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Utility of Atmospheric-Pressure Chemical Ionization and Photoionization Mass Spectrometry in Bottom-Up Proteomics
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SYSNO ASEP 0556186 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Utility of Atmospheric-Pressure Chemical Ionization and Photoionization Mass Spectrometry in Bottom-Up Proteomics Author(s) Sedláčková, Simona (UOCHB-X)
Hubálek, Martin (UOCHB-X) RID, ORCID
Vrkoslav, Vladimír (UOCHB-X) RID, ORCID
Blechová, Miroslava (UOCHB-X)
Cvačka, Josef (UOCHB-X) RID, ORCIDArticle number 42 Source Title Separations. - : MDPI
Roč. 9, č. 2 (2022)Number of pages 19 s. Language eng - English Country CH - Switzerland Keywords chemical ionization ; peptides ; photoionization ; proteomics OECD category Analytical chemistry R&D Projects GA20-09126S GA ČR - Czech Science Foundation (CSF) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 000767961800001 EID SCOPUS 85124237379 DOI 10.3390/separations9020042 Annotation In a typical bottom-up proteomics workflow, proteins are enzymatically cleaved, and the resulting peptides are analyzed by HPLC with electrospray ionization (ESI) tandem mass spectrometry. This approach is practical and widely applied. It has, however, limitations mostly related to less efficient or even inefficient ionization of some peptides in ESI sources. Gas-phase ionization methods like atmospheric-pressure chemical ionization (APCI) or atmospheric-pressure photoionization (APPI) offer alternative ways of detecting various analytes. This work is a systematic study of the ionization efficiencies of peptides in ESI, APCI, and APPI and the applicability of the mentioned ionizations in proteomics. A set of peptide standards and bovine serum albumin digests were examined using a high-resolution mass spectrometer coupled to an ultra HPLC system. Since the ionization efficiency in APCI and APPI depends strongly on experimental conditions, the ion source settings and mobile phase compositions were optimized for each ionization technique. As expected, tryptic peptides were best detected using ESI. The numbers of chymotrypsin peptides successfully detected by ESI, APPI, and APCI were comparable. In the case of Glu-C digest, APPI detected the highest number of peptides. The results suggest that gas-phase ionization techniques, particularly APPI, are an interesting alternative for detecting peptides and delivering complementary data in proteomics. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2023 Electronic address https://doi.org/10.3390/separations9020042
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