The 200 samples per day method
STANDARD METHOD

HIGH-THROUGHPUT ANALYSIS
The 200 SPD method is well suited for routine high-throughput applications. The timing and chromatographic performance is maximized to deliver an effective gradient elution of 82% and narrow peaks with FWHM of 1.7 seconds on average.
EXCELLENT SEQUENCING SPEED WITH MOST PEPTIDES PER MINUTE
Good chromatographic quality is essential, and especially to fully employ the high MS/MS scan rates on modern mass spectrometers.
The 200 SPD method provides excellent sequencing speed with most peptides per minute among our standard methods. This translates to a high number of proteins identified per minute.

Maximizing throughput, robustness and analytical depth for shotgun proteomics with the timsTOF Pro
This application note describes the timsTOF PASEF workflow, which enables sequencing speed of more than 120 Hz, high sensitivity and robustness as well as an additional dimension of separation (TIMS) for improved peak capacity. More than 1400 protein groups from 50 ng of HeLa digest were identified with the 200SPD method, all with outstanding robustness and reproducibility.
SENSITIVE ANALYSIS WITH FAIMS Pro coupled to an orbitrap exploris 480 ms
In this publication the Olsen Group from the Novo Nordisk Foundation Center for Protein Research, Denmark, showcase the sensitivity of the FAIMS Pro device on an Orbitrap Exploris 480 MS by using the 200 SPD method.
They analyzed 5 ng of tryptic peptides equivalent to approximately 30-50 HeLa cells. By using single compensation voltage (CV) of -50, they were able to identify more than 1000 proteins. FAIMS enhanced the reproducibility of DIA quantification, based on the number of proteins quantified with coefficient of variation below 20%.
USING ARTIFICIAL INTELLIGENCE ON ULTRAFAST LC-MS/MS DIA RUNS FOR BACTERIAL IDENTIFICATION IN URINE
In this seminar, Florence Roux-Dalvai presents a new strategy for bacterial species identification in urinary tract infection using artificial intelligence on ultrafast LCMS DIA runs with 200 samples per day.
The work is a collaboration between Québec Research Center, Canada, Thermo Scientific and Evosep Biosystems.
The six standard methods
With the six standard methods, the Evosep One covers a range of use cases from comprehensive proteome analysis with fractionation strategies to ultra high-throughput single-shot analysis. You decide how fast you want to go.
Standard method
The highest peak capacity per minute with our fastest method for ultra-high throughput analysis.
Standard method
Excellent sequencing speed with most peptides per minute among our standard methods.
Standard method
A great “in-between” method with run-to-run reproducibility of just one second.
Standard method
The most popular method with an excellent balance of throughput and depth.
Standard method
The longest method with a 44-minute gradient providing the best overall proteome coverage.
Explore more
Do you want to know how our users are already taking advantage of the 200 Samples per day method? Find all publications from users and collaboration partners here. Visit our Literature Room for a full overview.
