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Quantitative evaluation of peptide-extraction methods by HPLC-triple-quad MS-MS
Du, Yan1,2; Wu, Dapeng1; Wu, Qian1,2; Guan, Yafeng1
KeywordMethod Evaluation Peptide Extraction Plasma Triple-quad Ms-ms
Source PublicationANALYTICAL AND BIOANALYTICAL CHEMISTRY
2015-02-01
DOI10.1007/s00216-014-8389-0
Volume407Issue:6Pages:1595-1605
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordMOLECULAR-WEIGHT FRACTION ; MASS-SPECTROMETRY ; BIOMARKER DISCOVERY ; SHOTGUN PROTEOMICS ; ENRICHMENT METHODS ; SERUM SAMPLES ; PROTEINS ; CANCER ; BLOOD ; QUANTIFICATION
AbstractIn this study, the efficiency of five peptide-extraction methods-acetonitrile (ACN) precipitation, ultrafiltration, C18 solid-phase extraction (SPE), dispersed SPE with mesoporous carbon CMK-3, and mesoporous silica MCM-41-was quantitatively investigated. With 28 tryptic peptides as target analytes, these methods were evaluated on the basis of recovery and reproducibility by using high-performance liquid chromatography-triple-quad tandem mass spectrometry in selected-reaction-monitoring mode. Because of the distinct extraction mechanisms of the methods, their preferences for extracting peptides of different properties were revealed to be quite different, usually depending on the pI values or hydrophobicity of peptides. When target peptides were spiked in bovine serum albumin (BSA) solution, the extraction efficiency of all the methods except ACN precipitation changed significantly. The binding of BSA with target peptides and nonspecific adsorption on adsorbents were believed to be the ways through which BSA affected the extraction behavior. When spiked in plasma, the performance of all five methods deteriorated substantially, with the number of peptides having recoveries exceeding 70 % being 15 for ACN precipitation, and none for the other methods. Finally, the methods were evaluated in terms of the number of identified peptides for extraction of endogenous plasma peptides. Only ultrafiltration and CMK-3 dispersed SPE performed differently from the quantitative results with target peptides, and the wider distribution of the properties of endogenous peptides was believed to be the main reason.
Language英语
WOS IDWOS:000349337100008
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/145994
Collection中国科学院大连化学物理研究所
Affiliation1.Chinese Acad Sci, Key Lab Separat Sci Analyt Chem, Dept Instrumentat & Analyt Chem, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Dalian Inst Chem Phys, Beijing 100039, Peoples R China
Recommended Citation
GB/T 7714
Du, Yan,Wu, Dapeng,Wu, Qian,et al. Quantitative evaluation of peptide-extraction methods by HPLC-triple-quad MS-MS[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2015,407(6):1595-1605.
APA Du, Yan,Wu, Dapeng,Wu, Qian,&Guan, Yafeng.(2015).Quantitative evaluation of peptide-extraction methods by HPLC-triple-quad MS-MS.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,407(6),1595-1605.
MLA Du, Yan,et al."Quantitative evaluation of peptide-extraction methods by HPLC-triple-quad MS-MS".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 407.6(2015):1595-1605.
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