毛细管电泳法测定头孢曲松钠有关物质及含量

LC/MS Determination of Ceftriaxone Sodium Related Substances

目的:建立测定头孢曲松钠有关物质及含量的毛细管电泳方法。方法:采用非涂渍石英毛细管50 cm×75μm;运行缓冲液为含100 mmol·L^(-1)十二烷基硫酸钠的磷酸缓冲液(40 mmol·L^(-1),pH 7.0);运行电压为12 kV(电流40—50 μA);柱温为25℃;检测波长254 nm;用水制备样品溶液,相关物质测定时样品溶液浓度为5 mg·mL^(-1),含量测定时样品溶液的浓度为0.1 mg·mL^(-1);压力进样4 s(进样口压力3.447 kPa)。结果:头孢曲松钠中的主要杂质有21个,水解、酸解、碱解、氧化中产生的诸杂质彼此之间及与头孢曲松之间可得到有效的分离;线性范围为0.0125—0.4 mg·mL^(-1),y=1.0379x—0.004,r=0.9996;头孢曲松钠的最低检测限为5.2μg·mL^(-1),最低定量限为12.5μg·mL^(-1);与HPLC结果比较,含量测定的最大偏差为2.6%,有关物质测定,HPCE测得杂质数目较HPLC多,但两种方法测得的最大有关物质的含量基本一致;各杂质峰迁移时间的日内最大RSD为0.94%,日间最大RSD为1.15%;各杂质峰面积的日内最大RSD为17.0%,日间最大RSD为26.8%;且其杂质含量越低,峰面积的RSD越大。结论:毛细管电泳法分析头孢曲松钠有关物质与含量,方法准确,并与HPLC方法具有互补性,有助于企业分析产品中杂质产生的原因。

Objective: To develop a capillary electrophoresis method for determination of related substances in ceftriaxone sodium. Methods:The uncoated fused-silica capillary (50 cm×75μm) was used. The running buffer composed of 40 mmol·L^(-1) phosphate buffer containing 100 mmol·L^(-1) sodium dodecylsulphate. The separation voltage was 12 kV (40-50 μA) and the capillary temperature was 25℃. The detection wavelength was 254 nm. The sample was solved in water with a concentration of 5 mg·mL^(-1) and 0. 1 mg·mL^(-1) respectively for impurity determination and content assay,and was injected to the column by pressure(3. 447 kPa) for 4 s. Results:Twenty-one impurities in ceftriaxone sodium could be detected. Ceftriaxone sodium and the degradations obtained by acid hydrolysis, alkaline hydrolysis and oxidation could be separated well. The calibration curve of ceftriaxone sodium was linear in the range of 0.0125-0.4 mg·mL^(-1). The limit of detection and limit of quantitation were 5.2 μg·mL^(-1) and 12. 5 μg·mL^(-1) respectively. The number of impurities detected by CE was more than that by HPLC,however the biggest impurity content determined by CE was the same as that by HPLC. The values of intra-day and interday RSD of migration time of impurities were less than 0. 94% and 1. 15% respectively. The values of intra-day and inter-day RSD of peak area were less than 17.0% and 26. 8% respectively. The lower impurity content was, the bigger the RSD was. Conclusions: The HPCE method developed was accurate, and could be complement to HPLC for determination of the impurity and help pharmaceutical manufacturer to analysis the possible source of the impurity.