Evosep Proteomics / Evosep Eno
LC separation platform
Evosep Eno™
A standardized separation tool for proteomics. Ten characterized methods from 20 to 500 samples per day, retention time reproducibility measured in tenths of a second, and carryover below 0.05% — so the separation stops being the variable in your experiment.
Up to 500 samples/day
Up to 93% MS utilization
Six 96-tip racks on board
Five MS vendors
Evosep Eno is for General Laboratory Use.
9,240Proteins identified with the 30 SPD method on an Orbitrap Astral
~0.5 sIntra-instrument retention time SD; ~2 s across eight systems
<0.05%Carryover of total TIC for a 1 µg HeLa digest
1,056Samples processed from lysate to LC-MS result in ~2.5 days
2 amolDetection limit for standard peptides in targeted MRM assays
Why Evosep Eno
Separation as a constant, not a variable
Evosep Eno is a standardized LC separation platform built around Evotip Pure disposable trap columns. Samples arrive already desalted and ready to run, and partial elution from the Evotip means cleaner samples reach the analytical column — extending column lifetime and keeping the system robust across thousands of injections.
The patented architecture splits the work between a low-pressure subsystem, which minimises wear, and a high-pressure subsystem built for simplicity and reliability. Valves and pumps are controlled in real time by electronic hardware, which is where the retention time stability comes from — and it is that stability that makes tight scheduling and cross-site comparability possible.
Out of the box, not after optimisation. Fully characterized methods with specified columns mean consistent performance without method development. Most laboratories report installation and readiness within a few hours.
Reliability
Out-of-the-box consistent performance across systems, users and sites.
Standardization
Characterized methods with specified columns and documented performance metrics.
Low-maintenance design
Hybrid low/high-pressure architecture minimises wear; preventive maintenance once a year.
Sample handling
End-to-end workflows with room for six 96-tip Evotip racks on board.
Integration
Direct control through the mass spectrometer’s own software drivers.
Sample → digestion → Evotip Pure → Evosep Eno → mass spectrometer. Evosep standardizes the whole path rather than any single performance metric.
Standardized methods
Ten characterized methods, no method development
Six standard methods cover routine and high-throughput work; four Whisper™ Zoom methods cover high-sensitivity and low-input applications. Each ships with a documented description and expected performance metrics. Low overhead time keeps the mass spectrometer working — up to 93% utilization.
Standard methods
| Throughput | Cycle time | Gradient | Flow rate |
|---|---|---|---|
| 500 SPD | 2.90 min | 2.30 min | 4.0 µl/min |
| 300 SPD | 4.80 min | 3.95 min | 4.0 µl/min |
| 200 SPD | 7.20 min | 6.40 min | 2.0 µl/min |
| 100 SPD | 14.40 min | 12.95 min | 1.0 µl/min |
| 60 SPD | 24.00 min | 22.40 min | 0.6 µl/min |
| 30 SPD | 48.00 min | 45.40 min | 0.45 µl/min |
Whisper™ Zoom methods
| Throughput | Cycle time | Gradient | Flow rate |
|---|---|---|---|
| 120 SPD | 12.00 min | 10.50 min | 0.2 µl/min |
| 80 SPD | 18.00 min | 16.30 min | 0.2 µl/min |
| 40 SPD | 36.00 min | 32.80 min | 0.2 µl/min |
| 20 SPD | 72.00 min | 68.60 min | 0.2 µl/min |
Method selection balances speed against depth: moving from 300 to 200 SPD yields a 38% increase in proteome coverage for a 33% reduction in throughput. Peptide identifications stay evenly distributed across the hydrophobicity spectrum at every setting.
Performance
Coverage scales with the method, not with the operator
Standard methods evaluated with HeLa digest on an Orbitrap Astral in DIA mode, DIA-NN library-free analysis. Full conditions in application note AN-045C.
- 9,240Proteins identified with the 30 SPD method, at a mean protein CV of 3.6%.
- >8,000Proteins at 100 SPD throughput with an excellent protein CV of 4.8%.
- >6,000Proteins with the fastest 500 SPD method, mean CV 14.0% — CV stays below 15% across all standard methods at 200 ng input.
- >2,000Protein identifications per minute using the high-performance standardized methods.
- 2.4–5 sMedian peak width at half maximum across the method range.
Reproducibility that survives leaving the building
Intra-instrument retention time SD is approximately 0.5 seconds, measured on five diagnostic peptides across 15 injections. Across eight separate instruments, SD is approximately 2 seconds — the number that determines whether a scheduled assay transfers between labs.
