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

张俊朋 ¹王慧丽 ¹连莹 ¹朱永琴 ¹乔冲¹

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作者信息

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
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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=0.999 9),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.

Abstract

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 0.999 9,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

Key words

amino acids/fluorescence staining method/exogenous DNA

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出版年

2024

山东化工

山东省化工研究院山东省化工信息中心

山东化工

影响因子：0.249

ISSN：1008-021X

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