Jäger, Anna

Jønsson, Louise H.

Nyberg, Nils

Stærk, Dan

Tahtah, Yousof

English

Copenhagen University Research Information System

u n i ve r s i t y  o f  co pe n h ag e n  Københavns UniversitetHigh-resolution Assays Combined with HPLC-HRMS-SPE-ttNMR for Identification ofAntidiabetic Compounds in root of Scutellaria BaicalensisTahtah, Yousof; Jønsson, Louise H.; Nyberg, Nils; Jäger, Anna; Stærk, DanPublication date:2014Citation for published version (APA):Tahtah, Y., Jønsson, L. H., Nyberg, N., Jäger, A., & Stærk, D. (2014). High-resolution Assays Combined withHPLC-HRMS-SPE-ttNMR for Identification of Antidiabetic Compounds in root of Scutellaria Baicalensis . Postersession presented at 35th Danish NMR Meeting, Kobæk Strand, Denmark.Download date: 03. Feb. 2020U N I V E R S I T Y  O F  C O P E N H A G E N High-resolution Assays Combined with HPLC-HRMS-SPE-ttNMR for Identification of Antidiabetic Compounds in root of Scutellaria Baicalensis   Results Figure 2. High-resolution α-glucosidase inhibition and radical scavenging profiles of Radix Scutellaria with overlaid HPLC chromatogram at 280 nm. Peaks are numbered sequentially with increasing elution order. The radical scavenging and α-glucosidase inhibition profiles provide good resolution that allows disclosure of the individual metabolites responsible for the observed activities, that is, correlating peaks in the biochromatogram with individual peaks in the overlaid HPLC chromatograms.                    Yousof Tahtah, Louise H. Jønsson, Nils Nyberg, Anna K. Jäger, Dan Stærk  Natural Products Research, Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, Universitetsparken 2, DK-2100, Copenhagen.  E-mail: yousof.tahtah@sund.ku.dk     Abstract This work describes an analytical platform based on high-resolution radical scavenging and high-resolution α-glucosidase inhibition assays in combination with hyphenation of high-performance liquid chromatography, high-resolution mass spectrometry, solid-phase extraction, and tube-transfer nuclear magnetic resonance spectroscopy, i.e., HPLC-HRMS-SPE-ttNMR/high-resolution radical scavenging and high-resolution α-glucosidase assays. The platform enables fast screening of complex matrices for individual analytes with α-glucosidase inhibitory and radical scavenging activity, followed by structural indentification targeted the active analytes only.  Introduction Type 2 diabetes is one of the most prevalent diseases affecting 246 million worldwide and its incidence and serious complications continue to grow rapidly. Patients with type 2 diabetes suffer from a series of micro- and macrovascular complications such as visual impairment, blindness, neuropath, kidney failure and cardiovascular diseases.  Radix Scutellaria is the dried root of the medicinal plant Scutellaria baicalensis. Radix Scutellaria is officially listed in the Chinese Pharmacopeia and Japanese Pharmacopoeia, and exhibits a variety of therapeutic effects and has long history of application in traditional formulation and in modern herbal medication – and is also used as a food additive.            -3500 -2500 -1500 -500 500 1500 2500 3500 10 20 30 40 50 60 70 80 90 100 110 Absorbance (mAU) Minutes Relative intensities Concluding Remarks This work describe the development of a bioassay-coupled HPLC-HRMS-SPE-ttNMR platform for identification of radical scavengers and α-glucosidase inhibitors in an extract of Radix Scutellaria. The biochromatogram can be used to pinpoint HPLC-analytes with bioactivity.                Acknowledgments This work was supported by the Drug Research Academy, University of Copenhagen.                 Method Figure 1. Schematic represenation of HPLC-HRMS-SPE-ttNMR analysis of Radix Scutellaria extract guided by high-resolution ABTS˙+ reduction and α-glucosidase inhibition profiles. Path 1: microfractionation into six 96-well microplates followed by ABTS˙+ reduction assay of each microfraction to produce high-resolution ABTS˙+ reduction profile. Path 2: microfractionation into six 96-well microplates followed by α-glucosidase inhibition assay to produce high-resolution α-glucosidase inhibition profile. Path 3: HPLC-HRMS-SPE-ttNMR analysis targeting antioxidants and α-glucosidase inhibitory metabolites identified in the preceeding procedures.     1 23 4 5 6 78 9 10 11 UV, 280 nm ABTS˙+ reduction  α-glucosidase inhibition Peak 2. Viscidulin III Peak 4. Baicalin Peak 11. wogonin Peak 9. Baicalein 

High-resolution Assays Combined with HPLC-HRMS-SPE-ttNMR for Identification of Antidiabetic Compounds in root of <em>Scutellaria Baicalensis </em>

https://curis.ku.dk/ws/files/97000149/Danish_NMR_meeting_2014_poster_YT.pdf

High-resolution Assays Combined with HPLC-HRMS-SPE-ttNMR for Identification of Antidiabetic Compounds in root of <em>Scutellaria Baicalensis </em>

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