Multi-laboratory study (AN-043B). Three independent laboratories, each with their own liquid handler, Evosep Eno and Orbitrap Astral, prepared 84 replicate samples. Across all three sites, 5,795 protein groups were consistently identified, with roughly 75% quantified at CV below 20% — despite independently prepared lysates. Evotip blanks showed less than 0.05% carryover, and plate blanks confirmed the workflow was free from cross-well contamination.
Sources: AN-045C and AN-043B in the literature room.
Operating at scale
A thousand samples, start to finish, in about two and a half days
From application note AN-046A: eleven sample plates prepared in parallel on a Biomek i7 liquid handler, then analysed with the 500 SPD method on Evosep Eno.
1,056 samples
Processed from protein lysate to LC-MS result in ~2.5 days, with minimal hands-on time.
0.75 s RT SD
Retention time stability across all 1,056 consecutive injections.
2.5 bar
High-pressure pump stability (SD) across the full run — no drift over a thousand samples.
4,600 proteins
Identified from just 200 ng protein input; 5,500 from 1 µg. Inter-plate CVs of 16–18%.
Why it holds up. At 500 samples per day, any instability compounds fast — a single disruption costs a plate, not an injection. Consistent digestion efficiency of 75% was maintained across all eleven plates, and the LC held retention time and pressure stable throughout.
Whisper™ Zoom
High sensitivity without giving up throughput
Whisper Zoom combines optimised gradients, low-flow conditions and synchronized hardware to hold sensitivity down to single-cell-equivalent inputs — accessible to non-experts, not just specialists.
- ~7,000Proteins identified from just 5 ng of HeLa digest with Whisper Zoom 120 SPD.
- +500%Throughput increase moving from 20 SPD to 120 SPD, with only a modest reduction in protein identifications and CV around 8.5%.
- <3 sRetention time SD for the 120, 80 and 40 SPD methods, even at low sample input (~8 s at 20 SPD).
- 0.25 ngLowest sample load evaluated, up to 200 ng — with identifications evenly spread across the gradient at every load.
At low inputs the faster methods perform as well as the slow ones: longer gradients add little benefit and can reduce identifications. When sample is limited, there is no need to sacrifice throughput to protect sensitivity.
Whisper Zoom supports single-cell proteomics, immunopeptidomics, post-translational modification analysis and deep visual proteomics. Source: BR-013C.
Targeted workflows
Quantification you can defend, at plasma scale
A fully automated plasma workflow coupled to a scheduled MRM assay on the 200 SPD method, monitoring 14 peptides across 11 plasma proteins on a Waters Xevo TQ-Absolute (AN-044F).
- 2 amolSensitivity for spiked standard peptides, with a linear response spanning 4–5 orders of magnitude.
- 93–110%Measured against expected ratios across a matrix-matched dilution series, R² > 0.99, for early, middle and late-eluting peptides — comfortably inside the ±15% window bioanalytical methods are normally held to.
- <0.5 sRetention time SD across 200 consecutive plasma samples.
- 1.1Mean peak symmetry, with 2.4 s FWHM and 7 s full width at base — enabling tight scheduling without sacrificing data points per peak.
Evotip Pure — the sample introduction step. A high-efficiency disposable trap column combining desalting with LC-MS introduction: partial elution, overload protection and no carryover. It also decouples sample preparation from MS availability, with clean-up, concentration and long-term storage of ready-to-analyse samples. Source: BR-013C.
Compatibility
Connects to the mass spectrometers labs already run
Optimized emitters and zero dead-volume connectors, with control provided through integrated software drivers — no separate control layer to learn.
Evosep Eno with mass spectrometer platforms from Bruker, Thermo Scientific, Waters, Agilent and SCIEX.
