电泳分离,压力进样(1kPa×10s),检测波长254nm。对电泳条件各因素进行了讨论,如缓冲液的种类、浓度和pH值,有机改性剂的种类和浓度,分离电压和毛细管温度等。样品经0 45μm微孔滤膜过滤后直接进样;采用外标法峰面积定量。上述4种核苷回归曲线的相关系数均在0 9991以上,线性范围12 5～250mg/L,检出限分别为6 2(胞苷)、4 6(腺苷)、6 7(鸟苷)、9 0(尿苷)mg/L。4种核苷的三水平加样回收率为95 1%～102 3%,相对标准偏差(RSD)为0 3%～4 9%。方法简便快速,重现性和准确度令人满意,可作为控制板蓝根注射液中核苷类成分含量的有效方法。

The contents of cytidine, adenosine, guanosine and uridine in Banlangen Injections were determined by capillary zone electrophoresis. Shorter fused silica capillary (32.5 cm×50 μm i.d.with effective length of 23.5 cm) was used. The samples were analyzed with 60 mmol/L borate-10% (v/v) 2-propanol-20% (v/v) acetonitrile running buffer at 20 kV voltage (25 ℃ capillary temperature). Pressure injection of 1 kPa×10 s was employed, and the detection wavelength was 254 nm. Factors of the electrophoresis condition were investigated, such as the kinds and concentration of the electrolyte, buffer pH value, the type and concentration of organic modifier, separation voltage and temperature. The samples were filtered through 0.45 μm membrane, then direct injection was employed. External calibration of peak area versus concentration was used in the determination. Linear calibrations of four nucleosides were obtained within 12.5-250 mg/L (r>0.999 1). The limits of detection (mg/L) were 6.2 of cytidine, 4.6 of adenosine, 6.7 of guanosine, and 9.0 of uridin. The average recoveries of the four nucleosides were between 95.1%-102.3% with RSDs of 0.3%-4.9%. The method is simple, rapid, reproducible and accurate. It can be used for the routine analysis of the four nucleosides in Banlangen Injections.