PXD082365

PXD082365 is an original dataset announced via ProteomeXchange.

Title	Integrated NanoLC and UPLC-MS Platforms for Single Cell and Complex Mixture Analysis
Description	Background: Multiple Reaction Monitoring (MRM) using triple quadrupole mass spectrometry (QqQ-MS) has become essential for multiplexed protein quantification. This study presents a comprehensive optimization of Agilent 6495C QqQ-MS equipped with both nano-electrospray ionization (nanoESI) and ultra-performance liquid chromatography ESI (UPLC-ESI) sources for enhanced proteomics analysis. We systematically optimized source parameters and compared spectral intensities using a standard peptide mixture (6×5 LC-MS/MS Peptide Reference Mix). Results: The nanoLC system, operating at sub-microliter flow rates, demonstrated over 100-fold enhancement in spectral intensity compared to UPLC when analyzing equivalent sample amounts, highlighting its superior sensitivity for trace-level detection. Performance validation using 30 stable isotope-labeled peptides at varying concentrations confirmed the significant improvements achieved through systematic parameter optimization. Additionally, we evaluated the Multiple Heart-Cutting (MHC) 2D-LC system's separation capabilities using hyaluronic acid polymers. The MHC approach effectively resolved overlapping peaks by selectively transferring UV-detected peaks from the first dimension to a second dimension with orthogonal selectivity. Successful separation of hyaluronic acid tetramer [M+H]+ peaks within complex polymer mixtures validated the system's performance for challenging separations. Significance: These findings demonstrate that the integration of optimized nanoESI sources with nanoLC systems substantially enhances MRM performance for biomarker discovery and validation. The combination of enhanced sensitivity from nanoLC-QqQ optimization and superior separation capability of MHC 2D-LC presents a powerful analytical platform for precise quantification of target molecules in complex biological matrices, advancing proteomics research and clinical biomarker development.
HostingRepository	PanoramaPublic
AnnounceDate	2026-08-09
AnnouncementXML	Submission_2026-08-09_22:47:06.636.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	supported by repository but incomplete data and/or metadata
PrimarySubmitter	Hyunsoo Kim
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	Label:13C(3)15N(1); Label:13C(6)15N(4); Label:13C(6)15N(1); Label:13C(5)15N(1); Label:13C(6)15N(2); Label:13C(9)15N(1); mTRAQ:13C(3)15N(1)
Instrument	6495 Triple Quadrupole LC/MS

Revision	Datetime	Status	ChangeLog Entry
0	2026-08-09 22:40:51	ID requested
⏵ 1	2026-08-09 22:47:07	announced

no publication

submitter keyword: Targeted mass spectrometry, Multiple reaction monitoring, Integrated nanoLC-UPLC platform, Multiple heart-cutting 2D-LC

Hyunsoo Kim
contact affiliation	Chungnam National University
contact email	kimlab@cnu.ac.kr
lab head
Hyunsoo Kim
contact affiliation	Chungnam National University
contact email	kimlab@cnu.ac.kr
dataset submitter

Panorama Public dataset URI