High-temperature liquid chromatography coupled on-line to a continuous-flow biochemical screening assay with electrospray ionization mass spectrometric detection

A.R. de Boer; J.M. Alcaide-Hidalgo; J.G. Krabbe; J.J. Kolkman; C.N.V. Boas; W.M.A. Niessen; H. Lingeman; H. Irth

doi:10.1021/ac0510282

High-temperature liquid chromatography coupled on-line to a continuous-flow biochemical screening assay with electrospray ionization mass spectrometric detection

A.R. de Boer, J.M. Alcaide-Hidalgo, J.G. Krabbe, J.J. Kolkman, C.N.V. Boas, W.M.A. Niessen, H. Lingeman, H. Irth

BioAnalytical Chemistry

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

The potential of high-temperature liquid chromatography (HTLC) was investigated in an on-line combination with a screening system for bioactive compounds against the enzyme cathepsin B. Samples were separated by HTLC and subsequently analyzed by an on-line continuous-flow enzymatic assay. Detection was performed by electrospray ionization mass spectrometry, revealing both the bioactivity and the molecular mass of the bioactive compounds. Compared to conventional reversed-phase liquid chromatography, the amount of methanol necessary for separation could be decreased to only 10%, which improved the compatibility of LC with a biochemical assay. Sufficient preheating of the mobile phase prior to the separation and postcolumn cooling to prevent deactivation of the enzyme, even at column temperatures as high as 208°C, was achieved as indicated by the reliable peak shapes obtained. The sensitivity was comparable with previously described systems operating at ambient temperatures as similar IC

Original language	English
Pages (from-to)	7894-7900
Journal	Analytical Chemistry
Volume	77
Issue number	24
DOIs	https://doi.org/10.1021/ac0510282
Publication status	Published - 2005

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de Boer, A. R., Alcaide-Hidalgo, J. M., Krabbe, J. G., Kolkman, J. J., Boas, C. N. V., Niessen, W. M. A., Lingeman, H., & Irth, H. (2005). High-temperature liquid chromatography coupled on-line to a continuous-flow biochemical screening assay with electrospray ionization mass spectrometric detection. Analytical Chemistry, 77(24), 7894-7900. https://doi.org/10.1021/ac0510282

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abstract = "The potential of high-temperature liquid chromatography (HTLC) was investigated in an on-line combination with a screening system for bioactive compounds against the enzyme cathepsin B. Samples were separated by HTLC and subsequently analyzed by an on-line continuous-flow enzymatic assay. Detection was performed by electrospray ionization mass spectrometry, revealing both the bioactivity and the molecular mass of the bioactive compounds. Compared to conventional reversed-phase liquid chromatography, the amount of methanol necessary for separation could be decreased to only 10%, which improved the compatibility of LC with a biochemical assay. Sufficient preheating of the mobile phase prior to the separation and postcolumn cooling to prevent deactivation of the enzyme, even at column temperatures as high as 208°C, was achieved as indicated by the reliable peak shapes obtained. The sensitivity was comparable with previously described systems operating at ambient temperatures as similar IC",

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AU - de Boer, A.R.

AU - Alcaide-Hidalgo, J.M.

AU - Krabbe, J.G.

AU - Kolkman, J.J.

AU - Boas, C.N.V.

AU - Niessen, W.M.A.

AU - Lingeman, H.

AU - Irth, H.

PY - 2005

Y1 - 2005

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AB - The potential of high-temperature liquid chromatography (HTLC) was investigated in an on-line combination with a screening system for bioactive compounds against the enzyme cathepsin B. Samples were separated by HTLC and subsequently analyzed by an on-line continuous-flow enzymatic assay. Detection was performed by electrospray ionization mass spectrometry, revealing both the bioactivity and the molecular mass of the bioactive compounds. Compared to conventional reversed-phase liquid chromatography, the amount of methanol necessary for separation could be decreased to only 10%, which improved the compatibility of LC with a biochemical assay. Sufficient preheating of the mobile phase prior to the separation and postcolumn cooling to prevent deactivation of the enzyme, even at column temperatures as high as 208°C, was achieved as indicated by the reliable peak shapes obtained. The sensitivity was comparable with previously described systems operating at ambient temperatures as similar IC

U2 - 10.1021/ac0510282

DO - 10.1021/ac0510282

M3 - Article

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JO - Analytical Chemistry

JF - Analytical Chemistry

IS - 24

ER -

High-temperature liquid chromatography coupled on-line to a continuous-flow biochemical screening assay with electrospray ionization mass spectrometric detection

Abstract

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