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Preparation and characterization of methacrylate-based semi-micro monoliths for high-throughput bioanalysis

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Abstract

Hexyl methacrylate (HMA)-based monolithic semi-micro columns were prepared by in situ polymerization within the confines of 1.02-mm-i.d. silicosteel tubing for reversed-phase and/or precipitation–redissolution liquid chromatography. Practically useful monolithic columns with adequate separation efficiency, high permeability, and good mechanical strength were successfully obtained using a polymerization mixture comprising 24% hexyl methacrylate (HMA), 6% ethylene dimethacrylate (EDMA), 44.5% 1-propanol, and 25.5% 1,4-butanediol. The column performance was evaluated through the separations of a series of alkylbenzenes. At a normal flow rate of 50 μL min−1, the produced HMA-based monolithic columns typically exhibited 3,000 theoretical plates for a 20-cm-long column, and the pressure drop was generally less than 1 MPa per 20 cm. The monolithic columns were resistant to at least 15 MPa, and could be properly operated at 15–20 times higher flow rate than normal, reducing the separation time to 1/15–1/20. The HMA-based monolithic columns were applied to rapid and efficient separations of proteins such as ribonuclease A, cytochrome c, transferrin, and ovalbumin in the precipitation–redissolution mode. Using a CH3CN gradient elution at a flow rate of 1,000 μL min−1, four proteins were baseline separated within 20 s.

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Abbreviations

HMA:

hexyl methacrylate

EDMA:

ethylene dimethacrylate

AIBN:

2,2′-azobisisobutyronitrile

THF:

trifluoroacetic acid

RSDs:

relative standard deviations

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Acknowledgements

This work was supported by Grant-in-Aid for Young Scientists (A) (No.17685006) from the Ministry of Education, Culture, Sports, Science and Technology, Japan. The authors thank Y. Wakita and C. Ohi for their experimental assistance.

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Authors and Affiliations

  1. Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464–8603, Japan

    Tomonari Umemura & Hiroki Haraguchi

  2. Department of Chemistry, Faculty of Engineering, Gunma University, 1–5–1 Tenjin-cho, Kiryu, Gunma, 376–8515, Japan

    Yuji Ueki & Kin-ichi Tsunoda

  3. Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute, 1233 Watanuki, Takasaki, Gunma, 370–1292, Japan

    Akio Katakai & Masao Tamada

Authors
  1. Tomonari Umemura
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  2. Yuji Ueki
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  3. Kin-ichi Tsunoda
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  4. Akio Katakai
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  5. Masao Tamada
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  6. Hiroki Haraguchi
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Correspondence to Tomonari Umemura.

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Umemura, T., Ueki, Y., Tsunoda, Ki. et al. Preparation and characterization of methacrylate-based semi-micro monoliths for high-throughput bioanalysis. Anal Bioanal Chem 386, 566–571 (2006). https://doi.org/10.1007/s00216-006-0425-2

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  • DOI: https://doi.org/10.1007/s00216-006-0425-2

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