HPLC-ELSD分析硫酸异帕米星及其注射液

Determination of Isepamicin Sulfate and its Injection by HPLC-ELSD

关键词:  硫酸异帕米星  有关物质  硫酸异帕米星注射液  梯度洗脱  高效液相色谱-蒸发光散射检测法

Key words:  isepamicin sulfate  related substances  isepamicin sulfate injection  gradient  HPLC-ELSD

摘要:

目的 建立HPLC-ELSD测定硫酸异帕米星及其注射液的有关物质、硫酸盐和含量的方法。方法 采用Agilent SB-C₁₈柱(4.6 mm×250 mm，5 μm)，有关物质测定采用梯度洗脱法，流动相A为0.2 mol·L^-1三氟乙酸溶液，流动相B为甲醇。硫酸盐和含量测定采用等度洗脱法，流动相为0.2 mol·L^-1三氟乙酸溶液，流速为0.8 mL·min^-1。Alltech 3300 ELSD检测器，漂移管温度为50 ℃。结果 异帕米星与中间体、副产物完全分离。异帕米星、硫酸盐、庆大GE-B、庆大霉素B、3N-异帕米星在一定的浓度范围内呈较好的线性关系，硫酸异帕米星注射液含量测定的平均回收率为100.0% (RSD=0.8%，n=9)，有关物质测定庆大GE-B、3N-异帕米星、庆大霉素B的回收率分别为104.4%，97.6%和109.0%，异帕米星、庆大GE-B、庆大霉素B、3N-异帕米星的检测限分别为5.1，4.5，5.1 和5.0 μg·mL^-1。结论 本方法简便、灵敏、重复性好，可用于本品的质量控制。

Abstract:

OBJECTIVE To develop a new HPLC-ELSD method for the determination of isepamicin sulfate and its injection. METHODS The column was Agilent SB-C₁₈(4.6 mm×250 mm, 5 μm). The mobile phase for the determination of related substances consisted of 0.2 mmol·L^-1 trifluoroacetic acid (A) and methanol (B) with gradient program. The mobile phase for the determination of isepamicin and sulfate was 0.2 mol·L^-1 trifluoroacetic acid. The flow rate was 0.8 mL·min^-1. Detector used was an Alltech 3300 ELSD detector. The drift tube temperature was 50 ℃. RESULTS Good separation of isepamicin from main intermediates could be achieved. The standard curves of isepamicin, sulfate, HAPA-gentamine-B, 3N-isepamicin and gentamine-B were linear in the certain range. The average recovery of isepamicin for the determination of isepamicin sulfate injection was 100.0% (RSD=0.8%, n=9). The average recoveries of HAPA-gentamine-B, 3N-isepamicin and gentamine-B for the determination of related substances were 104.4%, 97.6% and 109.0%, respectively. The detection limits of isepamicin, HAPA-gentamine-B, gentamine-B and 3N-isepamicin were 5.1, 4.5, 5.1 and 5.0 μg·mL^-1, respectively. CONCLUSION The method is simple, sensitive and reproducible, and can be used for quality control of isepamicin sulfate and its injecton.