Title | Subject | Material | Year | Summary | Institute | Evosep method | MS instrumentation | Learn More |
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Chromatographic separation of peptides and proteins for characterization of proteomes | DIA, FAIMS, Peptidomics | Journal article | 2022 | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | |||
Slice-PASEF: fragmenting all ions for maximum sensitivity in proteomics | PASEF | Publication | 2022 | 200 SPD | Bruker timsTOF Pro | |||
High-throughput quantitative proteomics using Zeno SWATH data-independent analysis (DIA) and the Evosep One system | DIA, Peptidomics, Protein interaction | Application note | 2022 | 100 SPD, 200 SPD, 30 SPD, 60 SPD | SCIEX ZenoTOF 7600 | |||
A fast and sensitive absolute quantification assay for the detection of sars-cov-2 peptides using parallel reaction monitoring mass spectrometry | Covid-19, DDA | Publication | 2022 | 200 SPD | Thermo Q Exactive HF | |||
Fully Automated High-Throughput Host Cell Protein Analysis of Highly Diverse Samples | DDA, HCP | Application note | 2021 | Novozymes produces recombinant enzymes and microbial solutions for a plethora of bio-based industries. To meet regulatory demands, they have developed a highly efficient and streamlined HCP workflow, which translate raw samples to data in less than 24 hours, with less than 2 hours of hands-on time. Read more about their workflow in this application note. | 200 SPD | Bruker timsTOF | ||
AlphaTims: Indexing trapped ion mobility spectrometry – time of flight data for fast and easy accession and visualization | DDA, DIA, Technology | Publication | 2021 | This publication by the Mann group, introduces the open-source AlphaTims package with a stand-alone graphical user interface or as part of their AlphaPept ecosystem. With this package, unprocessed data are indexed, accessed, and visualized by a combination of tools. | 200 SPD, 60 SPD | Bruker timsTOF Pro | ||
AlphaPept, a modern and open framework for MS-based proteomics | DDA, Technology | Publication | 2021 | This publication by the Mann group, introduces AlphaPept, a Python-based open-source framework for efficient processing of large high resolution MS data sets. They used the 200 samples per day method on the Evosep One to generate test data for computational functionality. | 200 SPD | Bruker timsTOF Pro | ||
Unleashing the power of DIA combined with short gradients using Evosep One | DIA, Technology | Webinar | 2021 | This webinar is part of our Evosep webinar series and hosted by Nicolai Bache, Evosep. Florian Meier and Vadim Demichev discuss their use of the Evosep One, using novel DIA methodologies. | 100 SPD, 200 SPD, 60 SPD | Bruker timsTOF Pro | ||
Towards a Standardized Omics Platform with the 200 samples per day method | Technology | Application note | 2019 | This application note describes the 200 samples per day standard method. Get an overview of the method, the chromatographic performance and the results expected from Hela standard injections. | 200 SPD | Thermo Orbitrap Exploris 480 | ||
The proteomics dilemma – High throughput analysis versus proteome depth | Fractionation, Technology | Application note | 2019 | With the five standard methods, the Evosep One covers a range of use cases from comprehensive proteome analysis with fractionation strategies to ultra high-throughput single-shot analysis. | 100 SPD, 200 SPD, 30 SPD, 300 SPD, 60 SPD | |||
High sensitivity dia-PASEF proteomics with DIA-NN and FragPipe | diaPASEF, Technology development | Publication | 2021 | The Ralser and Nesvizhskii groups introduce the latest addition to the DIA-NN software with a neural network-based ion mobility module. DIA-NN was originally conceived to maximize the performance of fast proteomics experiments and here they demonstrate the identification of more than 5000 proteins from the 200 SPD method. | 100 SPD, 200 SPD, 60 SPD | Bruker timsTOF Pro | Download publication or learn more on our website | |
Rapid, robust and reproducible host-cell protein analysis with Evosep One | DDA, HCP | Webinar | 2021 | This webinar is part of our Evosep webinar series and hosted by Nicolai Bache, Evosep. Jiao Ma and Anders Giessing discuss their use of the Evosep One to perform rapid, robust and reproducible HCP analysis. | 200 SPD, 60 SPD | Bruker timsTOF Pro, Thermo Orbitrap Fusion Lumos | ||
