基于杂化颗粒色谱柱技术对欧洲药典中螺旋霉素有关物质检验方法的改进  

Improvement of the Determination Method for Spiramycin Related Substances in the European Pharmacopoeia Based on Hybrid Particle Chromatography Column Technology

作　　者：姚远 王倩 张志伟[3] 袁耀佐 YAO Yuan;WANG Qian;ZHANG Zhiwei;YUAN Yaozuo(NMPA Key Laboratory for Impurity Profile of Chemical Drugs,Jiangsu Institute for Food and Drug Control,Nanjing 210019,China;China Pharmaceutical University,Nanjing 210009,China;WuxiFortune Pharmaceutical Co.,Ltd.,Wuxi 214101,China)

机构地区：[1]江苏省食品药品监督检验研究院,国家药品监督管理局化学药品杂质谱研究重点实验室,南京210019 [2]中国药科大学,南京210009 [3]无锡福祈制药有限公司,江苏无锡214101

出　　处：《中国药学杂志》2024年第20期1956-1961,共6页Chinese Pharmaceutical Journal

基　　金：江苏省市场监督管理局科技计划项目资助(KJ2023027)。

摘　　要：目的建立并优化了螺旋霉素有关物质的分析方法,用于螺旋霉素有关物质的常规检验。方法采用基于杂化颗粒技术的Waters Xbridge shield RP C18(4.6 mm×250 mm,3.5μm)色谱柱,色谱条件为:水-0.2 mol·L^(-1)磷酸氢二钾溶液(用1 mol·L^(-1)的KOH溶液调节pH值至9.5)-乙腈-甲醇-(10∶60∶28.5∶1.5)作为流动相A,水-0.2 mol·L^(-1)磷酸氢二钾溶液(pH 9.5)-乙腈-甲醇(10∶30∶57∶3)作为流动相B,梯度程序洗脱。检测波长232 nm。结果所建立的检验方法可以将螺旋霉素各组分与其有关物质实现良好分离,螺旋霉素Ⅰ质量浓度在0.7~1200μg·mL^(-1)范围内,其峰面积与浓度呈良好线性关系(r=0.9993),检测限(LOD)为0.2μg·mL^(-1),定量限为(LOQ)为0.7μg·mL^(-1),重复性与精密度试验结果相对标准偏差(RSD)均<2.0%,供试品溶液在5℃下24 h稳定。结论与现行分析方法相比,该方法专属性强,灵敏度与稳定性均较好,且有效缩短检验时间,提升检验效率,可用于螺旋霉素有关物质的日常检验工作。OBJECTIVE To established and optimized an analytical method for spiramycin related substances for daily testing of spiramycin related substances.METHODS A Waters Xbridge shield RP18(4.6 mm×250 mm,3.5μm)column was used for the experiment.The chromatographic conditions were as follows:water:0.2 mol·L^(-1)dipotassium hydrogen phosphate(adjust the pH value to 9.5 using a 1 mol·L^(-1)KOH solution):acetonitrile:methanol,(10∶60∶28.5∶1.5)as mobile phase A,water:0.2 mol·L^(-1)dipotassium hydrogen phosphate(pH 9.5):acetonitrile:methanol,(10∶30∶57∶3)as mobile phase B,and gradient elution was performed.Detection wavelength 232 nm.RESULTS The established method can achieve good separation between the components of spiramycin and its related substances.The mass concentration of spiramycinⅠis in the range of 0.7-1200μg·mL^(-1),and its peak area shows a good linear relationship with concentration(r=0.9993).The limit of detection(LOD)is 0.2μg·mL^(-1),and the limit of quantification(LOQ)is 0.7μg·mL^(-1).The RSD of the repeatability and precision test is less than 2.0%,and the test solution is stable at 5℃for 24 h.CONCLUSION Compared with current analytical methods,this method has strong specificity,good sensitivity and stability,and effectively shortens the testing time and improves the testing efficiency.It can be used for the daily testing of related substances of spiramycin.

关 键 词：螺旋霉素 有关物质 杂化颗粒色谱柱 高效液相色谱法 质量控制 

分 类 号：R917[医药卫生—药物分析学]