高效毛细管电泳法同时分离检测肝素钠杂质

Separation and Determination of Impurities in Heparin Sodium Simultaneously by Capillary Electrophoresis

目的建立用高效毛细管电泳法同时分离检测肝素钠杂质的方法。方法采用高效毛细管电泳法,通过选择电极缓冲液的种类、浓度、pH值、检测波长等最佳实验条件,确定了一种检测肝素钠杂质的优化方法:800mmol.L-1磷酸-锂盐为运行缓冲液(pH3.5),熔融石英毛细管(25μm×60cm,有效长度50cm),分离电压-30kV,进样量:14kPa×50s,分离温度:20℃,检测波长200nm。结果此方法实现了肝素钠与各个杂质的分离,多硫酸软骨素(oversulfated chondroitin sulfate,OSCS)峰和肝素峰的分离度可达1.5,且线性、重复性良好,OSCS峰检测限为0.2mg.mL-1,相当于肝素钠质量的0.4%(Woscs/Whep-arin)。结论此方法准确、简便,优于美国药典(USP32)收载的方法,利于肝素钠的质量控制。

OBJECTIVE To establish a the HPCE method for determination of impurities in heparin sodium.METHODS The determination was performed by HPCE after establishing suitable separation conditions.It was carried out on a fused silica capillary of 25 μm×60 cm （50 cm effective length ）with the mobile phase consisting of 800 mmol·L-1 （ pH 3.5）phosphate-Li.The injection volume was 14 kPa×50 s.The applied voltage was-30 kV.The detection wavelength was set at 200 nm.RESULTS This method showed good linearity and repeatability.The resolution of oversulfated chondroitin sulfate（OSCS） and heparin was 1.5.The detection limits for OSCS was 0.2 mg·mL-1,0.4%（Woscs/Wheparin）.CONCLUSION Compared with the method documented in USP32,the present method provides good efficiency and resolution of heparin and impurities.