SPIN – Species by Proteome Investigation | Ancient proteomics, DIA | Publication | 2021 | This publication from the Olsen Group, introduces “Species by Proteome INvestigation” (SPIN), a proteomics workflow capable of querying over 150 mammalian species 200 times a day with the Evosep One. Genetic species determination is an indispensable tool in forensics, archaeology, ecology, and food authentication. | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | ||
diaPASEF: parallel accumulation–serial fragmentation combined with data-independent acquisition | diaPASEF, Technology | Publication | 2020 | A collaboration led by the Mann group at the Max Planck Institute for Biochemistry, Münich describes the development of the latest addition to the PASEF technology. The diaPASEF workflow combines the advantages of DIA such as high degree of reproducibility and data completeness with the effective PASEF technology. | 100 SPD, 200 SPD, 60 SPD | Bruker timsTOF Pro | Download publication or learn more on our website
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High throughput proteome and phosphorpoteome sample processing coupled to fast gradient DIA | DDA, DIA, FAIMS, Phosphorylation, PTM, Technology, Tissue, TMT | Publication | 2020 | High throughput workflows for proteomics | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | ||
New Orbitrap Exploris 480 Mass Spectrometer Coupled with Evosep One | DDA, DIA, FAIMS, Technology | Brochure | 2020 | Brochure of Exploris 480 with Evosep data from piece no 1 | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | ||
Online parallel accumulation – serial fragmentation (PASEF) with a novel trapped ion mobility mass spectrometer | DDA, Technology | Publication | 2018 | Description of timsTOF | 200 SPD | Bruker timsTOF Pro | ||
Improving proteome coverage and peptide identification rates in short LC gradients | DDA, DIA, FAIMS, Phosphorylation, PTM, Technology | Video | 2020 | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | |||
High-throughput proteomics with Evosep One | DDA, DIA, FAIMS, Phosphoproteomics, PTM, Technology | Video | 2020 | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | |||
Using Artificial Intelligence on Ultrafast LC-MSMS-DIA runs for Bacterial Identification in Urine | Bacteria, Clinical research, DIA | Video | 2019 | View the recording of our HUPO 2019 lunch seminar, where Florence Roux-Dalvai from Québec Research Center, Canada presents a new strategy for bacterial species identification in urinary tract infection using artificial intelligence on ultrafast LCMS DIA runs. | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | ||
High-throughput 4D-Proteomics – Application of dia-PASEF® and the Evosep One for short gradients | DIA, Technology | Application note | 2020 | diaPASEF app note | 100 SPD, 200 SPD, 300 SPD, 60 SPD | Bruker timsTOF Pro | ||
timsTOF Pro with PASEF and Evosep One: Maximizing throughput, robustness and analytical depth for shotgun proteomics | DDA, Technology | Application note | 2018 | Read this application note to learn more about the timsTOF Pro with PASEF and the Evosep One for maximizing throughput, robustness and analytical depth for shotgun proteomics. Increased peak capacity via trapped ion mobility spectrometry allows the analysis of 200 samples/day, at formidable analytical depth, with unparalleled robustness. | 100 SPD, 200 SPD, 60 SPD | Bruker timsTOF Pro | ||
Development of mass spectrometry-based targeted assay for direct detection of novel SARS-CoV-2 coronavirus from clinical specimens | Clinical research, Covid-19, DDA, Targeted workflow | Publication | 2020 | This publication by the Akhilesh Pandey group describes a targeted FAIMS-PRM assay on an Orbitrap Exploris 480 MS. More than 700 nasopharyngeal swab samples were analyzed with high specificity and comparable sensitivity to the gold standard RT-PCR method for the diagnosis of SARS-CoV-2. | 100 SPD, 200 SPD | Thermo Orbitrap Exploris 480 | ||
A Compact Quadrupole-Orbitrap Mass Spectrometer With Faims Interface Improves Proteome Coverage In Short Lc Gradients | Automation, DDA, DIA, FAIMS, Fractionation, Phosphoproteomics, PTM, Technology, Tissue, TMT | Publication | 2020 | Test of Orbitrap Exploris with DIA and TMT, proteome and phospho. | 100 SPD, 200 SPD, 60 SPD | Thermo Orbitrap Exploris 480 | ||
High-Throughput Proteomics Quantification Enabled by Fast LC Separation and Advanced PRM Acquisition | Phosphorylation, PTM, Targeted workflow | Poster | 2018 | High throughput LC-PRM to monitor the main protein components of AKT/mTOR signaling pathway. | 100 SPD, 200 SPD, 60 SPD | Thermo Q Exactive HF-X |
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