分析化学2025，Vol.53Issue(11)：1820-1827,中插1-中插3,11.DOI:10.19756/j.issn.0253-3820.251227
唾液多肽组单次分析结果的不确定性和可拓展性研究
Study on Uncertainty and Scalability in Single-Analysis Salivary Peptidome Studies
摘要
Abstract
Nano liquid chromatography-high resolution tandem mass spectrometry(LC-HRMS/MS)is widely used for body fluid peptidome analysis,yet the impact of replicate analyses remains overlooked.Using salivary peptidome,in this work,10 replicate analyses of the same sample were conducted.It was found that although m/z and molecular weight ranges were consistent across replicates,single runs identified only 348-576 unique peptides from 32-39 degraded proteins.Merging all replicates revealed 1237 peptides from 77 proteins,a 2.5 folds increase in peptides and 2 folds increase in proteins.Instrument stability was confirmed via intensity/retention time of 12 peptides.Merged peptides primarily derived from high-abundance proteins(e.g.,Statherin,PRP1/2).Analysis of 20 peptides showed that the some peptides were detected in single analysis but confidence varying(19%-99%),low-confidence peptides(<95%)exceeded 95%after replicate.signal intensity alone didn't determine confidence and peptides spanning the sequence region between the longest and the shortest detected fragments could be identified.The above findings suggested that single-run LC-MS peptidomics analysis carried inherent false negatives and uncertainties.However,by integrating the results of a single analysis with the mechanism of enzymatic hydrolysis for endogenous peptides,it was possible to make reasonable inferences and extrapolations regarding peptides derived from highly abundant degraded proteins.关键词
多肽组/唾液/假阴性/重复次数/高分辨串联质谱Key words
Peptidomics/Saliva/False negative/Replicates/High-resolution tandem mass spectrometry引用本文复制引用
FANG Xiu-Feng,CUI Bing-Jia,XU Jin-Ling,WANG Yong..唾液多肽组单次分析结果的不确定性和可拓展性研究[J].分析化学,2025,53(11):1820-1827,中插1-中插3,11.基金项目
深港脑科学创新研究院项目(No.2023SHIBS0003)资助. Supported by the Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions(No.2023SHIBS0003). （No.2023SHIBS0003）
