干旱区生态环境知识资源中心(Arid): Enrichment of nucleobase adducts from genomic DNA in the cytoplasm by solid-phase extraction

Arid

DOI	10.1016/j.jchromb.2022.123574
	Enrichment of nucleobase adducts from genomic DNA in the cytoplasm by solid-phase extraction
	Wu, Jiabin; Cai, Zongwei
通讯作者	Cai, ZW
来源期刊	JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
ISSN	1570-0232
EISSN	1873-376X
出版年	2023
卷号	1215
英文摘要	The exact levels of some DNA adducts, like N7-deoxyguanosine (N7-dG), can be under-calculated since these adducts may depurinate due to their chemical instability, leading to corresponding nucleobase adducts being released into the cytoplasm. To accurately quantify the levels of DNA adducts, it is necessary to consider those modified nucleobases. However, high levels and diversity of cytoplasmic small molecule metabolites (SMMs) can strongly interfere with the detection of adducts, and it is almost impossible to remove them with nucleobase adducts being well retained. Therefore, we aimed to establish an optimized enrichment method based on solid -phase extraction (SPE) to reduce the co-elution of SMMs with target analytes. In this vein, we employed three bisphenols (BPA, BPF, and BPAF) as examples, prepared corresponding N7-guanine (N7-Gua) adducts, loaded on an Oasis hydrophilic-lipophilic balance (R) (HLB) cartridge, used a series of mobile phases containing different percentage of methanol for elution, and evaluated the levels of these adducts in each eluent. First, we found that neutral samples led to the best retention for all three adducts compared with acidified or basified ones. We next employed normal distribution fitting model to characterize the elution of analytes from H2O/methanol with different pHs and observed that neutral mobile phases resulted in more hydrophobic elution for all three adducts. Besides, N7-BPA-Gua and N7-BPF-Gua obtained narrow fitted peaks at neutral pH, while N7-BPAF-Gua had minimized elution windows at low pH. After optimization, we exposed 293T cells to the aforementioned bisphenols and quantified the N7-Gua adducts in the cytoplasm and the corresponding N7-dG adducts in genomic DNA. The results revealed that with the same levels of BPs exposure, BPAF led to the highest levels of adducts in both cytoplasm and genomic DNA samples, followed by BPA and BPF in order. In summary, our research established an appropriate model to describe the elution of DNA adducts in the SPE, applied it to optimize the loading and elution conditions, and discussed the genotoxicity of bisphenols by accurate quantification of both cleaved and uncleaved N7-dG adducts.
英文关键词	Nucleobase adduct LC-MS Solid phase extraction Normal distribution fitting
类型	Article
语种	英语
收录类别	SCI-E
WOS记录号	WOS:000914067700001
WOS关键词	LIVER DNA ; METABOLITE ; CHROMATOGRAPHY ; BIOLOGY ; SAMPLES ; PH
WOS类目	Biochemical Research Methods ; Chemistry, Analytical
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/397211
推荐引用方式 GB/T 7714	Wu, Jiabin,Cai, Zongwei. Enrichment of nucleobase adducts from genomic DNA in the cytoplasm by solid-phase extraction[J],2023,1215.
APA	Wu, Jiabin,&Cai, Zongwei.(2023).Enrichment of nucleobase adducts from genomic DNA in the cytoplasm by solid-phase extraction.JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,1215.
MLA	Wu, Jiabin,et al."Enrichment of nucleobase adducts from genomic DNA in the cytoplasm by solid-phase extraction".JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 1215(2023).