Supported MS drivers
Bruker Compass® HyStar
Thermo Scientific™ Xcalibur™
Thermo Scientific™ Chromeleon™
SCIEX OS
Agilent MassHunter
CHRONOS (Xcalibur and Analyst)
Specifications
Technical detail
Analytical
| Retention time reproducibility | SD less than 2 seconds (200 SPD method), evaluated over 8 systems |
| Carryover | <0.05% total TIC, 1 µg HeLa digest <0.1% total TIC, 1 µg digested whole plasma |
| Throughput | Up to 500 samples/day; up to 93% MS utilization rate |
| Sample tray format | 6 boxes of Evotips (96 tips per box) |
| Solvents | A: 0.1% formic acid in water · B: 0.1% formic acid in acetonitrile. LC-MS grade only. |
| Preventive maintenance | Annually |
Physical and environmental
| Weight | 37 kg (81.6 lb) |
| Dimensions (working range) | 880 mm D × 690 mm W × 910 mm H (34.6 × 27.2 × 35.8 in) |
| Base unit | 440 mm D × 420 mm W (17.3 × 16.5 in) |
| Power | 2 × 100–240 V, 50–60 Hz; combined consumption 320 W max |
| Air conditioning load | 320 W (max) |
| Operating conditions | Indoor use, up to 2000 m altitude. 15–30 °C ambient; 22 ± 3 °C for analytical specifications, fluctuation <1 °C/hr. 20–80% RH, non-condensing. |
Ordering information
| Part number | Product | Notes |
|---|---|---|
| EV1500 | Evosep Eno | LC separation platform |
| EV1137 | Performance column — 30 SPD | Operated at 40 °C |
| EV1109 | Performance column — 60/100 SPD | Operated at 40 °C |
| EV1182 | Performance column — 200/300/500 SPD | 4 cm, operated at 40 °C |
| EV1187 | Evosep Pod column oven | Thermo EasySpray |
| EV1189 | Evosep Pod column oven | Waters NanoLockSpray |
| EV2011 / EV2013 / EV2015 / EV2018 | Evotip Pure | 1 box (96) / 10 boxes / 50 boxes / 100 boxes |
Evosep Pod column ovens are also available for Thermo NanoSpray Flex and Agilent NanoSpray, and integrate with all Evosep Performance columns. Contact us for pricing and availability.
Service coverage
| Included | Endurance | Performance | Excellence |
|---|---|---|---|
| Troubleshooting wizards and online tools | ✓ | ✓ | ✓ |
| Remote diagnosis and repair support | Priority | Priority | Priority |
| Software and method upgrades | ✓ | ✓ | ✓ |
| Preventive maintenance | 1 per year | 1 per year | 1 per year |
| Max. on-site repair response | 5 business days | 3 business days | 3 business days |
Contracts also cover user certification training, spare and wear parts and free service repair visits at higher tiers, and are available with Installation and Operational Qualification. Terms and conditions apply. 90% of support cases are solved remotely, and cases are answered within 24 hours.
For existing Evosep One users
What changes when you move to Evosep Eno
Evosep Eno is the next-generation platform designed to replace Evosep One, with the same standardized workflow philosophy and a short learning curve.
Up to 35%
Better peak symmetry, and roughly 20% sharper peaks at base — driving signal intensity and sensitivity.
Up to 80%
Better retention time reproducibility, enabling precise scheduling for targeted assays.
>50%
More precursor identifications at the fastest standard methods, at the same or faster throughput.
>95%
Accuracy in high-throughput targeted MRM assays, with tighter distributions.
Can I use my existing Evotips and columns?
Evosep Eno uses Evotip Pure consumables and Evosep Performance columns. The workflow is familiar, but columns are specifically characterized for Eno methods to ensure optimal performance.
Will my existing methods transfer easily?
Yes. Evosep Eno uses standardized methods similar in structure to Evosep One, with documented performance metrics and a minimal learning curve.
Does higher throughput compromise data quality?
No. Evosep Eno was specifically designed to improve chromatography at fast gradients, which results in higher identification rates and better quantitative accuracy at high throughput.
Is Evosep Eno suitable for clinical or regulated workflows?
The improved reproducibility, retention time stability and standardization make Evosep Eno well suited to clinical, translational and multi-site studies. Note that Evosep Eno is for General Laboratory Use.
How quickly can the system be installed and operational?
Most laboratories report installation and readiness within a few hours, with minimal optimisation required.
What is the main reason to upgrade?
Higher throughput, improved reproducibility, deeper proteome coverage, and compatibility with next-generation mass spectrometers.
Go deeper
The data behind these numbers
Application note · AN-045C
New standards for routine, scalable and high-performance LC-MS proteomics
The six standard methods characterised: proteome coverage against throughput, peak performance, and inter-instrument reproducibility across eight systems.
Read it → Application note · AN-043BReproducible, inter-laboratory, scalable and routine LC-MS proteomics
Three laboratories, three Evosep Eno systems, independently prepared lysates — and 5,795 protein groups identified consistently across all sites.
Read it → Literature roomAll Evosep literature
Whitepapers, application notes and product literature across Evosep Eno, Evosep Lupo, Evokit and Evotip Pure.
Browse the library →
Overview
Evosep Proteomics — the full workflow
How Evokit, Evosep Lupo, Evotip Pure and Evosep Eno combine into one controlled sample-to-separation system.
Explore the ecosystem → Product pageEvosep Lupo™ — sample preparation system
Specifications, performance data, the Evokit portfolio and ordering information.
Explore Evosep Lupo →Get started
Standardize the last step before detection
Talk to an Evosep specialist about connecting Evosep Eno to your mass spectrometer and the throughput methods that fit your studies.