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Reaction screening in multiwell plates: high-throughput optimization of a Buchwald–Hartwig amination


Chemical space is vast, and chemical reactions involve the complex interplay of multiple variables. As a consequence, reactions can fail for subtle reasons, necessitating screening of conditions. High-throughput experimentation (HTE) techniques enable a more comprehensive array of data to be obtained in a relatively short amount of time. Although HTE can be most efficiently achieved with automated robotic dispensing equipment, the benefits of running reaction microarrays can be accessed in any regularly equipped laboratory using inexpensive consumables. Herein, we present a cost-efficient approach to HTE, examining a Buchwald–Hartwig amination as our model reaction. Experiments are carried out in a machined aluminum 96-well plate, taking advantage of solid transfer scoops and pipettes to facilitate rapid reagent transfer. Reaction vials are simultaneously heated and mixed, using a magnetic stirrer, and worked up in parallel, using a plastic filter plate. Analysis by gas chromatography provides the chemist with 96 data points with minimal commitment of time and resources. The best-performing experiment can be selected for scale-up and isolation, or the data can be used for designing future optimization experiments.

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Fig. 1: Advantages of a high-throughput approach to optimizing reaction conditions.
Fig. 2
Fig. 3: Equipment used within this protocol.
Fig. 4
Fig. 5: Preparation for high-throughput experimentation.
Fig. 6: Essential equipment for high-throughput reaction setup.
Fig. 7: Layout for dosing the 96-well plate in a high-throughput evaluation of the Buchwald–Hartwig amination.
Fig. 8: Assembly of a 96-well plate for high-throughput experimentation.
Fig. 9: Equipment used during the analysis preparation of a high-throughput experiment.
Fig. 10: Depicting the results obtained from this protocol.

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Data availability

The authors declare that all the data supporting the findings of this study are available within the article and in the Supplementary Information files. All the data analysis was performed using published tools and packages and has been provided with the paper.


  1. Shevlin, M. Practical high-throughput experimentation for chemists. ACS Med. Chem. Lett. 8, 601–607 (2017).

    Article  CAS  Google Scholar 

  2. Vries, J. G. D. & Vries, A. H. M. D. The power of high-throughput experimentation in homogeneous catalysis research for fine chemicals. ChemInform 34, 799–811 (2003).

    Article  Google Scholar 

  3. Schmink, J. R., Bellomo, A. & Berritt, S. Scientist-led high-throughput experimentation and its utility in academia and industry. Aldrichimica Acta 46, 71–80 (2013).

    Google Scholar 

  4. Dreher, S. D. et al. Nanomole-scale high-throughput chemistry synthesis of complex molecules. Science 347, 49–53 (2015).

    Article  Google Scholar 

  5. Krska, S. W., DiRocco, D. A., Dreher, S. D. & Shevlin, M. The evolution of chemical high-throughput experimentation to address challenging problems in pharmaceutical synthesis. Acc. Chem. Res. 50, 2976–2985 (2017).

    Article  CAS  Google Scholar 

  6. Shah, A. A., Kelly, M. J. & Perkins, J. J. Access to unnatural α-amino acids via visible-light-mediated addition to dehydroalanine. Org. Lett. 22, 184–196 (2020).

    Article  Google Scholar 

  7. Collins, K. D., Gensch, T. & Glorius, F. Contemporary screening approaches to reaction discovery and development. Nat. Chem 6, 859–871 (2014).

    Article  CAS  Google Scholar 

  8. Shevlin, M. et al. Nickel-catalyzed asymmetric alkene hydrogenation of α,β-unsaturated esters: high-throughput experimentation-enabled reaction discovery, optimization and mechanistic elucidation. J. Am. Chem. Soc. 16, 3562–3569 (2016).

    Article  Google Scholar 

  9. Troshin, K. & Hartwig, J. F. Snap deconvolution: an informatics approach to high-throughput discovery of catalytic reactions. Science 357, 175–181 (2017).

    Article  CAS  Google Scholar 

  10. Robbins, D. W. & Hartwig, J. F. A simple, multidimensional approach to high-throughput discovery of catalytic reactions. Science 333, 1423–1427 (2011).

    Article  CAS  Google Scholar 

  11. McNally, A., Prier, C. K. & MacMillan, D. W. C. Discovery of an α-amino C-H arylation reaction using the strategy of accelerated serendipity. Science 334, 1114–1117 (2011).

    Article  CAS  Google Scholar 

  12. Collins, K. D. & Glorius, F. A robustness screen for the rapid assessment of chemical reactions. Nat. Chem. 5, 597–601 (2013).

    Article  CAS  Google Scholar 

  13. Granda, J. M., Donina, L., Dragone, V., Long, D.-L. & Cronin, L. Controlling an organic synthesis robot with machine learning to search for new reactivity. Nature 559, 377–381 (2018).

    Article  CAS  Google Scholar 

  14. Ahneman, D. T., Estrada, J. G., Lin, S., Dreher, S. D. & Doyle, A. G. Predicting reaction performance in C-N cross-coupling using machine learning. Science 360, 186–190 (2018).

    Article  CAS  Google Scholar 

  15. Mann, D. J. et al. High-throughput kinetic analysis for target-directed covalent ligand discovery. Angew. Chem. Int. Ed. 57, 5257–5261 (2018).

