Usability
Fraction collection of liquid chromatographic (LC) separations is used for analyte purification, but also for applications in which a detection technique cannot be coupled online to LC due to compatibility issues. It is most common to apply fractionation in environmental toxicant screening, drug discovery research and food chemistry, combining chemical analysis with bioassay testing for the identification of bioactive substances. In these types of studies, fractionation is required to reduce the sample complexity until preferably only one single compound is present in a fraction. Subsequently, each fraction is tested for biological activity and the detected bioactives are potentially identified.
Alternatively, this approach is used to purify sufficient amounts of material for typical off-line analyses such as NMR. This traditional process usually requires many orthogonal separations to obtain the pure compounds for chemical structure elucidation and furthermore many repeated fractionation cycles are required for analyte isolation.
All this results in a very costly process that is time consuming and prone to analyte losses. In environmental chemistry, this huge bottleneck has recently been solved by establishing High- Throughput Effect-Directed-Analysis (HT-EDA) which shifted traditional EDA towards high resolution small-volume fractionation.
Compounds in a sample are separated with liquid chromatography (LC). Subsequently, the eluent is split and one part is transferred by the innovative FractioMate spotting technology to a high-resolution bioassay (e.g. 96 or 384 well mammalian or yeast-based cellular format) to test the biological activity of eluting compounds. The other part is directed to a mass spectrometer. Peak shapes from ‘reconstructed bioassay chromatograms’ using the individually collected and bioassayed fractions are efficiently correlated to compound accurate masses from the parallel obtained chemical data (in most cases MS data).
Features
- Direct integration of biological and chemical lab.
- Lengthy EDA studies reduced to a few days.
- Capacity increase by orders of magnitude; resulting in 5 to 10 samples per week.
- Integration results in much faster correlation of bioactivity / toxicity to accurate mass.
Specifications
- Optimized for UHPLC
- Low dead volume spotting / dispensing
- Standard configuration with two well plates
- Suited for most liquid chromatography post-column fractionation applications
- Adjustable 1s > well fractionation time
- Dispense precision ≤ 4.4% CV
- Standard 96 and 384 well plate spotting for nanofractionation
- Flow rate 10μl/min – 1ml/min
- Spot volume 0.1μl- 1.7μl
- Spotting frequency 1-50Hz
Download
Download brochure
Credentials
The FractioMateTM is developed at the Vrije Universiteit Amsterdam by the BioMolecular Analysis group, Jeroen Kool en Marja Lamoree, the department of Environment & Health and the Electronics and Mechanics groups of the Faculty of Science in close cooperation with SPARK and Waterlaboratorium, Haarlem the Netherlands.
Read also the credential of Marja Lamoree of the BioMolecular Analyses Group Vrije Universiteit Amsterdam.
Related publications
Slagboom J, Derks RJE, Sadighi R, Somsen GW, Ulens C, Casewell NR, Kool J.
High-Throughput Venomics. Journal of Proteome Research, 2023, 22(6):1734–1746.
https://doi.org/10.1021/acs.jproteome.2c00780
van Thiel J, Alonso LL, Slagboom J, Dunstan N, Wouters RM, Modahl CM, Vonk FJ, Jackson TNW, Kool J.
Highly Evolvable: Investigating Interspecific and Intraspecific Venom Variation in Taipans (Oxyuranus spp.) and Brown Snakes (Pseudonaja spp.). Toxins, 2023, 15(1):74.
https://doi.org/10.3390/toxins15010074
Terzioglu S, Bittenbinder MA, Slagboom J, van de Velde B, Casewell NR, Kool J.
Analytical Size Exclusion Chromatography Coupled to Nanofractionation Analytics for Coagulopathic Venom Profiling. Toxins, 2023, 15(9):552.
https://doi.org/10.3390/toxins15090552
Weekers DJC, Alonso LL, Verstegen AX, Slagboom J, Kool J.
Qualitative Profiling of Venom Toxins in the Venoms of Several Bothrops Species Using High-Throughput Venomics and Coagulation Bioassaying. Toxins, 2024, 16(7):300.
https://doi.org/10.3390/toxins16070300
Xu H, Mastenbroek J, Krikke NTB, El-Asal S, Mutlaq R, Casewell NR, Slagboom J, Kool J.
Nanofractionation Analytics for Comparing MALDI-MS and ESI-MS Data of Viperidae Snake Venom Toxins. Toxins, 2024, 16(8):370.
https://doi.org/10.3390/toxins16080370
Palermo P, Schouten WM, Alonso LL, Ulens C, Kool J, Slagboom J.
Acetylcholine-Binding Protein Affinity Profiling of Neurotoxins in Snake Venoms with Parallel Toxin Identification. International Journal of Molecular Sciences, 2023, 24(23):16769.
https://doi.org/10.3390/ijms242316769
Houtman CJ, Janssen EH, Knol O, Lamoree MH, Leonards PEG, Kool J, Hamers T.
Characterisation of (anti-)progestogenic and (anti-)androgenic activities in wastewater using reporter gene bioassays after fractionation. Environment International, 2021, 152:106473.
https://doi.org/10.1016/j.envint.2021.106473
Xie C, Bittenbinder MA, Slagboom J, Arrahman A, Bruijns S, Somsen GW, Vonk FJ, Casewell NR, García-Vallejo JJ, Kool J.
Erythrocyte Haemotoxicity Profiling of Snake Venom Toxins after Nanofractionation. Journal of Chromatography B, 2021, 1176:122586.
https://doi.org/10.1016/j.jchromb.2021.122586
Xie C, Slagboom J, Albulescu LO, Somsen GW, Vonk FJ, Casewell NR, Kool J.
