Initial annotation of consistently detected features (n = 3 technical replicates) was conducted using CEU Mass Mediator v3.0 (mass accuracy < 10 ppm) and further refined against Human Metabolome Database (HMDB) (< 30 ppm) and LIPID MAPS (< 20 ppm). Tentative identities were verified by comparing chromatographic retention with standards and confirmed through MS/MS spectral matching.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative","Liquid Chromatography MS - - reverse phase"],"chromatography_protocol":["PROSPR-EV lipid extracts were subjected to Liquid Chromatography–Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry (LC-ESI-Q-TOF) analysis using an Agilent 1290 Infinity II UHPLC system, which was coupled to a 6545 Q-TOF mass spectrometer equipped with a dual ESI source (Agilent Technologies, Santa Clara, USA). The autosampler was maintained at a temperature of 4 °C, and 10 µL of each extract was injected onto a Waters ACQUITY HSS T3 column (100 × 2.1 mm, 1.8 µm; Waters, Milford, USA) maintained at 55 °C. Chromatographic separation was conducted at a flow rate of 0.40 mL/min using solvent A (10 mM ammonium acetate in ACN/water, 40:60 v/v) and solvent B (10 mM ammonium acetate in ACN/isopropanol, 10:90 v/v). A linear gradient was employed, transitioning from 40% to 100% solvent B over 10 minutes, followed by a 2-minute hold at 100% B, and a 3-minute re-equilibration at 60% B, resulting in a total run time of 15 minutes.
"],"publication":["Optimized Enrichment of Circulating Extracellular Vesicles from Whole Blood Samples Using PROSPR."],"submitter_affiliation":["IRBLleida"],"submitter_name":["Aida Serra"],"organism_part":["Plasma","Blood"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["For the extraction of lipids, 5 µL of PROSPR-enriched EVs were combined with 5 µL of Milli-Q water and 20 µL of ice-cold methanol (Optima LC-MS grade, Thermo Fisher Scientific, Waltham, USA). The samples were subjected to vigorous vortexing for 2 minutes, followed by the addition of 250 µL of methyl tert-butyl ether (MTBE) (Sigma-Aldrich, St. Louis, USA). The mixture underwent ultrasound-assisted extraction by immersion in a water bath sonicator (ATU Ultrasonidos, Valencia, Spain) operating at 40 kHz and 100 W for 30 minutes at 10 °C. After sonication, 25 µL of Milli-Q water was added to facilitate phase separation. The samples were then centrifuged at 1400 × g for 10 minutes at 10 °C. The resultant upper organic phase, containing the EV-derived lipids, was meticulously collected and transferred to glass autosampler vials for further analysis. Stock solutions were formulated by dissolving lipid standards in MTBE at a concentration of 1 mg/mL, and subsequently, working solutions were diluted to 2.5 µg/mL in MTBE.
"],"organism":["Homo sapiens"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS12801"],"author":["Aida Serra. Instituto de Investigación Biomédica de Lleida. Department of Medical Basic Sciences (CMB) University of Lleida (UdL) Biomedical Research Institute of Lleida – (IRBLLEIDA) +Pec Proteomics Research Group (+PPRG) 80, Av. Rovira Roure, 25198, Lleida, Spain. aida.serra@udl.cat.","Xavier Gallart-Palau. Instituto de Investigación Biomédica de Lleida. Biomedical Research Institute of Lleida – (IRBLLEIDA) +Pec Proteomics Research Group (+PPRG) - Neuroscience Area University Hospital Arnau de Vilanova (HUAV) 80, Av. Rovira Roure, 25198, Lleida, Spain. xgallart@irblleida.cat."],"data_transformation_protocol":["In the processing of lipidomics data, raw LC-ESI-Q-TOF files derived from PROSPR blood- and plasma-EV extracts were analyzed using MassHunter Qualitative Analysis B.07.00 (Agilent Technologies, Barcelona, Spain). The Molecular Feature Extractor (MFE) algorithm was employed to group co-eluting adducts of identical molecular entities. Molecular features comprising two or more ions were exported to MassHunter Mass Profiler Professional for untargeted statistical analysis. Features were aligned with a retention-time tolerance of 0.1% ± 0.25 min and a mass tolerance of 30.0 ppm ± 2.0 mDa; only those present in at least 70% of quality-control (QC) injections were retained. Signal drift was corrected using QC-based LOESS normalization, followed by the application of univariate ANOVA (p < 0.05) and multivariate PLS-DA to identify discriminatory lipids.
"],"study_factor":["Sample type"],"submitter_email":["aida.serra@udl.cat"],"sample_collection_protocol":["Whole blood and derivative plasma samples were obtained from elderly donors without relevant diagnosed pathologies (n = 5) and stored at -80°C, as provided by the Newcastle Biobank, Newcastle upon Tyne, United Kingdom. An independent validation cohort of whole blood was also collected at the University Hospital Arnau de Vilanova (HUAV), Lleida, Spain (whole blood, n = 5; EDTA-plasma, n = 10) and supplied by the HUAV Biobank. For whole blood samples, the raw blood was frozen and stored until further analysis. For plasma samples, immediately following venepuncture, heparin was added to each tube to achieve a final concentration of 20 IU/mL. The samples were then centrifuged at 4,200 × g for 10 minutes at room temperature. In some cases, the whole blood fraction obtained after plasma extraction by centrifugation was retained for further analyses. The resulting plasma was aliquoted, and both the plasma and corresponding whole-blood aliquots were snap-frozen and stored at −80°C until required for use.
"],"omics_type":["Metabolomics"],"study_design":["Multi-omics study","untargeted metabolites","electrospray ionisation quadrupole detection"],"curator_keywords":["Multi-omics study","untargeted metabolites","electrospray ionisation quadrupole detection"],"mass_spectrometry_protocol":["The Q-TOF mass spectrometer acquired full-scan spectra (m/z 100–3000, 2 GHz, 1 scan/s) in separate positive- and negative-ion modes, utilizing nitrogen as the nebulizer gas (5 L/min, 350 °C) with a capillary voltage of 3.5 kV. Continuous mass-axis calibration was achieved through dual-spray infusion of reference ions (m/z 121.050873 and 922.009798 in positive mode; m/z 119.036320 and 966.000725 in negative mode).
"],"metabolite_name":["L_P_548.6831_7.3840017"],"additional_accession":[]},"is_claimable":false,"name":"Characterization of Whole Blood EVs Using PRotein Organic Solvent PRecipitation","description":"Blood plays a vital role in diagnostic medicine by providing essential information for assessing health and diagnosing various conditions. Routine blood tests can reveal important biomarkers that support early diagnosis and timely intervention. Recently, extracellular vesicles (EVs) have emerged as promising biomarkers due to their role in intercellular communication and involvement in disease processes such as cancer and neurodegeneration. However, isolating EVs from plasma remains challenging, as residual blood cells, especially platelets, can interfere with vesicle release. In this study, we explore the use of the PRotein Organic Solvent PRecipitation (PROSPR) method, a straightforward and effective technique previously applied to plasma and brain samples, to isolate EVs directly from whole blood. We performed detailed analyses, including ultrastructural imaging, cytometry, and multi-omics profiling, to characterize the EVs obtained. Our findings demonstrate that PROSPR can successfully capture even the smallest exosomes (30–120 nm), which are often missed in plasma-based isolation. This approach represents a significant advancement in EV research and may enhance the identification of blood-based biomarkers for a range of diseases.
","dates":{"publication":"2026-08-24","submission":"2025-07-31"},"accession":"MTBLS12801","cross_references":{}}