    Article  Google Scholar 

  16. Campeau, L.-C. & Hazari, N. Cross-coupling and related reactions: connecting past successes to the development of new reactions for the future. Organometallics 38, 3–35 (2019).

    Article  CAS  Google Scholar 

  17. Hansen, E. C. et al. New ligands for nickel catalysis from diverse pharmaceutical heterocycle libraries. Nature Chem 8, 1126–1130 (2016).

    Article  CAS  Google Scholar 

  18. Larson, H., Schultz, D. & Kalyani, D. Ni-catalyzed C-H arylation of oxazoles and benzoxazoles using pharmaceutically relevant aryl chlorides and bromides. J. Org. Chem. 84, 13092–13103 (2019).

    Article  CAS  Google Scholar 

  19. Nelson, J. J. M. et al. High-throughput screening for discovery of benchtop separations systems for selected rare earth elements. Commun. Chem. 3, 1–6 (2020).

    Article  Google Scholar 

  20. Herrera, B. T. et al. Rapid optical determination of enantiomeric excess, diastereomeric excess, and total concentration using dynamic-covalent assemblies. A demonstration using 2-aminocyclohexanol and chemometrics. J. Am. Chem. Soc. 141, 11151–11160 (2019).

    Article  CAS  Google Scholar 

  21. Welch, C. J. et al. The enabling technologies consortium (ETC): fostering precompetitive collaborations on new enabling technologies for pharmaceutical research and development. Org. Process Res. Dev. 21, 414–419 (2017).

    Article  CAS  Google Scholar 

  22. Allen, C. L., Leitch, D. C., Anson, M. & Zajac, M. A. The power and accessibility of high-throughput methods for catalysis research. Nat. Catal. 2, 2–4 (2019).

    Article  Google Scholar 

  23. Vandavasi, J. K. & Newman, S. G. A high-throughput approach to discovery: Heck-type reactivity with aldehydes. Synlett 29, 2081–2086 (2018).

    Article  CAS  Google Scholar 

  24. Isbrandt, E. S., Sullivan, R. J. & Newman, S. G. High-throughput strategies for the discovery and optimization of catalytic reactions. Angew. Chem. Int. Ed. 58, 7180–7191 (2019).

    Article  CAS  Google Scholar 

  25. Kashani, S. K., Jessiman, J. & Newman, S. G. Exploring homogenous conditions for mild Buchwald-Hartwig amination in batch and flow. Org. Process Res. Dev. 24, 1948–1954 (2020).

    Article  CAS  Google Scholar 

  26. Yin, J. & Buchwald, S. L. Palladium-catalyzed intermolecular coupling of aryl halides and amides. Org. Lett. 2, 1101–1104 (2000).

    Article  CAS  Google Scholar 

  27. Ruiz-Castillo, P. & Buchwald, S. L. Applications of palladium-catalyzed C-N cross-coupling reactions. Chem. Rev. 116, 12564–12649 (2016).

    Article  CAS  Google Scholar 

  28. Vimolratana, M., Simard, J. L. & Brown, S. P. Palladium-catalyzed amidation of 2-chloropyrimidines. Tetrahedron Lett. 52, 1020–1022 (2011).

    Article  CAS  Google Scholar 

  29. Surry, D. S. & Buchwald, S. L. Dialkylbiaryl phosphines in Pd-catalyzed amination: a user’s guide. Chem. Sci. 2, 27–50 (2011).

    Article  CAS  Google Scholar 

  30. Wang, Y. et al. High-throughput reaction screening with nanomoles of solid reagents coated on glass beads. Angew. Chem. Int. Ed. 58, 7987–7991 (2019).

    Article  Google Scholar 

  31. Lee, J., Schmink, J. R. & Berritt, S. Introduction of low-barrier high-throughput experimentation in the undergraduate laboratory: Suzuki-Miyaura reaction. J. Chem. Educ. 97, 538–542 (2020).

    Article  CAS  Google Scholar 

  32. Schafer, W., Bu, X., Gong, X., Joyce, L. A. & Welch, C. J. High-throughput analysis for high-throughput experimentation in organic chemistry. in Comprehensive Organic Synthesis, Vol. 9 (ed Knochel, P.) 28–53 (Elsevier, 2014).

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Financial support for this work was provided by the University of Ottawa, the National Science and Engineering Research Council of Canada (NSERC), and the Canada Research Chair program. We thank the Canadian Foundation for Innovation (CFI) and the Ontario Ministry of Research, Innovation, & Science for essential infrastructure.

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Authors and Affiliations



A.C., R.C. and S.G.N. designed the experiments. A.C. performed the experiments. A.C., R.C. and S.G.N. wrote the manuscript.

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Correspondence to Stephen G. Newman.

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The authors declare no competing interests.

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Peer review information Nature Protocols thanks Frank Glorius and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Kashani, S. K., Jessiman, J. E. & Newman, S. G. Org. Process Res. Dev. 24, 1948–1954 (2020):

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Supplementary Procedure, Results, Figs. 1–18, Table 1 and Blueprints.

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Cook, A., Clément, R. & Newman, S.G. Reaction screening in multiwell plates: high-throughput optimization of a Buchwald–Hartwig amination. Nat Protoc 16, 1152–1169 (2021).

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