74th Annual ASMS Conference 2026

Recent advances in mass spectrometry have boosted both analytical speed and sensitivity, allowing for deeper proteome coverage and larger experimental cohorts. However, traditional nano-flow direct injection UHPLC workflows are still hindered by long sample loading times, which limit throughput and instrument use. Adding trap columns to UHPLC workflows is an effective way to solve these challenges, enabling fast, high-pressure loading at higher flow rates without sacrificing analytical quality.

Trap-and-elute performance was assessed using IonOpticks Aurora analytical columns in a high-throughput configuration (up to 200 samples per day). Trap-based workflows were directly compared with equivalent direct-injection runs to assess differences in chromatographic behavior and quantitative performance under rapid-sampling conditions. HeLa cell digests served as the test matrix and were loaded onto the system under controlled solvent and flow conditions prior to elution. Data were acquired using a Bruker nanoElute 2 UHPLC coupled to a Bruker timsTOF system, as well as an UltiMate 3000 UHPLC coupled to an Orbitrap Lumos mass spectrometer.

We developed a range of UHPLC workflows that integrate a novel trap-column design to minimise sample-loading overhead while maximising the time dedicated to chromatographic separation. In addition, these trap-based methods extend the longevity of analytical columns, reduce maintenance frequency, and deliver scalable solutions suitable for next-generation, high-throughput proteomic studies.

A 100 ng HeLa tryptic digest was analyzed using a reverse‑flush trap‑elute approach versus direct injection across two workflows: a 37‑min gradient on the Aurora® Ultimate™ CSI (25 cm × 75 μm) and a 14‑min gradient on the Aurora® Rapid™ CSI (8 cm × 75 μm). NanoShield trap column (5 cm x 100 μm, 3 μm C18 particles) produced less than a 1% reduction in protein identifications for both the 25 cm and the 8 cm columns; precursor identifications were reduced by 1–3% across the two column lengths. Similarly, a short NanoShield variation (1.5cm x 100 μm, 3 μm C18) showed similar direct-inject level performance in reverse flush orientation, and approximately a 2-5% reduction in identifications in forward flush orientation. Chromatographic quality remained excellent across all NanoShields tested, with only marginal increases in FWHM and minimal shifts in peak‑shape ratios relative to direct injection, and superior peak fidelity versus the alternative cartridge. These results demonstrate that NanoShield maintains deep proteome coverage and robust chromatographic performance in high‑throughput trap‑elute workflows.

For longevity testing, an Aurora Elite CSI (15 cm × 75 μm) column was paired with the NanoShield to evaluate long‑term performance under repeated use. Protein identifications, back-pressure, and retention times remain highly stable across more than 1,000 injections, demonstrating robust long-term performance.

The NanoShield® C18 trap column sets a new performance standard for trap-based proteomic workflows by capturing hydrophilic peptides and delivering near-direct injection performance while providing the essential benefits of in-line sample clean-up, analyte focusing and rapid sample loading.

Flexible trap-elute workflows achieve direct-injection-level performance while dramatically reducing loading time and enhancing high-throughput proteomics.