荧光染色法测定氨基酸中外源DNA残留含量  

Determination of Exogenous DNA Residual in Amino Acids by Fluorescence Staining

作　　者：张俊朋 王慧丽 连莹 朱永琴 乔冲 Zhang Junpeng;Wang Huili;Lian Ying;Zhu Yongqin;Qiao Chong(Henan Institute for Drug and Medical Device Inspection(Henan Vaccine Issuance Center),Zhengzhou Key Laboratory for Quality Research and Evaluation of Generic Drug,Zhengzhou 450018,China)

机构地区：[1]河南省药品医疗器械检验院(河南省疫苗批签中心),郑州市仿制药质量研究与评价重点实验室,河南郑州450018

出　　处：《山东化工》2024年第15期192-194,208,共4页Shandong Chemical Industry

基　　金：河南省市场监督管理局科技计划项目(2022sj80)。

摘　　要：目的:建立荧光染色法检测五种氨基酸原料中外源DNA残留量。方法:采用荧光染料与双链DNA特异结合后,使用荧光酶标仪测定,激发波长488 nm,检测波长520 nm。结果:DNA质量浓度在0.5~40 ng/mL范围内,浓度与荧光强度呈良好线性关系(r=0.9999),定量限0.55 ng/mL,不同品种氨基酸回收率在73%~132%。结论:该方法操作简便、快速,适用不同品种氨基酸,为企业开展氨基酸原辅料外源DNA残留量质量控制提供了技术基础。Objective:To establish a method for determination of DNA residual in 5 Amino acids by fluorescence staining.Methods:Determined by fluorescent enzyme labeling,fluorescent dye was specifically combined with double-stranded DNA,the excitation wavelength was 488 nm and the detection wavelength was 520 nm.Results:The assay was linear over the concentration range of 0.5~40 ng/mL with a correlation coefficient(r)of 0.9999,the limit of quantitation was 0.55 ng/mL,the recovery of different amino acids varieties ranged from 73%~132%(n=9).Conclusion:This method is simple,rapid and suitable for the determination of DNA residual in different amino acids,which provides the technical basis for quality control.

关 键 词：氨基酸 荧光染色法 外源DNA 

分 类 号：O657.3[理学—分析化学]