biostudies-literatureXu TMedical and Industry Joint Innovation ProgramsNational Natural Science Foundation of ChinaInnovation ProgramAffiliated Hospital of Dalian Medical University and DICPDICP, CASNational Key Research and Development Program of Chinae2306659https://www.ebi.ac.uk/biostudies/studies/S-EPMC11040340biostudies-literatureUnknown11(16)High-coverage mass spectrometry analysis of single-cell metabolomics remains challenging due to the extremely low abundance and wide polarity of metabolites and ultra-small volume in single cells. Herein, a novel concentric hybrid ionization source, nanoelectrospray ionization-atmospheric pressure chemical ionization (nanoESI-APCI), is ingeniously designed to detect polar and nonpolar metabolites simultaneously in single cells. The source is constructed by inserting a pulled glass capillary coaxially into a glass tube that acts as a dielectric barrier layer. Benefitting from the integrated advantages of nanoESI and APCI, its limit of detection is improved by one order of magnitude to 10 pg mL^-1. After the operational parameter optimization, 254 metabolites detected in nanoESI-APAdvanced science (Weinheim, Baden-Wurttemberg, Germany)Concentric Hybrid Nanoelectrospray Ionization-Atmospheric Pressure Chemical Ionization Source for High-Coverage Mass Spectrometry Analysis of Single-Cell Metabolomics.PMC11040340DMU-1&DICP UN20220322274156DICP I202141222041632022YFC340120021934006DMU-2&DICP UN20230222074144Li HTan GHu XDou PFeng DChen CXu GShi XHu CMu HZhang ZWang TXu TLuo YPu SfalseConcentric Hybrid Nanoelectrospray Ionization-Atmospheric Pressure Chemical Ionization Source for High-Coverage Mass Spectrometry Analysis of Single-Cell Metabolomics.High-coverage mass spectrometry analysis of single-cell metabolomics remains challenging due to the extremely low abundance and wide polarity of metabolites and ultra-small volume in single cells. Herein, a novel concentric hybrid ionization source, nanoelectrospray ionization-atmospheric pressure chemical ionization (nanoESI-APCI), is ingeniously designed to detect polar and nonpolar metabolites simultaneously in single cells. The source is constructed by inserting a pulled glass capillary coaxially into a glass tube that acts as a dielectric barrier layer. Benefitting from the integrated advantages of nanoESI and APCI, its limit of detection is improved by one order of magnitude to 10 pg mL^-1. After the operational parameter optimization, 254 metabolites detected in nanoESI-AP2024-01-01T00:00:00Z2024 Apr2025-04-22T20:44:37.323Z2025-04-06T03:15:17.898ZS-EPMC110403403835900510.1002/advs.202306659