摘要
目的:基于分析方法质量源于设计(analytical quality by design, AQbD)理念进行实验设计(design of experiment, DOE),建立硫酸新霉素药物中外源DNA荧光染色检测方法。方法:利用MODDE?Pro11软件进行DOE开发,对关键影响因素和模型参数进行考察和优化,再采用基于荧光染色原理的Qubit?DNA检测试剂盒进行方法建立。结果:建立了硫酸新霉素中外源DNA的通用性检测方法。该方法的关键影响因素为溶剂种类、溶剂浓度和供试液浓度;溶剂种类选择氯化钠,最优分析条件为溶剂浓度1.4 mol·L^(-1)、供试液浓度1.8 mg·mL^(-1);该方法线性关系良好(r=0.995 7,n=2),加样回收率>81.0%,耐用性实验中加样回收率均在优化标准80%~120%范围内。结论:该方法准确度高、稳固性强、通用性好,为建立发酵来源化学药品中的外源DNA方法提供了参考。
Objective:To establish a detection method for exogenous DNA in neomycin sulfate drugs based on the concept of analytical quality by design(AQbD).Methods:The MODDE Proll software was used to design the experiment,investigate the key influencing factors and optimize the parameters.Qubit DNA detection kit based on fluorescence staining method was employed for the determination of exogenous DNA.Results:The universal detection method of exogenous DNA in neomycin sulfate was established.The key effect factors are solvent type,solvent concentration and sample concentration.When NaCl was used as solvent,the optimal analysis conditions were solvent concentration at 1.4 mol·L^(-1) and sample concentration at 1.8 mg·mL^(-1).The linear range of DNA standard was 20~100 ng·mL^(-1)(r=0.9957,n=2).The accuracy was above 81.0%.The recoveries in ruggedness tests were 80%~120%.Conclusion:The established method is accurate,robust and universal.This study provides reference for the development of determination of exogenous DNA in aminoglycoside antibiotics.
作者
张培培
姚尚辰
李进
张夏
邹文博
许明哲
王琰
宁保明
ZAHNG Pei-pei;YAO Shang-chen;LI Jin;ZHANG Xia;ZOU Wen-bo;XU Ming-zhe;WANG Yan;NING Bao-ming(National Institutes for Food and Drug Control,Beijing 102629,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2023年第9期947-954,共8页
Chinese Journal of New Drugs
基金
国家药典委员会药品标准制修订研究课题资助项目(2021Y04)
国家药品监督管理局化学药品质量研究与评价重点实验室课题学科建设项目(2023HYZX19)。