Neutralising Effects of Small Molecule Toxin Inhibitors on Nanofractionated Coagulopathic Crotalinae Snake Venoms. Acta Pharmaceutica Sinica B, 2020, 10(10):1835–1845.
https://doi.org/10.1016/j.apsb.2020.07.010
Xie C, Albulescu LO, Bittenbinder MA, Somsen GW, Vonk FJ, Casewell NR, Kool J.
Antivenom Neutralization of Coagulopathic Snake Venom Toxins Assessed by Bioactivity Profiling Using Nanofractionation Analytics. Toxins, 2020, 12(1):53.
https://doi.org/10.3390/toxins12010053
Zwart N, Jonker W, ten Broek R, de Boer J, Somsen GW, Kool J, Hamers T, Houtman CJ, Lamoree MH.
Identification of Mutagenic and Endocrine Disrupting Compounds in Surface Water and Wastewater Treatment Plant Effluents Using High-Resolution Effect-Directed Analysis. Water Research, 2020, 168:115204.
https://doi.org/10.1016/j.watres.2019.115204
Slagboom J, Mladić M, Xie C, Kazandjian TD, Vonk FJ, Somsen GW, Casewell NR, Kool J.
High-throughput screening and identification of coagulopathic snake venom proteins and peptides using nanofractionation and proteomics approaches. PLOS Neglected Tropical Diseases, 2020, 14(4): e0007802.
https://doi.org/10.1371/journal.pntd.0007802
Bittenbinder MA, Capinha L, Da Costa Pereira D, Slagboom J, van de Velde B, Casewell NR, Jennings P, Vonk FJ, Kool J.
Development of a High-Throughput In Vitro Screening Method for the Assessment of Cell-Damaging Activities of Snake Venoms. PLOS Neglected Tropical Diseases, 2023, 17(9):e0011763.
https://doi.org/10.1371/journal.pntd.0011763
Bittenbinder MA, Wachtel E, Da Costa Pereira D, Slagboom J, Casewell NR, Jennings P, Vonk FJ, Kool J.
Development of a Membrane-Disruption Assay Using Phospholipid Vesicles as a Proxy for the Detection of Cellular Membrane Degradation. Toxicon X, 2024, 22:100197.
https://doi.org/10.1016/j.toxcx.2024.100197
Bittenbinder MA, van Thiel J, Cardoso FC, Casewell NR, Gutiérrez JM, Vonk FJ, Kool J.
Tissue Damaging Toxins in Snake Venoms: Mechanisms of Action, Pathophysiology and Treatment Strategies. Communications Biology, 2024, 7:358.
https://doi.org/10.1038/s42003-024-05852-y
Bittenbinder MA, Bergkamp ND, Slagboom J, Bebelman JPM, Casewell NR, Siderius MH, Smit MJ, Vonk FJ, Kool J.
Monitoring Snake Venom-Induced Extracellular Matrix Degradation and Identifying Proteolytically Active Venom Toxins Using Fluorescently Labeled Substrates. Biology (Basel), 2023, 12(6):765.
https://doi.org/10.3390/biology12060765
Alvarez-Mora I, FitzGerald R, O’Connell S, Williams GP, Riako D, Lamoree MH, Kool J, Hamers T.
High-Throughput Effect-Directed Analysis of Androgenic Compounds in Hospital Wastewater Using Reporter Gene Bioassays after FractioMate™-Based Fractionation. Environmental Science & Technology, 2025, 59(xx):xxxx–xxxx.
https://doi.org/10.1021/acs.est.4c09942
Langberg HA, van der Meer TP, Vermeirssen ELM, Houtman CJ, Leonards PEG, Lamoree MH, Hamers T.
Effect-Directed Analysis Based on Transthyretin Binding Activity of PFAS in a Contaminated Sediment Extract. Environmental Toxicology and Chemistry, 2024, 43(2):245–258.
https://doi.org/10.1002/etc.5777
Baygildiev T, Schneider A, Vermeirssen E, Brack W, Krauss M.
Identification of Polar Bioactive Substances in the Upper Rhine Using Effect-Directed Analysis with FractioMate™-Based Nanofractionation. Water Research, 2025, 268:122607.
https://doi.org/10.1016/j.watres.2024.122607
Menzies SK, Petras D, Albulescu LO, Still KBM, Kurian A, D’Souza R, Kamiguti A, Casewell NR.
In Vitro and In Vivo Preclinical Venom Inhibition Assays Identify Metalloproteinase-Inhibiting Drugs as Potential Treatments for Snakebite Envenoming by Dispholidus typus. Toxicon: X, 2022, 14:100118.
https://doi.org/10.1016/j.toxcx.2022.100118
Jonkers TJH, van der Meer TP, Houtman CJ, Hamers T, de Boer J, Lamoree MH.
High-Performance Data Processing Workflow Incorporating Effect-Directed Analysis for Feature Prioritization in Complex Environmental Samples. Environmental Science & Technology, 2022, 56(3):1639–1651.
https://doi.org/10.1021/acs.est.1c04168
Arrahman A, Xu H, Khan MA, Bos TS, Slagboom J, van der Velden GC, Nehrdich U, Casewell NR, Richardson MK, Tudorache C, Cardoso FC, Kool J.
Parallel In Vitro Ion Channel and In Vivo Zebrafish Assaying of Elapid Snake Venoms Following Chromatographic Separation of Toxin Components. SLAS Discovery, 2025, 30(3):100239.
https://doi.org/10.1016/j.slasd.2025.100239
More information
Precision Mechanics and Engineering Group VU
Joost Rosier
T +31 20 59 87439
E j.c.rosier@vu.nl