Microflow Liquid Chromatography Coupled to Multinozzle Electrospray Ionization for Improved Lipidomics Coverage of 3D Clear Cell Renal Cell Carcinoma

Article Article scientifique Article Microflow Liquid Chromatography Coupled to Multinozzle Electrospray Ionization for Improved Lipidomics Coverage of 3D Clear Cell Renal Cell Carcinoma eng 2025-02-25 2025-03-11 author Girel Sergey 0000-0002-8165-537X School of Pharmaceutical Sciences, University of Geneva, Geneva 4 1211, Switzerland.,Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4 1211, Switzerland. 941974 Section des sciences pharmaceutiques (FARMA) author Galmiche Mathieu 0000-0001-5840-4786 School of Pharmaceutical Sciences, University of Geneva, Geneva 4 1211, Switzerland.,Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4 1211, Switzerland.,Swiss Center of Applied Human Toxicology (SCAHT), Basel 4000, Switzerland. 406246 Section des sciences pharmaceutiques (FARMA) author Fiault Mathis 0009-0005-2068-9962 School of Pharmaceutical Sciences, University of Geneva, Geneva 4 1211, Switzerland.,Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4 1211, Switzerland.,Swiss Center of Applied Human Toxicology (SCAHT), Basel 4000, Switzerland. 416851 Section des sciences pharmaceutiques (FARMA) author Mieville Valentin 0009-0003-4626-0377 School of Pharmaceutical Sciences, University of Geneva, Geneva 4 1211, Switzerland.,Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4 1211, Switzerland. 855454 Section des sciences pharmaceutiques (FARMA) author Nowak-Sliwinska Patrycja School of Pharmaceutical Sciences, University of Geneva, Geneva 4 1211, Switzerland.,Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4 1211, Switzerland. 848409 Section des sciences pharmaceutiques (FARMA) author Rudaz Serge 0000-0002-4180-5417 School of Pharmaceutical Sciences, University of Geneva, Geneva 4 1211, Switzerland.,Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4 1211, Switzerland.,Swiss Center of Applied Human Toxicology (SCAHT), Basel 4000, Switzerland. 28978 Section des sciences pharmaceutiques (FARMA) author Meister Isabel 0000-0001-9063-0492 School of Pharmaceutical Sciences, University of Geneva, Geneva 4 1211, Switzerland.,Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4 1211, Switzerland.,Swiss Center of Applied Human Toxicology (SCAHT), Basel 4000, Switzerland. 968890 Section des sciences pharmaceutiques (FARMA) 5109-5117 10.1021/acs.analchem.4c06337 39998250 0003-2700 PMC11912133 https://pubs.acs.org/doi/10.1021/acs.analchem.4c06337 In most bioanalytical laboratories, high-resolution mass spectrometry (HRMS) systems with electrospray ionization (ESI) are hyphenated to liquid chromatography platforms. The latter typically operate under analytical flow (AF; 0.2-1 mL/min) regimes. Hence, AF/ESI-HRMS methods prioritize the detection of analytes of higher abundances or ionizability and tend to suffer from matrix effects or ion suppression. A far higher sensitivity can be obtained with electrospray at nanoflow (10-1000 nL/min) thanks to a better ionization efficiency and significant decrease in matrix effects. Both advantages are crucial to reliably accessing low-abundance compounds or weakly ionizable analytes. This work presents a microflow (μF) chromatographic setup coupled to a novel microfabricated multinozzle electrospray (mnESI) emitter with five nozzles spraying at 600 nL/min per nozzle for untargeted HRMS lipidomic profiling. With a runtime of 19 min, similar to our established analytical flow (AF/ESI) lipidomics platform, μF/mnESI produced a 16-fold median increase across 69 deuterated lipid standards. The performance of this new configuration was also evaluated in the context of the profiling of a 3D clear cell renal cell carcinoma (ccRCC) model exposed to a multidrug combination therapy. The processing of the acquired data resulted in 1270 (μF/mnESI) vs 752 (AF/ESI) MS<sup>2</sup> -annotated lipids. Among those, 762 achieved <10% variation on pooled QC samples for μF/mnESI compared to only 361 for the AF method. In addition, the measurements of ccRCC samples confirmed the improvements in ionization efficiency and adduct patterns observed with standards, enabling to annotate 79 oxidized triglycerides, 38 cholesterol esters (only five and four detected in AF/ESI, respectively), and 12 sitosterol esters, not yet reported in mammalian cell cultures. Dewey 615 Swiss Centre for Applied Human Toxicology 31 Section des sciences pharmaceutiques 17 false 881 Sciences analytiques 1325 Pharmacologie moléculaire Analytical chemistry 97 9 6e0756c9-22b6-4e6b-bc2a-ae491dc24fec Article (Published version) Public girel-et-al-2025-microflow-liquid-c.pdf 3300931 application/pdf CC BY-NC-ND-4.0 Restricted 2026-03-12 4b156f72e3c261b969fb0b15a3f6e7e52f62579aa5cdb2f1fa46445d85a50433 Vignette Public images_large_ac4c06337_0007.jpeg 131080 image/png openAccess Faculté des sciences / Section des sciences pharmaceutiques

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Microflow Liquid Chromatography Coupled to Multinozzle Electrospray Ionization for Improved Lipidomics Coverage of 3D Clear Cell Renal Cell Carcinoma	Analytical chemistry			2025	39	3

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