Quantitation of DNA by nuclease P1 digestion and UPLC-MS/MS to assess binding efficiency of pyrrolobenzodiazepine

作     者：Yong Ma Buyun Chen Donglu Zhang Yong Ma;Buyun Chen;Donglu Zhang

作者机构：Drug Metabolism and DispositionGenentech1 DNA WaySouth San FranciscoCA94080USA 

出 版 物：《Journal of Pharmaceutical Analysis》 (药物分析学报（英文版）)

年 卷 期：2020年第10卷第3期

页      面：247-252页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 1002[医学-临床医学] 1001[医学-基础医学(可授医学、理学学位)] 0817[工学-化学工程与技术] 0703[理学-化学] 0702[理学-物理学] 10[医学] 

主　　题：Nuclease P1 UPLC-MS/MS DNA quantitation DNA alkylation Pyrrolobenzodiazepine(PBD-Dimer) 

摘      要：Accurate DNA quantitation is a prerequisite in many biomedical and pharmaceutical *** we established a new DNA quantitation method by nuclease P1 digestion and UPLC-MS/MS *** fragments can be efficiently hydrolyzed to single deoxyribonucleotides by nuclease P1 in a short *** decent stabilities of all the four deoxyribonucleotides were confirmed under different *** monophosphate(dAMP)was selected as the surrogate for DNA quantitation because dAMP showed the highest sensitivity among the four deoxyribonucleotides in the UPLC-MS/MS *** linear range in DNA quantitation by this method is 1.2-5000 ng/*** the validation,the inter-day and intra-day accuracies were within 90%-110%,and the inter-day and intra-day precision were acceptable(RSD10%).The validated method was successfully applied to quantitate DNA isolated from tumors and organs of a mouse xenograft *** to the quantitation methods using UV absorbance,the reported method provides an enhanced sensitivity,and it allows for the accurate quantitation of isolated DNA with contamination of RNA and ribonucleotide.