HPLC测定替卡西林钠及注射用替卡西林钠克拉维酸钾中的有关物质

Determination of Related Substances in Ticarcillin Disodium and Potassium Clavulanate for Injection by HPLC with Gradient Elution

关键词:  梯度洗脱  高效液相色谱法  替卡西林钠  注射用替卡西林钠克拉维酸钾  有关物质

Key words:  gradient elution  HPLC  ticarcillin disodium  ticarcillin disodium and potassium clavulanate for injection  related substance

摘要:

目的 建立测定替卡西林钠及注射用替卡西林钠克拉维酸钾中有关物质的高效液相色谱梯度洗脱法。方法 采用HPLC梯度洗脱法，色谱柱为Waters XBridge^TM C₁₈柱（4.6 mm×250 mm，5 μm）；以0.01 mol·L^-1磷酸氢二铵溶液（pH 7.0）为流动相A，0.01 mol·L^-1磷酸氢二铵溶液（pH 7.0）-甲醇（50：50）为流动相B，梯度洗脱；流速为1.0 mL·min^-1；检测波长为220 nm；柱温为30℃。结果 与等度洗脱法比较，梯度洗脱法具有更强的分析杂质的能力，样品中各成分的分离度及检出灵敏度均满足有关物质测定要求。替卡西林杂质A与破坏条件下产生的降解杂质与主成分峰分离较好；替卡西林的最低检出限为4.8 ng，克拉维酸的最低检出限为7.3 ng；原料药与制剂样品中替卡西林杂质A均<1.5%，最大单个杂质均<2.0%，总杂质均<4.0%。结论 本方法专属性好，灵敏度高，可用于替卡西林钠原料药和注射用替卡西林钠克拉维酸钾中有关物质的测定。

Abstract:

OBJECTIVE To establish a gradient HPLC method for related substances of ticarcillin disodium-clavulanate potassium for injection. METHODS The column was Waters XBridge^TM C₁₈(4.6 mm×250 mm, 5 μm). Mobile phase A was 0.01 mol·L^-1 ammonium dibasic phosphate solution (pH 7.0), mobile phase B was 0.01 mol·L^-1 ammonium dibasic phosphate solution (pH 7.0)-menthol(50:50). The flow rate was 1.0 mL·min^-1, linear gradient elution. The column temperature was 30℃. The detection wavelength was 220 nm. RESULTS Contrast to isocratic elution, gradient elution was more widely applied on the separation of the related substance. The resolution and sensitivity of related substance was in accord with the demands. The impurities and impurity A were all separated from ticarcillin and clavulanate. Limit of detection for ticarcillin was 4.8 ng. Limit of detection for Clavulanate was 7.3 ng. The content of ticarcillin impurity A was less than 1.5%, the content of the single biggest impurity was less than 2.0%, the total content of related substance were less than 4.0% in samples of the materiel and preparation. CONCLUSION The proposed HPLC method was specific, sensitive and suitable for the determination of related substances of ticarcillin and ticarcillin disodium-clavulanate potassium for injection.