High-resolution α-amylase assay combined with high-performance liquid chromatography−solid-phase extraction−nuclear magnetic resonance spectroscopy for expedited identification of α-amylase inhibitors – proof of concept and α-amylase inhibitor in cinnamon.

Leyla Okutan; Kenneth Thermann Kongstad; Anna Jäger; Dan Stærk

doi:10.1021/jf5047283

High-resolution α-amylase assay combined with high-performance liquid chromatography−solid-phase extraction−nuclear magnetic resonance spectroscopy for expedited identification of α-amylase inhibitors – proof of concept and α-amylase inhibitor in cinnamon.

Leyla Okutan, Kenneth Thermann Kongstad, Anna Jäger, Dan Stærk

44 Citations (Scopus)

Abstract

Type 2 diabetes affects millions of people worldwide, and new improved drugs or functional foods containing selective α-amylase inhibitors are needed for improved management of blood glucose. In this article the development of a microplate-based high-resolution α-amylase inhibition assay with direct photometric measurement of α-amylase activity is described. The inhibition assay is based on porcine pancreatic α-amylase with 2-chloro-4-nitrophenyl-α-d-maltotriose as substrate, which this gives a stable, sensitive, and cheap inhibition assay as requested for high-resolution purposes. In combination with HPLC–HRMS–SPE–NMR, this provides an analytical platform that allows simultaneous chemical and biological profiling of α-amylase inhibitors in plant extracts. Proof-of-concept with an artificial mixture of six compounds—of which three are known α-amylase inhibitors—showed that the high-resolution α-amylase inhibition profiles allowed detection of sub-microgram amounts of the α-amylase inhibitors. Furthermore, the high-resolution α-amylase inhibition assay/HPLC–HRMS–SPE–NMR platform allowed identification of cinnamaldehyde as the α-amylase inhibitor in cinnamon (Cinnamomum verum Presl.).

Original language	English
Journal	Journal of Agricultural and Food Chemistry
Volume	62
Issue number	47
Pages (from-to)	11465-11471
Number of pages	7
ISSN	0021-8561
DOIs	https://doi.org/10.1021/jf5047283
Publication status	Published - 26 Nov 2014

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Okutan, L., Kongstad, K. T., Jäger, A., & Stærk, D. (2014). High-resolution α-amylase assay combined with high-performance liquid chromatography−solid-phase extraction−nuclear magnetic resonance spectroscopy for expedited identification of α-amylase inhibitors – proof of concept and α-amylase inhibitor in cinnamon. Journal of Agricultural and Food Chemistry, 62(47), 11465-11471. https://doi.org/10.1021/jf5047283

High-resolution α-amylase assay combined with high-performance liquid chromatography−solid-phase extraction−nuclear magnetic resonance spectroscopy for expedited identification of α-amylase inhibitors – proof of concept and α-amylase inhibitor in cinnamon. / Okutan, Leyla; Kongstad, Kenneth Thermann; Jäger, Anna et al.
In: Journal of Agricultural and Food Chemistry, Vol. 62, No. 47, 26.11.2014, p. 11465-11471.

Research output: Contribution to journal › Journal article › Research › peer-review

Okutan, L, Kongstad, KT, Jäger, A & Stærk, D 2014, 'High-resolution α-amylase assay combined with high-performance liquid chromatography−solid-phase extraction−nuclear magnetic resonance spectroscopy for expedited identification of α-amylase inhibitors – proof of concept and α-amylase inhibitor in cinnamon.', Journal of Agricultural and Food Chemistry, vol. 62, no. 47, pp. 11465-11471. https://doi.org/10.1021/jf5047283

Okutan L, Kongstad KT, Jäger A, Stærk D. High-resolution α-amylase assay combined with high-performance liquid chromatography−solid-phase extraction−nuclear magnetic resonance spectroscopy for expedited identification of α-amylase inhibitors – proof of concept and α-amylase inhibitor in cinnamon. Journal of Agricultural and Food Chemistry. 2014 Nov 26;62(47):11465-11471. doi: 10.1021/jf5047283

Okutan, Leyla ; Kongstad, Kenneth Thermann ; Jäger, Anna et al. / High-resolution α-amylase assay combined with high-performance liquid chromatography−solid-phase extraction−nuclear magnetic resonance spectroscopy for expedited identification of α-amylase inhibitors – proof of concept and α-amylase inhibitor in cinnamon. In: Journal of Agricultural and Food Chemistry. 2014 ; Vol. 62, No. 47. pp. 11465-11471.

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abstract = "Type 2 diabetes affects millions of people worldwide, and new improved drugs or functional foods containing selective α-amylase inhibitors are needed for improved management of blood glucose. In this article the development of a microplate-based high-resolution α-amylase inhibition assay with direct photometric measurement of α-amylase activity is described. The inhibition assay is based on porcine pancreatic α-amylase with 2-chloro-4-nitrophenyl-α-d-maltotriose as substrate, which this gives a stable, sensitive, and cheap inhibition assay as requested for high-resolution purposes. In combination with HPLC–HRMS–SPE–NMR, this provides an analytical platform that allows simultaneous chemical and biological profiling of α-amylase inhibitors in plant extracts. Proof-of-concept with an artificial mixture of six compounds—of which three are known α-amylase inhibitors—showed that the high-resolution α-amylase inhibition profiles allowed detection of sub-microgram amounts of the α-amylase inhibitors. Furthermore, the high-resolution α-amylase inhibition assay/HPLC–HRMS–SPE–NMR platform allowed identification of cinnamaldehyde as the α-amylase inhibitor in cinnamon (Cinnamomum verum Presl.).",

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AB - Type 2 diabetes affects millions of people worldwide, and new improved drugs or functional foods containing selective α-amylase inhibitors are needed for improved management of blood glucose. In this article the development of a microplate-based high-resolution α-amylase inhibition assay with direct photometric measurement of α-amylase activity is described. The inhibition assay is based on porcine pancreatic α-amylase with 2-chloro-4-nitrophenyl-α-d-maltotriose as substrate, which this gives a stable, sensitive, and cheap inhibition assay as requested for high-resolution purposes. In combination with HPLC–HRMS–SPE–NMR, this provides an analytical platform that allows simultaneous chemical and biological profiling of α-amylase inhibitors in plant extracts. Proof-of-concept with an artificial mixture of six compounds—of which three are known α-amylase inhibitors—showed that the high-resolution α-amylase inhibition profiles allowed detection of sub-microgram amounts of the α-amylase inhibitors. Furthermore, the high-resolution α-amylase inhibition assay/HPLC–HRMS–SPE–NMR platform allowed identification of cinnamaldehyde as the α-amylase inhibitor in cinnamon (Cinnamomum verum Presl.).

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