HPLC梯度洗脱法检测克林霉素磷酸酯及其注射剂的有关物质

HPLC Gradient Elution Determination of Related Substances of Clindamycin Phosphate and Its Injection

关键词:  高效液相色谱法  克林霉素磷酸酯  有关物质  特定杂质

Key words:  HPLC  clindamycin phosphate  related substances  specified impurities

摘要:

目的 探索建立改进的高效液相色谱法-梯度洗脱法检测克林霉素磷酸酯及其注射剂的有关物质。方法 色谱柱为Supelco Discovery C₁₈(4.6 mm×250 mm，5 mm)；流动相A为磷酸缓冲液(pH 3.9)-90%乙腈甲醇溶液(92∶8)，流动相B为磷酸缓冲液(pH 3.9)-90%乙腈甲醇溶液(52∶48)；柱温为40 ℃；检测波长为210 nm；流速为1.2 mL·min^-1。结果 克林霉素磷酸酯主峰与其相关物质峰分离良好，供试品中含林可霉素，克林霉素B磷酸酯，克林霉素及其他非特定杂质的量为0.006%~1.137%，0.016%~0.157%，0.005%~0.195%及0.016 3%~2.933%时，均具有良好的线性关系。林可霉素，克林霉素B磷酸酯，克林霉素以及非特定杂质(克林霉素磷酸酯)的LOD分别为0.170 5 mg·mL^-1，0.160 0 mg·mL^-1，0.146 2 mg·mL^-1和0.488 8 mg·mL^-1。林可霉素，克林霉素B磷酸酯，克林霉素各杂质的平均回收率分别104.0%(RSD=1.8%, n=9)，106.8% (RSD=1.8%，n=9)和104.9%(RSD=1.8%，n=9)。结论 改进方法的杂质色谱性能明显优于现行标准方法和USP个论方法，更适用于克林霉素磷酸酯及其注射剂的有关物质检测。

Abstract:

OBJECTIVE To establish a suitable high-performance liquid chromatography for the determination of the related substances of clindamycin phosphate and its injection. METHODS The HPLC method was carried out using a Supelco Discovery C₁₈ column (4.6 mm×250 mm, 5 mm); where the column temperature was 40 ℃ and the detection wavelength was 210 nm; mobile phase A was phosphate buffer-90% acetonitrile-methanol solution (92∶8), mobile phase B was phosphate buffer -90% acetonitrile-methanol solution (52∶48); the flow rate was 1.2 mL·min-1. RESULTS Clindamycin phosphate and related substances could be well isolated, the standard curves for lincomycin, clindamycin-B-phosphate clindamycin and other unspecified impurities were all linear in the range of 0.006%-1.137%, 0.016%-0.157%, 0.005%-0.195% and 0.0163%- 2.933%. The limit of detection (LOD) of lincomycin, clindamycin-B-phosphate, clindamycin and clindamycin phosphate were 0.170 5 mg·mL^-1, 0.160 0 mg·mL^-1, 0.146 2 mg·mL^-1 and 0.488 8 mg·mL^-1. The average recovery of lincomycin, clindamycin-B-phosphate and clindamycin were 104.0%(RSD=1.8%, n=9), 106.8%(RSD=1.8%, n=9) and 104.9%(RSD=1.8%, n=9). CONCLUSION The method shows much higher chromatographic performance and specificity than existing national standard method and USP monographs method. It is more suitable for determination of the related substances of clindamycin phosphate and its injection.